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Have fun its my last post god willing - http://www.hkgem.com/newlistings/prospectuses/e_8031pro-20020522chap14.pdf
Mingwan0-It is the same guy take a look - http://www.gis.a-star.edu.sg/homepage/cvdetailview.jsp?invid=14&alpha=
Ming i thought they were the same. Jianjum Liu - On the board of Sun City Industries; 2} is or was at the University of Alberta,Canada; 3}also at Columbia University; 4} Now attending tha Asia-Pacific conference.Looks like the same guy to me.Yes or No
Ming. not only is Tony going to be there but so is one of the members of the board who elect. him to be CEO of Sun. "Jianjun Liu"
BETTER READ THIS !!$$$$ - http://www.hkgem.com/newlistings/prospectuses/e_8031pro-20020522chap14.pdf
see post 153363 page 29 - Shaanxi Daiying Biological Engineering Co.LTD.
The Group entered into a letter of intent dated 30 June 2001 with Shaanxi Daiying Biological Engineering LTD. {Shaanxi Dailing} under which the parties agree to establish a screening system for screening Drugs for Hepatitis C in collaboration using the SimBIODas Tm Technology. Shaanxi Dailing an independentthird party, is engaged in research on biological Engineering and is specialised on the research and development of methods for treatment of Hepatitis C Virus.
Thankyou Ming and others for sharing your view you all may be right and everyday we learn something new. But thats not how i read it.There's nothing in that news release that says Dnap did only one test and if i were running a comp. and offered to do one test on 10,000 people for Several hundred dollars Then I would fire myself and hire someone with Brains.Thats not the way do do Business and run a company if you want to make money.
Dnap is like everyone else there in it to make money not lose money. My understanding is its $1000.00 for each test,Not less.If you go to our distributors of the test or do research on it from other sources, you will find it being sold for $1000.00 Now that $1000. for each of the 10,000 people tested,thats a lot of money but not for a whole country like England.Even if Dnap gave a discount because of the large number Dnap would still make a few Million on the deal.They went through 10,000 people what 10,000 Times $1000.=10 Million ecen at a 50% discount Dnap will still make 5 Million.
DNAPrint Helped Significantly Reduce the DNA Donor Sample Collections Required in a Major Criminal Investigation by New Scotland Yard and the London Metropolitan Police
Friday July 9, 5:19 pm ET
SARASOTA, Fla., July 9 /PRNewswire-FirstCall/ -- DNAPrint genomics, Inc. (OTC Bulletin Board: DNAP - News) announced that, according to the July 7, 2004 London Times article "1,000 men were asked to give DNA in hunt for serial rapist" by Madeleine Acey, "Detectives hunting one of Britain's worst serial sex attackers are inviting 1,000 men to provide DNA samples as they try to identify the rapist. Officers have narrowed down a list of possible suspects to 1,000 men who meet a geographical and racial profile. They started off with 10,000."
"The report DNAPrint provided to New Scotland Yard and the London Metropolitan Police clearly reduced the number of samples and possible leads that needed to be investigated," said Dr. Matthew Thomas, Senior Scientist at DNAPrint. "This shows the power of DNAWitness 2.5 when used in the early stages of an investigation. Saving time in a criminal investigation is critical in the proper allocation of investigative resources and, hopefully, preventing further crimes."
"Sometimes eyewitness reports and even reports and descriptions from the victims are not enough to narrow the focus of an investigation," said Richard Gabriel, CEO and President of DNAPrint genomics, Inc. "We were able, based on our database results, to narrow the field of possible donors. The agencies were able to trim their pool of possible donors from 10,000 to 1,000 individuals, using DNAWitness and other information," he said. "The reduction in possible donors represents a cost-savings to the agencies, not to mention the personnel time it would require to process 10,000 samples versus the time it takes for 1,000 samples. This not only saves money, but it also saves valuable time and helps speed up the investigation process," said Gabriel.
DNAWitness 2.5 is a genomic-based test that can determine an individual's relative amounts of Native-American, East-Asian, Sub-Saharan African and European genes. Without any information about the investigation or the crime, DNAWitness 2.5 can provide the investigator with necessary guidance to properly focus their investigations. DNAWitness results can provide the spark that ignites a new line of questioning in an investigation.
About DNAPrint genomics, Inc.
DNAPrint genomics Inc. uses proprietary human genome research methods to develop genomic-based services and products. The Company introduced ANCESTRYbyDNA in the consumer market and DNAWitness in the forensic market in 2003. DNAPrint is developing products in the pharmacogenomic market and has a disease gene discovery program. The Company is traded on the Nasdaq OTC Bulletin Board under the ticker symbol DNAP. For more information about the company, please visit http://www.dnaprint.com.
All statements in this press release that are not historical are forward- looking statements within the meaning of Section 21E of the Securities Exchange Act as amended. Such statements are subject to risks and uncertainties that could cause actual results to differ materially from those projected, including, but not limited to, uncertainties relating to technologies, product development, manufacturing, market acceptance, cost and pricing of DNAPrint's products, dependence on collaborations and partners, regulatory approvals, competition, intellectual property of others, and patent protection and litigation. DNAPrint genomics, Inc. expressly disclaims any obligation or undertaking to release publicly any updates or revisions to any forward-looking statements contained herein to reflect any change in DNAPrint's expectations with regard thereto or any change in events, conditions, or circumstances on which any such statements are based.
a $158. is not the price they would have to pay its much more. I think around $1,000. with around 20 free samples.
Officers have narrowed down a list of possible suspects
to 1,000 men who meet a geographical and racial profile.
That means Dnap did the work on 10,000 samples.IMO
stockboy - they found a 1000 matches out of 10,000.Thats 10k samples
What !!! PRB Pharmaceuticals working with University of South Florida !!!
see where it says - Plans are underway to create a new anti-viral, high throughput screening laboratory and a state-of-the-art drug discovery laboratory. Oh I wonder what that means !!!!
Check this out -
http://www.prbpharmaceuticals.com/RD/
RESEARCH AND DEVELOPMENT
Our approach to research and development is rigorously scientific, unbiased and open-minded.
In the United States, PRB Pharmaceuticals maintains extensive collaborative relationships with scientists at the University of Southern California and University of South Florida, and in Asia with the Chinese University of Hong Kong.
Our general approach is to screen thousands of compounds for their therapeutic properties. Candidate compounds are then formulated into OTC products or further fractionated, screened, and purified to produce lead molecules for ethical pharmaceutical development. This results in a rapid transformation of platform technologies into viable treatments and therapies.
We are currently conducting anti-viral research at the University of Southern California. We are also conducting laboratory research and clinical trials related to influenza virus, SARS coronavirus, and other viral diseases at the Prince of Wales Hospital in Hong Kong.
Plans are underway to create a new anti-viral, high throughput screening laboratory and a state-of-the-art drug discovery laboratory.
We ensure that all our treatments and therapies are rigorously tested and safe before releasing them on the market, and we provide extensive data in the Patient Product Information (PPI) distributed with our products.
PRB Pharmaceuticals' open-minded and scientific approach leaves no stone unturned in the pursuit of better solutions.
Remember Zengen has a Lic. agreement with PRB Pharmaceuticals who is working with Lee's Pharmaceuticals. Also Dragon Venture Inc. is working close with Zengen.
**********************************
WOODLAND HILLS, CALIF (October 10, 2002) — Zengen, Inc. announced today a strategic alliance with DragonVenture, Inc., a Silicon Valley based, cross-Pacific venture capital and consulting firm. DragonVenture will provide Zengen with services related to the Company’s international expansion into the Greater Chinese pharmaceutical and over-the-counter markets.
"This alliance further supports our overall corporate strategy to leverage our proprietary peptide technologies in the global healthcare arena," said R. Steven Davidson, Ph.D., president and CEO of Zengen. "Given their expertise and proven track record in the Asian marketplace, we believe that working with a premier organization like DragonVenture will help us successfully penetrate the world’s largest healthcare market and establish Zengen as a global biopharmaceutical player."
Under the terms of their agreement, DragonVenture will facilitate strategic alliances between Zengen and well-established healthcare businesses in Greater China. Zengen’s move builds on the Company’s recent research and development collaborations with companies including Abiogen Pharma S.p.A. in Italy and Lee’s Pharmaceutical Holdings, Ltd. headquartered in Hong Kong.
The Chairman and Co-Founder of DragonVenture is K. Bobby Chao, who happens to be a Director of Zengen. I think that one way in which Zengen/Lees might possibly figure in DNAP’s future is as a marketing and distribution mechanism for Greater China.
angelfund, the presentation on July 31 is "New tests for genomics-based physical profiling : case study" rather than presenting the published paper. My guess is that Tony will provide background on the science behind this and then go over the Louisiana case. He might provide details about some of the additional tests, i.e. eye and hair color. It would be very nice if he announced progress on tests for e.g. height, weight and facial characteristics. Of course, we do not know what other announcements, if any, before or at this meeting might have a bearing on his presentation...
Also by worktoplay - By: worktoplay
08 Jul 2004, 11:59 PM EDT
Msg. 304759 of 304845
(This msg. is a reply to 304755 by worktoplay.)
Jump to msg. #
Terry...One more thing...
http://www.prbpharmaceuticals.com/RD/
RESEARCH AND DEVELOPMENT
Our approach to research and development is rigorously scientific, unbiased and open-minded.
In the United States, PRB Pharmaceuticals maintains extensive collaborative relationships with scientists at the University of Southern California and University of South Florida, and in Asia with the Chinese University of Hong Kong.
Our general approach is to screen thousands of compounds for their therapeutic properties. Candidate compounds are then formulated into OTC products or further fractionated, screened, and purified to produce lead molecules for ethical pharmaceutical development. This results in a rapid transformation of platform technologies into viable treatments and therapies.
We are currently conducting anti-viral research at the University of Southern California. We are also conducting laboratory research and clinical trials related to influenza virus, SARS coronavirus, and other viral diseases at the Prince of Wales Hospital in Hong Kong.
Plans are underway to create a new anti-viral, high throughput screening laboratory and a state-of-the-art drug discovery laboratory.
We ensure that all our treatments and therapies are rigorously tested and safe before releasing them on the market, and we provide extensive data in the Patient Product Information (PPI) distributed with our products.
PRB Pharmaceuticals' open-minded and scientific approach leaves no stone unturned in the pursuit of better solutions.
This is from Arch's Research posted last night (Thanks GCBR):
Shaanxi Yangling Daiying Biological Engineering Ltd. The Group entered into a letter of intent dated 30 June 2001 with Shaanxi Yangling Daiying Biological Engineering Ltd.(“Shaanxi Daiying ”)under which the parties agreed to establish a screening system for screening drugs for Hepatitis C in collaboration using the SimBioDAS ™technology. Shaanxi Daiying, an independent third party, is engaged in research on biological engineering and is specialised on the research and development of methods for treatment of Hepatitis C virus. Further details of the collaboration are subject to further negotiation and mutual agreement.
And just so people understand what this SimBioDAS Technology does:
The main bottleneck in the development process before a drug candidate is introduced into humans is predicting the degree to which a drug candidate taken by mouth will be absorbed through the intestine and into the body. To help to solve this bottleneck, the Group has focused its research program on developing a platform technology called the SimBioDAS ™ technology to predict how well a drug candidate will be absorbed into the human body. This system will be used to screen drug candidates for their absorption properties as part of an automated screening program leading to decisions on which drug candidates to be brought forward for further development.
The need for PK/PD information is not restricted to western medicines. Natural products,including TCM and nutraceuticals,are comprised of complex mixtures of active ingredients,and ingredients with pharmacological activity must be absorbed to exert their activity in the body. In order to have an effective formulation based on natural products,the absorbable active ingredients should be identified in the raw herbs and then their activities and absorbability should be confirmed in the marketed product. This information is missing in regard to the majority of TCM products and nutraceuticals,so there is little basis for the quality control now demanded by consumers and regulatory agencies like the FDA.Identifying active ingredients has been a difficult task, but the Directors believe that with the proprietary technology being developed by the Group, the Group is poised to solve this major problem.
The emerging SimBioDAS ™technology is an in vitro screening system through which the Group aims to help shorten the drug development timeline,thereby reducing the cost and resources required for the pre-clinical drug development process.The amount of time that can be saved by using the SimBioDAS ™ technology in the drug development process cannot be generalized in any specific instance. It depends on various factors, including in which stage of the drug development process is the SimBioDAS ™technology being used and whether the SimBioDAS ™ technology is used in different stages in the process. The Directors believe that the SimBioDAS ™technology could shorten the drug development time by a few months to more than a year, depending on the situation. On the basis that it costs an average of US$500 million and 10 years to discover and develop a drug, every week is about US$1 million on average. Thus the saving of even a few weeks in the process could make using the SimBioDAS ™ technology cost effective.
The SimBioDAS ™ technology is intended to work in the following areas:
–to help pharmaceutical companies focus on developing drug candidates that are well-absorbed by humans;
–to predict pharmacokinetics,especially absorption,of drug candidates in animals using an in vitro system; and
–to predict possible pharmacokinetic and toxicity problems with drug candidates in humans by revealing the potential differences between animal and human absorption of screened compounds.
In summary,the SimBioDAS ™technology aims to contribute to the lead identification process and facilitate rapid decisions on compound selection for further development and on discontinuation of less well-absorbed candidates. The SimBioDAS ™technology is expected to be adapted to predict the absorption of ingredients in TCM in humans. The Group also intends to develop a related biotechnological method to effectively isolate the active ingredients in natural herbal products. Using the Group ’s expertise and technology,the Group aims to test and develop novel high quality TCM formulations as drug products for the world market.
Later,
W2P
By: worktoplay
08 Jul 2004, 11:33 PM EDT
Msg. 304755 of 304844
(This msg. is a reply to 304741 by terryhallinan.)
Jump to msg. #
terry...Speculation...
Are these events related in any way?
http://biz.yahoo.com/bw/040422/226216_1.html
Sun City Industries Enters into Reorganization Agreement with China Based Biotech & Pharmaceutical Company
Thursday April 22, 10:16 pm ET
Agreement Signed With Yangling Daiying Biological Engineering Co., Ltd.
APOPKA, Fla.--(BUSINESS WIRE)--April 22, 2004-- Sun City Industries, Inc. (OTCBB: SCII - News) has entered into a Reorganization Agreement with Yangling Daiying Biological Engineering Co., Ltd., a corporation organized under the laws of the People's Republic of China. Daiying has developed the world's very first Hepatitis C virus (HCV) culture that retains biological activity in vitro. The Reorganization Agreement is conditional upon the signing of a consulting agreement, warrant agreement and the completion of various schedules to be part of the Reorganization Agreement. A closing date has not yet been set but the parties are striving to close in April 2004. Sun City will be filing an 8K to satisfy its reporting requirements under Regulation FD and the Exchange Act of 1934 as amended. The Reorganization Agreement will be provided as an exhibit to the 8K.
And this:
http://www.prbpharmaceuticals.com/news/attachment/Lai_release.pdf
Dr. Lai has also been at the forefront of research on the hepatitis C virus. He lead the scientific team that discovered that the hepatitis virus can mimic one of the molecular targets of interferon, the body’s naturally occurring antiviral agent, blocking its ability to kill viruses. This finding has served as a cornerstone of research to develop effective therapies to combat hepatitis C.
I only mention it because of this:
http://www.prbpharmaceuticals.com/company/management.asp
Hector J. Gomez, M.D., Ph.D., Chief Medical Officer, Head of Research and Development.
Dr. Gomez has over twenty years of experience in the biopharmaceutical industry. From 1994-1999, as President and CEO of Transcend Therapeutics, a biotechnology company in Cambridge, MA, Dr. Gomez defined its strategic planning and oversaw its implementation. He led the transition from a private to a public company (IPO). He was also CEO of Zengen, Inc., a private Biotech Company in California. Before joining Transcend, he was Vice President of Medical Affairs for Vertex Pharmaceuticals in Cambridge, MA, where he was responsible for planning and implementing the Pre-Clinical, Clinical Research and Regulatory Affairs departments. Prior to that he was a senior executive for two large pharmaceutical companies: Executive Director, Cardiovascular for Ciba-Geigy in Summit, NJ, and before that, Senior Director, Cardiovascular Clinical Research for Merck in Rahway, NJ. Dr. Gomez received an M.D. from the National University of Colombia, and a Ph.D. from Marquette University and a Diploma in Clinical Pharmacology from Tulane University.
Most of the management of PRBPharmaceuticals were at one time associated with Zengen, a privately held California Biotech company conducting research into novel peptides. Dr. Gomez was at one time the CEO of Zengen. Interestingly, he was only CEO for about a six month period, and then left there. The month he left, he founded GemBiomics LLC with Richard Gabriel. A month after that he was named to DNAPrint's BOD. Some time in 2003 he reuinited with many of the former principals involved with Zengen, and became CMO of PRB (right around the time he was named CMO of DNAPrint).
Later,
W2P
BETTER READ THIS !!$$$$ - http://www.hkgem.com/newlistings/prospectuses/e_8031pro-20020522chap14.pdf
also
By: worktoplay
08 Jul 2004, 11:59 PM EDT
Msg. 304759 of 304780
(This msg. is a reply to 304755 by worktoplay.)
Jump to msg. #
Terry...One more thing...
http://www.prbpharmaceuticals.com/RD/
RESEARCH AND DEVELOPMENT
Our approach to research and development is rigorously scientific, unbiased and open-minded.
In the United States, PRB Pharmaceuticals maintains extensive collaborative relationships with scientists at the University of Southern California and University of South Florida, and in Asia with the Chinese University of Hong Kong.
Our general approach is to screen thousands of compounds for their therapeutic properties. Candidate compounds are then formulated into OTC products or further fractionated, screened, and purified to produce lead molecules for ethical pharmaceutical development. This results in a rapid transformation of platform technologies into viable treatments and therapies.
We are currently conducting anti-viral research at the University of Southern California. We are also conducting laboratory research and clinical trials related to influenza virus, SARS coronavirus, and other viral diseases at the Prince of Wales Hospital in Hong Kong.
Plans are underway to create a new anti-viral, high throughput screening laboratory and a state-of-the-art drug discovery laboratory.
We ensure that all our treatments and therapies are rigorously tested and safe before releasing them on the market, and we provide extensive data in the Patient Product Information (PPI) distributed with our products.
PRB Pharmaceuticals' open-minded and scientific approach leaves no stone unturned in the pursuit of better solutions.
This is from Arch's Research posted last night (Thanks GCBR):
Shaanxi Yangling Daiying Biological Engineering Ltd. The Group entered into a letter of intent dated 30 June 2001 with Shaanxi Yangling Daiying Biological Engineering Ltd.(“Shaanxi Daiying ”)under which the parties agreed to establish a screening system for screening drugs for Hepatitis C in collaboration using the {b}SimBioDAS ™technology{/b}. Shaanxi Daiying, an independent third party, is engaged in research on biological engineering and is specialised on the research and development of methods for treatment of Hepatitis C virus. Further details of the collaboration are subject to further negotiation and mutual agreement.
And just so people understand what this SimBioDAS Technology does:
The main bottleneck in the development process before a drug candidate is introduced into humans is predicting the degree to which a drug candidate taken by mouth will be absorbed through the intestine and into the body. To help to solve this bottleneck, the Group has focused its research program on developing a platform technology called the SimBioDAS ™ technology to predict how well a drug candidate will be absorbed into the human body. This system will be used to screen drug candidates for their absorption properties as part of an automated screening program leading to decisions on which drug candidates to be brought forward for further development.
The need for PK/PD information is not restricted to western medicines. Natural products,including TCM and nutraceuticals,are comprised of complex mixtures of active ingredients,and ingredients with pharmacological activity must be absorbed to exert their activity in the body. In order to have an effective formulation based on natural products,the absorbable active ingredients should be identified in the raw herbs and then their activities and absorbability should be confirmed in the marketed product. This information is missing in regard to the majority of TCM products and nutraceuticals,so there is little basis for the quality control now demanded by consumers and regulatory agencies like the FDA.Identifying active ingredients has been a difficult task, but the Directors believe that with the proprietary technology being developed by the Group, the Group is poised to solve this major problem.
The emerging SimBioDAS ™technology is an in vitro screening system through which the Group aims to help shorten the drug development timeline,thereby reducing the cost and resources required for the pre-clinical drug development process.The amount of time that can be saved by using the SimBioDAS ™ technology in the drug development process cannot be generalized in any specific instance. It depends on various factors, including in which stage of the drug development process is the SimBioDAS ™technology being used and whether the SimBioDAS ™ technology is used in different stages in the process. The Directors believe that the SimBioDAS ™technology could shorten the drug development time by a few months to more than a year, depending on the situation. On the basis that it costs an average of US$500 million and 10 years to discover and develop a drug, every week is about US$1 million on average. Thus the saving of even a few weeks in the process could make using the SimBioDAS ™ technology cost effective.
The SimBioDAS ™ technology is intended to work in the following areas:
–to help pharmaceutical companies focus on developing drug candidates that are well-absorbed by humans;
–to predict pharmacokinetics,especially absorption,of drug candidates in animals using an in vitro system; and
–to predict possible pharmacokinetic and toxicity problems with drug candidates in humans by revealing the potential differences between animal and human absorption of screened compounds.
In summary,the SimBioDAS ™technology aims to contribute to the lead identification process and facilitate rapid decisions on compound selection for further development and on discontinuation of less well-absorbed candidates. The SimBioDAS ™technology is expected to be adapted to predict the absorption of ingredients in TCM in humans. The Group also intends to develop a related biotechnological method to effectively isolate the active ingredients in natural herbal products. Using the Group ’s expertise and technology,the Group aims to test and develop novel high quality TCM formulations as drug products for the world market.
Later,
W2P
Heressome work done by Huimin Zhang {also on the Board} -
http://hyper.ahajournals.org/cgi/content/full/31/1/283
Angiotensin II Stimulates Synthesis of Endothelial Nitric Oxide Synthase
Bettye S. Hennington; Huimin Zhang; M. Todd Miller; Joey P. Granger; Jane F. Reckelhoff
From the Department of Physiology and Biophysics (B.S.H., H.Z., M.T.M., J.P.G., J.F.R.), University of Mississippi Medical Center, Jackson, MS.
Correspondence and reprint requests to Jane F. Reckelhoff, PhD, Department of Physiology and Biophysics, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 39216-4505. Email: jfr@fiona.umsmed.edu
Abstract
Top
Abstract
Introduction
Methods
Results
Discussion
References
Previous studies have suggested that NO may play an important role in protecting the renal vessels from angiotensin II (ANGII)-mediated vasoconstriction. One possible mechanism for this interaction is that ANGII could stimulate NO production in the kidney by increasing endothelial NO synthase (NOS III). The present studies were performed in rats to determine whether acute or chronic elevations in ANGII are associated with enhanced renal NOS III mRNA or protein synthesis. In both acute and chronic studies captopril (20 µg/kg/min) was given IV to inhibit endogenous ANGII production. Acute suprarenal infusion of ANGII (8 ng/kg/min) for 110 minutes had no effect on arterial pressure but decreased GFR and renal plasma flow by 20% and 30%, respectively, and increased renal vascular resistance by 70%. Acute ANGII increased renal NOS III mRNA by 70% (as determined by ribonuclease protection assay), but had no effect on renal NOS III protein concentration (as detected by Western blot analyses). In contrast, chronic infusion of ANGII (5 ng/kg/min) for 10 days, increased arterial pressure by 30% and tended to reduce GFR and renal plasma flow. Chronic ANGII had no effect on renal NOS III mRNA levels, but increased NOS III protein by 90%. These data suggest that ANGII can stimulate NOS III synthesis and suggest that this may be one of the mechanisms whereby AngII may enhance NO production.
Key Words: GFR • renal hemodynamics • mRNA • ribonuclease protection assay • Western blot
Abbreviations: ANGII = angiotensin II • FF = filtration fraction • NOS III = endothelial nitric oxide synthase isozyme III • GFR = glomerular filtration rate • MAP = mean arterial pressure • NO = nitric oxide • RPA = ribonuclease protection assays • RPF = renal plasma flow • RVR = renal vascular resistance
Introduction
Top
Abstract
Introduction
Methods
Results
Discussion
References
In recent years there has been considerable interest in the interaction between NO and angiotensin II (ANGII).1,2 While ANGII is known to play an important role in controlling renal hemodynamics via its actions on postglomerular vessels, recent studies have implied that ANGII may have an effect on preglomerular vessels that is modulated by the effects of nitric oxide (NO) or prostaglandins. Ito and colleagues, using isolated rabbit arterioles, demonstrated that production of NO is more important in modulation of afferent arteriolar tone rather than having an effect on the efferent arteriole.3 These investigators further found that NO modulated ANGII-induced vasoconstriction of the afferent arteriole but not the efferent arteriole.4 We have also recently reported that the acute effect of ANGII on preglomerular vessels in dogs is importantly modulated by nitric oxide.5 These data suggested that NO may play an important protective role in modulating the vasoconstrictor effect of ANGII on preglomerular vessels.
The mechanisms by which NO could protect the renal vasculature from vasoconstrictor ANGII have not been fully elucidated. One possible mechanism is that ANGII could enhance NO production and NO would offset a direct vasoconstrictor effect of ANGII. Whether ANGII is capable of stimulating renal NO production is unclear. Deng and colleagues found that acute infusion of pressor doses of ANGII increased urinary nitrate/nitrite excretion, metabolites of NO and indicators of NO production.6 However, chronic ANGII (5–6 days) had no effect on nitrate/nitrite excretion.6 Although this study suggests a role for ANGII in stimulating NO since urinary nitrate/nitrite excretion estimates whole body NO production, it is unclear whether ANGII stimulates renal NO production in these studies.
There are at least two mechanisms by which ANGII could increase renal NO production. First, ANGII could increase nitric oxide synthase enzyme activity by causing an increase in intracellular calcium concentration. Studies have been performed which document that ANGII is capable of increasing intracellular calcium in a concentration-dependent manner in endothelial cells in culture.7,8 Secondly, ANGII could increase NO by increasing endothelial NO synthase (NOS III) synthesis, either on a transcriptional level, typically by increasing and/or stabilizing the mRNA levels for NOS III, or on a translational level, and could thereby increase the amount of NOS III protein. Both transcriptional and translational increases in NOS III would increase NO production, if it is assumed that an increase in NOS III protein would be translated into an increase in NOS III activity in the cell and if the levels of substrate arginine and cofactors are not rate-limiting. A direct effect of ANGII on NOS III synthesis has not been previously investigated, however.
The present studies were performed to determine if acute intrarenal nonpressor doses of ANGII could stimulate NOS III synthesis. In addition, pressor doses of ANGII were given chronically for 10 days to determine the long-term effect of ANGII on transcription and translation of NOS III.
Methods
Top
Abstract
Introduction
Methods
Results
Discussion
References
Experimental Animals
For these studies, 53 male Sprague Dawley rats (Harlan SD, Indianapolis, Ind), aged 3 to 5 months, were used. The protocols were approved by the Institutional Animal Care and Use Committee of the University of Mississippi Medical Center. Rats were maintained on rat chow (Teklad) and tap water in a 12 hours/12 hours light/dark cycle until the time of study.
Short-Term Effect of ANGII on Renal Hemodynamics and NOS III Synthesis
The short-term effect of AngII on renal hemodynamics and NOS III synthesis was examined in rats pretreated with converting enzyme inhibitors. Prior to renal function studies, rats were placed on 4% NaCl diet for 24 to 48 hours, and during the experiment, converting enzyme inhibitors were given to all rats to block endogenous production of AngII. A diagram of the acute AngII Protocol is shown in Fig 1A. Rats were divided into two groups. Rats in group 1 (n = 17) were time controls and received intravenous infusions of captopril (20 µg/kg/min) and suprarenal infusions of isotonic saline in the first and second periods of the experiment. Rats in group 2 (n = 15) received intravenous captopril and suprarenal infusions of saline in the first period and captopril and suprarenal infusions of ANGII (8 ng/kg/min) in the second period of the experiment. Renal hemodynamics were measured in 6 control rats and 8 ANGII-treated rats).
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Figure 1. Diagrams for experimental protocols 1 and 2 to determine the acute (top) and chronic (bottom) effects of ANGII on renal and systemic hemodynamics and NOS III synthesis. In protocol 1 a total of 32 rats (17 control, 15 AngII-treated) were studied; hemodynamics were only measured in 14 rats (6 control, 8 AngII-treated). In protocol 2, 20 rats (9 control and 12 AngII-treated) were studied, and hemodynamics were measured in all rats.
Specifically, rats were anesthetized by intraperitoneal injection of the thiobarbiturate, Inactin (100 to 110 mg/kg body weight; RBI) and placed on a temperature-regulated surgery table to maintain rectal temperature at 36 to 38°C. Femoral and jugular catheters were placed, and a tracheostomy was performed as described previously.9,10 A catheter (PE-10) was also inserted below the bifurcation of the aorta on the left and advanced above the renal arteries for suprarenal infusion of isotonic saline or ANGII. A 23g needle connected to PE-50 tubing was inserted into the left renal vein in the retrograde direction for blood sampling for determination of extraction across the kidney for calculation of renal plasma flow. During catheter placement, the rats received an infusion of isoncotic artificial rat plasma (2.5g/dL bovine immunoglobulin, 2.5g/dL bovine serum albumin in Ringer’s solution) at 12.5 mL/kg/h for 45 minutes and thereafter at 1.5 mL/kg/h throughout the experimental period to maintain an euvolemic preparation.10,11 3H-inulin (15 to 20 µCi/ml saline; New England Nuclear), was infused at 1.5 mL/h.
As shown in Fig 1A, following a 50-minute equilibration period for the 3H-inulin and captopril (20 µg/kg/min), two 20-minute clearances were performed during the first (control) period. Following this control period, ANGII (8 ng/kg/min) was infused suprarenally for 50 minutes. After equilibration of the ANGII, two 30-minute clearances were performed (period 2). Following the experiments, kidneys were removed, weighed, and placed in liquid nitrogen for assessment of NOS III mRNA and protein.
Long-Term Effect of ANGII on Renal Hemodynamics and NOS III Synthesis
A diagram of the chronic ANGII protocol is shown in Fig 1B. Rats were divided into two groups. Rats in group 1 (time control) (n = 9) received captopril (20 µg/kg/min) IV throughout the control (4 days) and the experimental periods (10 days). Rats in group 2 (n = 12) received captopril during the control period (4 days) and captopril and ANGII (5 ng/kg/min) in the experimental period (10 days). Abdominal aortic and femoral venous catheters were implanted into rats under pentobarbital (50 mg/kg) anesthesia. A midline abdominal incision was made and the abdominal aorta was separated from the inferior vena cava. A small hole was made in the aorta with a 20g needle and a catheter of medical vinyl tubing (sz V/4, BOLAB) in the shape of an "s" was inserted into the aorta. The femoral vein was also cannulated with vinyl tubing (sz V/3, BOLAB). Both catheters were advanced subcutaneously along the back and exteriorized at the nape of the neck through a button sutured to the skin and connected to a spring. Rats were placed into individual metabolism cages, and the spring tether was connected to a two-channel hydraulic swivel above the cage. The femoral catheter was connected via the swivel to an infusion pump at a rate of 0.62 mL/h. The arterial catheter was connected via the swivel to a transducer (Cobe) connected to an A-D converter for 24-hour blood pressure recording. Rats were provided with normal sodium intake (2.2 mEq/d) via the combination of diet and saline infusion.
As shown in Fig 1B, after a 7 day recovery from surgery, the rats were given continuous infusions of captopril (15 µg/kg/min) via the femoral venous catheter. On day 3 of captopril infusion in 2 rats, bolus IV injections of angiotensin I (ANGI; 0.3 µg/kg) were given to verify adequate blockade of converting enzyme. Prior to captopril treatment, the ANGI bolus caused increases in systemic arterial pressure from 111 mm Hg to 150 mm Hg. On day 3 of captopril in the same animals, Ang I bolus caused no increase in systemic blood pressure (96 to 96 mm Hg).
On day 4 of the captopril infusion, GFR and estimated renal plasma flow (RPF) were measured by infusion of [125I]-iothalamate (Isotex Diagnostics; 0.05 µCi/kg/min) and 1% para-aminohippurate at a rate of 0.62 mL/h. After approximately 18 hours of infusion, blood samples were drawn. Following control GFR and RPF measurements, captopril and ANGII (5ng/kg/min) or saline vehicle was infused chronically for 10 days. On day 9, GFR and RPF measurements were repeated. Renal hemodynamics were measured in 9 control rats and 12 ANGII treated rats). On day 10, the rats were euthanized with Inactin (150 mg/kg), and the kidneys were removed, weighed, and placed in liquid nitrogen for assessment of eNOS mRNA and protein.
Ribonuclease Protection Assays (RPA)
Liquid nitrogen frozen whole kidney tissue was ground with a mortar and pestle, and total RNA was isolated by homogenization in RNAstat (Teltest), followed by two chloroform extractions, one isopropropyl precipitatation, and three 70% ethanol washes. The pellet was resuspended in DEPC-treated water, stored at -20°C, and used within 72 hours. Total RNA concentration and purity was determined spectrophotometrically. Rat endothelial NO synthase cDNA was a gift from Dr. Robert Star (University of Texas Southwestern Medical Center, Dallas). The cDNA was linearized with PST 1, and -32 P-UTP-labeled antisense RNA probes were made using the MAXIscript kit (Ambion), according to manufacturer’s directions. Full-length probes were gel-purified. RNA probes for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (template DNA from Ambion) were also made to serve as controls. Probe excess was determined by control experiments in which the amount of RNA added was increased twofold and decreased by one half to verify the linearity of the RNA/probe response. RPAs were performed using a commercially available RPA II kit according to the manufacturer’s instructions (Ambion). The protected fragments were separated by electrophoresis on 5% denaturing polyacrylamide gels. Quantitation was performed by phosphorimager system (Biorad).
Western Blot Analyses
Liquid nitrogen frozen kidneys were homogenized 20% (weight/volume) in 20 mmol/L Hepes, pH 7.5, containing 100 µM pepstatin, 100 µg/mL aprotinin, 10 mmol/L EDTA, 100 µg/mL leupeptin, 1 mmol/L phenanthroline, and 1 mmol/L E-64 (Sigma). Western blot analyses for NOS III were performed as previously described by us,12 using a mouse monoclonal anti-NOS III primary antibody (1:1000; Transduction Laboratories) and a horse radish-peroxidase-conjugated, goat anti-mouse IgG (1:1000) (Amersham) secondary antibody. The bound antibody was detected using the ECL kit (Amersham) and quantified by densitometry (Biorad).
Statistical Analyses
The renal functional data were analyzed by analyses of variance (ANOVA), using Statview 512 software for the Macintosh. Significance was defined as P<.05. All data values are expressed as mean±SEM. For RPAs and Western blots; differences between groups (controls and AngII-treated) were assessed by Student’s t test, P<.05, defined as significant.
Results
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Abstract
Introduction
Methods
Results
Discussion
References
Acute Response to ANGII
As shown in Table 1, there were no changes in mean arterial pressure, GFR, renal plasma flow (RPF), or renal vascular resistance within group 1 (control) rats. Suprarenal infusion of ANGII (group 2 rats) had no effect on mean arterial pressure, but caused GFR to decrease by 20%. RPF also decreased by 30% with ANGII, and renal vascular resistance increased by 70%. Renal NOS III mRNA was increased by 74% with acute suprarenal infusion of ANGII (Fig 2A). In contrast, there was no change in renal NOS III protein concentration in response to ANGII (Fig 2B).
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TABLE 1. Hemodynamic Response to Acute Angiotensin II Infusion in Male Sprague-Dawley Rats
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Figure 2. Effect of acute ANGII on renal mRNA and protein for endothelial NO synthase (NOS III). ANGII (8 ng/kg/min) was given for 110 minutes to anesthetized rats pretreated with captopril. A. Top: Representative ribonuclease protection assays (RPAs) taken from phosphorimaging scans: Lanes 1–5, ANGII-treated rats; lanes 6–11, control rats; lanes 12 and 13, controls containing yeast tRNA, labeled RNA probe in the presence (lane 12) and absence (lane 13) of RNase. Bottom: Densitometric analyses of RPAs (total n = 17 control and 15 ANGII-treated rats) of the acute effect of ANGII on renal NOS III mRNA. B. Top: Representative Western blots of renal NOS III in response to acute ANGII. Lanes 1–3, control; lanes 4–6, ANGII-treated. Bottom: Averages of densitometric scans (n = 7 control, 8 ANGII-treated rats) of Westerns. *,P<.05, in ANGII-treated group compared to control group.
Response to Chronic ANGII
As shown in Table 2, there was no effect of time on mean arterial pressure, GFR or RPF (group 1). In response to chronic ANGII infusion, mean arterial pressure increased by approximately 30% (group 2). There was a tendency for GFR and RPF to decrease, but not significantly so. Chronic infusion of pressor doses of ANGII for 10 days had no effect on renal NOS III mRNA levels (Fig 3A), but did increase NOS III protein in the kidney by 90% (Fig 3B).
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TABLE 2. Hemodynamics in Rats Given Chronic Angiotensin II
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Figure 3. Effect of chronic ANGII on renal mRNA and protein for endothelial NO synthase (NOS III). ANGII (5 ng/kg/min) was infused intravenously for 10 days in rats pretreated with captopril. A. Top: Representative RPA taken from phosphorimaging scans: Lane 1 and 2; ANGII-treated rats; lanes 3 and 4, control rats; lanes 5 and 6, controls containing yeast tRNA and labeled RNA probe in the presence (lane 5) and absence (lane 6) of RNase. Bottom: Densitometric analyses of RPAs (n = 9 control, 12 ANGII-treated rats) of the chronic effect of ANGII on renal NOSIII mRNA. B. Top: Representative Western blot for renal NOS III. Lanes 1–3, control; lanes 4–6, ANGII-treated; lane 7, NOS III standard. Bottom: Averages of densitometric scans of Westerns for NOS III in response to chronic AngII (n = 7 control, 8 ANGII-treated rats). *, P<.05, in ANGII-treated group compared to control group.
Discussion
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Abstract
Introduction
Methods
Results
Discussion
References
The present studies demonstrate that acute ANGII-induced reductions in renal hemodynamics are associated with increased renal NOS III mRNA production. In addition, we report that long-term elevations in ANGII result in increased NOS III protein synthesis, supporting the hypothesis that ANGII can chronically increase NO production in the kidney.
Acute ANGII infusions at nonpressor doses caused a reduction in GFR and renal plasma flow. In addition, ANGII increased the renal mRNA levels of NOS III. However, there was no effect on renal NOS III protein concentrations. The lack of an acute effect of ANGII to increase renal NOS III protein is not surprising since the ANGII infusion time was only 110 minutes and was probably not long enough to allow translation of NOS III protein to be upregulated or, if upregulated, to be detected using the technique of Western blot. These data suggest that it is unlikely that the increase in NOS III synthesis plays an important role in the acute ANGII-NO interaction. However, the fact that NOS III mRNA is increased suggests an important initiating step in the interaction between NO and ANGII which goes beyond the transient acute interactions.
In support of the latter hypothesis, chronic ANGII infusion for 10 days increased mean arterial pressure and tended to reduce GFR and renal plasma flow. Chronic ANGII infusion had no effect on renal NOS III mRNA levels. However, renal NOS III protein levels were increased significantly with ANGII. This pattern of increased protein in the absence of increases in mRNA is consistent with an increase in basal production of NOS III enzyme, perhaps due to stabilization of the message, thus not requiring an increase in mRNA to increase translation of the protein. Barring any change in L-arginine substrate or NO synthase cofactors, the increase in NOS III protein would cause increased production of NO in the kidney. These data suggest then that ANGII can stimulate renal NO production by increasing NOS III protein content. Consistent with our findings is a preliminary report by Zou and colleagues, who found the ANGII directly increases NO in the medulla of the kidney as measured by the oxyhemoglobin microdialysis technique.13
Although the present studies describe the effect of ANGII to increase NOS III synthesis, these studies do not explain the mechanism(s) by which NOS III synthesis could be increased by ANGII. Since there are shear stress elements in the 5' untranslated region of NOS III gene,14,15 it is possible that ANGII could cause an increase in NOS III synthesis due to an increase in shear stress. However, since Ito and colleagues have demonstrated an increased NO response to acute ANGII in isolated afferent arterioles, a preparation independent of flow,3 the mechanism for increased NOS III synthesis with ANGII may also be independent of shear stress.
It is also possible that ANGII could increase NO synthesis by directly affecting the second messenger systems in the endothelial cells and thereby increase synthesis of NOS III. Siragy and Carey have recently reported that the renal response of increased cGMP, an indicator of NO production, to acute ANGII was mediated by angiotensin AT2 receptors.16 Other investigators have shown that ANGII increases intracellular calcium concentrations in cultured endothelial cells in a dose-dependent manner.7,8 Since NOS III activity is calcium-dependent, this may also be a mechanism for the acute increase in NO production mediated by NOS III. However, future studies will be necessary to determine if the NOS III synthetic response to chronic ANGII is also mediated via the AT2 receptors, increases in intracellular calcium, or other second messenger systems.
In the present study, our results suggest that chronic infusion of ANGII increases NOS III activity. However, we have not directly measured the levels of NOS III activity. The major reason for this is that it is difficult to assess the activity of the NOS III isoform separately from the activity of the other NO synthase isoforms. There are no inhibitors specific for NOS III activity that do not also inhibit the activity of the other isoforms of NO synthase. Whether ANGII can indeed increase NOS III activity in the renal vasculature by increasing the NOS III protein concentration needs to be further investigated.
In summary, acute suprarenal infusion of nonpressor doses of ANGII increased renal mRNA for NOS III, but had no effect on renal NOS III protein concentration. In contrast, chronic ANGII infusion, at low pressor doses, had no effect on NOS III mRNA levels in the kidney, but significantly increased renal NOS III protein concentration. These data suggest that ANGII is capable of controlling local NO production in the renal vasculature, thus protecting against ANGII-induced vasoconstriction.
Acknowledgments
This work was supported by the American Heart Association, Mississippi Affiliate and by HL51971 from NIH.
Received September 17, 1997; first decision October 10, 1997; accepted October 24, 1997.
References
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Abstract
Introduction
Methods
Results
Discussion
References
Sigmon DH, Beierwaltes WH. Renal nitric oxide and angiotensin II interaction in renovascular hypertension. Hypertension. 1993; 22 : 237 –242.[Abstract]
Sigmon DH, Newman JM, Beierwaltes WH. Angiotensin II:endothelium-derived nitric oxide interaction in conscious rats. J Am Soc Nephrol. 1994; 4 : 1675 –1682.[Abstract]
Ito S, Johnson CS, Carretero OA. Modulation of angiotensin II-induced vasoconstriction by endothelium-derived relaxing factor in the isolated microperfused rabbit afferent arteriole. J Clin Invest. 1991; 87 : 1656 –1663.[Medline]
Ito S, Arima S, Ren YL, Juncos LA, Carretero OA. Endothelium-derived relaxing factor/nitric oxide modulates angiotensin II action in the isolated microperfused rabbit afferent but not efferent arteriole. J Clin Invest. 1993; 91 : 2012 –2019.[Medline]
Schnackenberg CG, Wilkins FC, Granger JP. Role of nitric oxide in modulationg the vasoconstrictor actions of angiotensin II in preglomerular and postglomeular vessels in dogs. Hypertension. 1995; 26 : 1024 –1029.[Abstract/Free Full Text]
Deng X, Welch WJ, Wilcox CS. Role of nitric oxide in short-term and prolonged effects of angiotensin II on renal hemodynamics. Hypertension. 1996; 27 : 1173 –1179.[Abstract/Free Full Text]
Saito S, Hirata Y, Emori T, Imai T, Marumo F. Angiotensin II activates endothelial constitutive nitric oxide synthase via AT1 receptors. Hypertens Res. 1996; 19 : 201 –206.[Medline]
Ko Y, Glodny B, Stier S, Totzke G, Nickenig G, Dusing R, Sachinidis A, Vetter H. Angiotensin type-1 (AT1) receptor gene expression in primarily cultured human arterial umbilical endothelial cells. Biochem Pharmacol. 1997; 53 : 417 –421.[Medline]
Reckelhoff JF, Kellum JA, Jr, Racusen LC, Hildebrandt DA. Long-term dietary supplementation with L-arginine prevents age-related reduction in renal function. Am J Physiol. 1997; 272 : R1768 –R1774.[Medline]
Reckelhoff JF, Manning RD, Jr. Role of endothelial-derived nitric oxide in the control of the renal micovasculature in aging male rats. Am J Physiol. 1993; 265 : R1126 –R1131.[Medline]
Ichikawa I, Maddox DA, Cogan MG, Brenner BM. Dynamics of glomerular ultrafiltration in euvolemic Munich Wistar rats. Renal Physiol. 1978; 1 : 121 –131.
Reckelhoff JF, Hennington BS, Moore AG, Blanchard EJ, Cameron J. Gender differences in renal NO system: dissociation between expression of endothelial NO synthase and renal hemodynamic response to NO synthase inhibition. Am J Hypertens. 1997 . (in press)
Zou A-P, Wu F, Cowley AW. Protective effect of angiotensin II-induced increase in nitric oxide in the renal medullary circulation. Hypertension. 1997 . (in press)
Buga GM, Gold LE, Fukuto J.M, Ignarro LJ. Shear stress-induced release of nitric oxide from endothelial cells grown on beads. Hypertension. 1991; 17 : 187 –193.[Abstract]
Sessa WC, Harrison JK, Luthin DR, Pollock JS, Lynch KR. Genomic analysis and expression patterns reveal distinct genes for endothelial and brain nitric oxide synthase. Hypertension. 1993; 21 : 934 –938.[Abstract]
Siragy H.M, Carey RM. The subtype 2 (AT2) angiotensin receptor mediates renal production of nitric oxide in conscious rats. J Clin Invest. 1997; 100 : 264 –269.[Abstract/Free Full Text]
Look at Sat. Feb.21 {Jianjum Liu} Second RECOMB Satellite Workshop on Computational Methods for SNPs and Haplotypes = "Comparison of Four Computational Methods for Haplotype Inference Problem" http://www.aladdin.cs.cmu.edu/workshops/recomb/
This guy is on the board
(iv) Change the name of the Company to CHINA BIOTECH & PHARMACEUTICAL CORP.; and
By worktoplay - What just happened...Back in April, a reorganization agreement was put in place, whereby Sun City would acquire the Chinese company in exchange for issuing 34,880,000 shares of common stock. That agreement was subject to the conditions listed below and as of this date has not closed.
Today's announcement moved Mannion out of the way and moved Charles Scimeca, the Consultant named below, into control of Sun City. So what should we expect to happen next?
ARTICLE THREE
SPECIAL CONDITIONS
3.1 Conditions to Closing
A. The obligations of each Party to this Agreement are subject to the condition precedent that the other Party's representations and warranties contained in this Agreement shall be true, correct and complete on and as of the date of Closing with the same effect as though such representations and warranties were made on and as of such date.
B. At the time of closing, all the original officers, directors, and employees of the Corporation shall have completed their legal resignations. (Done)
C. Prior to Closing, the Corporation's Director shall elect Dr. Tony N. Frudakis, Ph.D., Wenxia Guo, Peiyi Tian, Jianjun Liu, and Huimin Zhang to serve as members of the Board of Directors. Dr. Tony N. Frudakis shall serve as a director of the Board for the next two years without compensation. (Done)
D. The Corporation, as of the closing date, is fully reporting under the Securities Exchange Act of 1934, as amended and is currently trading on the over the counter bulletin board (OTC-BB). (Done)
E. Concurrent with the Closing, the Corporation shall:
(i) Transfer 34,880,000 shares of the outstanding shares of common stock of the Corporation to the Subscriber;
(ii) Transfer 1,400,000 shares to be held in escrow for the benefit of Consultant or its assigns by Anslow & Jaclin, LLP until completion of funding for the post reorganized corporation of at least Four Million USD ($4,000,000) within four (4) months of the effective date of the registration statement on Form SB-2, or any other acceptable registration statement, to be paid for by the Corporation. If the Consultant or its assigns fails to raise the above amount of funds for the reorganized corporation within the above stipulated time, it shall result in the unconditional transfer of the 1,400,000 shares to the Subscriber, and shall forfeit its right to an additional three million warrants set forth hereinafter in 3.1 E(iii). If the Consultant or its assigns fulfills the funding responsibilities for Four Million USD ($4,000,000) set forth herein within the stipulated time, the Consultant or its assigns shall be entitled to the release of the 1,400,000 shares as well as an additional three million warrants as set forth. Any of the participants related to this Agreement shall not be entitled to transfer, deposit or deal with this 1,400,000 shares without prior written consent from the Consultant and Corporation (post reorganization).
(iii) The Consultant shall be responsible for providing sources in raising the four million (USD) in capital and the Corporation shall be responsible for completing the registration of shares noted on Schedule B with the Securities and Exchange Commission to register the shares noted on Schedule B.
Financing method:
(1) If the financing of at least $4 million is arranged by Consultant in equity capital of 4 million shares of the Subscriber and the final financing amount reaches $5,000,000 USD, $1 million or any part thereof of that amount exceeding $4 million USD shall be paid to Consultant and exclusively used on behalf of the Corporation for its business activities related to this Agreement (such as promotions). This amount shall be paid to Consultant to perform such services set forth in the Consulting Agreement. Any financing amount exceeding $5 million shall be owned by the Corporation (post reorganization); (2) If the financing is fulfilled by the Consultant's sources in a loan of up to $10 million at the current prevailing market rate with 4 million shares of the Subscriber as collateral, 1.5% of the loan amount exceeding the $ 4 million financing amount shall be granted as a service fee to the Consultant in a lump sum payment. Neither the shareholders, nor the Corporation, nor the consultant shall be entitled to transfer or deal with the 4 million shares as collateral.
The Financing herein before shall be SEC compliant and shall not cause lawsuits to the Corporation (post reorganization). Four (4) million shares of Subscriber's stock shall be registered and allocated for this purpose. In no event, shall the Consultant act as a broker/dealer in connection with such funding obtained as a result of Consultant's sources on behalf of Subscriber.
After fulfillment of funding responsibilities set forth in 3.1E(ii) set hereinbefore, Consultant shall be entitled to an additional three million warrants as follows:
500,000 shares at the exercise price of $.75 per share, when the price of the Corporation's (post reorganized) shares of common stock close at or above $.75 within four (4) months of the effective date of the registration statement filed with the SEC, or such warrants will expire worthless.
800,000 shares at an exercise price of $1.50 per share when the price of the Corporation's (post reorganized) shares of common stock closes at or above $1.50 within six (6) months of the effective date of the registration statement filed with the SEC, or such warrants will expire worthless.
900,000 shares at an exercise price of $2.50 per share when the price of the Corporation's (post reorganized) shares of common stock closes at or above $2.50 within nine (9) months of the effective date of the registration statement filed with the SEC, or such warrants will expire worthless.
800,000 shares at an exercise price of $3.50 per share when the price of the Corporation's (post reorganized) shares of common stock closes at or above $3.50 within 12 months of execution hereof or such warrants will expire worthless.
All 3,000,000 warrants will be structured with cashless exercise language and the shares underlying the warrants will be registered in a registration statement filed with the SEC.
As set forth in Section 1.2 D., the reorganized Corporation shall execute a Consulting Agreement with the Consultant which shall relate to the Consultant obtaining financing for the Corporation (post reorganization).
(iv) Change the name of the Company to CHINA BIOTECH & PHARMACEUTICAL CORP.; and
(v) Obtain shareholder approval for all of the above.
F. The Corporation (post reorganization) shall take all corporate actions necessary to form a wholly owned subsidiary and to allow Consultant to complete a spin-off transaction whereby the Corporation will spin-off approximately 95% of the subsidiary and distribute approximately 5% of the subsidiary's common stock to the Corporation's shareholders on a pro-rata basis. Consultant shall have full authority to structure this transaction and will bear all related costs. This action shall not take place within the initial thirty (30) days after closing of this transaction. This action will comply with all the relevant rules and regulation and will not cause any liabilities and damage to the reorganized Corporation. This provision shall survive the closing of this transaction.
G. The reorganized Corporation shall execute a Warrant Agreement with Consultant.
BTW, according to the announcement, George Frudakis "loaned" the money to Coast to Coast Equity to make the transaction. He doesn't appear to have any other stake at this time. Charles Scimeca is the sole director, officer and shareholder of Coast to Coast Equity Group, Inc.
And as per the statement above, "C. Prior to Closing, the Corporation's Director shall elect Dr. Tony N. Frudakis, Ph.D., Wenxia Guo, Peiyi Tian, Jianjun Liu, and Huimin Zhang to serve as members of the Board of Directors. Dr. Tony N. Frudakis shall serve as a director of the Board for the next two years without compensation.", looks like that part of the agreement was taken care of today along with the resignation of the Officer and Director.
This company looks like it's going to be raising some cash and it looks like it will be SERIOUS cash. I'd be looking for the SEC filing indicating a close to the transaction, then the funding of the company within four months.
Anyone want to take a shot at explaining Item F. above, having to do with, "The Corporation (post reorganization) shall take all corporate actions necessary to form a wholly owned subsidiary and to allow Consultant to complete a spin-off transaction whereby the Corporation will spin-off approximately 95% of the subsidiary and distribute approximately 5% of the subsidiary's common stock to the Corporation's shareholders on a pro-rata basis..."???
Looks like one of those "big" moves to me, but I guess we'll have to just wait and see...
Later,
W2P
Bag8ger, sorry jumped the gun on that one,disregard that post.
There all the same 1}Yangling Daiying Biological Eng. Co.LTD.+ Sun City Industries + Coast to coast Equity Inc. are all the same and Tony is the CEO.
BETTER READ THIS !!$$$$ - http://www.hkgem.com/newlistings/prospectuses/e_8031pro-20020522chap14.pdf
REVERSE MERGER: {4-23-04} Our Profile List stock Sun City Industries, Inc. (OTCBB: SCII 1.01 x 4.00) issued an 8K announcing they entered into an agreement to acquire all of the authorized issued and outstanding stock of Yangling Daiying Biological Engineering Co. Ltd. (YDBE), a corporation organized under the laws of the People's Republic of China, in exchange for 34,880,000 shares of the corporation's common stock which upon issuance will constitute approximately 87.2% of the corporation's issued and outstanding common stock.
Hey if Gray Cary is involved than its a smart move.
You Must know the process of refinement.Test by there nature are subject to change.Nothing is real till the very end at that point only is it up for review.
Check this out on eye color - A genome scan for eye color in 502 twin families: most variation is due to a QTL on chromosome 15q.
Zhu G, Evans DM, Duffy DL, Montgomery GW, Medland SE, Gillespie NA, Ewen KR, Jewell M, Liew YW, Hayward NK, Sturm RA, Trent JM, Martin NG.
Queensland Institute of Medical Research, Brisbane, Australia.
We have rated eye color on a 3-point scale (1 = blue/grey, 2 = hazel/green, 3 = brown) in 502 twin families and carried out a 5-10 cM genome scan (400-757 markers). We analyzed eye color as a threshold trait and performed multipoint sib pair linkage analysis using variance components analysis in Mx. A lod of 19.2 was found at the marker D15S1002, less than 1 cM from OCA2, which has been previously implicated in eye color variation. We estimate that 74% of variance in eye color liability is due to this QTL and a further 18% due to polygenic effects. However, a large shoulder on this peak suggests that other loci affecting eye color may be telomeric of OCA2 and inflating the QTL estimate. No other peaks reached genome-wide significance, although lods > 2 were seen on 5p and 14q and lods >1 were additionally seen on chromosomes 2, 3, 6, 7, 8, 9, 17 and 18. Most of these secondary peaks were reduced or eliminated when we repeated the scan as a two locus analysis with the 15q linkage included, although this does not necessarily exclude them as false positives. We also estimated the interaction between the 15q QTL and the other marker locus but there was only minor evidence for additive x additive epistasis. Elaborating the analysis to the full two-locus model including non-additive main effects and interactions did not strengthen the evidence for epistasis. We conclude that most variation in eye color in Europeans is due to polymorphism in OCA2 but that there may be modifiers at several other loci.
gunn me to heres some more info on Dr.Sturm - Novel MC1R variants in Ligurian melanoma patients and controls.
Pastorino L, Cusano R, Bruno W, Lantieri F, Origone P, Barile M, Gliori S, Shepherd GA, Sturm RA, Scarra GB.
Dipartimento di Oncologia, Biologia e Genetica, Universita degli Studi di Genova, Vle Benedetto XV, 6, 16132 Genova, Italy.
Several variant forms of the melanocortin-1 receptor gene (MC1R) have been associated with red hair, fair skin and an increased risk for melanoma. Their involvement in melanoma susceptibility is apparently linked both to skin sensitivity and to non-pigmentary pathways. We investigated the frequency of the MC1R variants in the Italian region of Liguria, where the occurrence and penetrance of melanoma are low and primary susceptibility is characterized by prevalence of the CDKN2A c.301G>T [p.G101W] founder mutation. Additionally, we attempted to establish the frequency of the red hair/fair skin phenotype in our region. As predicted by anecdotal evidence, the frequency of red hair/phototype I was very low (0.7%). Screening of 17 red-haired individuals and their red-haired relatives, 207 controls and 214 melanoma patients unselected for hair color but all of Ligurian descent, led to the detection of 8 novel substitutions (c.133T>C [p.F45L], c.248C>T [p.S83L], c.332C>T [p. A111V], c.479G>A [p.R160Q], c.637C>T [p.R213W], c.793G>A [p. V265I], c.923C>T [p. T308M], c.943T>C [p.C315R]), 1 novel deletion (c.520_523delGTC [p.V174del]) and 3 novel synonymous variants (c.366G>C [p. V122V], c.684G>A [p. Q228Q], c.726C>T [p.T241T]). Preliminary genotype/phenotype correlation seems to indicate that other genes involved in the regulation of human pigmentation may mask the recessive action of high-penetrance MC1R alleles, thus determining the low frequency of at-risk phototypes and of incidence and/or penetrance of melanoma in Liguria. Copyright 2004 Wiley-Liss, Inc.
I wounder if Tony is going - http://www.waimr.uwa.edu.au/Genemappers2004/
Heres more he working on -
Merkel Cell Carcinoma
Merkel cell carcinoma (MCC) is an aggressive skin cancer with a poor prognosis (30% 3-year survival rates, more like small cell lung cancer than skin cancer) whose incidence is increasing. The cause is chronic long-term exposure to the sun, rather than acute exposure (sunburn) as seen with melanoma. The QRI laboratory has established a tumour bank and a number of cell lines from MCC biopsies, in order to study the biology and aetiology of this little-understood disease.
Our Research
Our investigations into this rare, aggressive skin cancer have shown that the expression of two developmental transcription factors, Brn-3c and HATH 1, can distinguish between cancers which still have a neuroendocrine phenotype and those that have lost expression of these markers. This suggests that these expression patterns may be prognostic in MCC. In addition, we have shown that normal human Merkel cells express Brn-3c and HATH 1. MCC cell lines which retain neuroendocrine phenotype, are slow growing in culture, grow in suspension and are thought to be less aggressive, have been shown to retain Brn-3c and HATH 1 expression. Variant suspension lines which no longer express neuroendocrine markers, retain Brn-3c expression but lack HATH 1. Further, Type IV Variant MCC lines which grow as adherent monolayers, have shorter doubling times, are more radiation resistant and have higher cloning efficiencies in soft agar (all thought to be indicative of aggressive tumours), lack expression of Brn-3c and HATH 1 transcription factors. (see pdf)
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Melanocytes and their precursors, melanoblasts
In collaboration with the laboratory of Dr. Rick Sturm, UQ, we have established a series of melanocyte strains which have all been genotyped for their melanocortin-1 receptor (MC1R) (Sturm, 2001). it has been shown that variants of this protein are responsible for red hair colour and fair skin, and people with this phenotype are more likely to get melanoma (Box et al, 1997, and Sturm, 2002). We have now established a bank of 300 strains of melanocytes to enable us to investigate the relative sensitivities of these strains to ultraviolet light.
In addition, we have recently developed the culture conditions required to grow melanocyte precursors or melanoblasts. The cultures will be useful in analysing the gene expression patterns and protein signal transduction pathways of melanoblasts which may be reactivated during the formation of melanoma tumour cells to form dysplastic melanocytes.
Melanoblasts (left panel) have many dendrites (processes)
while melanocytes (right panel) tend to have only two dendrites
Pancreatic Cancer
We have recently identified a chemical which causes pancreatic cells to undergo cell death by apoptosis. The mechanisms by which this occurs are being examined to determine whether the compound will be useful in the treatment of pancreatic cancer.
Mechanisms of lung cancer
Biopsy material from tumour and normal lung epithelium is being compared by cDNA array technology and 2-D PAGE (see image below) to determine the molecular changes which lead to lung cancer. In the past, normal lung has been used for such studies. The disadvantage of this is that it is not a diverse mix of tissue, whereas epithelium is the tissue of origin for most lung cancer.
Here's some more on the Dr.
Our Research
Our investigations into this rare, aggressive skin cancer have shown that the expression of two developmental transcription factors, Brn-3c and HATH 1, can distinguish between cancers which still have a neuroendocrine phenotype and those that have lost expression of these markers. This suggests that these expression patterns may be prognostic in MCC. In addition, we have shown that normal human Merkel cells express Brn-3c and HATH 1. MCC cell lines which retain neuroendocrine phenotype, are slow growing in culture, grow in suspension and are thought to be less aggressive, have been shown to retain Brn-3c and HATH 1 expression. Variant suspension lines which no longer express neuroendocrine markers, retain Brn-3c expression but lack HATH 1. Further, Type IV Variant MCC lines which grow as adherent monolayers, have shorter doubling times, are more radiation resistant and have higher cloning efficiencies in soft agar (all thought to be indicative of aggressive tumours), lack expression of Brn-3c and HATH 1 transcription factors. (see pdf)
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Melanocytes and their precursors, melanoblasts
In collaboration with the laboratory of Dr. Rick Sturm, UQ, we have established a series of melanocyte strains which have all been genotyped for their melanocortin-1 receptor (MC1R) (Sturm, 2001). it has been shown that variants of this protein are responsible for red hair colour and fair skin, and people with this phenotype are more likely to get melanoma (Box et al, 1997, and Sturm, 2002). We have now established a bank of 300 strains of melanocytes to enable us to investigate the relative sensitivities of these strains to ultraviolet light.
In addition, we have recently developed the culture conditions required to grow melanocyte precursors or melanoblasts. The cultures will be useful in analysing the gene expression patterns and protein signal transduction pathways of melanoblasts which may be reactivated during the formation of melanoma tumour cells to form dysplastic melanocytes.
Melanoblasts (left panel) have many dendrites (processes)
while melanocytes (right panel) tend to have only two dendrites
Pancreatic Cancer
We have recently identified a chemical which causes pancreatic cells to undergo cell death by apoptosis. The mechanisms by which this occurs are being examined to determine whether the compound will be useful in the treatment of pancreatic cancer.
Mechanisms of lung cancer
Biopsy material from tumour and normal lung epithelium is being compared by cDNA array technology and 2-D PAGE (see image below) to determine the molecular changes which lead to lung cancer. In the past, normal lung has been used for such studies. The disadvantage of this is that it is not a diverse mix of tissue, whereas epithelium is the tissue of origin for most lung cancer.
Did you know Dr. Sturm has not only done a great deal of work on Eye color but also Lung Cancer -
Proneural and proneuroendocrine transcription factor expression in cutaneous mechanoreceptor (Merkel) cells and Merkel cell carcinoma.
Leonard JH, Cook AL, Van Gele M, Boyle GM, Inglis KJ, Speleman F, Sturm RA.
Queensland Radium Institute Research Unit, Queensland Institute of Medical Research, Herston, Brisbane, Queensland, Australia. helenL@qimr.edu.au
Merkel cells form part of the peripheral neuroendocrine system of the skin and act as mechanoreceptors in touch response. Merkel cell carcinoma (MCC) is a rare, aggressive disease with similarities to small cell lung cancer (SCLC), which is also of neuroendocrine origin. We previously identified a novel DNA binding protein complex specific for MCC suspension cell lines, termed Merkel nuclear factor (MNF) by its binding to the POU-IV family DNA binding consensus sequence. Here we report that MNF contains the POU-IV family member Brn-3c and that Brn-3c is expressed in normal Merkel cells. Additionally, Brn-3c protein reactivity is restricted to a subset of MCC biopsies and is not seen in biopsies revealing adherent, variant cell lines lacking neuroendocrine markers. Recently, proper development of murine Merkel cells was shown to require the proneural basic helix-loop-helix transcription factor, atonal family member, MATH1. We demonstrate a correlation between Brn-3c and HATH1 reactivity in MCC biopsies and cell lines with retention of neuroendocrine phenotype. In SCLC, the related basic helix-loop-helix transcription factor HASH1 is responsible for neuroendocrine phenotype, but HASH1 transcripts were not detected in MCC cell lines. We propose that HATH1 and Brn-3c may form a transcriptional hierarchy responsible for determining neuroendocrine phenotype in Merkel cells and that lack of Brn-3c and/or HATH1 in MCC may indicate a more aggressive disease requiring closer patient follow-up. Copyright 2002 Wiley-Liss, Inc.
worktoplay,i was thinking about that my self perhaps i am getting old in me years but i'm glad to know it crossed some one else mind younger than myself.LOL
Partial 'Blue Gene' Systems Are Now Two of the Top Ten Most Powerful Supercomputers on Earth
June 21, 2004--For the first time, two IBM Blue Gene/L prototype systems appear on the Top 10 list of supercomputers. The Blue Gene/L prototype represents a radical new design for supercomputing. At 1/20th the physical size of existing machines of comparable power, Blue Gene/L enables dramatic reductions in power consumption, cost and space requirements for businesses requiring immense computing power. For a new architecture to produce so much compute power in such a small package is a stunning achievement, and provides a glimpse of the future of supercomputing.
The number four-ranked Blue Gene/L DD1 Prototype, with a sustained speed of 11.68 teraflops and a peak speed of 16 teraflops, uses more than 8,000 PowerPC processors packed into just four refrigerator-sized racks. This ground breaking system is only 1/16 of its planned final capacity and has skyrocketed to the 4th place from the 73rd spot on the list in November 2003. The eighth-ranked Blue Gene/L DD2 Prototype has a sustained speed of 8.66 teraflops and a peak speed of 11.47 teraflops. The DD2 system is based on the second generation of the Blue Gene/L chips, which are more powerful than those used in the DD1 prototype.
About IBM's Blue Gene Supercomputing Project
Blue Gene is an IBM supercomputing project dedicated to building a new family of supercomputers optimized for bandwidth, scalability and the ability to handle large amounts of data while consuming a fraction of the power and floor space required by today's fastest systems. The full Blue Gene/L machine is being built for the Lawrence Livermore National Laboratory in California, and will have a peak speed of 360 teraflops. When completed in 2005, IBM expects Blue Gene/L to lead the Top500 supercomputer list. A second Blue Gene/L machine is planned for ASTRON, a leading astronomy organization in the Netherlands. IBM and its partners are currently exploring a growing list of applications including hydrodynamics, quantum chemistry, molecular dynamics, climate modeling and financial modeling. Read more...
A genome scan for eye color in 502 twin families: most variation is due to a QTL on chromosome 15q.
Zhu G, Evans DM, Duffy DL, Montgomery GW, Medland SE, Gillespie NA, Ewen KR, Jewell M, Liew YW, Hayward NK, Sturm RA, Trent JM, Martin NG.
Queensland Institute of Medical Research, Brisbane, Australia.
We have rated eye color on a 3-point scale (1 = blue/grey, 2 = hazel/green, 3 = brown) in 502 twin families and carried out a 5-10 cM genome scan (400-757 markers). We analyzed eye color as a threshold trait and performed multipoint sib pair linkage analysis using variance components analysis in Mx. A lod of 19.2 was found at the marker D15S1002, less than 1 cM from OCA2, which has been previously implicated in eye color variation. We estimate that 74% of variance in eye color liability is due to this QTL and a further 18% due to polygenic effects. However, a large shoulder on this peak suggests that other loci affecting eye color may be telomeric of OCA2 and inflating the QTL estimate. No other peaks reached genome-wide significance, although lods > 2 were seen on 5p and 14q and lods >1 were additionally seen on chromosomes 2, 3, 6, 7, 8, 9, 17 and 18. Most of these secondary peaks were reduced or eliminated when we repeated the scan as a two locus analysis with the 15q linkage included, although this does not necessarily exclude them as false positives. We also estimated the interaction between the 15q QTL and the other marker locus but there was only minor evidence for additive x additive epistasis. Elaborating the analysis to the full two-locus model including non-additive main effects and interactions did not strengthen the evidence for epistasis. We conclude that most variation in eye color in Europeans is due to polymorphism in OCA2 but that there may be modifiers at several other loci.
Are we working with 2 different Dept. in IBM ? 1} in forensics with Dr.Sturm and 2} at H.Lee Moffit !!! 2 Different Dept. 2 different places with IBM ? Could there be 3 or 4, IBM MUST have both Eye Balls on us.
Power for mapping by admixture linkgage disequilibrium: a case/control simulation study. C.L Pfaff, M.D Shriver. Penn State University, University Park, PA.
Admixture between genetically distinct populations creates association between loci. After a few generations, the linkage disequilibrium between unlinked loci decays, and the remaining disequilibrium can be used to map genes (Mapping by Admixture Linkage Disequilibrium). In order to characterize the behavior of MALD, we designed a simulation program that determines the power for MALD depending on a set of parameter values (e.g. number of generations since admixture, proportion of admixture, recombination fraction (q), sample size, and marker and disease allele frequencies). This program examines the power of a case/control study to detect linkage disequilibrium between a marker and a disease allele. Our results show that microsatellite markers are, on average, more powerful than SNPs for MALD. The average power for a SNP marker is 56% (q=0.01, N=300 case, 300 control), whereas an average microsatellite marker achieves >90% power under the same conditions. This difference is mainly due to higher dC levels (the sum of all positive allele frequency differentials at a locus) at microsatellite loci than at SNP loci. We have found that a high dC value is the most important characteristic of a good marker for MALD. For example, a marker with a dC value of 0.71 achieves a power of almost 90% when q=0.06, whereas a marker with a dC value of 0.38 attains a power of only 57% at the same q. Similarly, markers with high dC levels demonstrate powers of 89% with only 5% admixture (q=0.001) whereas markers with lower dC values have powers of 48% under the same conditions. We estimate the average dC value for microsatellites to be 0.42. Values of dC above 0.20 generate power estimations >85% at q<0.01. Thus, for candidate gene mapping, most microsatellites and many SNPs will provide suitable power. However, for genome-scan mapping a panel of high d markers must be chosen to achieve sufficient power. The results of these simulations indicate that MALD can and should be an important technique for mapping complex disease genes, and has more statistical power than other mapping methods. The results of current efforts comparing the power of MALD to the power of affected sib-pair mapping will also be discussed.