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Biomaterials. 2014 Oct 4. pii: S0142-9612(14)01009-6. doi: 10.1016/j.biomaterials.2014.09.011. [Epub ahead of print]
Prediction of inflammatory responses induced by biomaterials in contact with human blood using protein fingerprint from plasma.
Engberg AE1, Nilsson PH2, Huang S3, Fromell K4, Hamad OA4, Mollnes TE5, Rosengren-Holmberg JP6, Sandholm K3, Teramura Y7, Nicholls IA8, Nilsson B4, Ekdahl KN9.
Author information
Abstract
Inappropriate complement activation is often responsible for incompatibility reactions that occur when biomaterials are used. Complement activation is therefore a criterion included in legislation regarding biomaterials testing. However, no consensus is yet available regarding appropriate complement-activation-related test parameters. We examined protein adsorption in plasma and complement activation/cytokine release in whole blood incubated with well-characterized polymers. Strong correlations were found between the ratio of C4 to its inhibitor C4BP and generation of 10 (mainly pro-inflammatory) cytokines, including IL-17, IFN-?, and IL-6. The levels of complement activation products correlated weakly (C3a) or not at all (C5a, sC5b-9), confirming their poor predictive values. We have demonstrated a direct correlation between downstream biological effects and the proteins initially adhering to an artificial surface after contact with blood. Consequently, we propose the C4/C4BP ratio as a robust, predictor of biocompatibility with superior specificity and sensitivity over the current gold standard.
Copyright © 2014 Elsevier Ltd. All rights reserved.
Int J Artif Organs. 2014 Oct 2;37(9):668-78. doi: 10.5301/ijao.5000354. Epub 2014 Sep 23.
Pore size - a key property for selective toxin removal in blood purification.
Harm S1, Falkenhagen D, Hartmann J.
Author information
Abstract
PURPOSE:
Extracorporeal blood purification systems based on combined membrane/adsorption technologies are used in acute liver failure to replace detoxification as well as to remove inflammatory mediators in sepsis patients. In addition to coating and chemical modification of the surface, pore size significantly controls the selectivity of adsorption materials.
METHODS:
This study addresses the adsorption of albumin bound liver toxins, cytokines, and representative plasma compounds on three adsorbents which differ only in pore size distribution. All three adsorbents are based on hydrophobic poly(styrene-divinylbenzene) copolymer matrices and have mean pore sizes of 15, 30, and 100 nm.
RESULTS:
The pores of adsorbents act as molecular sieves and prevent the entry of molecules that are larger than their molecular cut-off. The results of this study reveal that adsorbents based on styrene-divinylbenzene polymers with 15 nm pores are suitable for cytokine removal, and the same adsorbents with 30-40 nm pores are the best choice for the removal of albumin-bound toxins in the case of liver failure. Adsorbents with very large pores lack selectivity which leads to uncontrolled adsorption of all plasma proteins. Therefore, hydrophobic adsorbents with large pores offer inadequate plasma compatibility and do not fulfill the requirements for blood purification.
CONCLUSIONS:
Biocompatibility and efficiency of adsorbents used for blood purification can improved by fine tuning of adsorbent surface pore distributions.
Biomed Res Int. 2014;2014:238160. doi: 10.1155/2014/238160. Epub 2014 Aug 27.
Adsorption of the inflammatory mediator high-mobility group box 1 by polymers with different charge and porosity.
Tripisciano C1, Eichhorn T1, Harm S2, Weber V3.
Author information
Abstract
High-mobility group box 1 protein (HMGB1) is a conserved protein with a variety of biological functions inside as well as outside the cell. When released by activated immune cells, it acts as a proinflammatory cytokine. Its delayed release has sparked the interest in HMGB1 as a potential therapeutic target. Here, we studied the adsorption of HMGB1 to anionic methacrylate-based polymers as well as to neutral polystyrene-divinylbenzene copolymers. Both groups of adsorbents exhibited efficient binding of recombinant HMGB1 and of HMGB1 derived from lipopolysaccharide-stimulated peripheral blood mononuclear cells. The adsorption characteristics depended on particle size, porosity, accessibility of the pores, and charge of the polymers. In addition to these physicochemical parameters of the adsorbents, modifications of the molecule itself (e.g., acetylation, phosphorylation, and oxidation), interaction with other plasma proteins or anticoagulants (e.g., heparin), or association with extracellular microvesicles may influence the binding of HMGB1 to adsorbents and lead to preferential depletion of HMGB1 subsets with different biological activity.
PMID: 25243124 [PubMed - in process] PMCID: PMC4163473 Free PMC Article
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Liver Transpl. 2014 Sep 19. doi: 10.1002/lt.24005. [Epub ahead of print]
First description of SPAD combined with cytokine adsorption in fulminant liver failure and hemophagocytic syndrome due to generalized HSV-1 infection.
Frimmel S1, Schipper J, Henschel J, Tsui TY, Mitzner SR, Koball S.
Author information
KEYWORDS:
Cytosorb; Hemophagocytic lymphohistiocytosis; Herpes simplex virus; SPAD; acute liver failure
PMID: 25233991 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25233991
" transfusion of packed red blood cells (PRBCs) stored for >14 days is associated with increased rates of sepsis"
Background
Accumulating evidence suggests that transfusion of packed red blood cells (PRBCs) stored for >14 days is associated with increased rates of sepsis, multiple organ dysfunction, and mortality in human patients.
Objective
To determine if duration of PRBC storage has an effect on morbidity and mortality in dogs after transfusion.
Animals
Dogs admitted to the Matthew J Ryan Veterinary Hospital of the University of Pennsylvania.
Methods
A retrospective case review of dogs identified through blood bank logbooks that received PRBC transfusions (minimum, 5 mL/kg) between 2001 and 2010. Dogs were categorized according to major cause of anemia (eg, hemorrhage, hemolysis, ineffective erythropoiesis) for analysis.
Results
A total of 3,095 dogs received 5,412 PRBC units. Longer duration of PRBC storage was associated with development of new or progressive coagulation failure (P = .001) and thromboembolic disease (P = .005). There was no association between duration of PRBC storage and survival for all dogs overall. However, a logistic regression model indicated that for dogs with hemolysis, 90% of which had immune-mediated hemolytic anemia, longer duration of PRBC storage was a negative risk factor for survival. For every 7 day increase in storage, there was a 0.79 lesser odds of 30 day survival (95% CI, 0.64–0.97; P = .024).
Conclusions and Clinical Importance
Duration of PRBC storage does not appear to be a major contributing factor to mortality in the overall canine population. However, longer duration of PRBC storage may negatively impact outcome in dogs with immune-mediated hemolytic anemia, thus warranting further investigation with prospective studies.
http://onlinelibrary.wiley.com/doi/10.1111/jvim.12430/abstract;jsessionid=B8F99809609B95013BF1269A3BEDB7AE.f04t01?deniedAccessCustomisedMessage=&userIsAuthenticated=false
The reception of the HemoDefend technology at the AABB conference was excellent. We had steady booth traffic during the conference and received a lot of positive feedback on our unique approach from transfusion medicine specialists, possible research collaborators, and potential strategic partners. People compared HemoDefend to “washing blood without the time, expense and trouble of washing blood”. The “Beads in a Bag” configuration, where the HemoDefend beads are placed directly in the blood storage bag and allowed to contact blood directly for extended periods of time, drew much praise, as no one thought this approach could be possible, and that it enabled blood purification during the storage period in an easy to use manner without the need for any other handling or equipment. Many people were impressed that, in contrast to filter technologies, we could easily add further functionality to the HemoDefend platform by simply adding new beads optimized to remove something new to the bead mixture. Several commented that our technology was one of the few truly innovative technologies that they have seen in a long time. For potential partners, the idea of converting a commodity, low margin blood storage bag into a higher margin blood treatment system seemed to resonate with them. However, they also liked the flexibility to be able to use the beads in an in-line filter system as well, which is an additional configuration option of our HemoDefend beads. Given that our goal is to out-license this technology, we are pleased with the ongoing discussions and continued interest we have been receiving.
http://www.sec.gov/Archives/edgar/data/1175151/000114420412001976/v245290_ex99-1.htm
HemoDefend™ is a late-development stage technology platform that is available for out-licensing to potential partners.
http://02c390e.netsolhost.com/hemodefend.htm
"We continue to believe that our therapy may have applications beyond current uses, including the treatment of Ebola infection, particularly in the terminal phase that is dominated by cytokine storm."[color=red][/color]
http://finance.yahoo.com/news/preliminary-q3-2014-report-cytosorbents-104500370.html
Could CytoSorb® Help?
The 2014 Ebola epidemic in West Africa, called an “international emergency” by the WHO, continues to grow and has already infected nearly 1,900 people, and claimed more than 1,000 lives. Epidemic is expected to continue throughout the year. Our Strategy for outreach:
• WHO
• FDA
• CDC
• Government agencies such as USAMRIID (US Army Medical Research Institute of Infectious Diseases)
• Non-profit organizations
• Hospitals treating Ebola patients inside/outside of West Africa
•Europe
•U.S.
http://www.cytosorbents.com/pdf/CytoSorbents_Q2_2014_Presentation.pdf
Anybody have extra time to invistigate this site?
"Welcome to the Joint Deployable Waste to Energy (JDW2E) community! This site was established and made available to the general public so private sector participants can retrieve and download DoD requirements and solicitation information on this topic."
https://lite.apan.org/Group/Index/2484?pageSize=12
Something else to ponder...UCLA Link?
Administration of an adsorbent polymer for treatment of systemic inflammation
US 20110212172 A1
Inventors John A. Kellum, Mitchell P. Fink
BIO
In 1976, Mitchell P. Fink, M.D., graduated from the School of Medicine at Washington University in St. Louis. After completing residency training in general surgery at the Naval Hospital Bethesda, Dr. Fink began a long and successful career in academic medicine, which included posts as Chief of the Division of General Surgery at the University of Massachusetts, Director of Surgical Critical Care Medicine at the Massachusetts General Hospital, and Johnson & Johnson Professor of Surgery at Harvard Medical School and Surgeon-in-Chief at the Beth Israel Deaconess Medical Center. Until 2007, Dr. Fink was the Founding Chair of the Department of Critical Care Medicine at the University of Pittsburgh School of Medicine, the first such department at a medical school in the United States. Dr. Fink was also the Watson Professor of Surgery and the Associate Vice Chancellor for Translational Research and Entrepreneurial Development at the University of Pittsburgh.
Under Dr. Fink’s leadership, the Department of Critical Care Medicine at the University of Pittsburgh expanded rapidly. By the time that Dr. Fink left Pittsburgh to pursue other interests (see below), the Department of Critical Care Medicine included more than 70 fulltime faculty members, who provided “7 x 24” attending-level coverage for more than 200 adult and pediatric ICU beds. The clinical budget for the Department exceeded $14 million per year and research in the Department was supported by more $20 million per year in extramural grants or contracts from the National Institutes of Health (NIH), the Defense Advanced Projects Research Administration (DARPA), other governmental agencies and private industry. After Dr. Fink left the University of Pittsburgh (see below), the School of Medicine endowed the “Mitchell P. Fink Chair of Critical Care Medicine” in his honor.
The PubMed database of the National Library of Medicine lists more than 290 scientific publications, which have been authored or co-authored by Dr. Fink. Many of these research papers have appeared in very prestigious journals, including Nature, the New England Journal of Medicine, the Journal of Clinical Investigation, the Journal of Experimental Medicine, Proceedings of the National Academy of Sciences USA, Gastroenterology, the American Journal of Physiology, the Journal of Pharmacology and Experimental Therapeutics, JAMA, Annals of Surgery, Archives of Internal Medicine, the Journal of Infectious Diseases, and Critical Care Medicine. In addition to these journal articles, Dr. Fink is an author or co-author of numerous invited book chapters. Dr. Fink has been the editor or co-editor of 14 books. Dr. Fink has received numerous honors, including being named in Marquis Who’s Who in America every year since 2008.
As a researcher, Dr. Fink has been particularly interested in alterations in epithelial function due to inflammation, and the development of novel anti-inflammatory therapeutics. Dr. Fink is or has been a member of the Editorial Boards of numerous scientific publications, including the American Journal of Physiology, Biochemical Pharmacology, Intensive Care Medicine, and the Journal of Trauma. He is a Scientific Editor for Critical Care Medicine and is an Associate Editor of the Journal of Pharmacology and Experimental Therapeutic and the Journal of Leukocyte Biology.
Until he transitioned from the world of academic medicine to the world of entrepreneurial biotechnology, Dr. Fink’s research program was continuously funded by the National Institutes of Health (NIH) for more than 20 years. He is a Past-President of the Shock Society, a past Chair of the Program Committee for the Society of Critical Care Medicine, and a member of other prestigious scientific and medical societies, including the American Surgical Association, the James IV Association of Surgeons, and the American Physiological Society. Dr. Fink also has been on the Defense Sciences Research Council (DSRC) of DARPA and the Surgery, Anesthesiology and Trauma (SAT) study section of the NIH. In September 2004, Dr. Fink served as an ad-hoc member of the Council for the National Institutes of General Medical Sciences.
Dr. Fink is an inventor or co-inventor on more than a dozen issued or pending US and international patents.
In 2000, Dr. Fink co-founded Critical Therapeutics, Inc., a biotechnology company based in Lexington, Massachusetts. Dr. Fink and his two partners wrote the business plan for Critical Therapeutics and raised the initial funding (more than $10 million) from venture capitalists to launch the company. Critical Therapeutics, Inc. had a successful initial public offering in 2004.
In 2005, Dr. Fink co-founded Logical Therapeutics, Inc., a Boston-based early stage biotechnology company devoted to developing novel drugs to treat pain and inflammation. In June 2007, Logical Therapeutics closed a round of financing, raising $30 million from a syndicate of highly regarded venture funds, including SVLS, Burrill & Company, Novitas and Novo A/S. In July 2007, Dr. Fink resigned from his position as Chair of the Department of Critical Care Medicine at the University of Pittsburgh in order to assume the position of President and CEO of Logical Therapeutics. Under Dr. Fink’s leadership, Logical Therapeutics recruited a team of highly experienced drug developers, including experts in regulatory affairs, drug formulation and the conduct of clinical trials. This team with Dr. Fink as their leader organized and carried out a highly successful clinical trial of the company’s most advanced compound, LT-NS001. In October 2008, Dr Fink resigned from his leadership role at Logical Therapeutics, Inc. in order to live in California with his fiancé.
Dr. Fink has served or currently serves on the Boards of Directors of several biotechnology companies, including Critical Therapeutics, Logical Therapeutics, Inotek, Inc. (based in Beverly, Massachusetts) and Midway Pharmaceuticals, Inc. (based in Chicago, Illinois).
Currently, Dr. Fink is the Vice Chair for Critical Care in the Department of Surgery and Professor-in-Residence at the David Geffen School of Medicine at UCLA (Joint Appointments in Surgery and Anesthesiology).
Just incase you are woundering:
http://www.uclahealth.org/body.cfm?id=479&action=detail&ref=122432
Dont forget to add this...
Legal Events
Date Code Event Description
May 5, 2011 AS Assignment Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:KELLUM, JOHN A.;FINK, MITCHELL P.;SIGNING DATES FROM 20110421 TO 20110503;REEL/FRAME:026232/0827
Owner name: UNIVERSITY OF PITTSBURGH - OF THE COMMONWEALTH SYS
Apr 18, 2011 AS Assignment Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:KELLUM, JOHN A.;FINK, MITCHELL P.;SIGNING DATES FROM 20110413 TO 20110415;REEL/FRAME:026145/0136
Owner name: UNIVERSITY OF PITTSBURGH - OF THE COMMONWEALTH SYS
NGC owns the rights to the DGU for Government and Military, Cirque has the rights to everything else.
Metso Controls for Four New CNIM Waste to Energy & Biomass Plants in UK & France
Process systems control equipment manufacturer, Metso has been awarded four repeat orders from French firm, CNIM, which designs, produces and commissions waste to energy plants, to supply automation systems to new facilities in France and the UK.
According to Metso, it has now received a total of 12 orders for its process control systems for waste to energy and biomass plants from CNIM since 2011.
As part of the deal, the company said that it will supply an extensive Metso DNA automation system for all process controls and plant information management. The integrated process automation system covers all plant processes from boilers to turbines.
According to Metso, high-level reliability is ensured through an integrated and certified safety solution.
The company said that its solutions play a significant role in managing waste to energy plants successfully and help them reach their goals of high performance, as well as safe and reliable operation, with a high degree of integration.
Additionally, the systems are said to provide good information on the process performance.
At biomass to energy plants, Metso said that its automation systems contribute to maximising incineration capacity, keeping combustion and steam production stable while allowing high production process availability.
The facilities
In the UK, the control systems will be supplied to waste to energy plants in Battlefield, Shropshire, and in Leeds, as well as a biomass plant in Ridham Dock. In France systems will be supplied to a biomass to energy plant in Estrées-Mons.
The waste to energy plant in Leeds is owned and will be operated by Veolia Environmental Services.
It will have an electric output of 11 MW and is designed to process up to 20.5 tonnes of residual waste per hour and is expected to help the city to reach the target of recycling more than 60% of its waste. The plant is planned to go on line in 2016.
The Battlefield waste to energy (pictured) plant in Shropshire is also owned and operated by Veolia. Metso said that it is designed to process 45% of the municipal waste produced in the county and generate enough electricity to power 10,000 households. The plant is planned to be operational in 2015.
The biomas plant in Estrées-Mons is owned and will be operated by Nerea, a subsidiary of the French group Akuo Energy. Once on line at the end of 2014, it will have an electric output of 13 MW.
The Ridham Dock combined heat and power biomass power plant is owned by the German Mannheim energy company (MVV Energie). It is designed to have an electrical output of 23 MW and will process around 172,000 tonnes of waste timber per year and is scheduled to be operational in spring 2015.
CNIM is the lead contractor for all four facilities.
Evaluation of an experimental filter designed for improving the quality of red blood cells (RBCs) during storage.
“Slide 39: … And as you can see, there are companies like Haemonetics that cross over from blood transfusion to cardiac surgery. Maquet crosses over from cardiac surgery to critical care. Companies like Fresenius are on the dialysis side as well as on the blood transfusion side. You can see how CytoSorbents’ technology could be very synergistic to the larger businesses of companies like these, as an example. We address so many major markets that have billion dollar revenue potential that if we are successful, we can impact the top and bottom line growth of most major multi-national corporations, which is something that you cannot normally say of technologies from small companies such as ours.”
That being said…
Transfusion. 2014 Mar;54(3):592-601. doi: 10.1111/trf.12330. Epub 2013 Jul 9.
Evaluation of an experimental filter designed for improving the quality of red blood cells (RBCs) during storage by simultaneously removing white blood cells and immunomodulators and improving RBC viscoelasticity and Band 3 proteins.
Sowemimo-Coker SO.
Author information Haemonetics Corporation, Braintree, Massachusetts.
Abstract
BACKGROUND: Storage age of red blood cells (RBCs) has been reported to be associated with increased mortality and morbidity. During storage, RBCs undergo changes in biochemical and functional properties. Stored RBCs may also contain white blood cells (WBCs), activated platelets (PLTs), cytokines, immunoglobulin, and other bioactive proteins. Transfusion of these bioactive proteins and cells with RBCs has the potential to cause serious adverse effects. We evaluated the performances of an experimental filter (EF) designed to remove immunoglobulins, cytokines, and other bioactive proteins in RBCs.
STUDY DESIGN AND METHODS: Sixteen sets, each containing 3 units of ABO-identical RBCs in AS-3 were obtained from a blood bank. Three units of RBCs were combined together and then split into three equal aliquots, A, B, and C. Unit A was unfiltered while Units B and C were filtered with a leukoreduction filter and the EF, respectively. All the units were stored at 4°C in a blood bank refrigerator for 42 days. We measured RBC viscoelasticity, hemolysis, RBC adenosine triphosphate, Band 3 proteins, cytokines, PLTs, WBCs, and immunoglobulin before and after filtration and on Days 21 and 42 of storage. Data were analyzed by repeated-measures analysis of variance with Newman-Keuls multiple comparison test.
RESULTS: The EF significantly (p<0.05) reduced the levels of immunoglobulin (control IgG, 2.184 ± 1.918 mg/mL; BPF4, 2.216 ± 1.956 mg/mL; and EF, 0.363 ± 0.391 mg/mL), PLTs, cytokines, and improved viscoelastic properties when compared to either control or leukoreduced RBCs.
CONCLUSION: The EF achieved lower levels of WBCs, improved viscoelastic properties, and reduced levels of immunoglobulins and cytokines but significance will require clinical evaluation.
http://www.ncbi.nlm.nih.gov/pubmed/23834280
Interleukin 6 promotes endometrial cancer growth
Biochem Biophys Res Commun.
2014 Mar 28;446(1):167-72. doi: 10.1016/j.bbrc.2014.02.080. Epub 2014 Feb 25.
Interleukin 6 promotes endometrial cancer growth through an autocrine feedback loop involving ERK-NF-?B signaling pathway.
Che Q1, Liu BY1, Wang FY1, He YY1, Lu W1, Liao Y1, Gu W2, Wan XP3.
Author information 1Department of Obstetrics and Gynecology, Shanghai First People's Hospital Affiliated to Shanghai Jiao Tong University, Shanghai, China.2Department of Obstetrics and Gynecology, International Peace Maternity & Child Health Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China. Electronic address: krisgu70@163.com.3Department of Obstetrics and Gynecology, Shanghai First Maternity and Infant Hospital Affiliated to Tong Ji University, Shanghai, China. Electronic address: wanxp@sjtu.edu.cn.
Abstract
Interleukin (IL)-6 as an inflammation factor, has been proved to promote cancer proliferation in several human cancers. However, its role in endometrial cancer has not been studied clearly. Previously, we demonstrated that IL-6 promoted endometrial cancer progression through local estrogen biosynthesis. In this study, we proved that IL-6 could directly stimulate endometrial cancer cells proliferation and an autocrine feedback loop increased its production even after the withdrawal of IL-6 from the medium. Next, we analyzed the mechanism underlying IL-6 production in the feedback loop and found that its production and IL-6-stimulated cell proliferation were effectively blocked by pharmacologic inhibitors of nuclear factor-kappa B (NF-?B) and extra-cellular signal-regulated kinase (ERK). Importantly, activation of ERK was upstream of the NF-?B pathways, revealing the hierarchy of this event. Finally, we used an orthotopic nude endometrial carcinoma model to confirm the effects of IL-6 on the tumor progression. Taken together, these data indicate that IL-6 promotes endometrial carcinoma growth through an expanded autocrine regulatory loop and implicate the ERK-NF-?B pathway as a critical mediator of IL-6 production, implying IL-6 to be an important therapeutic target in endometrial carcinoma.
KEYWORDS: Autocrine; Endometrial carcinoma; IL-6; Proliferation
Did you sell your shares or those of your "friends"? Remember you don't own all those shares you claim... or did the story change again?
Per the blog:
"Testing and Certification
•Full-scale military and commercial prototype systems built "
I don't know why they did not put out a PR on this...
I have been here for at least 6 months. Just listening and watching while slowly aquireing the posisition. And of course doing DD...
Availability
Manufacturing Plan
•Gasifier and Gas Cleanup sub-systems manufactured by Cirque Energy
•Overall system manufacturing and final assembly by Northrop Grumman
Testing and Certification
•Full-scale military and commercial prototype systems built
•Extended duration testing and third party certification
•Air emissions testing and certification
•Military demonstrations and shakedowns
Commercial availability by 2nd quarter 2015
UPCOMING DOD ENERGY DEMONSTRATION
OPPORTUNITIES
DoD: Environmental Security Technology
Certification Program (ESTCP)
The ESTCP program issues annual solicitations for a
variety of technologies the DoD sees as ripe for
funding demonstrations using installations as a
proving ground. The program has served as a
launching point for microgrid, energy management,
waste-to-energy, and high-efficiency thermal
systems. With annual awards to experienced DoD
contractors, DoD and Department of Energy
laboratories, and innovative small businesses alike,
this program is a significant in-road to working on
base quickly and collaborating with DoD more
broadly. ESTCP Energy and Water topics are
expected in early 2014.
RENEWABLE ENERGY FOR MILITARY INSTALLATIONS:
2014 INDUSTRY REVIEW
MILITARY FACILITY ENERGY USE
Twenty-percent of the U.S. military’s energy
consumption occurs at its installations. DoD pays
around $4 billion annually to provide power to its
300,000 plus facilities in the U.S. and around the
world. DoD has made improvements in installation
energy systems and management a priority, driven
by the desire to provide maximum mission support
through improved security of supply and reduced
costs.
DoD is uniquely positioned to serve as a platform to
develop and install new technologies. The Army,
Navy, and Air Force are each implementing detailed
plans to achieve ambitious renewable energy and
energy efficiency targets at military installations
(Figure 1). These plans include a one gigawatt (GW)
per service target of renewable power capacity at
military facilities, primarily via third-party financing.
PURPOSE OF THIS REPORT
There are several lessons to be learned from the
private sector as DoD becomes an increasingly
significant player in the renewable energy industry.
This report contains a series of industry perspectives
about the business considerations associated with
renewable energy installations at military facilities,
including:
? Policy: State regulatory hurdles for military use
of renewables, and best practices in
collaboration with state energy offices
? Contracting and Financing: An overview of the
existing procurement landscape with insight and
recommendations about creating financeable
third-party-owned projects
? Siting and Technology: Siting effective
renewable energy projects at military bases and
making use of innovative technology options
? Energy Security and Microgrids: The importance
of energy-secure technologies, like microgrids,
to enhance the use of renewable energy at
military installations and ensure reliable power
for critical infrastructure
The report is intended to be a resource for the
Department of Defense and military service officials
as they design and amend renewable energy
programs, for renewable energy companies and
investors as they navigate the military’s
procurement processes, and for other interested
parties.
A group of prominent renewable energy developers,
energy service companies, financiers, law firms, and
other groups working with the military authored thetwelve articles in this report. The articles generally
provide insight about present business opportunities
for third-party-owned renewable electricity projects
at military installations. It should be noted that the
military employs renewable energy through a
number of diverse applications that continue to
expand, and this report does not attempt to give a
comprehensive overview of every renewable energy
technology or procurement option available.
Renewable Energy for Military Installations: 2014
Industry Review is a product of the American Council
On Renewable Energy’s (ACORE’s) National Defense
and Security Initiative, which defines the
opportunities for the expanded use of renewable
energy in support of national defense by drawing on
the collective expertise and experience of ACORE
members: renewable energy leaders involved in
manufacturing, project development, finance, end
use, and professional services.
The views and opinions expressed in this report are
those of the authors and do not necessarily reflect
the views of ACORE.
http://acore.org/files/pdfs/Renewable-Energy-for-Military-Installations.pdf
I have added 500k in the past few days as well
Cardiac intensive care unit in miniature formatBy Andrea BannertMany lives can be saved only in a special clinic. For patients who need a heart-lung machine, the transport is the biggest hurdle. A new portable device to change the© Sergey Nivens/Fotolia.comAt the Christmas party, Sabine H. is * suddenly bad. When she goes home, must pass to the 37-year-old, gets sharp pain in the chest. The diagnosis of Notarztes: Cardiogenic shock - the heart not pumping. "That I got through that, I owe to this device", she says, pointing to a large shoe box equipment rises from a bunch of hoses. The apparatus is a heart-lung machine in miniature format. The intensive care unit for on the road for the first time at the Munich University Clinic presented.The device takes over the functions of the heart and lungs. It sucks the blood from the body, enrich it with oxygen and pumps it back into the arteries. The helper moved up to six litres per minute.Shoe box versus chest of drawers© Frank bornThe new system fits into the small red bagSuch machines are not new. But usually they take as much space as a high chest of drawers. They are used in the hospital, to perform operations on the heart not beating, where heart and lungs need to be switched off. The 70 kilograms colossus then takes over their task. He can be difficult in an ambulance with himself - in the helicopter which is even impossible due to the large weight.The bio engineer Frank Born has developed the heart-lung machine in miniature format. She weighs only 10 kg - every helicopter can do it. "With the system patients who were previously not transportable, can be placed in special clinics", explained born. And it can save lives.Tragic death sentenceThe trigger for the idea with the portable life saver was a tragic experience: seven years ago, a young athlete with serious heart problems at the clinic was hospitalized where born at that time worked. Actually the patient would have to be laid as soon as possible in a specialized center. But because his own heart with a heart-lung machine support only worked, it was not possible. The 34-year-old died. If there was already the mini intensive care unit, he was maybe still alive.© Frank bornThe inventor of the heart-lung machine: Frank BornShort-term takeover of multiple organ functionSo far, there is the system only at the Hospital of the Maxi Milan University in Munich. There, a team of experts is available to carry out the transport of seriously ill patients. Most recently they brought a German patient by Kuwait in the German heart centre Berlin - flight time: seven hours. In addition to a heart-lung-therapy, the patient needed also a kidney replacement. Born for this installed a Hemofilter which assumes the function of the kidney. Also, the liver can be relieved with the system: A Cytosorb filter filter pollutants such as Interleukins and Cytokines in the blood. "With the mini heart-lung machine the hospital to patients such as Sabine H., who would have been otherwise celebrated their last Christmas", born says.© Frank bornThe mini intensive care unit in use* Name changed by the editors.Source: Ludwig-Maximilians University of Munich hospital
http://www.netdoktor.de/Magazin/Herzkrankheiten-Intensivstati-12710.html
Could be us, If we can catch up...
Exosome Diagnostics Enters Collaboration Agreement with Lilly for Exosome Blood-Based Biomarker Discovery
NEW YORK September 19, 2013 — Exosome Diagnostics today announced it has entered into a collaboration agreement with Eli Lilly and Company (NYSE: LLY) for biomarker discovery and validation using Exosome Diagnostics proprietary EXO50 nucleic acid extraction kit. Under the agreement, Lilly will gain early access to Exosome Diagnostics technology to help identify key gene mutations and expression levels in blood that may be correlated with drug response and disease recurrence. Financial terms were not disclosed.
"Exosome Diagnostics technology may provide a unique opportunity to gain insight into the biology of complex conditions such as cancer and immune disorders," said Andrew Schade, senior medical director, diagnostic and experimental pathology at Lilly. "Exosome technology enables biofluid molecular sampling and the ability to monitor disease progression in real time. As Lilly explores new ways to pursue patient tailoring, we'll continue to work with partners to expand our capabilities."
"Accessing high quality messenger and microRNA directly from frozen patient fluid samples offers a rapid, cost-effective route to identify and validate biomarkers, which may be correlated with drug response and disease recurrence," said James McCullough, chief executive officer of Exosome Diagnostics. "Lilly has accumulated an extensive and well annotated clinical blood sample biobank that provides a unique opportunity to track target biomarkers through the clinical trial process and help overcome the limitations of stored biopsy tissue."
Exosomes and other microvesicles are secreted by all cells into all biofluids, and provide a natural biological packaging and distribution mechanism for RNA and DNA. Exosome Diagnostics' rapid exosome isolation and extraction technology produces high-quality RNA and DNA, including full length mRNA and microRNA, from small volumes of patient biofluids, such as blood (serum and plasma), urine and cerebrospinal fluid, for analysis by standard PCR, array and sequencing instruments. Analysis can be performed on fresh or frozen fluid samples, allowing for broad, flexible and convenient analyses of clinical trial samples, both in real-time and retrospectively, with no special preservation methods required. Exosomes and their protected nucleic acid contents are being investigated in a broad range of diseases including cancer, CNS disorders such as Alzheimer's and Parkinson's disease, cardiovascular disease, maternal/fetal medicine, and chronic kidney disease, among others. In July, QIAGEN and Exosome Diagnostics signed an agreement for the creation of High-Performance Biofluid Sample Preparation Kits for Personalized Healthcare Research which covers the exclusive supply of these products upon availability in 2014.
About Exosome Diagnostics
Exosome Diagnostics is a leading developer of biofluid-based molecular diagnostic tests for use in personalized medicine. Exosomes are packaged and shed into all biofluids, including blood, urine and CSF, providing a stable source for intact, cell-specific nucleic acids. The Company's proprietary exosome technology makes use of the presence and natural stability of RNA in exosomes to detect and measure levels of genes responsible for cancer and other diseases. The Company is commercializing in vitro diagnostic tests for use in companion diagnostic applications and real-time monitoring of disease
Exosomes & Custom Biomarker Development
Exosome technology allows for biomarker discovery, development and validation in biofluid clinical samples, including plasma, serum, CSF and urine. As opposed to using cell culture models, detecting biomarkers in exosomes allows for direct access to clinically relevant, unmanipulated disease information. Cells throughout the body shed exosomes into fluids as a natural delivery system to other cells. Exosomes carry RNA (full length, stable mRNA as well as miRNA and other non-coding RNAs), as well as proteins and DNA, responsible for commanding disease growth that can be accessed easily and repeatedly from a biofluid for analysis by array, PCR or next-generation sequencing.
Exosomes are remarkably stable; fresh or frozen archival samples can be used with equivalent results. Required sample volumes range from 500µL to 30mL, depending on the biofluid, downstream application and project scope (single biomarker vs. full scale array or sequencing panel). All preservation methods (EDTA, Heparin, etc.) are compatible.
Exosome technology has the potential to allow doctors, researchers and drug developers to take repeated genetic snapshots of disease for diagnostic and therapeutic use without the need for a surgical tissue biopsy.
Exosome Diagnostics partners with diagnostic and pharmaceutical companies to develop custom exosome-based biofluid assays for use via qPCR, sequencing and array instruments for use with frozen archival and prospective clinical samples.
Not ESI But looks like the same... Circa 2013?
Exosome Diagnostics Presents Data Demonstrating Clinical Applications of Exosome Technology in Brain Cancer, Prostate Cancer, and Alzheimer's Disease at the International Society for Extracellular Vesicles 2013 Annual Meeting
NEW YORK April 16, 2013 /PRNewswire/ — Exosome Diagnostics, a developer of biofluid-based molecular diagnostic products for use in personalized medicine, today announced data demonstrating exosome technology performance in clinical applications which will be presented at the International Society for Extracellular Vesicles 2013 (ISEV 2013) annual meeting, April 16–20, in Boston. The ISEV meeting is the preeminent international conference in the rapidly growing field of exosome and microvesicle research.
"The ability to deliver full-length, high quality mRNA and microRNA from biofluids is critical to development of new research tools and clinical grade in vitro diagnostic tests not subject to the limitations of tissue biopsy," said Dr. Johan Skog, chief science officer of Exosome Diagnostics. "Our Alzheimer's disease findings are especially pertinent to advancing a molecular diagnostic test capable of detecting CNS disorders years before the onset of clinical symptoms when therapeutic intervention could provide the most benefit."
Dr. Skog will give two oral presentations. The first is entitled "Characterization of Exosome RNA as a Tool for Molecular Diagnostics", and will be given at 9:30 a.m. EDT on April 16 during the Frye Halloran Educational Symposium. The latest data on Exosome Diagnostics' proprietary biofluid RNA capture technology for disease biomarkers in urine, blood and cerebrospinal fluid (CSF) will be presented. The second, to be given at 9:45 a.m. EDT on April 20, is entitled "The Use Of CSF Microvesicle RNA For Diagnosis Of Alzheimer's Disease". This presentation describes differences in CSF exosome microRNA expression associated with Alzheimer's disease and patients with subjective memory complaints but no evidence of Alzheimer's disease.
James McCullough, chief executive officer of Exosome, said, "We are committed to enabling the research community with high-performance exosome technology RNA isolation kits beginning in 2014 for biomarker discovery from fresh and frozen biofluids. In addition, we are continuing to advance multi-center in vitro diagnostic development programs in genitourinary (GU) Oncology and CSN disorders."
Exosome Diagnostics' proprietary technology provides access to a previously unexplored source of genetic information directly from diseased cell populations. Cells throughout the body shed exosomes into fluids as a natural package delivery system to other cells. Exosomes carry key genes (mRNA and miRNA transcript) responsible for commanding disease growth that can be accessed quickly and repeatedly from a biofluid for analysis on PCR or next-generation sequencing instruments. Exosome technology has the potential to allow doctors, researchers and drug developers to take repeated, accurate genetic "snapshots" of disease for diagnostic and therapeutic use without the need for a surgical tissue biopsy.
Exosome Diagnostics will also be presenting a poster on an additional study:
Abstract #37, 8-5:30pm EDT, Wednesday, April 17th: Utility of Urinary Exosomes/Microvesicles to Non-Invasively Analyze Gene Expression in Prostate Cancer
L.M. Russo, A.N. Scott, P. Motamedinia, K.L. Bate, N. Sadeghi, G. Salazar, M. Lipsky, J. Lin, G. Hruby, K.K. Badani, D.P Petrylak, J.M McKiernan
About Exosome Diagnostics
Exosome Diagnostics is a leading developer of biofluid-based molecular diagnostic tests for use in personalized medicine. Exosomes are packaged and shed into all biofluids, including blood, urine and CSF, providing a stable source for intact, cell-specific nucleic acids. The Company's proprietary exosome technology makes use of the presence and natural stability of RNA in exosomes to detect and measure levels of genes responsible for cancer and other diseases. The Company is commercializing exosome technology products for use in both the life sciences and clinical settings.
Deaths from Alzheimer’s underreported, study says
Alzheimer’s disease is currently the sixth leading causes of death in the United States, according to the Centers for Disease Control and Prevention (CDC). But now, new research published in Neurology suggests that Alzheimer’s disease may actually be responsible for as many deaths each year as heart disease or cancer – the two leading causes of death in the U.S.
The disease, the most common form of dementia, tends to be underreported as a cause of death because it leads to other conditions that are then listed on death reports.
“The most common thing people with Alzheimer’s die of is pneumonia— the part of the brain that controls breathing and eating starts to dysfunction and eventually more infections [occur] in the lungs through food particles or just not being able to breath properly,” researcher Bryan D. James, an assistant professor and epidemiologist at the Rush Alzheimer’s Disease Center at Rush University Medical Center in Chicago, told FoxNews.com.
While about one-third of Alzheimer’s patients die of pneumonia, nearly that many die from heart failure, as the brain’s ability to control the heart and other organs starts to slowly shut down.
To more accurately understand death rates from Alzheimer’s, James and his team conducted an observational study of 2,566 volunteers, all age 65 or older. At the start of the study, participants underwent cognitive testing and were then re-tested annually. Researchers used those tests, combined with family history, to identify participants with dementia. They then followed participants for an average of eight years, though they observed some for up to almost 20 years.
A total of 559 participants developed Alzheimer’s disease during the course of the study. Participants with Alzheimer's who were between ages 75 to 84 had a death rate that was more than four times higher than participants without Alzheimer's disease. Furthermore, participants over age 85 with Alzheimer’s had a death rate nearly three times higher compared to the rest of the group. Overall, Alzheimer’s disease contributed to more than one-third of all deaths in participants over age 75.
According to researchers, their findings translates into an estimated 503,400 deaths from Alzheimer’s in Americans over age 75 in 2010 – five to six times higher than the 83,494 deaths due to Alzheimer’s reported by the CDC.
Researchers say their data also points to the fact that a single cause of death is not always accurate for older adults. While a diagnosis of Alzheimer’s disease is usually associated with loss of memory— misplacing keys, forgetting names— as the disease progresses, it affects other parts of the brain that control unconscious functions such as breathing, swallowing and heart rate. Eventually, it begins to affect the part of the brain that controls basic function, which can lead to complications like pneumonia.
“Really what’s happening is you’re slowly deteriorating from this disease in your brain,” James said. “It’s a fatal disease. Until we come up with a cure or an effective treatment, eventually people will succumb to the disease or pass away. [Patients have] 3 to 10 years, average, to live, after diagnosis.”
The findings emphasize the need for public awareness about the disease.
“From a public health point of view, it points to the fact that this is a major risk factor for our society and because it’s so linked to the aging process and we know baby boomers are getting older now and we’re going to have a rapidly expanding aging population, this is going to be a more important issue in the future, as well,” Dr. James Leverenz, director of the Cleveland Clinic Lou Ruvo Center for Brain Health in Las Vegas, who was not involved in the study, told FoxNews.com. “[This study] is something that really puts the highlight on how important Alzheimer’s is to our public health.”
While it may seem morbid that study participants volunteered for research on their own mortality, James said their efforts will pay off for future generations.
“[They’re] giving back to science, giving back to their grandchildren,” he said. “Everyone knows we may not find anything to help prevent disease or anyone currently in their 80s, but for their grandchildren, this could be extremely helpful.”
www.foxnews.com/health/2014/03/06/deaths-from-alzheimers-underreported-study-says/?intcmp=obnetwork
TITLE: IL-6 Receptor Isoforms and Ovarian Cancer
PRINCIPAL INVESTIGATOR: Susan E. Waltz, Ph.D.
CONTRACTING ORGANIZATION: University of Cincinnati
Cincinnati, OH 45221
REPORT DATE: January 2013
TYPE OF REPORT: Final
PREPARED FOR: U.S. Army Medical Research and Materiel
Award Number: W81XWH-09-1-0673
Introduction
The overall goal of the proposed study has been to test the hypothesis that membrane and soluble IL6R play distinct roles in driving tumor progression. Two major Tasks were funded as
outlined in the Statement of Work, which encompasses examining IL6R expression in ovarian patient tumor samples and performing experiments to define the function of IL6R in the tumor
proper and host microenvironment.
Body
The overall goals have been accomplished and the data published related to Task 1 (1). Task 1 entailed the “Identification of IL6R isoforms in patient samples.” In the reported studies,
elevated levels of IL6, total IL6R, differential spliced forms of IL6R, ADAM10 and ADAM17 were found in ovarian tumors. In addition, these studies further demonstrated that both tumor and
host cells contribute to soluble IL6R expression in the tumor microenvironment. Task 2 entailed “In vivo experiments to determine functions of IL6R isoforms and tumor:stroma
relationships. For this task, mouse breeding colonies and genotyping procedures were set up.
A recent paper was also published related to the characterization and general phenotype of these mice (2). Xenograft studies using ovarian cancer cell lines in SCID mice demonstrated that host IL6R may be an important means by which IL6R levels are increased in the tumor microenvironment along with the importance of IL6R in the tumor proper (1). Further studies have been published which also show that overexpression of IL6R and to a more modest extent IL6, in ovarian cancer cells promotes their colonization on the omentum ex vivo and increased have been published which also show that overexpression of IL6R and to a more modest extent
IL6, in ovarian cancer cells promotes their colonization on the omentum ex vivo and increased
their adherence to plastic as well as plates coated with laminin and collagens I and II (3). To test the significance of host derived tumor growth, ovarian tumor cells were next injected
intraperitoneally (IP) into mice deficient in either IL6 or IL6R. In examining tumor adherence to the omentum following IP injection, mice with a complete loss of IL6 or IL6R exhibited less
ovarian cancer cell adherence to the omentum. To obtain information as to the host cell type in which IL6R expression is needed for tumor adherence, ovarian cancer cells were injected into mice with conditional losses of IL6R in either hepatocytes or in myeloid cells (monocytes/granulocytes). Interestingly, tumor adherence was reduced in both hosts with IL6R loss, although IL6R loss in host myeloid cells lead to a greater inhibition of tumor adherence to the omentum compared to loss in IL6R from hepatocytes. At this point, the project was halted
as the initial PI left the institution. A few months later, a new PI was designated (Susan Waltz) and the mouse breeding colonies were re-initiated. Unfortunately, having to regenerate the animal colonies entailed approximately 8-12 months to obtain the complex IL6R-/- mice and the IL6R conditional mutants as the original colonies were sacrificed prior to the designation of a new PI. Over the course of the past year, the focus of the new PI has been to generate the appropriate mouse colonies and cell lines to continue experiments designated in Task 2. The primary goal of these final studies has been to examine the significance and potential mechanisms associated with host IL6R on ovarian cancer growth and metastasis. The following is a synopsis of the final data that has been obtained since the new PI was designated through completion of the funded studies:
have been published which also show that overexpression of IL6R and to a more modest extent IL6, in ovarian cancer cells promotes their colonization on the omentum ex vivo and increased
their adherence to plastic as well as plates coated with laminin and collagens I and II (3). To test the significance of host derived tumor growth, ovarian tumor cells were next injected
intraperitoneally (IP) into mice deficient in either IL6 or IL6R. In examining tumor adherence to the omentum following IP injection, mice with a complete loss of IL6 or IL6R exhibited less
ovarian cancer cell adherence to the omentum. To obtain information as to the host cell type in which IL6R expression is needed for tumor adherence, ovarian cancer cells were injected into mice with conditional losses of IL6R in either hepatocytes or in myeloid cells (monocytes/granulocytes). Interestingly, tumor adherence was reduced in both hosts with IL6R loss, although IL6R loss in host myeloid cells lead to a greater inhibition of tumor adherence to the omentum compared to loss in IL6R from hepatocytes. At this point, the project was halted
as the initial PI left the institution. A few months later, a new PI was designated (Susan Waltz) and the mouse breeding colonies were re-initiated. Unfortunately, having to regenerate the
animal colonies entailed approximately 8-12 months to obtain the complex IL6R-/- mice and the IL6R conditional mutants as the original colonies were sacrificed prior to the designation of a
new PI. Over the course of the past year, the focus of the new PI has been to generate the appropriate mouse colonies and cell lines to continue experiments designated in Task 2. The primary goal of these final studies has been to examine the significance and potential mechanisms associated with host IL6R on ovarian cancer growth and metastasis. The following is a synopsis of the final data that has been obtained since the new PI was designated through completion of the funded studies:
IL6 and IL6R stimulate migration in ovarian overexpressing IL6R exhibited increased migration/invasion within the peritoneal
cavity (Figure 1). To follow up on increased migratory
ability of tumor cells overexpressing IL6R, we next sought to determine whether IL6 signaling regulates tumor cell migration. To accomplish this, ES-2 human ovarian cancer cells, as well as
ES-2 cells overexpressing IL6 or IL6R were plated on tissue culture plates and scratch wounds were made. Over the course of the experiment, photographs of the cultures were taken temporally and the percent migration was measured based on wound closure. As depicted in Figure 2, both IL6 and IL6R overexpression lead to significantly more cell migration compared to control cells. Slight increases, although not significant, were observed following the addition of exogenous IL6 or soluble IL6R. This data suggests that tumor cell derived IL6/IL6R signaling may be an important factor regulating the local migration/invasive ability of ovarian caner cells and that further supplementation of IL6 or IL6R may only marginally be required beyond that present in the tumor cell proper to increase this effect.
Jump to the end...
The combination of the specific tumor microenvironment,
ascitic dissemination pathways, and early omentalcolonization may create a situation in which LY75 expression favors the colonization of ovarian tumors. Increased expression of IL6 and IL6Ra in patients with ovarian cancer may enhance LY75 expression in tumor cells and facilitate tumor adherence and colonization of the omentum. Indeed, we found a previously unreported correlation
between IL6Ra and LY75 expression in patients with ovarian cancer in large public databases, providing some clinical significance for our findings. Our study therefore demonstrates a novel mechanism by which IL6 signaling may contribute to tumor progression and suggests an opportunity for reducing the mortality associated with ovarian cancer.
http://www.dtic.mil/dtic/tr/fulltext/u2/a575873.pdf
Interleukin-6 Cytokine: A Multifunctional Glycoprotein for Cancer
www.omicsonline.com/open-access/1745-7580/1745-7580-9-062.pdf
Abstract
Interleukin 6 is a multifunctional cytokine. Its increased levels have been associated with elevated cancer risk,
and also these levels have been found to be a prognostic factor for several cancer types. In addition, increased
levels have been found in coronary heart disease, insulin resistant patients, advance stage cancer patients, atopy/
asthma and in patients with blood circulating micrometastasis. Additionally several studies with different types of
cancers have been performed to identify the correlation between interleukin-6 levels, stage, treatment response and
severity of symptoms. The influence of interleukin-6 is performed mainly through the janus kinase-signal transducer
and activator of transcription-zinc finger protein 1-2 signaling pathway. As a result, the increased levels of interleukin-6
are responsible for enhanced neo-angiogenesis, inhibition of cancer cell apoptosis and deregulation of the control
mechanisms in the microenvironment. In addition, increased levels of inteleukin-6 have been found to increase the
production of collagen and a-actin which induce interstitial lung disease. In the current mini review we will present
information regarding the interleukin-6 and published results in several cancer studies and finally we will comment in
future treatment approaches blocking this cytokine in cancer patients.
Introduction
Interleukin-6 (IL-6) is a multifunctional cytokine that influences
the activity of cancer cells. It is a glycoprotein which consists of 184
amino acids and has 26 kilodaltons (kDa) as a molecular weight [1]. It
is involved in tumor growth, malignant differentiation of cancer cells
and microenvironment immunomodulation [2]. These properties are a
result of enhanced neo-angiogenesis, inhibition of cancer cell apoptosis
and acquired cell resistance [3]. Again these results are mediated through
several signaling pathways, however; the most important ones are the
transcription activator 3 and the signal transducer [4-9]. Interleukin-6
levels have been observed elevated in advanced stage cancer [10]
and again elevated levels have been associated with increased risk of
cancer [11,12]. Moreover; IL-6 has been found to be a cancer disease
prognostic marker [13] and also it was increased in patients with cancer
induced cachexia [14]. Interleukin-6 has been found overexpressed in
cancer cells lines expressing epidermal growth factor receptor (EGFR)
mutation [4], however; in another study further investigation of the
levels of IL-6 after tyrosine kinase inhibitors (TKIs) indicated that
TKIs induce apoptosis, but at the same time increase IL-6 levels [15].
During the cancer, cell apoptosis growth factors, which stimulate the
IL-6 release, circulate in the extracellular matrix. The increased levels
of interleukin-6 subsequently induced collagen and a-actin production
and interstitial lung disease pattern was observed. In addition, there are
few data whether IL-6 levels are associated with treatment effectiveness
or cancer stage for patients with EGFR mutations [10]. IL-6 receptor
mutation has been investigated in mice and it was observed that the
animals carrying this mutation had additionally overexpression of
signal transducer and activator of transcription 3 (STAT3) activities. In
these animals the extent of the infiltrates was directly associated with
the overexpression of the stat-3 signaling pathway [16]. Furthermore;
increased levels of IL-6 have been associated with increased lung cancer
risk in lifetime non-smoking women with asthma/atopy [17]. Increased
levels of IL-6 have been found in patients with coronary heart disease
[18] and insulin resistance has been reported from increased levels of
IL-6 in a lung cancer patient [19]. Several trials have been performed
investigating the IL-6 levels and the effects on the prognosis, stage,
treatment outcome and cancer relapse. Regarding surgically resected
patients the higher the interleukin-6 levels after the first day, the sooner
was the relapse [20]. These studies can be summarized according to
each cancer type as follows: a) ocular [21], b) lung EGFR mutation
positive adenocarcinomas [4,15], c) colorectal [22], d) small cell lung
cancer [3], e) brain [2,23], f) cancer stem cells [24,9], g) head and neck
[25], h) hepatocellular [26], i) breast [27], j) melanoma [28], k) bone
[29] and l) nasopharyngeal cancer cells [30].
Search Methods
We performed an electronic article search through PubMed,
Google Scholar, Medscape, and Scopus databases, using combinations
of the following keywords: interleukin-6, cancer, angiogenesis, and
cytokine. All types of articles (randomized controlled trials, clinical
observational cohort studies, review articles, case reports) were
included. Selected references from identified articles were searched for
further consideration, without language limitation.
IL-6 targeted treatment
Targeting the signaling pathway of the IL-6 is a novel target. The
tissue inhibitor of metalloproteinase-3 (TIMP-3) is a possible target.
It has been previously observed that low TIMP-3 and high IL-6
levels patients had lower survival [33]. The novel (2E,5E)-2,5-bis(4-
(3(dimethylamino)-propoxy) benzylidene) cyclopentanone (A13) has
been observed to reduce nitric oxide (NO), TNF-a, HGF and IL-6. In
addition, it reduces damages to the lung and reduced pain in several
induced inflammatory models [34]. Neutralizing IL-6 antibodies have
been used to block the JAK1-STAT3 signaling pathway in a lung cancer
model [5]. Tocilizumab an anti-IL-6R antibody has been used and
apart from reducing the lung cancer tumorigenic activities it was also
observed to reduce the cancer related cachexia [14]. Toll like receptor 2
is a novel molecule under consideration as a future immunomodulatory
therapy for targeting tumor inflammation. Previous positive results
have been published in a melanoma model [35]. Moreover, another
novel IL-6R blocker which is under investigation is siltuximab [36].
This novel agent has been used also in combination with docetaxel
chemotherapy [37]. The IL-6R antibodies will be soon replaced by
nonantibody-based inhibitors called avimers. Their main advantage
is that they do not induce allergic reactions and have more prolonged
activity in comparison to the antibodies (> two weeks). Currently the
AMG-220 is being developed by AVIDA [38].
Discussion
Inflammation cytokines such as: a) C-reactive protein, b) IL-6 and
IL-8have been identified as lung cancer markers. Increased plasma
levels of these cytokines have been associated with increased for risk
lung cancer [39,12,11]. However, only IL-8 has been identified as
an early lung cancer biomarker (more than 2 years whereas IL-6 for
<2 years) [11]. Also in never smokers, different polymorphisms of
inflammatory pathway genes are also related with increased lung cancer
risk, especially IL-6, IL-1ß and IL-1RN [40]. Increased levels of IL-6
have been observed in advanced stage cancer in several cancer types
[41]. Caregivers of cancer patients with advanced stage lung cancer
have also high levels of IL-6 and other inflammatory markers, which
are reduced when the cancer patient is treated efficiently [41]. A joint
effect of asthma/atopy and IL-6 gene polymorphism in lung cancer
has been identified in lifetime non-smoking Chinese women [17]. The
IL-6 levels have been also associated with coronary heart disease and
efforts have been made to target the IL-6 receptor as a treatment for
coronary heart disease [18]. Moreover, IL-6 levels have been identified
as a marker for EMT malignant transition in lymph nodes [25]. Insulin
allergy and immunologic insulin resistance has been identified in a
patient with lung cancer in the pre-chemotherapy period. In specific
due to excessive IL-6 plasma levels >96.4 pg/ml (reference range <4 pg/
ml) local skin irritation (erythema) was observed in the site of insulin
administration and the HbA1c levels were deregulated. In addition,
increased IgG anti-insulin antibodies were observed. All the excessive
laboratory parameters return to normal after chemotherapy initiation [19].
Moreover, different pro-inflammatory and fibrogenic cytokines
have been investigated in correlation to radiation induced lung toxicity
(RILT) and it was observed that low pretreatment levels of IL-8 were
correlated with the development of RILT. In addition, if IL-8 and
TGF-ß1 levels were combined, then an independent prognostic cluster
was formed for RILT development [42]. Increased levels of IL-6,
collagen and a-actin have been found after TKI inhibitor treatment in
lung cancer cell lines and tongue cancer HSC-3 cells. The treatment
efficiency was associated with the fibrogenic activity [15]. Again in
the study by O`Donoghue in interstitial fibrosis patients elevated
levels of IL-6 were correlated with overexpression of STAT3 signaling
pathway and enhanced fibrogenic activity. Therefore, fibrotic lesions are
expected when IL-6 levels are elevated; however, no clear association
has been made between the levels and extent of the lesions. There is
however literature for stage IIIB and IV lung cancer patients regarding
their treatment response and IL-6 serum levels. It has been observed
that increased levels of circulating IL-6 are a prognostic factor for poor
treatment response [10]. The raltitrexed, which is a specific thymidylate
synthase inhibitor when administered in colorectal cancer patients
with or without chemotherapy activated a cascade of cytokines and
chemokines and the identification of infection in these patients was
impossible. Again, we would like to have data regarding the fibrogenic
activation in these patients with radiologic findings [22]. Neutralizing
IL-6 antibodies are on the market and they have been observed to
reduce cancer related cachexia [14]. However, we would like to have a
study with sleep disturbances and IL-6R antibody treatment. Regarding
the factors that are mediated by IL-6 in several cancer types, we can
summarize them to the following. a) Multiple myeloma (MyC regulator
gene, STAT3, fibroblast growth factor (FGFR))[43], b) lung cancer
(EGFR, STAT3) [4], c) colon cancer (STAT3, c-Myc) [44], d) prostate
cancer (IGF-1R, Human Epidermal Growth Factor Receptor 2 (Erb2))
[45], e) breast cancer (Notch, Rat sarcoma (Ras), Human Epidermal
Growth Factor Receptor 2 (HER2)) [46], f) ovarian cancer (Stat3,
VEGF) [47] and g) bladder cancer nuclear factor kappa-light-chainenhancer
of activated B cells (Nf-kappaB) [48].
Finally, the immune system has a dual role, it can either enhance
the tumor growth and tumor metastasis, however, it can also protect
the body from carcinogenesis and metastasis [49]. Therefore, a balance
is necessary since not all cytokines are responsible for enhancing the
tumor microenvironment and tumor activities.
CytoSorbents to Exhibit at the 43rd Annual Meeting of the German Society for Thoracic, Heart and Vascular Surgery.
CytoSorbents Corporation
55 minutes ago
....MONMOUTH JUNCTION, NJ--(Marketwired - Feb 5, 2014) - CytoSorbents Corporation (OTCQB: CTSO), a critical care immunotherapy company, announced that it will exhibit at the 43rd Annual Meeting of the German Society for Thoracic, Heart and Vascular Surgery in Freiburg, Germany from February 9-12, 2014. The conference is in cooperation with the Austrian and Swiss Societies for Thoracic and Cardiovascular Surgery.
Cardiothoracic surgery accounts for approximately 1.5 million open-heart surgeries performed worldwide each year, for procedures such as coronary artery bypass graft surgery (for blocked coronary arteries), valve replacement surgery, left ventricular assist device implantation (for end-stage heart failure), and heart and/or lung transplant. Former President Bill Clinton and comedian David Letterman are amongst those notable for having open-heart surgery for heart disease.
In order to operate on the heart, the heart is typically stopped, with blood shunted away from the heart and lungs, to a cardiopulmonary bypass (CPB) machine, also known as a heart-lung machine, outside of the body. The machine oxygenates the blood and then pumps the blood at high pressure back to the rest of the body. Patients can develop serious post-operative complications such as kidney and lung failure due to the production of inflammatory cytokines, and the release of free hemoglobin by damaged red blood cells.
CytoSorb® is the only specifically approved extracorporeal cytokine filter in the European Union that can be easily installed into a bypass circuit of the CPB machine without the need for an additional blood pump. CytoSorb® directly removes a broad range of cytokines and free hemoglobin from blood, and is a superior drop-in replacement for leukoreduction filters, an existing commercial product that tries, but cannot remove cytokines or free hemoglobin directly. CytoSorb® is being used today in a growing number of major heart centers throughout Germany and Austria, in both regular clinical usage and in investigator initiated studies. It is used both intra-operatively, as well as during the recovery phase in patients that develop a post-operative systemic inflammatory response syndrome (SIRS). Cardiac surgery represents a total addressable market for CytoSorb® of $0.5 - 1.0 billion in Europe and the U.S. alone.
Dr. Christian Steiner, MD, Vice President of Sales and Marketing of CytoSorbents, stated, "This conference is the most important cardiothoracic surgery meeting in the German speaking community. It attracts cardiac surgeons and heart-lung machine perfusionists from across the region. In order to accelerate sales of CytoSorb® for this segment, we recently hired a very experienced cardiac surgery sales representative with extensive key opinion leader contacts in our direct markets. Her focus will significantly add to the momentum generated by the rest of the sales team in this application. We benefit from encouraging early clinical data generated by our collaborators, a unique competitive position, and a strong value proposition. We see cardiac surgery as a very robust additional market to the critical care applications that we continue to pursue aggressively."
About CytoSorbents Corporation
CytoSorbents is a critical care focused immunotherapy company using blood purification to modulate inflammation -- with the goal of preventing or treating multiple organ failure in life-threatening illnesses. Organ failure is the cause of nearly half of all deaths in the intensive care unit, with little to improve clinical outcome. CytoSorb®, the Company's flagship product, is approved in the European Union as a safe and effective extracorporeal cytokine filter, designed to reduce the "cytokine storm" that could otherwise cause massive inflammation, organ failure and death in common critical illnesses such as sepsis, burn injury, trauma, lung injury, and pancreatitis. These are conditions where the mortality is extremely high, yet no effective treatments exist. In addition, CytoSorb® can be used in other inflammatory conditions such as cardiac surgery and autoimmune disease flares. CytoSorbents' purification technologies are based on biocompatible, highly porous polymer beads that can actively remove toxic substances from blood and other bodily fluids by pore capture and surface adsorption. CytoSorbents has numerous products under development based upon this unique blood purification technology, protected by 32 issued US patents and multiple applications pending, including HemoDefend™, ContrastSorb, DrugSorb, and others. Additional information is available for download on the Company's website: http://www.cytosorbents.com/
Forward-Looking Statements
This press release includes forward-looking statements intended to qualify for the safe harbor from liability established by the Private Securities Litigation Reform Act of 1995. Forward-looking statements in this press release are not promises or guarantees and are subject to risks and uncertainties that could cause our actual results to differ materially from those anticipated. These statements are based on management's current expectations and assumptions and are naturally subject to uncertainty and changes in circumstances. We caution you not to place undue reliance upon any such forward-looking statements. Actual results may differ materially from those expressed or implied by the statements herein. Risk factors are detailed in the Company's Form 10-K filed with the SEC on April 3, 2013, which is available at http://www.sec.gov.
Health Care IndustryHealthCardiothoracic surgeryred blood cellsvalve replacement surgeryEuropean Union
Contact:
Company
CytoSorbents Corporation
Dr. Phillip Chan
Chief Executive Officer
(732) 329-8885 ext. *823
Its all about the NOC deal:
3. Exclusivity of Agreement
In view of the close cooperation which shall be required, the expenditures which shall be incurred and the necessity to exchange confidential and proprietary business and technical information for the purpose of this Agreement, CIRQUE agrees that during the term of this Agreement, CIRQUE shall not participate in any manner in the design, manufacture, assembly, test and commission of a prototype gasifier unit similar in specification, design or intent, or any part thereof, by itself or by any third party, relating to or competitive with the Program, or provide any services, data, information or other assistance to any third party in furtherance thereof or enter into any agreement with any third party for the provision of services or equipment relating to or competitive with the Program.
How does this part of the NGC agreement affect the DPS contracts?
3. Exclusivity of Agreement
In view of the close cooperation which shall be required, the expenditures which shall be incurred and the necessity to exchange confidential and proprietary business and technical information for the purpose of this Agreement, CIRQUE agrees that during the term of this Agreement, CIRQUE shall not participate in any manner in the design, manufacture, assembly, test and commission of a prototype gasifier unit similar in specification, design or intent, or any part thereof, by itself or by any third party, relating to or competitive with the Program, or provide any services, data, information or other assistance to any third party in furtherance thereof or enter into any agreement with any third party for the provision of services or equipment relating to or competitive with the Program.
Joint Development Agreement
This JOINT DEVELOPMENT AGREEMENT, made and entered into as of the date last executed, by and between Northrop Grumman Systems Corporation, a Delaware corporation, acting through its Information Systems Sector, Defense Systems Division, with a place of business at 201 Electronics Blvd., Huntsville, Alabama 35824 (hereinafter “Northrop Grumman”) and Cirque Energy, Inc., with a place of business at 243 W. Congress St, Suite 350, Detroit, MI 48226 (hereinafter referred to as “CIRQUE”).
Northrop Grumman has certain technical knowledge and trade secrets that were used by Cirque Energy, Inc., a subcontractor, to create a conceptual Deployable Gasification Unit (DGU) capability on behalf of Northrop Grumman. Now, Northrop Grumman wishes to continue technology development for the DGU and towards this end, desires to use the capabilities of Cirque Energy in order to realize a working DGU prototype for testing and demonstration. In consideration for Cirque Energy to fund and develop a test unit, NG wishes to permit its Intellectual Property to be used by Cirque Energy; and provided a prototype is a success and the device is deemed viable, a division of responsibilities, obligations and customer sales privileges is contemplated and is discussed within Exhibit A.
WITNESSETH:
WHEREAS, Both Parties are working on a Mobile Energy / Power Generation Initiative to develop a Deployable Gasification Unit (DGU) capable of producing electricity at tactical field locations or in support of disaster relief and humanitarian assistance operations (hereinafter referred to as the “Program”); and
WHEREAS, Both Parties desire to construct a working prototype of a Deployable Gasification Unit (DGU) to demonstrate such a capability is not only feasible, but more efficient than existing tactical power generation when relating output power to input fuel rates; and
WHEREAS, Northrop Grumman has certain rights and ownership, capabilities and certain know-how regarding a Deployable Gasification Unit (DGU), whereby Northrop Grumman has invested significant research and development funding and labor to design a tactical DGU capable of converting waste byproducts into electricity using largely off-the-shelf technology and components.
WHEREAS, Northrop Grumman further conceptually developed and documented the DGU design in the “Proof of Concept Study” dated December 7, 2012 (Exhibit C) with CIRQUE as a subcontractor to Northrop Grumman in order to establish the requirements for the Program, and the Parties agreed that the Intellectual Property documented in the proof of concept study (Exhibit C) are owned by Northrop Grumman; and
WHEREAS, CIRQUE has certain capabilities and expertise in the area of energy development as it relates to the procurement, processing and transporting of biomass materials from multiple waste environments (including landfills), material handling and feedstock preparation systems necessary for both Static and Mobile Systems of various types of gasification technologies; with the most recent advancements in gasification technology including; demonstration units, technical papers and research that are necessary to meet the requirements for the Program; and
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WHEREAS, CIRQUE has resources to fund the manufacturing and test of a working Prototype DGU and support demonstrations; and
WHEREAS, neither Northrop Grumman nor CIRQUE individually possesses all of the capabilities, resources, expertise or existing capacity to address all of the requirements of the Program without assistance from each other; and
WHEREAS, the combination of the capabilities, resources, expertise and existing capacity of Northrop Grumman and CIRQUE should result in a working DGU prototype best meeting the technology identified in Exhibit C which supports the Program; and
WHEREAS, Northrop Grumman and CIRQUE desire to form a Joint Development Agreement that is not in derogation of the applicable antitrust laws and does not prejudice the Program in any way with respect to any action it may take in procuring goods or services on the basis of awarding of contracts on a leader/follower or other type basis; and
WHEREAS, Northrop Grumman and CIRQUE may need to exchange significant proprietary and sensitive information in the process of CIRQUE manufacturing and testing the prototype for the Program, which exchange of information would not be possible if Northrop Grumman and CIRQUE were working with other companies in support of the Program;
NOW, THEREFORE, Northrop Grumman and CIRQUE hereby agree as follows:
1. Definitions
1.1. “Agreement” shall mean this Joint Development Agreement, together with Exhibit A, Exhibit B, Exhibit C, and other documents incorporated by reference and any amendments thereto.
1.2. “Parties” shall mean Northrop Grumman and CIRQUE collectively, and “Party” shall refer to either Northrop Grumman or CIRQUE individually.
2. Activities
2.1 Each Party will exert all reasonable efforts to develop a DGU prototype which will meet the details (including cost objectives) of Exhibit C.
2.2 CIRQUE will lead the design, manufacturing, development, testing, site location (including transportation thereof), and prototype development and demonstration process. This will include involvement by Northrop Grumman as they desire.
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2.3 CIRQUE will cooperate with Northrop Grumman by providing such assistance as may be required during the manufacturing of the DGU Prototype, including, but not limited to: (i) furnishing a status within reasonable timeframes of CIRQUE established goals and achievements of those goals (to include testing and test results); (ii) assuring the availability of management, pricing, and technical personnel for potential program related meetings; (iii) submitting a Rough Order of Magnitude estimate for the material required to manufacture the DGU; and (iv) a written manufacturing process that was utilized for the prototype to make it a repeatable process for production. CIRQUE will provide 100 percent (100%) of the funds required to accomplish the above and demonstrate to Northrop Grumman the DGU Prototype.
2.4 In consideration for CIRQUE to fund and develop a test unit, Northrop Grumman wishes to permit its Intellectual Property to be used by CIRQUE for DGU development. Further, if the DGU Prototype is determined acceptable and suitable by Northrop Grumman, Exhibit A hereof shall be used as the basis to draft additional agreements for the DGU.
2.5. The Parties recognize that, during the term of this Agreement, conditions relating to the Program may change such as to dictate a change in the scope of the work set forth in Exhibit A or in the continuation of the Program by either Party.
Therefore, prior to any revisions to Exhibit A by Northrop Grumman, CIRQUE will, upon request, enter into good faith negotiations with Northrop Grumman to revise Exhibit A hereof to increase, decrease or clarify the work hereunder. Northrop Grumman agrees not to initiate such request, unless it has a good faith belief that such is necessary, and will in such event advice CIRQUE of the basis for such belief. In the event, after such Northrop Grumman request, the Parties are unable to reach mutual agreement as to an appropriate revision to Exhibit A, either Party may, upon ten (10) days prior notice to the other Party, terminate this Agreement, unless within such ten (10) day period Northrop Grumman withdraws the request or mutual agreement upon a revision is reached.
3. Exclusivity of Agreement
In view of the close cooperation which shall be required, the expenditures which shall be incurred and the necessity to exchange confidential and proprietary business and technical information for the purpose of this Agreement, CIRQUE agrees that during the term of this Agreement, CIRQUE shall not participate in any manner in the design, manufacture, assembly, test and commission of a prototype gasifier unit similar in specification, design or intent, or any part thereof, by itself or by any third party, relating to or competitive with the Program, or provide any services, data, information or other assistance to any third party in furtherance thereof or enter into any agreement with any third party for the provision of services or equipment relating to or competitive with the Program.
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4. Acceptance & Suitability of DGU Prototype
Provided the DGU Prototype is determined to be commercially viable by Northrop Grumman, Exhibit A hereof is provided to document additional agreements for the DGU.
Either Party will, in the event of an acceptable and suitable DGU Prototype, accept a subcontract to perform work and render services in accordance with this Agreement and Exhibit A hereto, subject to mutual agreement on prices and other terms and conditions. In the event mutually acceptable prices and other subcontract terms and conditions or a Letter Subcontract cannot be negotiated by the Parties within a reasonable time, and in any event within 60 days after award of a contract to a Party, the other Party shall have the right, upon ten (10) days prior notice to terminate this Agreement and make other arrangements for the performance of the work in Exhibit A, in which case the rights and obligations of both Parties under this Agreement will terminate pursuant to Article 5.4 below. This right is in addition to other rights the Parties may have hereunder or under applicable law.
Any such resulting subcontract or changes to this agreement, shall be subject to applicable laws, regulations and required or reasonably implied flow-down terms and conditions of the contract, mutual agreement on pricing and other subcontract terms and conditions, and prior approval of Customer, if required.
5. Termination
This Agreement and all rights and duties hereunder, will terminate upon the first of the following events to occur:
5.1. Any material change to CIRQUE capabilities or other attributes (e.g., size status; financial stability) that is significant to the basis on which Northrop Grumman has entered into this Agreement, as reflected in the recitals to this Agreement;
5.2. The failure of the CIRQUE to provide the effort defined in Article 4, above, or provide such assistance in a timely manner or of acceptable quality;
5.3. Mutual consent of both Parties by execution of a rescission agreement;
5.4. Ten days after the issuance of the written notice required by Article 4 when the Parties fail to consummate a subcontract as contemplated herein;
5.5. Written notification of a decision by either Party not to continue with the Program;
5.6. A filing of a bankruptcy petition or other material adverse financial change by either Party;
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5.7. A written determination by Northrop Grumman that an actual or potential Organizational Conflict of Interest (OCI) for either Party pursuant to Article 24 has failed to be mitigated;
5.8. A written determination that either Party is ineligible to receive an award by the Government by being listed on the Consolidated List of Debarred, Suspended and Ineligible Contractors; or
5.9. The expiration of two (2) years from the effective date hereof, provided, however, if the Program is still under consideration by Northrop Grumman senior management upon the expiration of the (2) year period, this Agreement shall continue in force until terminated pursuant to one of the foregoing conditions.
6. Proprietary Information
During the course of this Agreement, the Parties shall disclose and protect all proprietary and/or sensitive information in accordance with the Non-Disclosure Agreement executed between the Parties, which is attached as Exhibit B and is hereby incorporated herein. As a result of the execution of this Agreement, the Parties further agree that Northrop Grumman may disclose information disclosed under such Non-Disclosure Agreement to the U.S. Government in furtherance of the Program, subject to the appropriate restrictive legends set forth in the FAR or DFARS, and, on a need-to-know basis, to their other subcontractors and potential subcontractors on the Program subject to the same restrictions on use and disclosure as are set forth in the Non-Disclosure Agreement. The obligations of the Non-Disclosure Agreement shall survive the expiration or termination of this Agreement.
7. Classified Information
To the extent the obligations of the Parties hereunder require the handling or the access to classified U.S. Government security information; the same shall be subject to the requirements of the Department of Defense National Industrial Security Program Operating Manual (NISPOM).
8. Intellectual Property
Intellectual property developed under this agreement shall remain the property of the originating Party. All rights, title and interest in the technology and documentation, and any right, title or interest to any intellectual property rights in the DGU as expressed within Exhibit C shall remain at all times with Northrop Grumman, and CIRQUE obtains only the rights expressly set forth in this Agreement.
CIRQUE recognizes and agrees that any modifications, alterations, revisions, upgrades, changes, adaptations, and/or improvements to the baseline DGU system and/or technology (not including CIRQUE/CIRQUE “Auger Gasifier” and “Syngas Scrubber”) is considered under the ownership of and the intellectual property of Northrop Grumman.
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CIRQUE recognizes and agrees that at no time during prototype and/or product development does Northrop Grumman release any of its rights to CIRQUE in the engineering or design of hardware and/or software which includes the effort necessary to meet specific mission objectives, specifications, or performance requirements to provide a capability that did not previously exist.
CIRQUE recognizes and agrees that all Northrop Grumman intellectual property is provided "as-is," without any express or implied warranty. In no event shall Northrop Grumman be held liable for any damages arising from CIRQUE use of the Northrop Grumman intellectual property.
In the event of joint development, the Parties shall establish their respective rights in the intellectual property by negotiations between the Parties. Each Party shall grant to the other Party the right to use its intellectual property necessary for the other Party to support the prototype development and demonstration. It is recognized and agreed that the Parties may be required to, and shall, grant licenses or other rights to inventions, data, software and information as specified in United States (U.S.) Government Solicitations or as required by law. Such rights, which shall be identified in any resulting subcontract, shall not exceed those required by the U.S. Government contract.
9. Expenses
Except as otherwise set forth herein, or as may be mutually agreed by the Parties, and except for the compensation which may be paid to the Parties in accordance with any such contracts and subcontracts, each Party shall bear all of its own risks and expenses incurred in connection with this Agreement and the Program referred to herein including, without limitation, its marketing, sales and proposal activities.
10. Relationship
The cooperation of the Parties contemplated by this Agreement is for the purpose of complementing their respective capabilities for the Program objectives. This Agreement shall not constitute, create, or in any way be interpreted as a partnership, joint venture or formal business organization of any kind. This Agreement does not establish any relationship of principal or agent; and neither Party shall have any power or authority to accept on behalf of the other any offer, agreement, or contract, or to make, incur, contract or create any claim, promise, guarantee, debt, obligation, expense or liability of any kind whatsoever in the name of, on behalf of or for the account of the other Party. Nothing in this Agreement shall be construed as providing for the sharing of profits and losses of either or both of the Parties. Except to the extent of a breach of this Agreement, neither Party shall acquire, by virtue of this Agreement, any liability to the other Party for expenses, risks or liabilities incurred by the other Party.
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11. Scope of Agreement
This Agreement shall relate only to the Program specified herein, and nothing herein shall be deemed to:
11.1. Confer any right or impose any obligation or restriction on either Party with respect to any other program effort or marketing activity at any time undertaken by either Party hereto, jointly or separately; or
11.2. Preclude either Party hereto from soliciting or accepting a contract or subcontract from any third party contractor (or subcontractor of any tier) under any other program or under this Program after termination of this Agreement; or
11.3. Limit the rights of either Party to promote, market, sell, lease, license or otherwise dispose of its standard products or services, except where such would conflict with the obligations of the Party under this Agreement.
12. Condition on Obligations
The obligations of either Party hereunder, including without limitation the obligations to prepare and submit any proposal and to award or accept any subcontract, are subject to the following conditions:
12.1. There shall be no litigation or proceeding pending or threatened against the Party or any of its officers or employees (i) which is for the purpose of enjoining or otherwise restricting the activities contemplated by this Agreement, or otherwise claiming that any such activity is improper, (ii) which would adversely affect the rights and/or capabilities of the Party in respect of such activities, or (iii) which, in the judgment of an officer of either Party, would make the continuation of such activities inadvisable.
12.2. Prior to the submission of any proposal or the award of any subcontract, there shall have been no material adverse change in the financial condition or operational capabilities of either Party relating to the activities contemplated by this Agreement, and there shall not have been any occurrence, circumstance or combination thereof which might reasonably be expected to result in any material adverse change in the ability of either Party to perform the work covered by such proposal or contemplated subcontract.
12.3. In the event of any occurrence or circumstance as set forth in Articles 12.1 and 12.2 above, each Party shall provide written notice to the other within ten (10) working days of knowledge of such occurrence or circumstance.
13. Indemnity
The employees of Northrop Grumman and CIRQUE shall obey all pertinent rules and regulations of the other Party while on the premises of the other Party, including those relating to the safeguarding of classified information. Each Party agrees to indemnify and save harmless the other Party from and against all claims for damage to, or loss of use of, the other Party’s tangible property; and injury or death of any of the other Party’s employees or agents, to the extent any such damage, injury or death is caused by any negligent act or omission to act of the indemnifying Party’s employees or agents in connection with performance under this Agreement.
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14. Compliance with Law
The Parties shall comply with all federal, state and local laws and regulations including Executive Orders of the President of the United States applicable to the effort contemplated under this Agreement. Technical data exchanged hereunder may be subject to U.S. export control laws and regulations. Accordingly, the Parties shall not transfer technical data received under this Agreement to any foreign person, country, foreign subsidiary or parent corporation, without specific authorization from the disclosing Party and pursuant to an appropriate U.S. Government agency license.
15. Governing Law
This Agreement shall be construed in all respects in accordance with, and any dispute arising hereunder shall be governed by, the substantive and procedural laws of Virginia except, however, that choice of law provisions shall not apply. The Parties waive any right to a jury trial.
16. Disputes
The Parties will attempt in good faith to resolve through good faith negotiations of any dispute, claim or controversy arising out of or relating to this Agreement. If the Parties are unable to resolve such dispute, claim or controversy through good faith discussions at the Program level, higher level management representatives from each Party shall meet and attempt to resolve the dispute. Parties may exercise any right available under the law if the disputes cannot be resolved through good faith negotiations. No provision of this Agreement shall prevent either Party from exercising any right available under the law if (a) good faith efforts to resolve the dispute have been unsuccessful, or (b) interim relief from a court or other adjudicative body is necessary to prevent serious and (or) irreparable injury.
17. Limitation of Liability
Neither Party shall be liable to the other for any indirect, incidental, special or consequential damages, however caused, whether as a consequence of the negligence of the one Party or otherwise.
18. Publicity
No releases shall be made to the news media or the general public relating to this Agreement without the prior written approval of the other Party, which approval shall not be unreasonably withheld. The Parties further agree that news releases made by either of them shall recognize the participation and contributions of the other Party.
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19. Non-solicitation of Employees
Northrop Grumman’s obligations and commitments under this Agreement, including the non-solicitation provision below, apply only to its Information Systems Sector, Defense Systems Division (DSD).
The parties agree that during the term of this Agreement and for a period of one (1) year after the expiration or termination of this Agreement, neither party shall, without the advance written consent of the other, actively solicit for employment any person or persons employed by the other who is working on or with respect to the parties contract and/or subcontracts under the Program. The hire of individuals responding to general public marketing and recruiting advertisements and events shall not be a violation of this provision; only active, targeted solicitation is prohibited.
20. Assignments
Neither Party shall assign or transfer any of its rights or obligations hereunder in whole or part without the prior written consent of the other Party, except to another U.S. corporate division or affiliate of the Party so long as sufficient assets, personnel and other resources necessary to perform the obligations hereunder remain available. Any consent required under this clause shall not be unreasonably withheld.
21. Severability
If any provision of this Agreement or part of such provision is or becomes invalid or unenforceable, then the remaining provisions hereof shall continue to be effective. Nothing in this Agreement shall be construed as requiring any Party to take any action which is prohibited under any applicable governmental laws or regulations, or as prohibiting any Party from complying with such laws or regulations.
22. Waivers
No waiver by a Party of any of its rights or remedies shall be construed as a waiver by such Party of any other rights or remedies that such Party may have under this Agreement.
23. Notices
All notices or communications (other than normal business communications) required by this Agreement or desired to be given hereunder, shall be in writing addressed as follows, and given by certified or registered mail, return receipt requested, or an overnight mail service that confirms delivery and shall be deemed to be given when received.
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If To: Northrop Grumman:
Northrop Grumman Systems Corporation,
201 Electronics Blvd.
Huntsville, AL 35824
Attention: Carl V. Ehler
If To: CIRQUE:
Cirque Energy, Inc.
243 W. Congress St, Suite 350
Detroit, MI 48226
Attention: Richard L. Fosgitt, PE
24. Entire Agreement
This Agreement contains the entire understanding between the Parties and is the complete and exclusive expression of the Agreement between the Parties with respect to the Program. This Agreement supersedes all prior or contemporaneous communications, agreements or understandings between the Parties on the subject matter of this Agreement. A modification to this Agreement may only be made in writing and must be signed by authorized representatives of both Parties.
The Parties hereto, by and through their duly authorized representatives, execute this Agreement.
CIRQUE ENERGY, INC.
By
Name Joseph L. DuRant
Title CEO
Date November 13, 2013
NORTHROP GRUMMAN SYSTEMS CORPORATION
DEFENSE SYSTEMS DIVISION (DSD)
By
Name
Title
Date
During the manual randomization phase:
"Our CRO did not have electronic randomization capability and instead distributed sealed envelopes containing patient randomization assignments to each site. During the manual randomization phase, the Company noted a large number of patients with renal failure being entered into the treatment arm versus control (57% vs 17% control, p=0.05) resulting in a much sicker treatment arm. Envelopes were audited and two sites most responsible for this bias were also found to have had envelopes
opened with no corresponding patient enrolled."
"After a thorough review by the SAB and external DSMB, they concluded that data from the manual randomization phase was biased by a compromise of randomization and unanimously recommended to reset the clock with electronic randomization. It was believed that the remaining 47 patients to be enrolled under electronic randomization could be sufficient to achieve the goals of the study. Statistically, electronic randomization corrected the randomization issue. Except for safety data, only data from the electronic randomization phase will be presented here."
This is the real story lhat is never told...
What is even more interesting is your post regarding the messed up trial. One could argue that everything rises and falls on leadership. The truth seems to be lost every time someone talks about the trial. The Doctors had the proper protocols in place to conduct the trial. The doctors chose to bypass the protocols to ensure their patients received the filter as the mode of treatment. The doctors saw firsthand the results of the filter and chose to make sure their patient survived the illness. This behavior by the trial doctors skewed the randomization of the first 50+/- patients. Was this the fault of Cytosorbents? On the top level of course as a management oversight situation, but it was the protocol doctors that did it to save their patients.
"MONMOUTH JUNCTION, NJ--(September 7, 2011)- CytoSorbents Corporation (OTC.BB: CTSO.OB - News), a critical care-focused company using blood purification to treat life-threatening illnesses, announced additional data from its recently completed European Sepsis Trial using CytoSorb™, a first-in-class extracorporeal cytokine filter approved for use in the European Union. The Germany-based, randomized, controlled, multi-center trial evaluated the use of CytoSorb™ plus standard of care therapy versus standard of care therapy alone in patients with severe sepsis or septic shock in the setting of lung injury. Data from 100 patients were used to evaluate the safety of CytoSorb™ treatment. Due to a patient randomization issue that was subsequently corrected by institution of electronic randomization, the last 43 patients that completed the trial were used to evaluate clinical efficacy. At the time of enrollment, most of these 43 patients (25 control, 18 treated) had septic shock (94% vs. 100% control) and acute respiratory distress syndrome (67% vs. 56% control), while many had renal failure requiring renal replacement therapy (39% vs. 24% control) before entering the trial. The presence of multiple organ failure reflects the severity of illness in these patients and confers a high expected rate of mortality. "
CytoSorbents Corporation (OTCBB: CTSO)
2013 Shareholder Update Conference Call Transcript
August 21, 2013 @ 4:15 pm Eastern
Operator:
Good day, ladies and gentlemen. Thank you for standing by. Welcome to the CytoSorbents 2013 Shareholder Update Conference Call. During today's presentation, all parties will be in a listen-only mode. This conference is being recorded today, August 21, 2013.
I would now like to turn the conference over to our host, Mr. Alan Sheinwald of Alliance Advisors. Please go ahead, sir.
Alan Sheinwald – Alliance Advisors – Moderator:
Thank you operator and good afternoon. Welcome to CytoSorbents 2013 Shareholder Update Conference Call. With us today are Dr. Phillip Chan, Chief Executive Officer and President, Kathleen Bloch, Chief Financial Officer, Vincent Capponi, Chief Operating Officer, Christian Steiner, VP of Sales and Marketing from Germany, and Chris Cramer, VP of Business Development
Before I turn the call over to Dr. Chan, I’d like to remind listeners that during the call, management's prepared remarks may contain forward-looking statements which are subject to risks and uncertainties. Management may make additional forward-looking statements in response to your questions today. Therefore, the Company claims protection under Safe Harbor for forward-looking statements contained in the Private Securities Litigation Reform Act of 1995. Actual results may differ from results discussed today and therefore, we refer you to a more detailed discussion of these risks and uncertainties in the Company's filings with the SEC. Any projections as to the Company's future performance represented by management include estimates today as of August 21, 2013 and the Company assumes no obligation to update these projections in the future as market conditions change.
Well during today’s conference call we have done our best to take everyone’s submitted questions and answer within our prepared remarks as best we can. We received many, many questions and thank everyone for participating. If we do not answer your question we would ask you to either contact myself and my team at Alliance Advisors or the Company directly after the call today.
At this time, I would like to turn the call over to Dr. Phillip Chan. Phil, the floor is yours, please go ahead.
Phillip Chan - CEO:
Thanks very much Alan and thanks to everyone who’s joined our call today.
During today’s call we’ll provide an overview of CytoSorbents for those of you who may be new to the story and update everyone on where we stand today. Along the way we plan to address many of the questions that have been submitted by investors. So thank you very much to those who submitted questions to us.
CytoSorbents is a critical care focused medical device company using blood purification to treat life-threatening illnesses. In a relatively short period of time, CytoSorbents has grown to 27 full time employees and part-time consultants, both here in New Jersey and at our subsidiary in Berlin, Germany. And we are very fortunate to have an excellent management team to lead this rapid growth. After quickly introducing myself, I’d like to ask each member of the management team to give a quick summary of his or her background.
As for myself, I have been the CEO of CytoSorbents Corporation for nearly 5 years. By background, I am a Board-certified internal medicine physician with a strong background in clinical medicine and research, having received my MD and PhD from Yale University School of Medicine and completed my residency at the Harvard Medical School at the Beth Israel Deaconess Medical Center. I am also an alum of Cornell University, where I received a BS in cell and molecular biology. Prior to joining CytoSorbents, I led healthcare and life science investments as a Partner for the $80 million NJTC Venture Fund, which is a loyal shareholder of the company. During my 5-year tenure at NJTC Venture Fund, I was responsible for numerous investments covering the gamut of therapeutic, medical device and diagnostics companies. I also co-founded Andrew Technologies, a venture-backed medical device company currently commercializing its FDA approved HydraSolve™ lipoplasty system. Vince?
Vincent Capponi - COO:
Hello everyone. My name is Vince Capponi, Chief Operating Officer of CytoSorbents. I have been with the company for more than a dozen years and have more than 20 years of management experience in medical device, pharmaceutical and imaging equipment companies including Baxter, Upjohn, Sims Deltec and Sabratek I led the regulatory approval of the first heparin flush syringe now used ubiquitously in hospitals around the world to keep IV’s patent, as well as the manufacturing of more than a million units a week for this device. At CytoSorbents, I led the CE Mark approval of CytoSorb®, the recent signing of Hitit Medical as one of our first distributors, and am heavily involved in all aspects of the company, including manufacturing of CytoSorb® and new product development. Kathy?
Kathleen Bloch - CFO:
Good afternoon everyone. This is Kathleen Bloch and I am the Chief Financial Officer of CytoSorbents. I bring more than 20 years of executive financial experience at both public and private companies to help drive the success of CytoSorbents. Before joining the company, I was the CFO of Laureate Biopharmaceuticals, a leading contract manufacturer of biologic drugs and my background includes significant experience in rapidly-growing medical device and manufacturing companies. I am a Certified Public Accountant with an MBA in finance. Chris?
Chris Cramer – VP Business Development:
Thank you Kathy. Hi, I am Chris Cramer, Vice President of Business Development for CytoSorbents. I have over 15+ years of business development and commercial experience, most recently at Johnson & Johnson, where over the course of six years, I held various leadership roles in the medical device division, most recently as Senior Director of New Venture Development. I also spent more than eight years in management consulting at PWC Consulting, working with leading health care companies across the world. Christian?
Christian Steiner – VP Sales and Marketing:
Hello, I am Dr. Christian Steiner, Vice President of Sales and Marketing of CytoSorbents and Co-Managing Director of CytoSorbents Europe GmbH. I manage all direct sales and marketing efforts of CytoSorb® in the German speaking countries of Germany, Austria and Switzerland, and am working with my colleagues in the US to expand distribution of CytoSorb® in Europe. I am a German medical physician by background and have spent more than 13 years in the sales and marketing of critical care therapies, particularly extracorporeal therapies such as the MARS liver dialysis system which was eventually acquired by Gambro, and non-invasive hemodynamic monitoring systems by Pulsion Medical. During this time, I have established a significant network of key opinion leader contacts throughout Germany, but also elsewhere in Europe and the US. So it is a pleasure to be on the call with you today.
Alan Sheinwald - Moderator:
Great. Dr. Chan – to start out, could you please give listener’s a quick overview on CytoSorbents?
Phillip Chan - CEO:
Absolutely. CytoSorbents is an emerging growth opportunity aiming to revolutionize critical care medicine with a powerful blood purification technology called CytoSorb ® , now approved in the European Union. The goal of CytoSorb ® is to prevent or treat organ failure, the cause of nearly half of all deaths in the ICU for which no therapies exist.
The underlying blood purification technology is based on highly biocompatible, porous polymer beads that act like tiny sponges to remove harmful substances from blood. When these beads are placed into a cartridge roughly the size of a drinking glass, blood can be pumped outside of the body with a standard hemodialysis machine, through this cartridge where toxic materials are removed, and that purified blood is simply recirculated back into the body. These beads are some of the highest purity sorbent materials commercially available for medical use, and are protected by 32 issued US patents with multiple applications pending.
I mentioned before that we focus on critical care medicine. Millions of people are admitted to the intensive care unit in hospitals all over the world each year for life-threatening conditions such as sepsis, trauma, burn injury, lung injury and many other conditions where inflammation plays a serious detrimental role. You may have known people who have landed in the ICU after a serious car crash, or after a bout of severe pneumonia, or after a major burn injury from a fire. These patients are extremely sick, and frequently develop organ failure where vital organs stop working. Organ failure puts these patients at extremely high risk of death.
The problem with critical care medicine today is that although we can help patients stay alive when their organs fail with supportive care therapies like mechanical ventilation or dialysis – often called “life support” - there is in fact very little that can be done to prevent or treat organ failure and actually help these patient get better except for the passage of time. In fact, keeping a patient alive until his or her own body heals itself is a major strategy on how many critically ill patients are treated today.
But because of the general inability to improve clinical outcome, patients often linger for days to weeks at a time at a cost of $2000-3000 a day in the United States, and comparable numbers in Germany and other places in Europe. If they don’t die first, they often develop complications such as infections, medical errors, and other problems which make them even more difficult to treat. So it’s no wonder that the US spends nearly 1% of its gross domestic product, or almost $80-90 billion every single year on critical care medicine and why hospitals lose billions of dollars annually.
With organ failure such a major unmet medical need, we have approached this by going after one of the most important root causes of organ failure – which is uncontrolled inflammation. Inflammation is the immune system’s normal way of dealing with infection and injury and is driven by small proteins made by the immune system called cytokines. But in life threatening illnesses, the immune system releases a massive amount of cytokines, often called cytokine storm, that unleashes deadly widespread inflammation, that can then lead to organ failure, and frequently death.
Our solution, CytoSorb®, is the only specifically approved extracorporeal cytokine filter in the European Union specifically targeting the prevention or treatment of organ failure by reducing cytokine storm and controlling potentially deadly inflammation. CytoSorb® is indicated and approved for use in any situation in the European Union where cytokines are elevated and it allows for extensive “on-label” use for many different applications. The device is plug and play with a hospital’s existing hemodialysis infrastructure and can treat your entire blood volume 20-30 times in a 6 hour period. Each treatment uses a new cartridge. And we now have more than 1,000 human treatments where there have been no serious device related adverse events so it’s been relatively safe.
CytoSorb® is a very powerful new tool for physicians – giving them the potential ability to modulate the immune response for the first time in many different diseases, and to address organ failure at its root cause. Since the launch of CytoSorb® in the second half of 2012 in our direct territories of Germany, Austria and Switzerland, we have focused on building our key opinion leader network, and now have approximately 100 key opinion leaders throughout Germany, Austria as well as the United Kingdom. CytoSorb is in the majority of University hospitals in the major cities in Germany with a growing presence in Austria. And we also now have 18 investigator initiated studies getting ready to start or being planned, and many successful case report studies also.
With approximately $400,000 in initial CytoSorb sales from June 2012 to June 2013, we believe we are just scratching at the surface of the commercial potential for CytoSorb®. With a multi-billion dollar critical care opportunity with little to no competition, an outstanding product that targets the key reason why patients die, and a very profitable razorblade in someone else’s razor business model with current 60+% gross margins that are expected to increase with volume over time, this has the potential to not only revolutionize critical care medicine by saving lives and reducing costs, but also the potential to make CytoSorbents a commercial success. The key is how we plan to get there, which is what we will discuss today.
Our technology has also received recognition from DARPA, the US Army, and the US Air Force. DARPA, or the Defense Advanced Research Projects Agency, that was responsible for funding the development of the internet, Global Positioning Satellites and robotic surgery, believes that blood purification is the right approach to treat sepsis. To this end, they awarded us a $3.8 million 5-year contract in 2012 as part of its “Dialysis-Like Therapeutics” program to treat sepsis, to remove cytokines and bacterial toxins from blood. The U.S. Army also awarded us SBIR contracts for trauma and burn injury totaling $1.15 million. In addition, the U.S. Air Force is funding a U.S.-based, 30-patient randomized, controlled pilot study to use CytoSorb to treat rhabdomyolysis in trauma patients. The FDA has granted approval, under an IDE, to begin this study, expected this year. These grants and contracts validate and advance our technologies and significantly offset operating expenses as we further commercialize CytoSorb.
Alan Sheinwald - Moderator:
Thank you Phil. Before we get into the details of the business, I think it would be helpful for investors to know what the company looks like from a financial perspective. Kathy, could you give us a snapshot of where CytoSorbents is from a financial standpoint?
Kathleen Bloch - CFO:
Certainly Alan. We have worked hard as a company to develop multiple sources of revenue. These are divided primarily into CytoSorb product revenue, of which we have both direct sales and distributor sales, and also grant income. Clearly product revenues are very important, but at this early stage in our commercialization cycle, grant income is extremely helpful as well, because it helps to covers a good portion of our R&D headcount and helps to offset many of our fixed expenses. By leveraging non-dilutive grant and contract funding, we have been able to significantly advance our technology platform without needing to raise more capital to fund this development, two things which directly benefit the Company and our shareholders.
For the six months ended June 30, 2013, we generated revenue of approximately $663,000 as compared to revenues of approximately $133,000 for the same period in 2012, which is an increase of 497%. Revenue from product sales was approximately $304,000 in the first half of 2013, as compared to $53,000 in the first half of 2012, an increase of about 600%. This increase was driven primarily by our direct sales force, with some initial sales to distributors. Product gross margins were approximately 60% for the six months ended June 30, 2013. Revenue from grants was approximately $359,000 in the first half of 2013, as compared to approximately $83,000 in the first half of 2012. Investors can find more detail in our second quarter 10-Q report filed on August 14, 2013 at http://www.sec.gov.
We were pleased with the significant growth in revenue in the first half of this year, compared to the same period a year ago and we would encourage investors to similarly look at our future progress in this manner. At our current early stage of commercialization, our sequential quarter-to-quarter results are subject to many different variables such as the timing of direct sales and reorders, and unique events such as initial stocking orders by distributors. First quarter 2013 was a particularly strong quarter that benefited from many new orders and reorders from key opinion leaders, as well as initial stocking orders from both of our distributors, where they wanted to have inventory on hand to be able to pre-market CytoSorb® to potential customers. In terms of direct sales, we have been at the key opinion leader stage, and have observed that the key opinion leaders personally want to oversee treatment with CytoSorb® to see for themselves how it is working. Despite being extremely busy, they have made time to be directly involved in patient selection, treatment, and oversight of these patients. So far the level of enthusiasm and interest remains very positive. Obviously, this level of attention is a good thing, but given the time restraints of these key opinion leaders, it means that CytoSorb is being used intermittently, rather than regularly. Our second quarter 2013 appeared to be too soon to get the usage and re-orders that we were expecting, but we have been seeing an encouraging level of activity from specific accounts already in the third quarter, despite it being the summer vacation months in Europe. Later in the call, Phil and Christian will discuss how we are planning to move from the key opinion leader stage, and begin to drive usage and demand from the rest of the physicians in the department. Until this occurs, we may have significant variability in our quarter over quarter product sales. We encourage investors to look at revenue growth from the big picture and be patient while we continue to execute our sales plan, which we believe will be successful.
I’d also like to comment on our current cash situation and how we intend to fund our operations moving forward. As of June 30, 2013, CytoSorbents had approximately $1.9 million in cash. The Company receives funds from a variety of sources, all of which are independent from each other. First, we receive revenue from government grants. Over the next six months we are expecting to achieve milestones that will permit us to receive more than $1.0M in grant funding. Second, as our sales and marketing efforts take hold, revenue growth is expected to provide additional operating funds, particularly with blended gross margins from direct sales and distributors expected to remain well above 50%. Third, we are seeking traditional commercial debt to help finance working capital expansion needs. Fourth, as we have in the past, we will seek funding in the market to support the needs of our strategic plan. Recently, for example, in June, we raised $1.1 million in convertible notes to fund our product commercialization efforts. And, lastly, in the event we need additional funding, we have an arrangement with Lincoln Park Capital to provide financing.
Alan Sheinwald - Moderator:
Thank you Kathy, that was very helpful. Dr. Chan, in your past presentations you have talked about building CytoSorbents on three pillars of value: First, commercialization of CytoSorb®. Second, Clinical trials, product development and R&D. And third, Business Development.
Let’s focus on each of these areas because I think it will provide investors with a good 360 degree view of your business opportunities. Let’s take commercialization of CytoSorb® first. I think the market recognizes the blockbuster potential of CytoSorb® as a critical care therapy. So how do we get there?
Phillip Chan - CEO:
Thanks Alan. I think its worthwhile to reiterate exactly where we are and then to turn it over to Christian, our VP of Sales and Marketing, who is on the front lines and seeing the tremendous amount of activity related to CytoSorb first hand.
CE Mark approval was a pivotal event for us because, with the notably exception of the US and Japan, it has opened up most of the world as potential markets for CytoSorb. This includes all 28 countries of the European Union where we already have regulatory approval to sell CytoSorb®, as well as the Middle East, Israel, Canada, India, Brazil, Australia, and many other countries that will accept CE Mark approval, with the appropriate product registrations. We have elected to focus on Germany, Austria and Switzerland ourselves, while we work with critical care focused distributors and strategic partners to expand our geographic footprint to other major countries and territories. This is a tried and true approach to maximizing the revenue potential of medical products.
But the important thing to note is that if we were only successful in Germany, we could make CytoSorbents into a very successful company. The critical care market in Germany alone for CytoSorb is approximately $1 billion. To this end, we have established a subsidiary in Berlin, Germany with a 4 person sales force that is targeting direct sales in Germany, Austria and Switzerland. We have established reimbursement for CytoSorb® in Germany and Austria at more than $500 per cartridge. We have also established ISO 13485 certified manufacturing at our facility in NJ, a requisite to sell medical devices into Europe. Looking at a map of Germany, our team of four salespeople have created a broad foothold with product sales, new clinical studies, case reports, and ongoing discussions in the university and public hospitals across most of the major cities in Germany and a growing number of hospitals in Austria as well.
As a small company, we have approached commercialization of CytoSorb® very methodically and conservatively to ensure that there is demand for CytoSorb before incurring costs of manufacturing and our sales force. To date, our strategy has been to cast the net wide, to create awareness, interest, and usage amongst key opinion leaders in our direct sales territories. And we have been remarkably successful at it, despite only 4 sales people in the field. We now count approximately 100 key opinion leaders, primarily in Germany, but also in Austria and the UK, who are either using or committed to using CytoSorb® in clinical use or in clinical studies, and now we have 18 investigator initiated studies being planned with two that are now registered and several others that will start soon. These studies are primarily in sepsis and cardiac surgery but include, for example, acute kidney injury, liver failure, lung injury, and pancreatitis.
Now that we see this demand and have support of the key opinion leaders, the goal is to now transition the sales strategy to not go wide, but rather to go deep into these accounts and drive adoption of CytoSorb amongst the rest of the physicians in the department, and to other ICUs in the same hospital. The goal is to generate the end-user demand that is expected to drive revenues from current hundreds of thousands of dollars in this key opinion leader phase, to really millions in sales in the “Departmental” phase. We want to target doctors who are admitting patients in the middle of the night and get them thinking about how to use CytoSorb for their patients. We have already started doing this at certain key accounts that should become reference centers for us in the near future.
The message to these junior and senior doctors is simple. The message is if you do just standard of care, the chance your patient dies, despite your best efforts, is often greater than 1 in every 3 patients. CytoSorb® is the only specifically approved cytokine filter in the EU, that has been demonstrated to safely reduce cytokine storm, with the goal of reducing inflammation and preventing or treating organ failure. The head of your department wants you to use it, the hospital is already ordering it, and the device is reimbursed. Lastly, we now have more than 1,000 human treatments where the device has been used safely, meeting the first tenet of medicine – which is, of course, first do no harm.
So these numbers can get big very quickly. If we assume 15 patients treated a quarter at an average revenue of $5,000 per patient, that single intensive care unit would produce CytoSorb sales in excess of $300K. That’s our entire sales for the first half of this year. With an average of 300 to 600 patients alone that are admitted to one of these large ICUs each year with the diagnosis of sepsis, we believe these are absolutely achievable numbers.
But the opportunity is clearly too large for a four person sales force to effectively manage, which is why our goal is to continue to expand our sales force in a responsible way but to aggressively pursue these exciting opportunities.
Now I like to turn it over to Christian to get his comments as well.
Christian Steiner – VP Sales and Marketing:
Following Phil’s comments, I’d like to give you some color about what we are seeing in the field. In Germany, I have been involved in the launch of a number of critical care products and the enthusiasm behind CytoSorb by doctors continues to impress me. Although there are other older competitive technologies in the European market from such well known companies as Fresenius Medical Care, Gambro and Bellco, these technologies are more difficult to use, do not have the flexibility of our technology, and are not commonly used by the majority of hospitals we are visiting. When we travel to critical care conferences or talk to key opinion leaders about new innovations, we also see nothing else competitive in the near term. It suggests to me that we have a unique product, with real potential, as Europe’s only specifically approved cytokine filter.
Excessive inflammation and cytokine injury is such a repetitive theme in medicine that I am also still pleasantly surprised by many of the new application ideas that are proposed by the doctors we talk to. We appear to be at the right place at the right time with this technology. Outside of critical care, an area that we are seeing growing interest is in cardiac surgery, and the use of our CytoSorb filter in a parallel bypass circuit. The interest in using CytoSorb in this application is because many things specific to the surgery and the heart-lung bypass machine can generate cytokines, free hemoglobin and inflammation – all of which can be bad for the patients and cause complications. This could be a rapid future area of growth for CytoSorb.
In general, the physicians using CytoSorb have expressed their excitement to me and our sales team with the results they have seen with the technology, and this has led to an increase in activity with more usage, more reorders, more case report studies being prepared, more investigator initiated studies being started, and more discussions with other doctors. This has led to introductions to other ICUs in the same and other outside hospitals, and more opportunities for us.
I can imagine that it is very difficult for investors to appreciate all of this activity, but as these relationships become more public we will be able to discuss them more openly, as we did in the last earnings release and today’s press release on the dosing study. We will also have 4-5 major reference accounts in the future that we will be able to talk about. And as we have more success in converting junior and senior doctors in each department, our sales numbers will begin to reflect this as well. We are confident that our efforts to drill down within each department to sell to the junior and senior physicians will be successful, as we have done this many times before. We just need more manpower, which we are working on, to be able to execute this strategy.
Finally some notable recent successes. We have seen some additional remarkable reports. For example, as reported from our distributor in Turkey, a man was accidentally electrocuted by 34,000 volts and was featured on the evening news after surviving severe external and internal burn injuries after treatment with CytoSorb. Another patient in Germany survived a serious case of Legionnaire’s disease and muscle breakdown after being treated by our therapy. And yet another patient with chronic liver disease survived septic shock with our therapy. This adds to a growing number of success stories with CytoSorb. So what we are planning is to establish a patient registry database in the near future to capture more of these data and to show more of these success stories.
Alan Sheinwald – Moderator:
Thanks Christian, that was very informative. Vince, the press release issued today on the expansion of the dosing study to now 8 major clinical sites in Germany, touches specifically on the second pillar of value which is the generation of clinical data. Having played an integral role in managing the first European Sepsis Trial, the ongoing sepsis dosing trial, and CE Mark approval for CytoSorb, can you please give listeners a quick background on sepsis, what the follow-up dosing study is intended to do, and why these particular sites are important to the company?
Vincent Capponi – COO:
Thanks Alan. CytoSorb, as a platform technology, has the potential to treat many different life threatening conditions in the ICU that include sepsis, acute respiratory distress syndrome, burn injury, trauma, pancreatitis, and many other inflammatory conditions. We have decided to focus on the treatment of sepsis, an unmet medical need, which is the result of an overzealous immune response to a severe infection and is driven by “cytokine storm”. Sepsis afflicts 18 million people worldwide every year and is increasing in incidence. There are no approved therapies to treat sepsis in the US or Europe, only a handful of therapies are being tested in Phase 3 trials, and as the literature points out, despite the best medical treatment which includes antibiotics, severe sepsis has a mortality of 25-30%, while that of septic shock is 40-50%. It accounts for 10-20% of all ICU admissions and is a top ten killer, killing more people in the US than either heart attacks, strokes, or any single type of cancer. At an average cost of more than $45,000 to treat, hospitals lose billions of dollars annually.
Despite sepsis being very difficult to treat, we believe that we have one of the most comprehensive solutions to date to treat it. Unlike other therapies that have tried and failed to stop sepsis by using very directed therapies, such as antibody therapies, CytoSorb uses a broad spectrum approach.
As Phil points out in his scientific presentation, CytoSorb® has many ways it can improve outcome in sepsis. First, it can reduce inflammatory cytokines that are directly responsible for driving deadly inflammation that can result in organ failure. Second, it can reduce excessive anti-inflammatory cytokines that can lead to immune suppression and infection – a primary driver of death in late sepsis. Third, CytoSorb® can remove many different deadly bacterial toxins as shown in vitro , such as that associated with MRSA, or methicillin resistant Staph aureus, a common hospital acquired drug resistant bacteria. And fourth, CytoSorb® can help to redirect the dangerous activated white cell component of the immune system to target the true area of infection, and to avoid innocent “bystander” organs, thereby preventing inadvertent organ injury.
Fast forward to our European Sepsis Trial. During this 47 patient randomized controlled multi-center trial, CytoSorb® was clinically proven to safely and broadly reduce cytokine storm and key cytokines in septic shock patients with lung failure, by 30-50%. Further post-hoc analysis of the data suggested that two subgroups really benefited the most from therapy – those patients with extremely high cytokine levels, and patients older than 65 years of age - where reductions in 28-day and 14-day mortality, respectively, were statistically significant. This was achieved by treating for 6 hours a day for 7 days, each day with a new device.
The data suggested that longer or more aggressive treatment could be beneficial and we set out to do a dose ranging study, what we call the “Dosing Study”, to evaluate the safety and preliminary efficacy of extended treatment, either continuous treatment for up to 7 days, which is currently ongoing, or 6 hours of treatment a day for up to 14 days, which is currently planned, each day with a new device. As we mentioned in the press release, the dosing study was designed to evaluate the safety and preliminary efficacy of extended CytoSorb® treatment, in order to give physicians more flexibility on how to treat critically ill patients with CytoSorb®. In addition, the study is not intended to produce pivotal data, but rather is intended to support the design of a pivotal trial protocol in the United States. The pivotal trial study in sepsis is expected to start in 2014, pending successful discussions with the FDA, with the goal of definitively demonstrating the effectiveness of CytoSorb® for the treatment of sepsis, and to seek U.S. regulatory approval.
We started the dosing study with three of our top performing sites from the European Sepsis Trial – University of Goettingen, Helios Erfurt, and the University of Aachen, but were approached by a number of other leading institutions to participate in our trial. As you saw in the press release, these included University of Jena, University of Hamburg-Eppendorf, University of Greifswald, University of Leipzig, and European Medical School of Oldenburg. We slowed down the trial to get these sites on board and although it took longer than expected, due to the time needed for contract negotiations, ethics committee review, and site training, we are excited that nearly every site is now eligible to enroll patients into the dosing study. As we were bringing new sites on, we conducted antibiotic compatibility testing with CytoSorb® and have provided investigators with guidance on which antibiotics are best to use with our therapy.
Regarding the new sites, these are all great sites and investigators. For example, Dr. Michael Quintel, Chief of the Department of Anesthesiology, Emergency Medicine and Critical Care at Goettingen. He is the PI of this trial, a leader in critical care research and the president of the DIVI society, one of the two major intensivist societies in Germany. Dr. Konrad Reinhart, Director of the Department of Anesthesiology and Intensive Care Medicine at the University of Jena, is the Chairman of the Global Sepsis Alliance and founding President of the German Sepsis Society, and the head of the SepNet sepsis trials network – a consortium of leading hospitals and intensivists throughout Germany who organized and ran government funded sepsis trials. And the list goes on.
We believe the addition of these sites will accelerate the completion of the Dosing Study and is key in furthering the development of the German market and raising awareness of CytoSorb for the treatment of sepsis. As data becomes available from the dosing study, we will be reviewing with our Data Safety Monitoring Board and discussing interim results when appropriate, with the goal of having at least interim data by the end of this year. Alan, back to you.
Alan Sheinwald – Moderator:
Thanks Vince, that was very informative. Turning to Strategic Partnerships as the third pillar of value, Chris – can you please give us an overview of your thoughts on the potential to do partnerships at CytoSorbents?
Chris Cramer – VP Business Development:
Yes Alan, it’s a very exciting time to be joining the company. Throughout my career I’ve seen hundreds of different medical technologies. CytoSorbents, however, clearly stands out from the rest. What’s so compelling about the technology is how flexible the platform is, and how many high value therapeutic applications it may address. For CytoSorb alone, there are numerous partnership opportunities. But I also believe the rest of the development pipeline, including HemoDefend™, ContrastSorb, DrugSorb, and several others that have not yet been publicly disclosed, offers the Company significant future growth opportunities as they are monetized.
As the head of business development, my goal is to establish one or more partnerships by the end of the year. Today we are actively building a robust business development pipeline. Over the past quarter we have been working to access key decision makers and establish relationships with leading medical device and pharmaceutical companies. Our ideal partner, and focus of our business development efforts, is a large multi-national company with strong commercial and product development capabilities that can help to advance our pipeline and accelerate product distribution.
Currently, we are aggressively focused on establishing strategic partnerships for our two most advanced assets, CytoSorb® and HemoDefend™.
For CytoSorb, we are looking to establish distribution agreements for the various applications that Phil described earlier on today’s call. Given the versatility of the platform, we are targeting a wide range of potential partners such as equipment manufacturers with strong commercial capabilities in the intensive care unit, cardiac surgery companies with heart-lung bypass equipment, and multi-national pharmaceuticals with complementary drugs for sepsis to name a few. For CytoSorb, we are seeking favorable transfer pricing and, where appropriate, upfront payments for market exclusivity and working capital for initial product stock and capacity expansion.
For HemoDefend, we are targeting leading players involved in the blood collection, processing, and transfusion businesses. We are seeking partnerships that involve collaborative development to advance our HemoDefend In-Line Filter and HemoDefend Beads-in-a-Bag products in exchange for an option for future rights to commercial distribution.
To date we’ve made significant progress in filling the business development funnel with high quality partnership opportunities. Overall, I am pleased with the strong reception and high level of interest that we are receiving from potential partners. In addition to several opportunities that were ongoing when I arrived here, we have been successful in advancing discussions with new targets. While BD deals can take some time to develop, I am confident we are laying the groundwork for future success.
Although I cannot comment on specific deals, I can give you some color commentary on the business development opportunity for CytoSorbents and the progress that we are making. Overall for CytoSorb and HemoDefend combined, we’ve identified approximately sixty potential target partners that we are aware of today. This is a very healthy number and one that has the potential to grow as we continue to explore the market for strategic partners. Of these sixty, we are actively engaged in discussions with about 1/3 of those targets. Of the targets that we are talking with, I would characterize roughly 80% to be early stage, exploratory and roughly 20% to mid to late stage in nature. Our goal is to fill the funnel with new prospects, convert new prospects into serious partnership discussions, and ultimately turn those late stage discussions into formal partnerships.
In closing, I am very happy to be joining CytoSorbents at this very exciting time. I am optimistic about the business development opportunities that are currently available and look forward to successfully driving one or more partnerships to completion by the end of the year.
Thank you. I’d now like to turn the call back over to Phil.
Phillip Chan - CEO:
Thanks very much Chris. Before concluding today’s call I’d like to leave our investors with potential catalysts in the next 6-9 months.
We believe there are some major drivers of value in our business. The first is increasing revenues, which demonstrate product adoption and acceptance in the market of our CytoSorb® technology. We plan to demonstrate significant year over year improvements, particularly as our strategy to focus heavily on the department physicians takes hold. We are also targeting the signing of at least one significant strategic partnership this year. In addition, we plan to have more clinical data, either from company-sponsored trials, military funded trials, case report studies, or through investigator initiated studies. We are also working to initiate a pivotal study with CytoSorb next year that will provide the data for either US approval or greater usage in Europe. And finally we plan to continue communicating our positive results to our shareholders and a larger investment community.
And finally, to summarize our investment opportunity in a few key points.
· First of all, we believe we are at the ground floor of commercialization, poised to create significant value with an approved product that may revolutionize critical care medicine, save lives, and reduce costs. This is the holy grail of modern medicine today.
· We are operating in a massive market with some of the biggest unmet medical needs in medicine.
· We have received validation of our company and technology from many fronts including DARPA, the US Army, the US Air Force, scientific advisors, as well as analysts.
· We have a unique, highly profitable product and pipeline with little to no competition, and an outstanding business model.
· We are led by an experienced and responsible management team.
· And there are potential major catalysts in the next 6-9 months including revenue growth, partnerships, clinical data, and other positive milestones being reached and communicated to our investors.
With that, let me thank everyone for taking the time to attend today’s call. We look forward to providing you another update in the near future. And if you have any additional questions, feel free to forward them to Alliance Advisors. With that, have a great evening.
Operator: Ladies and gentlemen, this concludes the CytoSorbents 2013 Shareholder Update Conference Call. If you would like to listen to a replay of today’s conference call, please dial 1-877-870-5176 and international 1-858-384-5517. We thank you for your participation and you may now disconnect.
2013 Shareholder Update Conference Call Transcript
August 21, 2013 @ 4:15 pm Eastern
Operator:
Good day, ladies and gentlemen. Thank you for standing by. Welcome to the CytoSorbents 2013 Shareholder Update Conference Call. During today's presentation, all parties will be in a listen-only mode. This conference is being recorded today, August 21, 2013.
I would now like to turn the conference over to our host, Mr. Alan Sheinwald of Alliance Advisors. Please go ahead, sir.
Alan Sheinwald – Alliance Advisors – Moderator:
Thank you operator and good afternoon. Welcome to CytoSorbents 2013 Shareholder Update Conference Call. With us today are Dr. Phillip Chan, Chief Executive Officer and President, Kathleen Bloch, Chief Financial Officer, Vincent Capponi, Chief Operating Officer, Christian Steiner, VP of Sales and Marketing from Germany, and Chris Cramer, VP of Business Development
Before I turn the call over to Dr. Chan, I’d like to remind listeners that during the call, management's prepared remarks may contain forward-looking statements which are subject to risks and uncertainties. Management may make additional forward-looking statements in response to your questions today. Therefore, the Company claims protection under Safe Harbor for forward-looking statements contained in the Private Securities Litigation Reform Act of 1995. Actual results may differ from results discussed today and therefore, we refer you to a more detailed discussion of these risks and uncertainties in the Company's filings with the SEC. Any projections as to the Company's future performance represented by management include estimates today as of August 21, 2013 and the Company assumes no obligation to update these projections in the future as market conditions change.
Well during today’s conference call we have done our best to take everyone’s submitted questions and answer within our prepared remarks as best we can. We received many, many questions and thank everyone for participating. If we do not answer your question we would ask you to either contact myself and my team at Alliance Advisors or the Company directly after the call today.
At this time, I would like to turn the call over to Dr. Phillip Chan. Phil, the floor is yours, please go ahead.
Phillip Chan - CEO:
Thanks very much Alan and thanks to everyone who’s joined our call today.
During today’s call we’ll provide an overview of CytoSorbents for those of you who may be new to the story and update everyone on where we stand today. Along the way we plan to address many of the questions that have been submitted by investors. So thank you very much to those who submitted questions to us.
CytoSorbents is a critical care focused medical device company using blood purification to treat life-threatening illnesses. In a relatively short period of time, CytoSorbents has grown to 27 full time employees and part-time consultants, both here in New Jersey and at our subsidiary in Berlin, Germany. And we are very fortunate to have an excellent management team to lead this rapid growth. After quickly introducing myself, I’d like to ask each member of the management team to give a quick summary of his or her background.
As for myself, I have been the CEO of CytoSorbents Corporation for nearly 5 years. By background, I am a Board-certified internal medicine physician with a strong background in clinical medicine and research, having received my MD and PhD from Yale University School of Medicine and completed my residency at the Harvard Medical School at the Beth Israel Deaconess Medical Center. I am also an alum of Cornell University, where I received a BS in cell and molecular biology. Prior to joining CytoSorbents, I led healthcare and life science investments as a Partner for the $80 million NJTC Venture Fund, which is a loyal shareholder of the company. During my 5-year tenure at NJTC Venture Fund, I was responsible for numerous investments covering the gamut of therapeutic, medical device and diagnostics companies. I also co-founded Andrew Technologies, a venture-backed medical device company currently commercializing its FDA approved HydraSolve™ lipoplasty system. Vince?
Vincent Capponi - COO:
Hello everyone. My name is Vince Capponi, Chief Operating Officer of CytoSorbents. I have been with the company for more than a dozen years and have more than 20 years of management experience in medical device, pharmaceutical and imaging equipment companies including Baxter, Upjohn, Sims Deltec and Sabratek I led the regulatory approval of the first heparin flush syringe now used ubiquitously in hospitals around the world to keep IV’s patent, as well as the manufacturing of more than a million units a week for this device. At CytoSorbents, I led the CE Mark approval of CytoSorb®, the recent signing of Hitit Medical as one of our first distributors, and am heavily involved in all aspects of the company, including manufacturing of CytoSorb® and new product development. Kathy?
Kathleen Bloch - CFO:
Good afternoon everyone. This is Kathleen Bloch and I am the Chief Financial Officer of CytoSorbents. I bring more than 20 years of executive financial experience at both public and private companies to help drive the success of CytoSorbents. Before joining the company, I was the CFO of Laureate Biopharmaceuticals, a leading contract manufacturer of biologic drugs and my background includes significant experience in rapidly-growing medical device and manufacturing companies. I am a Certified Public Accountant with an MBA in finance. Chris?
Chris Cramer – VP Business Development:
Thank you Kathy. Hi, I am Chris Cramer, Vice President of Business Development for CytoSorbents. I have over 15+ years of business development and commercial experience, most recently at Johnson & Johnson, where over the course of six years, I held various leadership roles in the medical device division, most recently as Senior Director of New Venture Development. I also spent more than eight years in management consulting at PWC Consulting, working with leading health care companies across the world. Christian?
Christian Steiner – VP Sales and Marketing:
Hello, I am Dr. Christian Steiner, Vice President of Sales and Marketing of CytoSorbents and Co-Managing Director of CytoSorbents Europe GmbH. I manage all direct sales and marketing efforts of CytoSorb® in the German speaking countries of Germany, Austria and Switzerland, and am working with my colleagues in the US to expand distribution of CytoSorb® in Europe. I am a German medical physician by background and have spent more than 13 years in the sales and marketing of critical care therapies, particularly extracorporeal therapies such as the MARS liver dialysis system which was eventually acquired by Gambro, and non-invasive hemodynamic monitoring systems by Pulsion Medical. During this time, I have established a significant network of key opinion leader contacts throughout Germany, but also elsewhere in Europe and the US. So it is a pleasure to be on the call with you today.
Alan Sheinwald - Moderator:
Great. Dr. Chan – to start out, could you please give listener’s a quick overview on CytoSorbents?
Phillip Chan - CEO:
Absolutely. CytoSorbents is an emerging growth opportunity aiming to revolutionize critical care medicine with a powerful blood purification technology called CytoSorb ® , now approved in the European Union. The goal of CytoSorb ® is to prevent or treat organ failure, the cause of nearly half of all deaths in the ICU for which no therapies exist.
The underlying blood purification technology is based on highly biocompatible, porous polymer beads that act like tiny sponges to remove harmful substances from blood. When these beads are placed into a cartridge roughly the size of a drinking glass, blood can be pumped outside of the body with a standard hemodialysis machine, through this cartridge where toxic materials are removed, and that purified blood is simply recirculated back into the body. These beads are some of the highest purity sorbent materials commercially available for medical use, and are protected by 32 issued US patents with multiple applications pending.
I mentioned before that we focus on critical care medicine. Millions of people are admitted to the intensive care unit in hospitals all over the world each year for life-threatening conditions such as sepsis, trauma, burn injury, lung injury and many other conditions where inflammation plays a serious detrimental role. You may have known people who have landed in the ICU after a serious car crash, or after a bout of severe pneumonia, or after a major burn injury from a fire. These patients are extremely sick, and frequently develop organ failure where vital organs stop working. Organ failure puts these patients at extremely high risk of death.
The problem with critical care medicine today is that although we can help patients stay alive when their organs fail with supportive care therapies like mechanical ventilation or dialysis – often called “life support” - there is in fact very little that can be done to prevent or treat organ failure and actually help these patient get better except for the passage of time. In fact, keeping a patient alive until his or her own body heals itself is a major strategy on how many critically ill patients are treated today.
But because of the general inability to improve clinical outcome, patients often linger for days to weeks at a time at a cost of $2000-3000 a day in the United States, and comparable numbers in Germany and other places in Europe. If they don’t die first, they often develop complications such as infections, medical errors, and other problems which make them even more difficult to treat. So it’s no wonder that the US spends nearly 1% of its gross domestic product, or almost $80-90 billion every single year on critical care medicine and why hospitals lose billions of dollars annually.
With organ failure such a major unmet medical need, we have approached this by going after one of the most important root causes of organ failure – which is uncontrolled inflammation. Inflammation is the immune system’s normal way of dealing with infection and injury and is driven by small proteins made by the immune system called cytokines. But in life threatening illnesses, the immune system releases a massive amount of cytokines, often called cytokine storm, that unleashes deadly widespread inflammation, that can then lead to organ failure, and frequently death.
Our solution, CytoSorb®, is the only specifically approved extracorporeal cytokine filter in the European Union specifically targeting the prevention or treatment of organ failure by reducing cytokine storm and controlling potentially deadly inflammation. CytoSorb® is indicated and approved for use in any situation in the European Union where cytokines are elevated and it allows for extensive “on-label” use for many different applications. The device is plug and play with a hospital’s existing hemodialysis infrastructure and can treat your entire blood volume 20-30 times in a 6 hour period. Each treatment uses a new cartridge. And we now have more than 1,000 human treatments where there have been no serious device related adverse events so it’s been relatively safe.
CytoSorb® is a very powerful new tool for physicians – giving them the potential ability to modulate the immune response for the first time in many different diseases, and to address organ failure at its root cause. Since the launch of CytoSorb® in the second half of 2012 in our direct territories of Germany, Austria and Switzerland, we have focused on building our key opinion leader network, and now have approximately 100 key opinion leaders throughout Germany, Austria as well as the United Kingdom. CytoSorb is in the majority of University hospitals in the major cities in Germany with a growing presence in Austria. And we also now have 18 investigator initiated studies getting ready to start or being planned, and many successful case report studies also.
With approximately $400,000 in initial CytoSorb sales from June 2012 to June 2013, we believe we are just scratching at the surface of the commercial potential for CytoSorb®. With a multi-billion dollar critical care opportunity with little to no competition, an outstanding product that targets the key reason why patients die, and a very profitable razorblade in someone else’s razor business model with current 60+% gross margins that are expected to increase with volume over time, this has the potential to not only revolutionize critical care medicine by saving lives and reducing costs, but also the potential to make CytoSorbents a commercial success. The key is how we plan to get there, which is what we will discuss today.
Our technology has also received recognition from DARPA, the US Army, and the US Air Force. DARPA, or the Defense Advanced Research Projects Agency, that was responsible for funding the development of the internet, Global Positioning Satellites and robotic surgery, believes that blood purification is the right approach to treat sepsis. To this end, they awarded us a $3.8 million 5-year contract in 2012 as part of its “Dialysis-Like Therapeutics” program to treat sepsis, to remove cytokines and bacterial toxins from blood. The U.S. Army also awarded us SBIR contracts for trauma and burn injury totaling $1.15 million. In addition, the U.S. Air Force is funding a U.S.-based, 30-patient randomized, controlled pilot study to use CytoSorb to treat rhabdomyolysis in trauma patients. The FDA has granted approval, under an IDE, to begin this study, expected this year. These grants and contracts validate and advance our technologies and significantly offset operating expenses as we further commercialize CytoSorb.
Alan Sheinwald - Moderator:
Thank you Phil. Before we get into the details of the business, I think it would be helpful for investors to know what the company looks like from a financial perspective. Kathy, could you give us a snapshot of where CytoSorbents is from a financial standpoint?
Kathleen Bloch - CFO:
Certainly Alan. We have worked hard as a company to develop multiple sources of revenue. These are divided primarily into CytoSorb product revenue, of which we have both direct sales and distributor sales, and also grant income. Clearly product revenues are very important, but at this early stage in our commercialization cycle, grant income is extremely helpful as well, because it helps to covers a good portion of our R&D headcount and helps to offset many of our fixed expenses. By leveraging non-dilutive grant and contract funding, we have been able to significantly advance our technology platform without needing to raise more capital to fund this development, two things which directly benefit the Company and our shareholders.
For the six months ended June 30, 2013, we generated revenue of approximately $663,000 as compared to revenues of approximately $133,000 for the same period in 2012, which is an increase of 497%. Revenue from product sales was approximately $304,000 in the first half of 2013, as compared to $53,000 in the first half of 2012, an increase of about 600%. This increase was driven primarily by our direct sales force, with some initial sales to distributors. Product gross margins were approximately 60% for the six months ended June 30, 2013. Revenue from grants was approximately $359,000 in the first half of 2013, as compared to approximately $83,000 in the first half of 2012. Investors can find more detail in our second quarter 10-Q report filed on August 14, 2013 at http://www.sec.gov.
We were pleased with the significant growth in revenue in the first half of this year, compared to the same period a year ago and we would encourage investors to similarly look at our future progress in this manner. At our current early stage of commercialization, our sequential quarter-to-quarter results are subject to many different variables such as the timing of direct sales and reorders, and unique events such as initial stocking orders by distributors. First quarter 2013 was a particularly strong quarter that benefited from many new orders and reorders from key opinion leaders, as well as initial stocking orders from both of our distributors, where they wanted to have inventory on hand to be able to pre-market CytoSorb® to potential customers. In terms of direct sales, we have been at the key opinion leader stage, and have observed that the key opinion leaders personally want to oversee treatment with CytoSorb® to see for themselves how it is working. Despite being extremely busy, they have made time to be directly involved in patient selection, treatment, and oversight of these patients. So far the level of enthusiasm and interest remains very positive. Obviously, this level of attention is a good thing, but given the time restraints of these key opinion leaders, it means that CytoSorb is being used intermittently, rather than regularly. Our second quarter 2013 appeared to be too soon to get the usage and re-orders that we were expecting, but we have been seeing an encouraging level of activity from specific accounts already in the third quarter, despite it being the summer vacation months in Europe. Later in the call, Phil and Christian will discuss how we are planning to move from the key opinion leader stage, and begin to drive usage and demand from the rest of the physicians in the department. Until this occurs, we may have significant variability in our quarter over quarter product sales. We encourage investors to look at revenue growth from the big picture and be patient while we continue to execute our sales plan, which we believe will be successful.
I’d also like to comment on our current cash situation and how we intend to fund our operations moving forward. As of June 30, 2013, CytoSorbents had approximately $1.9 million in cash. The Company receives funds from a variety of sources, all of which are independent from each other. First, we receive revenue from government grants. Over the next six months we are expecting to achieve milestones that will permit us to receive more than $1.0M in grant funding. Second, as our sales and marketing efforts take hold, revenue growth is expected to provide additional operating funds, particularly with blended gross margins from direct sales and distributors expected to remain well above 50%. Third, we are seeking traditional commercial debt to help finance working capital expansion needs. Fourth, as we have in the past, we will seek funding in the market to support the needs of our strategic plan. Recently, for example, in June, we raised $1.1 million in convertible notes to fund our product commercialization efforts. And, lastly, in the event we need additional funding, we have an arrangement with Lincoln Park Capital to provide financing.
Alan Sheinwald - Moderator:
Thank you Kathy, that was very helpful. Dr. Chan, in your past presentations you have talked about building CytoSorbents on three pillars of value: First, commercialization of CytoSorb®. Second, Clinical trials, product development and R&D. And third, Business Development.
Let’s focus on each of these areas because I think it will provide investors with a good 360 degree view of your business opportunities. Let’s take commercialization of CytoSorb® first. I think the market recognizes the blockbuster potential of CytoSorb® as a critical care therapy. So how do we get there?
Phillip Chan - CEO:
Thanks Alan. I think its worthwhile to reiterate exactly where we are and then to turn it over to Christian, our VP of Sales and Marketing, who is on the front lines and seeing the tremendous amount of activity related to CytoSorb first hand.
CE Mark approval was a pivotal event for us because, with the notably exception of the US and Japan, it has opened up most of the world as potential markets for CytoSorb. This includes all 28 countries of the European Union where we already have regulatory approval to sell CytoSorb®, as well as the Middle East, Israel, Canada, India, Brazil, Australia, and many other countries that will accept CE Mark approval, with the appropriate product registrations. We have elected to focus on Germany, Austria and Switzerland ourselves, while we work with critical care focused distributors and strategic partners to expand our geographic footprint to other major countries and territories. This is a tried and true approach to maximizing the revenue potential of medical products.
But the important thing to note is that if we were only successful in Germany, we could make CytoSorbents into a very successful company. The critical care market in Germany alone for CytoSorb is approximately $1 billion. To this end, we have established a subsidiary in Berlin, Germany with a 4 person sales force that is targeting direct sales in Germany, Austria and Switzerland. We have established reimbursement for CytoSorb® in Germany and Austria at more than $500 per cartridge. We have also established ISO 13485 certified manufacturing at our facility in NJ, a requisite to sell medical devices into Europe. Looking at a map of Germany, our team of four salespeople have created a broad foothold with product sales, new clinical studies, case reports, and ongoing discussions in the university and public hospitals across most of the major cities in Germany and a growing number of hospitals in Austria as well.
As a small company, we have approached commercialization of CytoSorb® very methodically and conservatively to ensure that there is demand for CytoSorb before incurring costs of manufacturing and our sales force. To date, our strategy has been to cast the net wide, to create awareness, interest, and usage amongst key opinion leaders in our direct sales territories. And we have been remarkably successful at it, despite only 4 sales people in the field. We now count approximately 100 key opinion leaders, primarily in Germany, but also in Austria and the UK, who are either using or committed to using CytoSorb® in clinical use or in clinical studies, and now we have 18 investigator initiated studies being planned with two that are now registered and several others that will start soon. These studies are primarily in sepsis and cardiac surgery but include, for example, acute kidney injury, liver failure, lung injury, and pancreatitis.
Now that we see this demand and have support of the key opinion leaders, the goal is to now transition the sales strategy to not go wide, but rather to go deep into these accounts and drive adoption of CytoSorb amongst the rest of the physicians in the department, and to other ICUs in the same hospital. The goal is to generate the end-user demand that is expected to drive revenues from current hundreds of thousands of dollars in this key opinion leader phase, to really millions in sales in the “Departmental” phase. We want to target doctors who are admitting patients in the middle of the night and get them thinking about how to use CytoSorb for their patients. We have already started doing this at certain key accounts that should become reference centers for us in the near future.
The message to these junior and senior doctors is simple. The message is if you do just standard of care, the chance your patient dies, despite your best efforts, is often greater than 1 in every 3 patients. CytoSorb® is the only specifically approved cytokine filter in the EU, that has been demonstrated to safely reduce cytokine storm, with the goal of reducing inflammation and preventing or treating organ failure. The head of your department wants you to use it, the hospital is already ordering it, and the device is reimbursed. Lastly, we now have more than 1,000 human treatments where the device has been used safely, meeting the first tenet of medicine – which is, of course, first do no harm.
So these numbers can get big very quickly. If we assume 15 patients treated a quarter at an average revenue of $5,000 per patient, that single intensive care unit would produce CytoSorb sales in excess of $300K. That’s our entire sales for the first half of this year. With an average of 300 to 600 patients alone that are admitted to one of these large ICUs each year with the diagnosis of sepsis, we believe these are absolutely achievable numbers.
But the opportunity is clearly too large for a four person sales force to effectively manage, which is why our goal is to continue to expand our sales force in a responsible way but to aggressively pursue these exciting opportunities.
Now I like to turn it over to Christian to get his comments as well.
Christian Steiner – VP Sales and Marketing:
Following Phil’s comments, I’d like to give you some color about what we are seeing in the field. In Germany, I have been involved in the launch of a number of critical care products and the enthusiasm behind CytoSorb by doctors continues to impress me. Although there are other older competitive technologies in the European market from such well known companies as Fresenius Medical Care, Gambro and Bellco, these technologies are more difficult to use, do not have the flexibility of our technology, and are not commonly used by the majority of hospitals we are visiting. When we travel to critical care conferences or talk to key opinion leaders about new innovations, we also see nothing else competitive in the near term. It suggests to me that we have a unique product, with real potential, as Europe’s only specifically approved cytokine filter.
Excessive inflammation and cytokine injury is such a repetitive theme in medicine that I am also still pleasantly surprised by many of the new application ideas that are proposed by the doctors we talk to. We appear to be at the right place at the right time with this technology. Outside of critical care, an area that we are seeing growing interest is in cardiac surgery, and the use of our CytoSorb filter in a parallel bypass circuit. The interest in using CytoSorb in this application is because many things specific to the surgery and the heart-lung bypass machine can generate cytokines, free hemoglobin and inflammation – all of which can be bad for the patients and cause complications. This could be a rapid future area of growth for CytoSorb.
In general, the physicians using CytoSorb have expressed their excitement to me and our sales team with the results they have seen with the technology, and this has led to an increase in activity with more usage, more reorders, more case report studies being prepared, more investigator initiated studies being started, and more discussions with other doctors. This has led to introductions to other ICUs in the same and other outside hospitals, and more opportunities for us.
I can imagine that it is very difficult for investors to appreciate all of this activity, but as these relationships become more public we will be able to discuss them more openly, as we did in the last earnings release and today’s press release on the dosing study. We will also have 4-5 major reference accounts in the future that we will be able to talk about. And as we have more success in converting junior and senior doctors in each department, our sales numbers will begin to reflect this as well. We are confident that our efforts to drill down within each department to sell to the junior and senior physicians will be successful, as we have done this many times before. We just need more manpower, which we are working on, to be able to execute this strategy.
Finally some notable recent successes. We have seen some additional remarkable reports. For example, as reported from our distributor in Turkey, a man was accidentally electrocuted by 34,000 volts and was featured on the evening news after surviving severe external and internal burn injuries after treatment with CytoSorb. Another patient in Germany survived a serious case of Legionnaire’s disease and muscle breakdown after being treated by our therapy. And yet another patient with chronic liver disease survived septic shock with our therapy. This adds to a growing number of success stories with CytoSorb. So what we are planning is to establish a patient registry database in the near future to capture more of these data and to show more of these success stories.
Alan Sheinwald – Moderator:
Thanks Christian, that was very informative. Vince, the press release issued today on the expansion of the dosing study to now 8 major clinical sites in Germany, touches specifically on the second pillar of value which is the generation of clinical data. Having played an integral role in managing the first European Sepsis Trial, the ongoing sepsis dosing trial, and CE Mark approval for CytoSorb, can you please give listeners a quick background on sepsis, what the follow-up dosing study is intended to do, and why these particular sites are important to the company?
Vincent Capponi – COO:
Thanks Alan. CytoSorb, as a platform technology, has the potential to treat many different life threatening conditions in the ICU that include sepsis, acute respiratory distress syndrome, burn injury, trauma, pancreatitis, and many other inflammatory conditions. We have decided to focus on the treatment of sepsis, an unmet medical need, which is the result of an overzealous immune response to a severe infection and is driven by “cytokine storm”. Sepsis afflicts 18 million people worldwide every year and is increasing in incidence. There are no approved therapies to treat sepsis in the US or Europe, only a handful of therapies are being tested in Phase 3 trials, and as the literature points out, despite the best medical treatment which includes antibiotics, severe sepsis has a mortality of 25-30%, while that of septic shock is 40-50%. It accounts for 10-20% of all ICU admissions and is a top ten killer, killing more people in the US than either heart attacks, strokes, or any single type of cancer. At an average cost of more than $45,000 to treat, hospitals lose billions of dollars annually.
Despite sepsis being very difficult to treat, we believe that we have one of the most comprehensive solutions to date to treat it. Unlike other therapies that have tried and failed to stop sepsis by using very directed therapies, such as antibody therapies, CytoSorb uses a broad spectrum approach.
As Phil points out in his scientific presentation, CytoSorb® has many ways it can improve outcome in sepsis. First, it can reduce inflammatory cytokines that are directly responsible for driving deadly inflammation that can result in organ failure. Second, it can reduce excessive anti-inflammatory cytokines that can lead to immune suppression and infection – a primary driver of death in late sepsis. Third, CytoSorb® can remove many different deadly bacterial toxins as shown in vitro , such as that associated with MRSA, or methicillin resistant Staph aureus, a common hospital acquired drug resistant bacteria. And fourth, CytoSorb® can help to redirect the dangerous activated white cell component of the immune system to target the true area of infection, and to avoid innocent “bystander” organs, thereby preventing inadvertent organ injury.
Fast forward to our European Sepsis Trial. During this 47 patient randomized controlled multi-center trial, CytoSorb® was clinically proven to safely and broadly reduce cytokine storm and key cytokines in septic shock patients with lung failure, by 30-50%. Further post-hoc analysis of the data suggested that two subgroups really benefited the most from therapy – those patients with extremely high cytokine levels, and patients older than 65 years of age - where reductions in 28-day and 14-day mortality, respectively, were statistically significant. This was achieved by treating for 6 hours a day for 7 days, each day with a new device.
The data suggested that longer or more aggressive treatment could be beneficial and we set out to do a dose ranging study, what we call the “Dosing Study”, to evaluate the safety and preliminary efficacy of extended treatment, either continuous treatment for up to 7 days, which is currently ongoing, or 6 hours of treatment a day for up to 14 days, which is currently planned, each day with a new device. As we mentioned in the press release, the dosing study was designed to evaluate the safety and preliminary efficacy of extended CytoSorb® treatment, in order to give physicians more flexibility on how to treat critically ill patients with CytoSorb®. In addition, the study is not intended to produce pivotal data, but rather is intended to support the design of a pivotal trial protocol in the United States. The pivotal trial study in sepsis is expected to start in 2014, pending successful discussions with the FDA, with the goal of definitively demonstrating the effectiveness of CytoSorb® for the treatment of sepsis, and to seek U.S. regulatory approval.
We started the dosing study with three of our top performing sites from the European Sepsis Trial – University of Goettingen, Helios Erfurt, and the University of Aachen, but were approached by a number of other leading institutions to participate in our trial. As you saw in the press release, these included University of Jena, University of Hamburg-Eppendorf, University of Greifswald, University of Leipzig, and European Medical School of Oldenburg. We slowed down the trial to get these sites on board and although it took longer than expected, due to the time needed for contract negotiations, ethics committee review, and site training, we are excited that nearly every site is now eligible to enroll patients into the dosing study. As we were bringing new sites on, we conducted antibiotic compatibility testing with CytoSorb® and have provided investigators with guidance on which antibiotics are best to use with our therapy.
Regarding the new sites, these are all great sites and investigators. For example, Dr. Michael Quintel, Chief of the Department of Anesthesiology, Emergency Medicine and Critical Care at Goettingen. He is the PI of this trial, a leader in critical care research and the president of the DIVI society, one of the two major intensivist societies in Germany. Dr. Konrad Reinhart, Director of the Department of Anesthesiology and Intensive Care Medicine at the University of Jena, is the Chairman of the Global Sepsis Alliance and founding President of the German Sepsis Society, and the head of the SepNet sepsis trials network – a consortium of leading hospitals and intensivists throughout Germany who organized and ran government funded sepsis trials. And the list goes on.
We believe the addition of these sites will accelerate the completion of the Dosing Study and is key in furthering the development of the German market and raising awareness of CytoSorb for the treatment of sepsis. As data becomes available from the dosing study, we will be reviewing with our Data Safety Monitoring Board and discussing interim results when appropriate, with the goal of having at least interim data by the end of this year. Alan, back to you.
Alan Sheinwald – Moderator:
Thanks Vince, that was very informative. Turning to Strategic Partnerships as the third pillar of value, Chris – can you please give us an overview of your thoughts on the potential to do partnerships at CytoSorbents?
Chris Cramer – VP Business Development:
Yes Alan, it’s a very exciting time to be joining the company. Throughout my career I’ve seen hundreds of different medical technologies. CytoSorbents, however, clearly stands out from the rest. What’s so compelling about the technology is how flexible the platform is, and how many high value therapeutic applications it may address. For CytoSorb alone, there are numerous partnership opportunities. But I also believe the rest of the development pipeline, including HemoDefend™, ContrastSorb, DrugSorb, and several others that have not yet been publicly disclosed, offers the Company significant future growth opportunities as they are monetized.
As the head of business development, my goal is to establish one or more partnerships by the end of the year. Today we are actively building a robust business development pipeline. Over the past quarter we have been working to access key decision makers and establish relationships with leading medical device and pharmaceutical companies. Our ideal partner, and focus of our business development efforts, is a large multi-national company with strong commercial and product development capabilities that can help to advance our pipeline and accelerate product distribution.
Currently, we are aggressively focused on establishing strategic partnerships for our two most advanced assets, CytoSorb® and HemoDefend™.
For CytoSorb, we are looking to establish distribution agreements for the various applications that Phil described earlier on today’s call. Given the versatility of the platform, we are targeting a wide range of potential partners such as equipment manufacturers with strong commercial capabilities in the intensive care unit, cardiac surgery companies with heart-lung bypass equipment, and multi-national pharmaceuticals with complementary drugs for sepsis to name a few. For CytoSorb, we are seeking favorable transfer pricing and, where appropriate, upfront payments for market exclusivity and working capital for initial product stock and capacity expansion.
For HemoDefend, we are targeting leading players involved in the blood collection, processing, and transfusion businesses. We are seeking partnerships that involve collaborative development to advance our HemoDefend In-Line Filter and HemoDefend Beads-in-a-Bag products in exchange for an option for future rights to commercial distribution.
To date we’ve made significant progress in filling the business development funnel with high quality partnership opportunities. Overall, I am pleased with the strong reception and high level of interest that we are receiving from potential partners. In addition to several opportunities that were ongoing when I arrived here, we have been successful in advancing discussions with new targets. While BD deals can take some time to develop, I am confident we are laying the groundwork for future success.
Although I cannot comment on specific deals, I can give you some color commentary on the business development opportunity for CytoSorbents and the progress that we are making. Overall for CytoSorb and HemoDefend combined, we’ve identified approximately sixty potential target partners that we are aware of today. This is a very healthy number and one that has the potential to grow as we continue to explore the market for strategic partners. Of these sixty, we are actively engaged in discussions with about 1/3 of those targets. Of the targets that we are talking with, I would characterize roughly 80% to be early stage, exploratory and roughly 20% to mid to late stage in nature. Our goal is to fill the funnel with new prospects, convert new prospects into serious partnership discussions, and ultimately turn those late stage discussions into formal partnerships.
In closing, I am very happy to be joining CytoSorbents at this very exciting time. I am optimistic about the business development opportunities that are currently available and look forward to successfully driving one or more partnerships to completion by the end of the year.
Thank you. I’d now like to turn the call back over to Phil.
Phillip Chan - CEO:
Thanks very much Chris. Before concluding today’s call I’d like to leave our investors with potential catalysts in the next 6-9 months.
We believe there are some major drivers of value in our business. The first is increasing revenues, which demonstrate product adoption and acceptance in the market of our CytoSorb® technology. We plan to demonstrate significant year over year improvements, particularly as our strategy to focus heavily on the department physicians takes hold. We are also targeting the signing of at least one significant strategic partnership this year. In addition, we plan to have more clinical data, either from company-sponsored trials, military funded trials, case report studies, or through investigator initiated studies. We are also working to initiate a pivotal study with CytoSorb next year that will provide the data for either US approval or greater usage in Europe. And finally we plan to continue communicating our positive results to our shareholders and a larger investment community.
And finally, to summarize our investment opportunity in a few key points.
· First of all, we believe we are at the ground floor of commercialization, poised to create significant value with an approved product that may revolutionize critical care medicine, save lives, and reduce costs. This is the holy grail of modern medicine today.
· We are operating in a massive market with some of the biggest unmet medical needs in medicine.
· We have received validation of our company and technology from many fronts including DARPA, the US Army, the US Air Force, scientific advisors, as well as analysts.
· We have a unique, highly profitable product and pipeline with little to no competition, and an outstanding business model.
· We are led by an experienced and responsible management team.
· And there are potential major catalysts in the next 6-9 months including revenue growth, partnerships, clinical data, and other positive milestones being reached and communicated to our investors.
With that, let me thank everyone for taking the time to attend today’s call. We look forward to providing you another update in the near future. And if you have any additional questions, feel free to forward them to Alliance Advisors. With that, have a great evening.
Operator: Ladies and gentlemen, this concludes the CytoSorbents 2013 Shareholder Update Conference Call. If you would like to listen to a replay of today’s conference call, please dial 1-877-870-5176 and international 1-858-384-5517. We thank you for your participation and you may now disconnect.
Mediators of Inflammation
Volume 2013 (2013), Article ID 697972, 10 pages
http://dx.doi.org/10.1155/2013/697972
Research Article
Monocytes, Peripheral Blood Mononuclear Cells, and THP-1 Cells Exhibit Different Cytokine Expression Patterns following Stimulation with Lipopolysaccharide
Anita Schildberger,1,2 Eva Rossmanith,1 Tanja Eichhorn,1,2 Katharina Strassl,1 and Viktoria Weber1,2
1Department for Health Sciences and Biomedicine, Center for Biomedical Technology, Danube University Krems, Dr.-Karl-Dorrek-Straße 30, 3500 Krems, Austria
2Christian Doppler Laboratory for Innovative Therapy Approaches in Sepsis, Danube University Krems, Dr.-Karl-Dorrek-Straße 30, 3500 Krems, Austria
Received 8 February 2013; Revised 23 March 2013; Accepted 25 March 2013
Academic Editor: Eduardo López-Collazo
Copyright © 2013 Anita Schildberger et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
THP-1 cells are widely applied to mimic monocytes in cell culture models. In this study, we compared the cytokine release from THP-1, peripheral blood mononuclear cells (PBMC), monocytes, or whole blood after stimulation with lipopolysaccharide (LPS) and investigated the consequences of different cytokine profiles on human umbilical vein endothelial cell (HUVEC) activation. While Pseudomonas aeruginosa-stimulated (10?ng/mL) THP-1 secreted similar amounts of tumor necrosis factor alpha (TNF-a) as monocytes and PBMC, they produced lower amounts of interleukin(IL)-8 and no IL-6 and IL-10. Whole blood required a higher concentration of Pseudomonas aeruginosa (1000?ng/mL) to induce cytokine release than isolated monocytes or PBMC (10?ng/mL). HUVEC secreted more IL-6 and IL-8 after stimulation with conditioned medium derived from whole blood than from THP-1, despite equal concentrations of TNF-a in both media. Specific adsorption of TNF-a or selective cytokine adsorption from the conditioned media prior to HUVEC stimulation significantly reduced HUVEC activation. Our findings show that THP-1 differ from monocytes, PBMC, and whole blood with respect to cytokine release after stimulation with LPS. Additionally, we could demonstrate that adsorption of inflammatory mediators results in reduced endothelial activation, which supports the concept of extracorporeal mediator modulation as supportive therapy for sepsis.
1. Introduction
As a barrier between the blood stream and the surrounding tissues, the endothelium is involved in regulation of blood flow, vascular tone, thrombosis, thrombolysis, adherence of platelets, and extravasation of circulating leukocytes [1, 2]. In infection, endothelial cells are activated either directly by pathogen-associated molecular patterns, such as lipopolysaccharide (LPS) from Gram-negative bacteria, or by host-derived mediators, such as chemokines, cytokines, complement, and serine proteases [3, 4]. The endothelium responds to these mediators by switching to a proinflammatory and procoagulant state, which is associated with enhanced adhesion of platelets, monocytes, and neutrophils. Endothelial injury and endothelial dysfunction are involved in a variety of disease processes, including atherosclerosis, inflammatory syndromes, sepsis, and multiple organ failure [5].
Sepsis and sepsis-associated multiple organ failure arise in response to severe microbial infection with extensive tissue damage due to overactivation of the innate immune system and the proinflammatory cascade [6, 7]. Worldwide, sepsis is one of the leading causes of morbidity and mortality, and its incidence continues to increase [8]. The development of targeted therapies for sepsis remains a major challenge due to the extreme heterogeneity of septic patients and due to the complex network of inflammatory mediators involved in the septic process. Numerous clinical trials using specific antagonists, such as antibodies or soluble receptor constructs, to target individual inflammatory mediators were performed in the last two decades, but none of these trials did result in convincingly improved survival rates [9]. Extracorporeal blood purification techniques, such as hemofiltration or apheresis, have been proposed as possible strategies to modulate the multiple inflammatory mediators in sepsis. A potential advantage of extracorporeal approaches is that they affect only excess circulating pools of inflammatory mediators, while systemic administration of specific antagonists leads to a complete blockade of their targets also in tissues [10, 11], which may actually be detrimental.
To assess the effect of mediator modulation on endothelial activation and thus to support the preclinical development of extracorporeal adsorption therapies, we have previously established a cell culture model based on stimulation of monocytic THP-1 cells with lipopolysaccharide (LPS) in media containing human plasma. Culture supernatants derived from the stimulated THP-1 cells containing LPS and mediators secreted by THP-1 cells in response to stimulation were used to activate human umbilical vein endothelial cells (HUVEC). This model allows assessing the effect of mediator modulation with adsorbent polymers on subsequent endothelial activation [12, 13].
Due to their uniform genetic background, THP-1 cells are frequently used as a model system for monocytes. They have been shown to respond with a similar transcriptional pattern as PBMC-derived macrophages after stimulation with LPS from E. coli [14]. Since the comparability of LPS-induced cytokine secretion between THP-1 cells and PBMCs or monocytes is crucial for all cell culture models employing these cell types, we aimed to compare the LPS-induced cytokine secretion patterns of THP-1 cells, PBMC, monocytes, or whole blood and to assess their influence on subsequent activation of endothelial cells.
2. Materials and Methods
2.1. Cell Culture Media and Reagents
Medium 199 (M199), RPMI-1640 (RPMI), phosphate-buffered saline (PBS), bovine serum albumin (BSA), ethylene diamine tetraacetic acid disodium salt (EDTA), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), penicillin-streptomycin (PS), and LPS from P. aeruginosa or E. coli (O55:B5) were purchased from Sigma-Aldrich (St Louis, MO, USA). Fetal bovine serum (FBS) was obtained from PAA Laboratories GmbH (Pasching, Austria). Heparin (5000?IU/mL) was from Baxter (Vienna, Austria).
2.2. Blood and Plasma
Blood was freshly drawn from healthy volunteers after informed written consent and anticoagulated with sodium citrate at a final concentration of 12.9?mM (Mayrhofer Pharmazeutika, Leonding, Austria). Human plasma was obtained from a local plasma donation center.
2.3. Cells and Cell Culture
The human monocytic cell line THP-1 was obtained from the American Type Culture Collection (Nr. TIB-202) and was maintained as described in [12]. Isolation of PBMC from human blood was performed by density gradient centrifugation on Ficoll-Paque PLUS (GE Healthcare, Uppsala, Sweden). Briefly, 300?mL of freshly drawn blood was mixed at a ratio of 1?:?1 (vol/vol) with PBS at room temperature and 30?mL of the mixture was layered to 15?mL Ficoll-Paque PLUS in 50?mL tubes (NALGENE polycarbonate round-bottom tubes, Thermo Fisher Scientific, Waltham, MA, USA). The tubes were pretreated with Sigmacote (Sigma-Aldrich, St Louis, MO, USA) and sterilized before use. After centrifugation at 1180?g for 30?min at 20°C without break, buffy coats were collected, pooled, resuspended in PBS and centrifuged at 1180?g for 20?min at 20°C without break. The pellets were resuspended in 60?mL of PBS and again layered onto Ficoll-Paque PLUS as described previously. After centrifugation at 160?g for 20?min at 20°C without break, the top 15?mL of platelet-rich solution was discarded and the rest was recentrifuged at 350?g for 20?min at 20°C without break. Buffy coats were aspirated and washed with ice-cold PBS containing 0.1?wt% BSA and 2?mM EDTA and centrifuging at 400?g for 8?min at 4°C without break. One half of the harvested PBMCs was cultured overnight in RPMI-1640 supplemented with 0.02?M HEPES, 100?µM PS, and 10?vol% heat-inactivated FBS in HydroCell Surface 24-well plates (Thermo Fisher Scientific, Waltham, Massachusetts, USA) in humidified atmosphere (5?vol% CO2, 37°C). The other half of the harvested PBMCs was used to isolate monocytes using the Dynabeads Untouched Human Monocytes Kit (Invitrogen, Lofer, Austria) according to the manufacturer’s protocol. Isolated monocytes were cultured overnight under the same conditions as PBMCs. Isolation and maintenance of primary HUVEC were performed as described before [12].
2.4. Stimulation of Monocytes, PBMC, or THP-1 with LPS
Cells were suspended in M199 containing 0.02?M HEPES, 100?µM penicillin-streptomycin, 6?IU/mL heparin, and 10?vol% human plasma at a concentration of cells/mL and stimulated with 10?ng/mL LPS from P. aeruginosa. Stimulations were carried out in HydroCell Surface 24-well plates (1?mL/well) for 1, 2, 4, 6, 8, and 24?h in humidified atmosphere (5?vol% CO2, 37°C). Afterwards, the cell suspensions were pelleted by centrifugation, the supernatants (conditioned media) were harvested, aliquoted, and stored at -70°C until quantification of cytokines or HUVEC stimulation (Figure 1).
697972.fig.001
Figure 1: Scheme of the cell culture model. THP-1 cells, peripheral blood mononuclear cells (PBMC), monocytes, or whole blood are stimulated with lipopolysaccharide (LPS). The harvested conditioned medium containing LPS and cytokines is treated with adsorbents to modulate mediators of inflammation and is subsequently used to stimulate human umbilical vein endothelial cells (HUVEC).
2.5. Stimulation of HUVEC with Conditioned Media from Monocytes, PBMC, or THP-1
Endothelial cells were suspended in M199 supplemented with 0.02?M HEPES, 100?µM penicillin streptomycin, 6?IU/mL heparin, 10?µg/mL ECGS, and 20?vol% FBS and seeded into 96-well plates at a density of 18000 cells/well (Greiner Bio-One, Kremsmünster, Austria). After an overnight incubation, HUVEC were washed with M199 and stimulated with conditioned media which were derived from a 4, 8, or 24?h stimulation of THP-1, PMBC, or monocytes. HUVEC stimulation was performed using 200?µL of conditioned media per well for 16?h in a humidified atmosphere. Thereafter, supernatants were aspirated, centrifuged at 15500?g (5?min, 4°C), and stored at -70°C until quantification of cytokines.
2.6. Stimulation of Whole Blood with LPS
Aliquots of 1.5?mL of freshly drawn heparin-anticoagulated human blood (5?IU/mL blood) from healthy volunteers were spiked with either 10 or 1000?ng/mL LPS from P. aeruginosa or E. coli. Samples without LPS served as control. After stimulation for 1, 2, 4, 6, 8, or 24?h at 37°C with gentle shaking, the samples were centrifuged at 1500?g (5?min, 4°C), and the plasma was stored at -70°C until further use.
2.7. Adsorption of Mediators of Inflammation
Blood was stimulated with 100?ng/mL LPS from E. coli for 4?h as described previously. The plasma resulting from this stimulation was diluted tenfold with M199 to obtain conditioned medium derived from blood (CMB). THP-1 cells were stimulated with 10?ng/mL LPS from E. coli for 4?h as described previously to obtain conditioned medium derived from THP-1 cells (CMT). An adsorbent for the specific binding of TNF-a was prepared by covalent binding of a chimeric human-mouse monoclonal anti-TNF-a antibody (Infliximab, Centocor, Leiden, The Netherlands) onto cyanogen bromide-activated Sepharose 4B (GE Healthcare, Uppsala, Sweden). A polystyrene-divinylbenzene (PS-DVB) copolymer (Amberchrom HPR10; Dow Chemical, Midland, MI, USA) was used as a selective adsorbent to bind cytokines and complement factors [15–17]. The PS-DVB copolymer was coated with human serum albumin (Octapharma, Vienna, Austria) to improve its blood compatibility. The conditioned media derived from blood or THP-1 stimulation were incubated with the adsorbents for 1?h at 37°C with gentle shaking. A ratio of 1?vol% (TNF-a adsorbent) or 10?vol% (PS-DVB) of adsorbent to medium was used. After treatment with the adsorbents, aliquots of 1.5?mL of the media were applied to HUVEC (400000 cells per well of a 6-well culture plate) for 15?h in humidified atmosphere (5?vol% CO2, 37°C). Thereafter, culture supernatants were aspirated, centrifuged at 15500?g for 5?min at 4°C, aliquoted, and stored at -70°C until quantification of cytokines. Surface expression of the adhesion molecules E-selectin and ICAM-1 was determined with flow cytometry.
2.8. Quantification of Cytokines
Concentrations of TNF-a, IL-1ß, IL-6, IL-8, and IL-10 were determined using the Bio-Plex 200 system (Bio-Rad, Vienna, Austria).
2.9. Flow Cytometric Analysis
The purity of monocytes was assessed by determining the CD14-positive cell population. After monocyte isolation, cells were washed twice with ice-cold PBS and stained with FITC-conjugated anti-CD14 or with the respective IgG control antibody (Becton Dickinson, Vienna, Austria) in PBS supplemented with 2?vol% FBS on ice for 30?min. After two washing steps, cells were analyzed on a FACScan flow cytometer and data were analyzed using the CellQuest software (Becton Dickinson, Vienna, Austria). For detection of the surface expression of the adhesion molecules E-selectin and ICAM-1, HUVEC were detached using 0.02?wt% EDTA and washed with ice-cold PBS containing 0.1?wt% sodium azide (Sigma-Aldrich, St Louis, MO, USA). Staining of surface markers was performed by incubation with PE-conjugated anti-E-selectin, PE-Cy5-conjugated anti-ICAM-1, or the respective IgG control antibodies (Becton Dickinson, Vienna, Austria) in PBS supplemented with 2?vol% FBS on ice for 30?min. After two washing steps, 10000 gated cells were analyzed on a FC 500 flow cytometer and data were analyzed using the CXP software (Beckman Coulter, Vienna, Austria).
2.10. Statistical Analysis
Statistical analysis was performed using the software package SPSS Statistics for Windows, version 18.0 (SPSS Inc., Chicago, IL, USA). When comparing two groups, data were analysed by the nonparametric Wilcoxon rank sum test. Data are expressed as means ± SD. Significance was accepted at .
3. Results
3.1. Cytokine Release upon Stimulation of Monocytes, PBMC, and THP-1 Cells with LPS
Monocytes were 95% pure according to flow cytometry. The stimulation of THP-1 cells, PMBC, and monocytes with LPS resulted in comparable TNF-a expression patterns with a peak at 4?h and a subsequent decrease over time (Figure 2). Monocytes secreted higher amounts of TNF-a than PBMC and THP-1 cells (?pg/mL versus ?pg/mL versus ?pg/mL at 4?h, resp.). After 24?h of stimulation, TNF-a levels declined to 460?pg/mL (monocytes), 790?pg/mL (PBMC), and 110?pg/mL (THP-1 cells), respectively. THP-1 cells did not secrete IL-6 and IL-10, while PBMC and monocytes produced increasing amounts of IL-6 (?pg/mL and ?pg/mL after 24?h) and IL-10 (?pg/mL and ?pg/mL after 24?h) over time. THP-1 cells secreted 900 ± 830?pg/mL of IL-8 after 24?h of stimulation, whereas PBMC and monocytes exhibited much higher IL-8 release (?pg/mL and ?pg/mL). In summary, monocytes and PBMC secreted comparable amounts of TNF-a, IL-6, IL-8, and IL-10 over time. THP-1 showed a secretion of TNF-a that was comparable to monocytes and PBMC but released by far less IL-8 than PBMC or monocytes and failed to secrete IL-6 and IL-10.
697972.fig.002
Figure 2: Cytokine release upon stimulation of THP-1, PBMC, and monocytes with lipopolysaccharide. Cultures of one million cells per?mL of medium containing 10?vol% plasma were stimulated with 10?ng per?mL of LPS from P. aeruginosa (black circles). Unstimulated cultures served as control (white circles). Concentrations of TNF-a, IL-6, IL-8, IL-10, and IL-1beta are given as mean ± SD ().
3.2. Stimulation of HUVEC with Conditioned Media
Conditioned media derived from THP-1 cells, PBMC, or monocytes after 4, 8, or 24?h of stimulation with LPS were used to stimulate HUVEC for 16?h (Figure 3, values for 8?h are shown). HUVEC secreted less IL-6 with conditioned media derived from PBMC and monocytes than with conditioned media derived from THP-1 cells. IL-8 levels increased after HUVEC stimulation with conditioned media from THP-1 cells but decreased after stimulation with conditioned media from PBMC and monocytes. For all three conditioned media, IL-10 concentrations remained stable after HUVEC stimulation, while TNF-a concentrations decreased.
697972.fig.003
Figure 3: Effect of conditioned media on HUVEC. Conditioned media (white bars) derived from an 8?h stimulation of THP-1, PBMC, or monocytes with LPS were applied onto HUVEC and the cytokine release was measured. Concentrations of IL-6, IL-8, IL-10 and TNF-a are expressed as mean ± SD (). Black bars indicate control media without LPS.
3.3. Stimulation of Whole Blood with LPS
In addition to THP-1 cells, PBMC, or monocytes, whole blood was stimulated with LPS. Freshly drawn blood anticoagulated with heparin (5?IU/mL blood) was treated with LPS for up to 24?h. In contrast to the stimulation of THP-1 cells, PBMC, or monocytes, 10?ng/mL LPS from P. aeruginosa did not lead to cytokine secretion from whole blood (Figure 4). However, 1000?ng/mL LPS from P. aeruginosa lead to significant secretion of TNF-a, IL-1ß, IL-6, IL-8, and IL-10. LPS from E. coli showed a higher stimulatory potential than P. aeruginosa, and 10?ng/mL was sufficient to elicit secretion of TNF-a, IL-1ß, IL-6, IL-8, and IL-10.
fig4
Figure 4: Effect of LPS on cytokine secretion from whole blood. Freshly drawn blood was stimulated with 10 or 1000?ng/mL LPS from P. aeruginosa or E. coli. Samples without LPS served as control. Concentrations of TNF-a, IL-1ß, IL-6, IL-8, and IL-10 are expressed as mean ± SD ().
3.4. Effect of Cytokine Adsorption on HUVEC Activation
Given the different cytokine expression patterns of THP-1 cells and whole blood, we aimed to investigate whether these differences would also have an effect on subsequent HUVEC activation. Therefore, whole blood was stimulated with 100?ng/mL LPS from E. coli for 4?h and the plasma was diluted tenfold with M199 to obtain conditioned medium derived from blood (CMB), which contained ?pg/mL TNF-a, ?pg/mL IL-6, and ?pg/mL IL-8 (Figure 5). In parallel, THP-1 cells were stimulated with 10?ng/mL LPS from E. coli for 4?h in medium M199 supplemented with 10?vol% plasma to yield conditioned medium derived from THP-1 cells (CMT) which contained ?pg/mL TNF-a but no IL-6 and lower concentrations of IL-8 (?pg/mL). The conditioned media were treated with either a specific adsorbent for TNF-a or with a selective cytokine adsorbent. Treatment with the specific TNF-a adsorbent resulted in complete removal of TNF-a from CMB and CMT. The selective cytokine adsorbent completely removed IL-6 and IL-8 and decreased TNF-a concentrations by 95%. After specific adsorption of TNF-a or selective cytokine adsorption from CMB or CMT, HUVEC were stimulated with the conditioned media and the secretion of IL-6 and IL-8 and surface expression of the adhesion molecules E-selectin and ICAM-1 were monitored. Secretion of IL-6 ( versus ?pg/mL) and IL-8 ( versus ?pg/mL) was higher for CMB than for CMT despite equal TNF-a concentrations in both media (Figure 5), indicating the presence of additional stimulatory factors in CMB next to TNF-a. Pretreatment of conditioned media with the specific TNF-a adsorbent or the selective cytokine adsorbent resulted in decreased secretion of IL-6 and IL-8 from HUVEC. For conditioned medium derived from blood, the release of IL-6 was reduced to 63% (not significant) and 1% () for specific TNF-a adsorption and selective cytokine adsorption, respectively, while IL-8 release was reduced to 54% and 4% ( in both cases). For conditioned medium derived from THP-1 cells, the release of IL-6 was reduced to 33% (not significant) and 6% (), and IL-8 release was reduced to 12% () and 2% () for specific adsorption of TNF-a versus selective cytokine adsorption. Thus, selective cytokine adsorption had a much stronger influence on HUVEC activation as compared to specific TNF-a adsorption. Regarding the expression of surface adhesion molecules, HUVEC exhibited significantly higher expression of E-selectin after stimulation with conditioned medium derived from blood as compared to conditioned medium derived from THP-1 cells. Adsorption of TNF-a resulted in significantly decreased or completely abolished E-selectin expression for CMB and CMT, respectively. Selective cytokine adsorption completely abolished E-selectin expression with both stimulation media (Figure 6).
697972.fig.005
Figure 5: Effect of mediator modulation on HUVEC activation. Whole blood was stimulated with 100?ng/mL LPS from E. coli for 4?h and diluted tenfold with M199 to obtain conditioned medium derived from blood (CMB). THP-1 cells were stimulated with 10?ng/mL LPS from E. coli for 4?h in medium M199 supplemented with 10?vol% human plasma to yield conditioned medium derived from THP-1 cells (CMT). HUVEC were stimulated with CMB and CMT, respectively. Concentrations of TNF-a, IL-6 and IL-8 prior to mediator modulation (CM) and after specific TNF-a adsorption or selective cytokine adsorption (PS-DVB) are shown. Data are given as mean of three experiments ± standard deviation. Significance was accepted at . n.s.: not significant.
fig6
Figure 6: Effect of mediator modulation on adhesion molecule expression. The experimental setup was identical to Figure 5. HUVEC surface expression of the adhesion molecules E-selectin (a) and ICAM-1 (b) is expressed as mean fluorescent intensity (mfi) minus basal expression of adhesion molecules on HUVEC. Data are given as mean of three experiments ± standard deviation. Significance was accepted at . n.s.: not significant.
4. Discussion
Extracorporeal modulation of inflammatory mediators, such as cytokines, with filters or adsorbents is regarded as a promising supportive therapy for sepsis. During preclinical development of such extracorporeal approaches, cell culture models allow to assess the biological effect of mediator modulation. We have previously established a cell culture model based on stimulation of monocytic THP-1 cells with lipopolysaccharide and subsequent activation of endothelial cells with the conditioned medium [12]. THP-1 cells are widely used to study the function of monocytes [18]. One of their major advantages over primary monocytes is their homogenous genetic background, which abolishes donor variability. Further, they are easily accessible and can be obtained without contamination with other blood components, while the availability of primary human monocytes is limited. In several studies, THP-1 have been shown to represent a more mature monocytic phenotype than other immortalized human monocyte cell lines, such as U937 cells [19], and it has been demonstrated that the interaction between THP-1 and endothelial cells is comparable to human primary monocytes [20–22]. Gene expression profiles of THP-1 after LPS stimulation are very similar to primary PBMC-derived macrophages [14]. Still, the extent to which THP-1 cells mimic monocytes is not fully elucidated, which prompted us to compare THP-1 cells to freshly isolated human peripheral blood mononuclear cells (PBMC) or monocytes. We found that upon stimulation with LPS, primary human monocytes and PBMC secreted comparable amounts of TNF-a, IL-6, IL-8, and IL-10 over time, while THP-1 cells secreted similar amounts of TNF-a but did not secrete IL-6 and IL-10. Moreover, their release of IL-8 was much lower than the observed one for primary human monocytes and PBMC under identical experimental conditions.
The expression pattern of TNF-a was in accordance with previously published data showing that monocytes reacted to an LPS stimulus by secretion of TNF-a in vivo or in vitro within the first hours after stimulation and that TNF-a concentration declined after the initial peak [23]. In contrast to our findings, THP-1 cells have been reported in the literature to secrete IL-6 and IL-10 [24, 25], albeit under different experimental conditions, as THP-1 cells were differentiated to macrophages and higher LPS concentrations were used in the published studies.
In addition to monocytes, PBMC, and THP-1, we stimulated whole blood with lipopolysaccharide. During whole blood stimulation, cells are not stressed by isolation and cultivation procedures. In addition, whole blood stimulation experiments are faster and cheaper to perform. In our study, a concentration of 10?ng/mL LPS from P. aeruginosa was sufficient to stimulate isolated cells, while 1000?ng/mL was needed to activate whole blood. In accordance with literature [26], lipopolysaccharide from E. coli showed a higher stimulatory potential and 10?ng/mL elicited cytokine secretion from whole blood. Thus, isolated blood cells show higher sensitivity to stimulation with lipopolysaccharide as compared to whole blood, which may be due to interaction of LPS with other blood components, such as lipoproteins.
As shown previously, endothelial cells react to culture supernatants from LPS-activated THP-1 cells by increased gene expression of inflammation-related factors [27], by increased activity of NF-?B, by increased secretion of cytokines and plasminogen activator inhibitor, and by enhanced surface expression of adhesion molecules such as ICAM-1 and E-selectin. In this study, we chose secretion of IL-6 and IL-8 as activation markers and showed that conditioned media derived from PBMC and monocytes resulted in comparable IL-6 secretion from HUVEC, whereas no IL-8 was secreted, in contrast to the use of conditioned media from THP-1 cells. Conditioned medium derived from whole blood stimulation activated HUVEC even to a higher extent than conditioned medium derived from THP-1 cells despite comparable concentrations of LPS and TNF-a in both media. Mediator modulation with either a specific adsorbent for TNF-a or with a selective polystyrene divinylbenzene copolymer, which binds to a range of cytokines, significantly reduced subsequent HUVEC activation. While the specific adsorbent resulted in reduction of HUVEC cytokine release to at least 50%, the effect of selective cytokine adsorption was even more pronounced with a reduction of cytokine secretion by more than 90%, indicating that factors in addition to TNF-a are relevant for HUVEC stimulation. These findings support the concept of selective mediator modulation as supportive therapy for sepsis rather than the specific targeting of individual factors.
Conflict of Interests
The authors have no conflict of interests to declare.
Acknowledgments
The excellent technical support by Ingrid Linsberger is gratefully acknowledged. This work was funded by the Christian Doppler Society (Christian Doppler Laboratory for Innovative Therapy Approaches in Sepsis).
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53 Employees? This is Huge...
Frank Clark
Partner at Game On 365
Co-Founder and Executive Vice President CytoSorbents Public Company; 11-50 employees; CTSO.OB; Medical Devices industry
August 1993 – March 2004 (10 years 8 months) New York, NY
Actively working with the company on several projects 2010-2013.
Co-founded and raised $50 million for startup medical device company from institutional and high net worth individual investors. Managed road shows and presented at venture capital and investment banking conferences internationally.
Managed investor relations and business development. Negotiated agreements with Fortune 500 companies. Created educational events for industry thought leaders, physicians, scientists and investors. Presented at industry events (business development) and venture investor conferences.
Company now public (CTSO) and the device received the CE Mark in March 2011 for application of blood purification in sepsis patients in a critical care setting. Device is now being sold in Europe for blood purification to treat sepsis and other critical care applications in which cytokine storm plays a substantial role. These may include severe burns, Acute Lung Injury, Acute Respiratory Distress Syndrome, pancreatitis, and improving outcomes in cardio pulmonary bypass surgeries and possibly improving organ transplant outcomes.
Built the management team and oversaw growth from 2 people to 53 people in three locations. Oversaw construction of manufacturing facility in NJ.
http://www.linkedin.com/in/frankclark