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Now if I could just patch things up with Neuro, though I wouldn't blame him for putting me on permanent ignore. I was definitely way out of line with my PARS comments. I've been acting strangely lately since losing my job, though that isn't any excuse for my being an ahole..
Dew, Cortex hasn't had much institutional/mutual support yet, so once that starts kicking in at around ~$5.00, that should help propel the stock. There are a lot of warrants out there though, in the $2.55, $3.00, and $3.25 areas, which could be a barrier. I figure many of those may not be excercised until they have some decent profits (above $4-5/share). I see the $3.50 area, give or take, as a likely upper pre-decision area for the stock, though that's just a guess.
Yes, I've been a pretty good contrary indicator over the last 12 months, even though I haven't owned any shares over that period. I had the ADHD results wrong (though I still think that trial had a needlessly risky design, with no intermediate dose cohort. If the 800 mg BID hadn't been well tolerated (it was after all the BID Max Tol Dose), then those trial results would have been completely worthless). Then I had the DARPA results wrong (though I did qualify my bullish stance with the caveat that DARPA might find some way to screw things up). And now with this latest episode with the histo finding I was also wrong (though getting good electron microscopic results was by no means a given). But being out of the stock altogether, I missed the clinical hold meltdown entirely, which was pure luck, so I guess it all evens out to some extent.
After watching these bio stocks for a number of years, I think there's a lot more luck involved than we'd all like to admit. I don't know, I just want one reasonably big score and I'll hopefully be out of this sector for good :o)
Brigrajac, One of the good things about a straight arrow like Dr. Stoll is that he's never exaggerated, pumped, or overstated. So when he gets really confident and enthusiastic it means something.
Dew, I think one of the reasons Dr. Stoll sounded wired was because they had just gotten the electron microscopic data that day (ahead of schedule). After all they've been through in the last year, he must have felt vindicated.
Dew, The handwriting is on the wall now (IMO), this baby's going to the moon. It won't be straight up, I figure we'll gravitate into the $2.50-3.50 area pre-decision, and then go up from there. I won't jinx us by saying slam dunk, but I'd say the odds of a positive FDA decision that will allow ADHD dosing is around 85% or better.
The ironic thing is that a tiny little blemish in those electron microscope results and the short/long term outcome for Cortex as a company would have been ugly. But those results were clean, and Dr. Stoll left no doubt that what they were seeing was clearly post-mortem - no ifs, ands, or buts about it. While the FDA can be unpredictable, I'm now willing to place a good size bet on the outcome, if I can just get the funds together and into my account..
I never thought I'd be rolling the bio dice again, but I now like the odds here a lot. OK, now that I'm on board, you can all start worrying :o)
Haysaw, I'm committed alright, right to the looney bin :o) I guess this stock has done a dance on my brain a few too many times. One way or another I always seem to miss the boat..
American Dairy (2-07) to Present at the ICR XChange Investor Conference
This company also presented at the recent Roth Conference, right before Cortex's presentation. There were a lot of small/mid cap China related companies presenting at that conference. It's difficult to find much info on this company however, and a mutual fund is probably be the best way to play China. The China market has already had a very large move over the last several years -
>>> Thursday January 11, 8:30 am ET
- Presentation Takes Place at 11:00 am PT -
BEIJING, Jan. 11 /PRNewswire-FirstCall/ -- American Dairy, Inc. (NYSE Arca: ADY), one of the leading producers and distributors of milk powder and soybean products in China, today announced that the Company will present at the Ninth Annual ICR XChange Investor Conference at the St. Regis Monarch Beach Resort & Spa in Dana Point, California, on January 11, 2007. The Company will present to investors at 11:00 am PT and will meet with institutional investors throughout the day. Please note that this presentation timeslot has changed from the previously announced 2:45 pm PT timeslot.
Listeners may access a live webcast of the presentation by visiting the ICR XChange webcast link at http://www.icr-online.com/xchange/x9/attendee/webcast.htm. For more information on the Ninth Annual ICR XChange, or to meet with American Dairy at the conference, please contact ICR at 203 682 8200.
About American Dairy, Inc.
American Dairy, Inc. conducts operations in The People's Republic of China ("China") through its wholly owned subsidiary, Feihe Dairy. Founded in 1962, Feihe Dairy is one of the leading producers and distributors of milk powder and soybean products in China. Feihe Dairy is located in Kedong County, China, and has been in operation since 2001. American Dairy also has a milk powder processing plant, BaiQuan Feihe Dairy in Kedong County, and also has a milk powder processing plant in the city of QiQiHaEr, Heilongjiang Province. http://www.feihe.com/ <<<
Elbit Systems (2-07) Awarded a PFI Contract to Establish and Operate Israeli Air Force "Tzofit" (Beechcraft) Training Center
Tuesday February 20, 4:20 am ET
Potential Revenues Expected to Exceed $15 Million
Training Center can Also Serve Civil Aircraft Pilots
HAIFA, Israel, February 20 /PRNewswire-FirstCall/ -- Elbit Systems Ltd. (NASDAQ: ESLT - News) was awarded a contract to establish a training centre for "Tzofit" (King Air B200 Beechcraft) for the Israeli Air Force. Elbit Systems will serve as the project's prime contractor, while Arkia and the Canadian Mechtronix will serve as sub-contractors.
The training centre will operate through a PFI (Private Financing Initiative) program, with Elbit System providing the Israeli Air Force a turn-key solution including the establishment of the training centre, its operation and the supply of simulators, training services and maintenance for a 10-year period. The MoD will purchase flight training hours for the Air Force from Elbit Systems. Potential revenues from the project are expected to exceed $15 million. The training center will be established on a civilian property outside military bases.
Furthermore, for the first time in Israel, it will be possible to open a training centre for civilian pilots providing them training in accordance with international aviation requirements while using local flight simulators and facilities.
Joseph Ackerman, President and CEO of Elbit Systems, said: "There is a wider global demand for use of civilian and military simulators in all areas of operations in order to achieve better performance with the same level of training hours.
The "Tzofit" training centre project coupled with the company's on-going Israel Air Force cadets training project, strengthen Elbit Systems' position as Israeli Air Force's leading contractor in the field of simulators in general and in flight simulator in particular. Mr. Ackerman added that the prospect of civilian pilots training constitutes an important addition to Elbit Systems' ever growing portfolio directed at the civilian aviation market.
Elbit Systems operates within the global simulator market and supplies systems for all types of platforms, including those for air, land and naval applications, as well as interdisciplinary simulators allowing for joint forces training. Elbit Systems utilizes its advanced technological capabilities, particularly in developing software for complex system programs, and its global leadership in upgrading defense and civilian platforms.
About the Tzofit (Beechcraft)
Received by the Israeli Air Force in 1991, the Tzofit is a civilian business jet manufactured by Beechcraft which has been converted for military use. The first aircraft was manufactured in 1973 and is used worldwide for transportation, passengers' transfer, weather exploration and communication and marine reconnaissance missions. The Israeli Air Force employs the aircraft in various missions.
The Tzofit's maximal cruise altitude is 31,000 feet, its maximal range 3,254 Km and its maximal cruise speed is 289 knots.
Following its acquisition by the Israeli Air Force, advanced systems, manufactured in Israel, were installed on board the aircraft which enhance IAF's capabilities, significantly prolong the aircraft's cruise range and facilitate its special missions.
(Quote from an article in IAF Magazine, Issue # 130, December 1999).
About Elbit Systems
Elbit Systems Ltd. is an international defense electronics company engaged in a wide range of defense-related programs throughout the world. The Elbit Systems Group, which includes the company and its subsidiaries, operates in the areas of aerospace, land and naval systems, command, control, communications, computers, intelligence, surveillance and reconnaissance ("C4ISR"), advanced electro-optic and space technologies, EW suites, airborne warning systems, ELINT systems, data links and military communications systems and equipment. The Group also focuses on the upgrading of existing military platforms and developing new technologies for defense and homeland security applications. <<<
Graco (2-07) Stock Offers Price Appreciation, Dividend
I'm adding Graco (GGG) to my virtual portfolio. Check out their long term chart, a thing of beauty to be sure, plus the company originally started out a maker of grease guns - the perfect Peter Lynch type stock ("dull and disgusting") :o) -
>>> By Jack Hough
February 6, 2007
FEARS OF A PROLONGED slump in home building have sent Graco (GGG) shares $7 lower to $41 and change since April. The Minneapolis company makes commercial sprayers, so its fortunes depend in part on a steady supply of big things that need paint. Home builders account for more than a third of its sales.
The selloff might be an overreaction, for three reasons. First, recent data suggests home builders might be cutting prices and settling for smaller profits, but they're still putting up plenty of houses. Second, paint is flying nicely in Europe and Asia, judging by the company's fourth-quarter results. Third, Graco is still cashing in on a world-wide manufacturing boom, since lubricants and industrial chemicals need spraying, too.
The stock turned up recently in our Not Just Income screen. It searches for companies that pay handsome dividends but also look likely to produce ample share-price gains. Graco has paid plentiful dividends since 2003 and now yields about 1.7%, but longtime shareholders are perhaps more impressed with the stock having increased more than sixfold in value over the past decade.
Spotlight Stock
Graco (GGG)
Supplies technology and expertise for the management of fluids in both industrial and commercial applications. It designs, manufactures and markets systems and equipment to move, measure, control, dispense and spray fluid materials.
Tuesday's Close $41.52
Market Value $2.8 billion
Trailing 12-Month Sales $817 million
2007 P/E 17
Proj. Long-Term EPS Growth Rate 18%
Our screen isn't greedy for yield; anything over 1.5% will do. But companies must pay out no more than half their profits as dividends. That keeps payments safe, while leaving funds available to invest in expansion. Sales and earnings must have increased by at least 15% a year over the past five years. Stocks must be modestly priced, with PEG ratios — price/earnings multiple divided by earnings growth forecast — below the broad market's average. And debt levels must be manageable.
Graco, not to be confused with the brand of baby strollers11 owned by Newell Rubbermaid (NWL12), started as a grease gun manufacturer. Russell Gray developed an air-powered model in 1926 and his brother Leil went about selling it to service stations. The company added paint products in 1958. Today it has machines that squirt muffin batter, ricotta cheese, glue, ink, car gloss and more. The company still leads the lubrication business with a 60% market share, but such equipment now accounts for just 11% of total sales.
Operating profits for the company — what's left after paying manufacturing costs and corporate expenses like salaries and ad spending — totaled just five to seven cents on the dollar in the early 1990s. Today the company keeps about 28 cents. The improvements have come from a focus on cutting manufacturing costs and focusing on niche markets. (Did I mention potato salad pumpers?)
Last year, sales for the company increased 12% to $816.5 million. Earnings per share jumped 21%. A housing slowdown in the second half of 2006 tempered fourth-quarter results. Sales increased 10% and earnings, 13%. Contractor sales declined 2.3%. Sales to contractors in Europe and Asia, though, increased 30% and 21%, respectively. Analysts say the company gained market share overseas and that more builders switched from brushes to sprayers. Lubrication sales jumped more than 40%, but about 35 of those percentage points were owed to the company's acquisition last year of Warrensville Heights, Ohio-based Lubriquip. Operating profits for the unit increased more than 16% as Graco found ways to cut costs after the acquisition.
Housing is still a concern, but on Jan. 18 the Commerce Department said housing starts unexpectedly increased 4.5% in December from November. Building permits jumped 5.5%, the most in four years.
Graco doesn't provide earnings guidance, but Wall Street reckons the company will increase its earnings by 12% this year and by 18%, on average, over the next five years. That makes the stock, at 17 times forecast 2007 earnings, seem a bargain. The numbers make for a PEG ratio of just under 1.0. That's a discount of more than a third to the broad stock market.
<<<
Way OT - Judy, I wasn't able to reply privately since I don't have a premium I-Hub membership, but private donations are always welcome :o) But seriously, as much as I could use the money, I would most likely just lose it as before, since my destiny seems to be the soup kitchen/breadline. Thanks for the kind words though, Rick :o)
Cortex (2-07) - Monster move -
Here's the comment from Dr. Tracy's NeuroInvestment website -
>>> Online comment 2/22/07
Cortex was surprisingly definitive in their presentation at Roth Capital yesterday: the cellular 'signal' which has hamstrung the CX717 program since last April is a post-mortem artifact--it only occurs in tissue which is being prepared with a formalin-based fixative. If the same target tissue is instead frozen, the cellular signal is not seen, even when examined via electron microscope. Even the FDA will find it near-impossible to object to a resumption of full-dosage ADHD trials when the signal of concern only occurs after death. CEO Roger Stoll said quite bluntly--"I think we'll be able to get the dose limitation lifted by the FDA." No hedging with anything like 'of course, the final decision will be up to the FDA...' Cortex will submit the full package to the FDA in mid-March, and we expect a 'go' decision from the FDA in May. It has been a long wait, but it appears that CX717, which has now been more thoroughly vetted than any other Phase II drug, is in fact safe, and will go back into ADHD, where the initial findings were so positive. It was also timely in that this occurred on the day that the FDA announced that all current ADHD drugs would have to include inserts detailing their safety risks. CX717 appears to have none of their side effect baggage. This will add to the program's lustre as a partnerhip candidate, in addition to the high-impact group, where a partnership is expected midyear. Cortex is now in a position to resume the upward trajectory that was so abruptly interrupted last April--which for those who lost track during the interim, had the share price over 5. We expect it back in--and perhaps above--that range during 2007. <<<
Fearfrost, I like the Vanguard Target Retirement group of funds. Their Target 2045 fund has approx 79% US and 21% International exposure. T.Rowe Price has a similar group of funds, but the Vanguard funds have a 0.20% expense ratio, vrs TR Price's 0.65%, so Vanguard has the advantage. The 21% International exposure in the Vanguard fund includes the European Index (11%), Pacific Rim Index (5%), Emerging Markets Index (2%), and Total International Index (3%). The rest is in the Total US Market Index (69%), with some in the Total Bond Index (10%). The fund automatically reconfigures the allocation gradually as you get closer to retirement. This type fund is extremely boring but since you are young it's the surest way to safely reach your financial goals. You can always spice things up by keeping 10% or so in individual stocks to play with.
Real estate is nother asset class you should have. A home is usually sufficient there, but additional investments in a REIT fund or index gives added exposure to commercial real estate.
BTW, the above allocation strategies can also be accomplished using Exchange Traded Funds. I like the Index fund approach better however, since there is a lot less temptation to trade than in a brokerage account using ETFs. Don't try to time the market, you need to stay the course and dollar cost average in on a regular basis.
An interesting part of today's action is that one didn't need to be in the stock at all before the event to participate in almost all the upside. One could have bought as much stock as desired between $1.20-1.30 early in the day (I would have done just that if I had any money in my brokerage account). Of course part of the delayed upside in the morning was probably from the unloading of the recent PIPE shares, and that kept the lid on the stock for a couple hours, but that was just long enough for folks to load up.
Oh well, live and learn. Fwiw, I'm going to assemble what funds I can muster and then consider a possible entry point in the next couple weeks, depending on where the price is.
Neuro had this situation nailed, and I should have considered his analysis more closely. However, if the electron microscope data hadn't been favorable, today's action would have been considerably different. Objectively, I still think the situation prior to Stoll's presentation was extremely dicey.
Bio-Imaging (2-07) - Support Product Development in the Pharmaceutical, Biotechnology and Medical Device Industries With Bio-Imaging Technologies Live on MN1.com
Thursday February 22, 1:16 pm ET
NEWTOWN, PA--(MARKET WIRE)--Feb 22, 2007 -- Mark L. Weinstein, President and Chief Executive Officer of Bio-Imaging Technologies, Inc. (NasdaqGM:BITI - News), will be joining Market News First (www.mn1.com) for an exclusive live interview with MN1's Saul Albom. The interview is scheduled for Thursday, Feb. 22, 2007, at 12:30pm CDT.
Operation in the U.S., Bio-Imaging provides services that support product development process of pharmaceutical, biotechnology and medical device industries. It operates in two segments: Pharmaceutical Contract Services and CapMed division. Pharmaceutical contract service segment provides services that support the product development process of the pharmaceutical, biotechnology and medical device industries. CapMed segment provides software application that enables users to manage and store personal health information, like the medical images, drug information, patient education and disease guidelines.
During the interview, Mr. Weinstein will be discussing Bio-Imaging's recently released Fourth Quarter and Year-End 2006 Financial Results. "In 2006 we solidified our core Clinical Trials Business and made good progress in our CapMed Division. We returned the company to profitability and ended the year with a record backlog," stated Mr. Weinstein. "Fourth quarter 2006 service revenues of $8.6 million represented a 7% sequential quarter over quarter increase. The increased profitability on these revenues is a testament to our leverageable expense base that generally provides increasing margins as we grow our revenues."
Join Mr. Weinstein to learn more about the goals of the company, as well as its position in the stock market.
About Bio-Imaging Technologies
Bio-Imaging Technologies, Inc. is a healthcare contract service organization providing services that support the product development process of the pharmaceutical, biotechnology and medical device industries. The Company has specialized in assisting its clients in the design and management of the medical-imaging component of clinical trials since 1990. Bio-Imaging serves its clients on a global basis through its US Core Lab in Newtown, PA, its European Core Lab in Leiden, The Netherlands and its recent acquisition of Theralys SA in Lyon, France. Through its CapMed division, Bio-Imaging provides the Personal HealthKey(TM) technology and the Personal Health Record (PHR) software allowing patients to better monitor and manage their health care information. Copies of Bio-Imaging Technologies' press releases and other information may be obtained through Bio-Imaging's web site at www.bioimaging.com.
About MN1.com
Market News First is an online, market news provider that brings investors current news on the market. Market News First is the only online, live radio web site that brings real market news to investors and features live interaction with companies from the AMEX, NYSE, NASDAQ, Over the Counter Bulletin Board and the Pink Sheets.
Through daily, live commentary we keep you up to date on the companies you invest in along with what's going on in the stock markets by the second.
BioMarin (2-07) Pullback Offers New Opportunity
By Marc Lichtenfeld
Senior Columnist
2/22/2007 11:09 AM EST
URL: http://www.thestreet.com/newsanalysis/investing/10340086.html
Despite BioMarin's (BMRN) solid quarterly report on Tuesday, investors were discouraged that hypertension drug candidate 6R-BH4 failed to show efficacy in phase II trials. The stock opened sharply down Tuesday but pared those losses somewhat to finish the session at $18, 12% lower than Friday's close.
Perhaps investors realized that while a marketable hypertension drug would certainly have been a positive, the stock is more reflective of the company's two marketed drugs and Phenoptin, a treatment for mild to moderate phenylketonuria (PKU) that could hit the market later this year.
Tuesday morning, BioMarin said there was no statistically significant or clinically meaningful effect of 6R-BH4 relative to placebo in a study of 116 patients over eight weeks. This was a bit of a surprise, because the drug did show efficacy in an earlier trial. Management said it would halt its research in hypertension but would continue studying the drug in other indications, such as peripheral arterial disease and sickle cell anemia.
While the results were clearly disappointing, investors can take some solace in the fact that there were no safety issues raised. This is significant because 6R-BH4 is the active ingredient in Phenoptin, so the hypertension trial should have little impact on whether Phenoptin receives approval for PKU at the end of the year.
Morningstar's Karen Andersen said she only modeled for a 30% chance of approval of 6R-BH4 for hypertension. While investors would have liked to have seen success in treating hypertension, Andersen adds, "rare diseases are their [BioMarin's] bread and butter."
In fact, BioMarin's products for the rare disorder Mucopolysaccharidosis (MPS) performed extremely well in the fourth quarter, and further strength is projected for 2007.
Aldurazyme, which treats MPS I, brought in $26.5 million in sales, up from $21.2 million in the year-ago period and higher by $1.5 million from the third quarter. The company, which splits sales of Aldurazyme with Genzyme (GENZ) , forecasts a roughly 25% increase in sales this year.
Naglazyme, for the treatment of MPS VI, is expected to rack up sales of $74 million to $78 million in 2007. BioMarin, which owns the full rights to the drug, recorded Naglazyme sales of $46.5 million in 2006. In the case of Naglazyme, success breeds success. As patients live longer, they gain weight. Because dosage is based on weight, the more a patient thrives, the more drug he or she will require.
The Technical Side
The stock has had an impressive run, trending upward since early 2005. John Hughes of technical research firm Epiphany Equity Research says, "The stock was previously in a strong uptrend and moving higher with supporting volume." Tuesday's weakness "brought the stock back to a key intermediate-term support level in the $17 area."
Hughes suggests that a break below $17 indicates the advance is ending, with further price vulnerability to the $14-$15 level. He concludes, "The longer-term weekly configuration remains positive and suggests that any weakness can eventually be used as a buying opportunity." Hughes and his firm have no position in BioMarin. Epiphany does not engage in investment banking.
Based on both the fundamentals and technicals, I agree with Hughes' last assertion. Currently, BioMarin is expected to turn profitable in 2008. The company's balance sheet, while not perfect, is solid. And management has proved that it can successfully bring products to market.
Setbacks are an unfortunate part of biotech investing. Sometimes a failed trial is a binary event than can sink a stock. Other times, it is a disappointment, as it is in this occurrence, but does not significantly alter the company's prospects.
Fearfrost, If it's only for 3 months, I'd just park it in a money market fund. The Vanguard Prime Money Market is the one I use, and their rate is currently around what a CD would be.
Concerning zeros, I assume you mean zero coupon bonds/CDs. These are bonds stripped of their coupons and discounted way back to what their theoretical core value would be based on the time to maturity. I owned some of these back in the 1980s when rates were very high and falling. They're very sensitive to interest rate fluctuations. One might be priced at say $350, and it matures at $1000 in X number of years. You don't get any current interest income from it, but get all your money at the end. Zeros are best suited for a tax-deferred account like an IRA since you have to pay tax on what the imputed interest would have been, even though you're not getting it (unless it's a zero coupon municipal bond). Aggressive investors can use zeros to take advantage of drops in interest rates since they move faster in price than would a traditional bond. They're also good for long term capital appreciation when held to maturity. Back in the 1980's I had some 12-13% zeros, and also some zero municipals that were over 10%. Rates are low now though, and at your age you're better off in equities for the long haul. Just put the bulk of your retirement money in a very broad index fund (one that ideally includes some international exposure, perhaps 20%), and let it ride. Don't worry about the market fluctuations, just keep adding every month/quarter without fail. After 20-30 years you'll have a big pile of money.
Wish I had some money in my brokerage account, but I guess I'll just have to watch this one as the train leaves the station..
Lookin good folks! I think we might be in the clear, with some luck. He said the electron microscopic data was clean and sounded very confident that it is a post-mortem phenomenon. I'm considering picking up some shares now, believe it or not! :o)
Well, I'm out of posts til midnight so I'll read the board discussion. Neuro, your 70% looks pretty close now, and maybe a little on the low side :o)
I still would like to eventually hear an explanation for why this phenomenon is unique to CX-717, but it does sound like some type of artifact.
Yes, that makes sense, since it's apparently an effect not previously seen. Since Lilly had the ataxia problem though, presumably a cerebellum effect, the FDA must have put CX-717's original 3 month tox data under extreme scrutiny. In doing so, perhaps they found something mysterious in another area of the brain.
Striaterminalis, as I understand it, using the traditional fixation method (vrs immediate freezing) there is a period of time (hours I think) between death and fixation. That's why I figured that some subtle cellular change that wouldn't be easily visible at death might cause the cells to begin their normal post-death degeneration at a faster rate than usual, which would make them visible after fixation.
Oh well, this may just be something we investors will never have an answer to, and going into the FDA decision period will just require a major leap of faith.
Here's a theory on what might be going on -
The FDA must have known about Lilly's ataxia problem in the LY-451395 trial. Once Cortex submitted the original 3 month tox data on CX-717 to the FDA (Q1-06 period), the FDA immediately focused in on the cerebellum tissue slides to see if there was a similar effect to Lilly's. They found something in one primate, halted the drug, and asked for more tox studies.
So let's assume for a second that the histo "phenomenon" is in fact in the cerebellum. Why would it not appear after fast freezing vrs after a delay/fixative? Since CX-717 doesn't induce anywhere near the same amount of damage to the Perkinje cells as Lilly's compound does (thus no clinical manifestations), there isn't actual cellular death or severe damage. Some subtle damage is there, but to a much lesser degree than with Lilly's compound. So at the time of death, the subtle damage might be visible under an electron microscope (maybe), but not under a regular light micropscope. However in the hours after death, the mildly damaged Purkinje cells start to deteriorate faster than do completely healthy cells, and when the fixative is applied, the cellular damage is clearly visible under even a normal light microscope.
Striaterminalis, Ataxia. Well, things may be getting clearer now (maybe). I assume the Lilly trial that Oldies was referring to was the ill-fated LY-451395 AD Phase 2 which was stopped reportedly due to excitotoxicity related side effects (what we heard from the grapevine). As you said, if ataxia was seen, then there was probably some destructive effect in the cerebellum, with the Perkinje cells. I'll repost that "nightmare" paper again (see below) that links AMPA receptor activation-elicited excitotoxicity to calpain mediated destruction of Perkinje cells. This is not good folks. If ataxia was happening in the Lilly LY-451395 trial, then the FDA probably had CX-717 under suspicion/guilt by association. I'd be interested in hearing Neuro's take on this -
>>> : Eur J Pharmacol. 2007 Feb 28;557(2-3):106-14. Epub 2006 Nov 22. Links
Involvement of calpain in AMPA-induced toxicity to rat cerebellar Purkinje neurons.
Mansouri B, Henne WM, Oomman SK, Bliss R, Attridge J, Finckbone V, Zeitouni T, Hoffman T, Bahr BA, Strahlendorf HK, Strahlendorf JC.
Department of Physiology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
AMPA receptor-elicited excitotoxicity is manifested as both a type of programmed cell death termed dark cell degeneration and edematous necrosis, both of which are linked to increased intracellular Ca(2+) concentration. The appearance of marked cytoskeletal changes in response to abusive AMPA receptor activation, coupled with increased intracellular Ca(2+) concentration suggests activation of various destructive enzymes such as calpains, a family of Ca(2+)-dependent cysteine proteases. Since calpains and AMPA have been linked to both necrotic cell death and programmed cell death, we sought to determine the role of calpains in mediating both types of AMPA-mediated toxicity in Purkinje neurons of the cerebellum. These studies employed immunohistochemistry for cytoskeletal breakdown products of calpain activity coupled with confocal microscopy and pharmacological interventions with calpain and AMPA receptor antagonists. The present study identifies an early involvement of calpains in mediating AMPA-induced dark cell degeneration, but not edematous necrosis, based upon the effectiveness of AMPA to generate calpain-derived alpha-spectrin cleavage products in cerebellar Purkinje neurons that express dark cell degeneration, and the effectiveness of calpain antagonists, PD150606 and MDL28170, to attenuate AMPA-induced dark cell degeneration. Moreover, the AMPA receptor antagonist CNQX, a proven inhibitor of AMPA-elicited dark cell degeneration, also blocked AMPA-induced increases in alpha-spectrin, further suggesting interplay between abusive AMPA receptor activation, calpain activation and dark cell degeneration. Since AMPA-induced dark cell degeneration possesses morphological changes that resemble those that occur following brain ischemia in vivo, hypoglycemia, or extended seizure episodes, the involvement of calpains as mediators of cell death is potentially far reaching and has widespread therapeutic implications in numerous CNS disorders.
PMID: 17188264 [PubMed - in process] <<<
Striaterminalis, Maybe not that obvious - the various published calpain/spectrin papers concerning CX-614 (Baudry and Lynch had the paper I posted, and Baudry had several without Lynch) all said that the calpain/spectrin activity that was induced by CX-614 was apparently not associated with cellular damage. While that might conceivably change after longer periods of dosing and/or higher doses, it indicates that the cell changes could be a lot more subtle than bursting/apoptotic cells (and if cells were dying all over the place, I doubt the FDA would have lifted the hold at all).
Because of the apparent lack of cell damage, in his paper Lynch theorized that the calpain/spectrin activity induced by CX-614 is probably due to the upregulation of LTP, something one would normally expect from an Ampakine. If so, the calpain activity could be relatively benign, with no clinical manifestations ever seen in a clinical setting. But seeing subtle microscopic cellular changes that had never been seen before might be very unsettling, especially to the FDA.
Calpain/spectrin activity could explain many of the observations we see with our histo "phenomenon", including why it happens with CX-717 and not with other low impacts, why it is dose level and duration dependent, why there are no clincal manifestations seen. Of course it doesn't explain why it isn't seen with fast freezing but only when a fixative is used. But it could explain why there is an apparent ceiling effect to the phenomenon (not getting worse between 2 weeks and 3 months), since if the phenomenon is associated with LTP upregulation, that upregulation likely reaches an upper ceiling level and tops out with no additional upregulation possible after a couple weeks. It would also explain why the "phenomenon" gradually disappears after dosing is stopped, since the upregulation of LTP would also diminish, reverting back to normal.
That would leave the freezing vrs fixative observation as unexplained. Since the finding is only seen when fixative is used and not with freezing (pending confirmation from the electron microscope data), it could be due to an interaction between CX-717 and the fixative. But it would have to be an interaction that is unique to CX-717, since it doesn't occur with other compounds. If it is unique to CX-717, the question then is WHY is it unique to only that compound? That's something Cortex will need to explain to the FDA. The puzzle continues..
Spectrum (2-07) Expanded Access Program Launched for Satraplatin
Wednesday February 21, 8:00 am ET
IRVINE, Calif., Feb. 21 /PRNewswire-FirstCall/ -- Spectrum Pharmaceuticals, Inc. (Nasdaq: SPPI - News) today announced that the Satraplatin Expanded Rapid Access protocol (SPERA) has been launched by the company's partner, GPC Biotech. Expanded Access Programs (EAPs) are intended to give patients access to investigational drugs to treat serious or life-threatening diseases or conditions for which there are limited treatment options available. Under the SPERA program, satraplatin will be provided to patients free of charge.
"Prostate cancer patients who have failed first line therapy have very limited treatment options available to them," said Rajesh C. Shrotriya, Spectrum's Chairman, President, and Chief Executive Officer. "The SPERA program provides these patients with an additional treatment option."
U.S. physicians interested in receiving more information about SPERA can contact 1-800-349-8086 or www.speratrial.com.
About Prostate Cancer
Prostate cancer is the most common cancer among men in the U.S. and Europe. Approximately 234,000 men in the U.S. are expected to be diagnosed with the disease in 2006 and more than 27,000 men are expected to die from the disease. In the European Union, over 200,000 new cases are expected to be diagnosed, and more than 60,000 patients are expected to die each year. Since the incidence of prostate cancer increases with age, the aging of the overall population is expected to further increase the number of prostate cancer patients. Most patients diagnosed with prostate cancer initially receive surgery or radiation therapy, and some of these patients are cured. For many others, though, the disease recurs. Recurrent disease is treated with hormone therapy, and most patients initially respond well to this treatment. Eventually, however, the tumor cells become resistant to the hormones -- or hormone-refractory -- and the tumor again progresses. Increasingly, chemotherapy is being used as an effective first-line treatment for HRPC. However, it is not a cure. Consequently, there is a growing need for effective therapeutic options, such as second-line chemotherapy treatments, for patients once they have progressed.
About Satraplatin
Satraplatin, a fourth-generation, oral investigational drug, is a member of the platinum family of compounds. Over the past three decades, platinum-based drugs have become a critical part of modern chemotherapy treatments and are used to treat a wide variety of cancers. Unlike the platinum drugs currently on the market, all of which require intravenous administration, satraplatin is an oral compound and is given as capsules that patients can take at home. In addition to HRPC, satraplatin has been studied in clinical trials involving a range of tumors, and Phase 2 trials have been completed in ovarian cancer and small cell lung cancer. Other trials evaluating the effects of satraplatin in combination with radiation therapy, in combination with other cancer therapies and in various other cancers are underway or planned.
In September 2006, topline results were announced from a double-blind, randomized Phase 3 trial, the SPARC trial. The trial evaluated satraplatin plus prednisone versus placebo plus prednisone as a second-line treatment in 950 patients with hormone-refractory prostate cancer.
In 2002, Spectrum licensed the global rights to GPC Biotech (Frankfurt Stock Exchange: GPC; TecDAX index; Nasdaq: GPCB). GPC Biotech is responsible for all costs associated with the development and regulatory filings of satraplatin. GPC Biotech has a co-development and license agreement with Pharmion GmbH, a wholly owned subsidiary of Pharmion Corporation, under which Pharmion has been granted exclusive commercialization rights to satraplatin for Europe and certain other territories.
About Spectrum Pharmaceuticals
Spectrum Pharmaceuticals is acquiring and advancing a diversified portfolio of oncology drug candidates that meet critical health challenges for which there are few other treatment options. Spectrum's expertise lies in identifying undervalued drugs with demonstrated safety and efficacy, and adding value through further clinical development and selection of the most viable and low-risk methods of commercialization. The company's pipeline includes promising early and late-stage drug candidates with unique formulations and mechanisms of action that address the needs of seriously ill patients, such as at-home chemotherapy and new treatment regimens for refractory disease. For more information, please visit our website at www.spectrumpharm.com.
Henry Schein (2-07) fourth-quarter profit jumps 30%
By Michael Baron
Last Update: 7:35 AM ET Feb 21, 2007
NEW YORK (MarketWatch) -- Henry Schein Inc.'s (HSIC ) fourth-quarter net income rose 30% to $63 million, or 70 cents a share, from $48.6 million, or 55 cents a share, a year earlier. The provider of healthcare products said Wednesday the latest quarter includes one extra week of business. The Melville, N.Y., company's fourth-quarter sales increased 12% to $1.5 billion from $1.34 billion. The company said it experienced sales growth in each of its business groups. Analysts polled by Thomson First Call expected, on average, fourth-quarter earnings of 67 cents a share and revenue of $1.57 billion. For 2007, Henry Schein expects to earn $2.51 to $2.57 a share and to distribute about 20 million doses of flu vaccine.
Sepracor (2-7) Announces Publication of Additional Data from Lunesta(R) Insomnia-Depression Study in the Journal of Clinical Sleep Medicine
Wednesday February 21, 7:00 am ET
MARLBOROUGH, Mass.--(BUSINESS WIRE)--Sepracor Inc. (Nasdaq: SEPR - News) today announced that additional data from its Phase IIIB/IV, 545-patient study of LUNESTA® brand eszopiclone in patients with insomnia and co-existing major depressive disorder (MDD) have been published in the February 2007 edition of the Journal of Clinical Sleep Medicine. This manuscript contains results from the two-week, single-blind, placebo run-out period of the study that evaluated hypnotic discontinuation effects following an eight-week, placebo-controlled study of LUNESTA and PROZAC® brand fluoxetine co-therapy in patients with insomnia and co-existing MDD.
The double-blind, placebo-controlled study evaluated the efficacy and safety of LUNESTA in patients who met DSM-IV®(1) criteria for both insomnia and MDD (either newly diagnosed or patients who had a new recurrence of MDD). Patients were randomized to receive open-label fluoxetine each morning and either double-blind LUNESTA 3 mg (n=270) or matching placebo (n=275) nightly for the first eight weeks, followed by a two-week period during which all patients received single-blind placebo treatment and continued receiving fluoxetine.
Journal of Clinical Sleep Medicine Published Results
The study endpoints contained in the Journal of Clinical Sleep Medicine manuscript were the evaluation of withdrawal effects during the two-week discontinuation period, which is also referred to as the run-out period, by examining: 1) the prevalence of new or worsening CNS (central nervous system)-related adverse events, 2) rebound insomnia, and 3) worsening of MDD symptoms. The rebound insomnia variables were assessed using an interactive voice response system (IVRS) during the run-out period and they included sleep onset, wake time after sleep onset (WASO), total sleep time (TST), and measures of daytime functioning.
Sleep and daytime function were also assessed subjectively during the run-out period using the insomnia severity index (ISI) scale, which consists of self-ratings of difficulties with sleep onset, sleep maintenance, and early morning awakenings, as well as daytime function, degree of impairment, and concern and satisfaction with current sleep pattern. Depressive symptoms were assessed during the run-out period using the HAM-D17 scale (Hamilton Depression Rating Scale, the standard scale used by clinicians in research studies to assess depression; the HAM-D17 is a list of symptoms commonly associated with depression; a lower score indicates fewer symptoms of depression). The HAM-D17 was also used to evaluate antidepressant response and remission rates. In addition, at Weeks 8 and 10, antidepressant efficacy was evaluated using Clinical Global Impression Improvement (CGI-I) and Severity (CGI-S) scales (used by clinicians to assess improvement in a patient's MDD symptoms and the severity of their depression at various time points, respectively).
In this study, patients discontinued from LUNESTA maintained improvements from baseline in sleep onset, WASO, and TST during the two-week discontinuation period (p less than 0.05). LUNESTA discontinuation did not result in significant CNS withdrawal adverse events, rebound insomnia or rebound depression, and improvements in sleep and depressive symptoms were maintained. The improvements in the HAM-D17 scale scores with and without the three sleep items observed with co-therapy versus monotherapy at Week 8 (p less than 0.02) were maintained at Week 10 (p less than 0.002) in the group initially randomized to co-therapy.
Previously Published Results From the Insomnia-Depression Study
The results of the double-blind, placebo-controlled insomnia-depression study of LUNESTA were published in the June 2006 edition of the journal, Biological Psychiatry. The primary endpoint for this study was WASO, and secondary endpoints included additional sleep parameters and overall HAM-D17 scores. Other key secondary endpoints included 30% antidepressant response (30% reduction from baseline) and time to 30% antidepressant response based on Bech and Maier subscales of patient-reported HAM-D17, which do not contain sleep-related items.
Throughout the double-blind treatment period, which was the first eight weeks of the study, patients treated with LUNESTA plus fluoxetine showed improvements in time to sleep onset (p less than or equal to 0.0001), WASO (p less than or equal to 0.002) and TST (p less than or equal to 0.0004), compared to those patients taking placebo plus fluoxetine. In this study, there was no evidence of tolerance to sleep efficacy (no waning of effect) across the eight weeks of treatment, and there was no rebound insomnia upon discontinuation of LUNESTA. Patients receiving LUNESTA plus fluoxetine also reported improvements in sleep quality (p less than or equal to 0.0002) and depth of sleep (p less than or equal to 0.0007) relative to patients receiving placebo plus fluoxetine. Averaged over the double-blind period, patients receiving LUNESTA plus fluoxetine also reported improved daytime alertness (p=0.03), ability to think clearly and concentrate (p=0.02) and ability to function (p=0.007) relative to patients receiving placebo plus fluoxetine.
Patients co-administered LUNESTA with fluoxetine demonstrated reductions (p=0.01) in overall HAM-D17 scores at Week 4, and there was a progressive improvement at Week 8 (p=0.002), versus patients in the placebo-fluoxetine group. After removing insomnia-specific questions from HAM-D17, improvements in scores remained significant at Week 8 (p=0.04). At Week 8, 59% of LUNESTA-fluoxetine-treated patients were responders (patients experiencing a 50 percent or greater decrease in their HAM-D17 scores; p=0.009) versus 48% of patients in the placebo-fluoxetine group, and 42% of the LUNESTA-fluoxetine-treated patients were remitters (patients who have HAM-D17 scores of 7 or lower and are therefore considered to no longer be depressed; p=0.03) versus 33% of patients in the placebo-fluoxetine group. Patients in the LUNESTA-fluoxetine group also demonstrated shorter times to antidepressant response based on CGI-I (p=0.0002) and CGI-S (p=0.01) as compared to the placebo-fluoxetine group. Fewer patients in the LUNESTA-fluoxetine group (44.7%; p=0.04) required titration to a higher dose of fluoxetine as compared to those in the placebo-fluoxetine control group (53.7%). The LUNESTA-fluoxetine treatment group did not show a worsening of depression as compared to the placebo-fluoxetine group, and the LUNESTA-fluoxetine combination appeared to result in greater improvement in HAM-D17 scores than was observed in the placebo-fluoxetine group. There were no significant differences between the treatment groups at any assessment point on patient-reported Bech and Maier subscales of the HAM-D17, and there were no significant differences in the 30% antidepressant response or time to onset of a 30% response with patient-reported Bech and Maier subscales. However, clinician-rated versions of the Bech and Maier subscales were improved in the LUNESTA-fluoxetine treatment group at both assessment points (Week 4, p less than 0.05; Week 8, p less than 0.05) during the double-blind period.
Study completion rates were similar in both the placebo- and LUNESTA-treated groups, and LUNESTA and fluoxetine co-administration was well tolerated.
An estimated one third of all adults in the U.S. suffer from either chronic or occasional insomnia.(2) Symptoms of insomnia include difficulty falling asleep, awakening frequently during the night, waking up too early, an inability to fall back to sleep, or awakening feeling unrefreshed.
Important Safety Information
LUNESTA is indicated for the treatment of insomnia. LUNESTA is not indicated for the treatment of depression. LUNESTA works quickly and should only be taken immediately before bedtime. Patients should have at least eight hours to devote to sleep before becoming active. Patients should not engage in any activity after taking LUNESTA that requires complete alertness, such as driving a car or operating machinery. Patients should use extreme care when engaging in these activities the morning after taking LUNESTA. Patients should not use alcohol while taking any sleep medicine. Most sleep medicines carry some risk of dependency. Patients should not use sleep medicines for extended periods without first talking to their doctor. Patients should see their doctor if they experience unusual changes in thinking or behavior, or if sleep problems do not improve in 7 to 10 days as this may be due to another medical condition. Side effects may include unpleasant taste, headache, drowsiness and dizziness.
About Sepracor
Sepracor Inc. is a research-based pharmaceutical company dedicated to treating and preventing human disease by discovering, developing and commercializing innovative pharmaceutical products that are directed toward serving unmet medical needs. Sepracor's drug development program has yielded a portfolio of pharmaceutical products and candidates with a focus on respiratory and central nervous system disorders. Sepracor's corporate headquarters are located in Marlborough, Massachusetts.
Bio-Imaging Technologies (2-07) Announces Fourth Quarter and Year-End 2006 Financial Results
Wednesday February 21, 7:00 am ET
EPS of $0.05 per share in Q4 2006; Record backlog of $75.2 Million
NEWTOWN, Pa.--(BUSINESS WIRE)--Bio-Imaging Technologies, Inc. ("Bio-Imaging") (NASDAQ: BITI - News) today announced its financial results for the quarter and year ended December 31, 2006.
Financial highlights for the fourth quarter and year ended December 31, 2006 include:
Service revenues increased 28.6% to $8.6 million for fourth quarter fiscal 2006 as compared to $6.7 million for the same period in fiscal 2005. Full year 2006 service revenues increased to $31.9 million, as compared to $23.7 million in fiscal 2005.
Operating income for the fourth quarter fiscal 2006 was $861,000 compared to operating loss of $556,000 for fourth quarter 2005. For full year 2006, operating income was $1.1 million compared to operating loss of $4.3 million for fiscal 2005.
Net income for fourth quarter fiscal 2006 was $613,000, or $0.05 per fully diluted share, versus a net loss of $305,000, or $0.03 per fully diluted share, in the fourth quarter of 2005. For the full year 2006, net income was $1.0 million, or $0.08 per fully diluted share, versus net loss for fiscal 2005 of $2.5 million, or $0.23 per fully diluted share.
Backlog increased 28.8% to $75.2 million, as of December 31, 2006, as compared to $58.4 million as of December 31, 2005.
Mark L. Weinstein, President and Chief Executive Officer of Bio-Imaging, said, "In 2006 we solidified our core Clinical Trials Business and made good progress in our CapMed Division. We returned the company to profitability and ended the year with a record backlog. Full year 2006 service revenues of $31.9 million exceeded our 2006 guidance of $29-$31 million and fully diluted earnings per share of $0.08 met our guidance of being profitable both for the fourth quarter and for the full year. Based on the strength of our record backlog and the positive trends in service revenue and operating income, we are increasing our full year 2007 guidance to service revenue in the range of $36-$38 million and earnings per share of $0.12 to $0.16. Previous 2007 guidance in November 2006 was service revenue in the range of $35-$37 million and earnings per share of $0.11 to $0.15.
He added, "Fourth quarter 2006 service revenues of $8.6 million represented a 7% sequential quarter over quarter increase. The increased profitability on these revenues is a testament to our leverageable expense base that generally provides increasing margins as we grow our revenues."
Fourth quarter results include CapMed revenue of $32,000 and operating expenses of $395,000, respectively, and for the full year 2006, revenue of $262,000 and operating expenses of $1,817,000 respectively. Mr. Weinstein remarked, "Although the reported revenues from CapMed were below expectations, we continue to pursue potential market opportunities in the electronic personal health records space. CapMed is involved in a number of initiatives that are demonstrating the value of having a patient centric personal record that is focused on interoperability through supporting all of the emerging standards. We are currently working on pilot projects with industry associations, hospitals, large employers and payors to test various scenarios in search of the right sustainable revenue model. Once the results of these pilot studies are analyzed, we believe there will be larger follow-on projects for CapMed.
"As announced previously, we closed the acquisition of Theralys SA in February 2007. Led by Chahin Pachai, Ph.D., Theralys is a global provider of customized imaging services in the field of central nervous system disorders ("CNS") and Neurovascular diseases. Theralys' proprietary image processing software enables the introduction of quantitative imaging markers in the design of multicenter clinical trials for Neurovascular and CNS disorders, including stroke, secondary prevention drugs, multiple sclerosis and dementia, including Alzheimer's disease (AD). The addition of Theraly expands our addressable market by strengthening our expertise in the CNS and Neurovascular area. We are now actively marketing Theralys' capabilities to our current pharmaceutical and biotech clients."
Mr. Weinstein concluded, "We are pleased that we returned to profitability in 2006 and are looking forward to continued progress in our operating results in 2007."
Management of Bio-Imaging Technologies, Inc. will host a conference call today at 11:00 A.M. EST to discuss the company's financial results and achievements. Those who wish to participate in the conference call may telephone (888) 335-6674 from the US or (973) 321-1100 for international callers, passcode #8442569 approximately 15 minutes before the call. There will be a simultaneous webcast under "Investor Relations" at www.bioimaging.com. A digital replay will be available by telephone for two weeks and may be accessed by dialing (877) 519-4471, from the US, or (973) 341-3080, for international callers, passcode #8442569. The replay will also be available for two weeks under "Investor Relations" at www.bioimaging.com.
Bio-Imaging Technologies, Inc. is a healthcare contract service organization providing services that support the product development process of the pharmaceutical, biotechnology and medical device industries. The Company has specialized in assisting its clients in the design and management of the medical-imaging component of clinical trials since 1990. Bio-Imaging serves its clients on a global basis through its US Core Lab in Newtown, PA, its European Core Lab in Leiden, The Netherlands and its recent acquisition of Theralys SA in Lyon, France. Through its CapMed division, Bio-Imaging provides the Personal HealthKey(TM) technology and the Personal Health Record (PHR) software allowing patients to better monitor and manage their health care information. Copies of Bio-Imaging Technologies' press releases and other information may be obtained through Bio-Imaging's web site at www.bioimaging.com.
Certain matters discussed in this press release are "forward-looking statements" intended to qualify for the safe harbors from liability established by the Private Securities Litigation Reform Act of 1995. In particular, the Company's statements regarding trends in the marketplace and potential future results are examples of such forward-looking statements. The forward-looking statements include risks and uncertainties, including, but not limited to, the consummation and the successful integration of current and proposed acquisitions, the timing of projects due to the variability in size, scope and duration of projects, estimates made by management with respect to the Company's financial results, backlog, critical accounting policies, regulatory delays, clinical study results which lead to reductions or cancellations of projects, and other factors, including general economic conditions and regulatory developments, not within the Company's control. The factors discussed herein and expressed from time to time in the Company's filings with the Securities and Exchange Commission could cause actual results and developments to be materially different from those expressed in or implied by such statements. The forward-looking statements are made only as of the date of this press release and the Company undertakes no obligation to publicly update such forward-looking statements to reflect subsequent events or circumstance. You should review the Company's filings, especially risk factors contained in the Form 10-K and the recent form 10-Q.
BIO-IMAGING TECHNOLOGIES, INC. AND SUBSIDIARIES
Consolidated Statements of Operations
(In thousands, except per share data)
(unaudited)
For the Three Months For the Twelve Months
Ended Ended
--------------------- ---------------------
12/31/06 12/31/05 12/31/06 12/31/05
--------- --------- --------- ---------
Service revenues 8,603 6,691 31,857 23,712
Reimbursement revenues 2,085 1,983 8,662 6,774
---------- ---------- ---------- ----------
Total revenues $ 10,688 $ 8,674 $ 40,519 $ 30,486
---------- ---------- ---------- ----------
Costs and expenses:
Cost of revenues 6,993 6,705 28,157 25,088
General & admin.
expenses 1,342 1,262 5,508 4,960
Sales & marketing
expenses 1,492 1,263 5,739 4,772
---------- ---------- ---------- ----------
Total cost and
expenses 9,827 9,230 39,404 34,820
---------- ---------- ---------- ----------
(Loss) income from
operations 861 (556) 1,115 (4,334)
Interest income (expense)
- net 151 39 504 83
---------- ---------- ---------- ----------
Income before taxes 1,012 (517) 1,619 (4,251)
Income tax (benefit)
provision 399 (212) 615 (1,706)
---------- ---------- ---------- ----------
Net (loss) income 613 (305) 1,004 (2,545)
========== ========== ========== ==========
Basic (loss) earnings per
share $ 0.05 $ (0.03) $ 0.09 $ (0.23)
========== ========== ========== ==========
Weighted average number of
shares - basic 11,283 11,157 11,219 11,114
Diluted (loss) earnings
per share $ 0.05 $ (0.03) $ 0.08 $ (0.23)
========== ========== ========== ==========
Weighted average number of
shares - diluted 12,372 11,157 12,187 11,114
BIO-IMAGING TECHNOLOGIES, INC. AND SUBSIDIARIES
Consolidated Balance Sheets
(In thousands)
(unaudited)
ASSETS
December 31, December 31,
2006 2005
------------- -------------
Current assets:
Cash and cash equivalents $ 16,166 $ 10,554
Accounts receivable 5,481 6,631
Prepaid expenses and other current
assets 1,238 992
Deferred income taxes 2,211 715
------------- -------------
Total current assets 25,096 18,892
Property & equipment, net 5,908 5,109
Intangibles & goodwill 2,227 2,519
Deferred income taxes 273 1,844
Other assets 520 427
------------- -------------
Total assets $ 34,024 $ 28,791
============= =============
LIABILITIES AND STOCKHOLDERS' EQUITY
Current liabilities:
Accounts payable $ 1,721 $ 1,681
Accrued expenses and other current
liabilities 3,335 2,027
Deferred revenue 9,367 6,255
Current maturities of capital lease
obligations 454 874
------------- -------------
Total current liabilities 14,877 10,837
Long-term capital lease obligations 97 551
Other liability 208 206
------------- -------------
Total liabilities 15,182 11,594
------------- -------------
Stockholders' equity:
Common stock 3 3
Additional paid-in capital 22,865 22,303
Accumulated other comprehensive gain
(loss) 17 (62)
Accumulated deficit (4,043) (5,047)
------------- -------------
Total stockholders' equity 18,842 17,197
------------- -------------
Total liabilities & stockholders'
equity $ 34,024 $ 28,791
============= =============
BIO-IMAGING TECHNOLOGIES, INC. AND SUBSIDIARIES
Consolidated Statements of Cash Flows
(In thousands)
(unaudited)
For the Twelve Months Ended
12/31/06 12/31/05
------------- --------------
Cash flows from operating activities:
Net income (loss) 1,004 (2,545)
Adjustments to reconcile net income to
net cash provided by Operating
activities:
Depreciation and amortization 2,035 2,312
Provision (benefit) for deferred
income taxes 25 (1,817)
Sales leaseback gains -- 17
Bad debt provision (benefit) 16 (11)
Non-cash stock based compensation
expense 357 72
Loss on foreign currency options 82 29
Changes in operating assets and
liabilities:
Decrease in accounts receivable 1,134 1,337
Increase in prepaid expenses and
other current assets (234) (92)
Increase in other assets (93) (109)
(Decrease) increase in accounts
payable (271) 411
Increase in accrued expenses and
other current liabilities 1,359 262
Increase in deferred revenue 3,113 3,178
Increase in other liabilities 2 74
------------- --------------
Net cash provided by operating
activities $ 8,529 $ 3,118
------------- --------------
Cash flows from investing activities:
Purchases of property and equipment (2,233) (1,870)
------------- --------------
Net cash used in investing
activities $ (2,233) $ (1,870)
------------- --------------
Cash flows from financing activities:
Payments under equipment lease
obligations (874) (826)
Purchase of foreign currency options (14) (118)
Proceeds from exercise of stock options 153 93
Excess tax benefit related to stock
options 51 --
Proceeds from sales leaseback -- 507
------------- --------------
Net cash used in financing
activities $ (684) $ (344)
------------- --------------
Net increase in cash and cash equivalents 5,612 904
Cash and cash equivalents at beginning of
period 10,554 9,650
------------- --------------
Cash and cash equivalents at end of
period $ 16,166 $ 10,554
============= ==============
Fearfrost, If you've got an oversized position and some profits, then trim it back and put that profit into a different promising stock. Say if you start out with a $2500 position, the stock goes up to be worth $4000, then trim it back to $2500 again. That way you still participate in further upside, but you book a nice profit, and size-wise the position is back into your comfort zone. Plus it also depends on whether/if you're going to need the money for something else. In his excellent book "One Up On Wall Street", the brilliant Peter Lynch says to not even think about owning stocks until you own a home, which is sound advice. Also, never buy stocks with borrowed money or on margin. Just don't do it - ever.
Since you're just starting out with stocks, you're in for a lot of grief. You should just skip the whole trading/excitement thing and plunk most of your money into a broad index fund like the Vanguard Total Stock Market Index Fund and their Total International Index Fund (they also have retirement funds that combine both of these into a single 80%/20% fund). Dollar cost average in every month or quarter and just let it ride until retirement. It's boring as hell, but do that religiously and you'll be able to retire by age 40-50 or so, no problem. That's the way to accumulate a nice big nest egg. If you want excitement, set aside maybe 5-10% for individual stocks, enough to play with but not enough to put you in the poor house if you lose it.
You're still in the "this is fun" stage, but let me tell you, if you continue in biotech or, God forbid, options, or margin, this little hobby known as investing will become one really bad dream. Think of that experience you told us about on saliva divinorum (?) where the floor cracked open and was swallowing you and your friend. Investing can turn into a real nightmare if you're not very careful and disciplined.
Striaterminalis, Something tells me that we investors will never be told the full details of this histo "phenomenon". Cortex will submit everything to the FDA, the verdict will come down, and either way it goes Dr. Stoll still won't tell us all the details of what was happening on a cellular level. That would really blow, but he'll probably continue to say that he doesn't want to help Cortex's competitors. Maybe if the high impact approach works someday and Drs. Lynch and Rogers get Nobel prizes, we'll read about it in their memoirs :o)
Fearfrost, Biomarin is clearly a good long term hold IMO, and I would have a modest position as part of a diversified portfolio. That way you don't have to worry about trading or timing, and won't get an ulcer either :o) When you own too much and things are out of balance is when you'll feel anxiety. Be in balance and everything feels cool. It's the same old moderation / balance / diversification mantra.
Striaterminalis, But it (calpain activity) wouldn't be scattered all across the CNS, it would primarily be happening in brain areas where AMPA receptors are the most plentiful, and where Ampakines have the most activity, like the hippocampus and cerebral cortex. Ampakines have very little effect in lower brain areas that control autonomic functions like respiration, heart rate, etc.
Looking at - 1) Excitotoxicity - with the massive doses being used in the animal tox trials, it's hard to imagine there not being at least some degree of excitotoxicity being induced. I don't know if they've induced actual seizures in the animals, but they have to be getting fairly close to something approaching seizures, one would think.
2) Calpain activity - this process is calcium dependent and related to excitotoxicity. It's also thought to be associated with the upregulation of LTP and memory formation (breaking down and reestablishing/strengthening neural connections). So it's not surprising that an Ampakine would induce calpain activity, especially at extremely high doses over longer periods.
3) Neurotrophin upregulation - even a low impact like CX-717 induces some BDNF upregulation (CX-727 was found to upregulate BDNF by ~30% or so). Perhaps the upregulation of BDNF by CX-717 causes some structural cellular changes that constitute or contribute to the histo "phenomenon". This is quite possible since we know BDNF stimulates neuro repair, regeneration, neurogenesis, etc.
Striaterminalis, As I indicated, if the level of AMPA receptor-elicited excitotoxicity had been happening to the extent described in that first paper, it would have been clearly visible in frozen slices and without using an electron microscope. So that degree of excitotoxicity obviously isn't occurring with CX-717. In the 2nd paper however, CX-614 induced calpain activity had much more subtle effects. That's what I'm suggesting might conceivably be happening with CX-717.
All I'm saying is that we need to consider the most obvious possibilities, the things that we know AMPA upmodulation can cause, the "usual suspects" as it were -
1) Excitotoxicity
2) Activation of calpain
3) Neurotrophin upregulation
Drs. Lynch and Baudry were credited on this paper (see below). If you scroll down to section 3.3, in addition to CX-614 they tested CX-546 and cyclothiazide. The results were very interesting (CX-614 induced calpain activity, cyclothiazide didn't induce it, and CX-546 reduced calpain activity). The 2nd paragraph in the Discussion section (part 4) discusses possible reasons for these differences, the deactivation vrs desensitization mechanisms, etc. With the incredibly massive doses of CX-717 that Cortex is giving in the animal tox studies, one can't rule out the involvement of this calpain/spectrin effect as a possible source of our histo phenomenon (IMO) -
>>> Effects of positive AMPA receptor modulators on calpain-mediated spectrin degradation in cultured hippocampal slices
Hussam Jourdi,ab Ted Yanagihara,b Ulises Martinez,b Xiaoning Bi,b Gary Lynch,b and Michel Baudrya*
aNeuroscience Program, University of Southern California, Los Angeles, CA 90089-2520, USA
bDepartment of Psychiatry and Human Behavior, University of California at Irvine Medical School, Irvine, CA 92697-1695, USA
*Corresponding author. Tel.: +1 213 740 9188; fax: +1 213 740 E-mail address: baudry@neuro.usc.edu.
The publisher's final edited version of this article is available at Neurochem Int.
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Abstract
1. Introduction
References Abstract
Positive modulators of AMPA receptors (AMPAr), also known as ampakines, are allosteric effectors of the receptors and have been extensively studied in past years due to their potential use as treatment for various diseases and ailments of the central nervous system such as mild cognitive impairment, schizophrenia, and Alzheimer’s disease. Ampakines have been shown to improve performance on memory tasks in animals and in human subjects, an effect linked to their ability to increase agonist-mediated ion influx through AMPAr, thus leading to enhanced synaptic responses and facilitation of long-term potentiation (LTP) induction at glutamatergic synapses. As LTP is associated with calpain activation and spectrin degradation, we determined the effects of ampakine treatment of cultured hippocampal slices on spectrin degradation. Calpain activation was evaluated by determining the levels of the 145-150 kDa degradation products of spectrin. Our data indicated that incubation of hippocampal slices with some, but not all positive modulators of AMPA receptors resulted in enhanced spectrin degradation, an effect that was blocked by a calpain inhibitor. In addition, an antagonist of AMPAr but not of NMDAr blocked ampakine-induced spectrin degradation. These results indicate that prolonged treatment with selected ampakines leads to spectrin degradation mediated by activation of the calcium-dependent protease calpain.
Keywords: Ampakine, Calpain, Cytoskeleton, Glutamate, Hippocampus, Spectrin, synapse
Top
Abstract
1. Introduction
References 1. Introduction
Calpain is a neutral Ca2+-activated cysteine protease present in postsynaptic structures where it has a wide range of substrates (Lynch and Baudry, 1987; Goll et al., 2003). This protease is essential for the induction of synaptic long-term potentiation (LTP) (Oliver et al., 1989; Muller et al., 1995; Bednarski et al., 1995; Vanderklish et al., 1996; Caba et al., 2002). One of the preferred calpain substrates in brain is brain spectrin; brain spectrin, initially referred to as fodrin, is a prominent component of the membrane-associated cytoskeleton. Substantial evidence implicates spectrin in the regulation of cellular morphology in red blood cells and muscle sarcomeres, and in the clustering of acetylcholine receptors at neuromuscular junctions (Bloch and Morrow, 1989). Distinct isoforms of spectrin are expressed in neurons (Hesketh et al., 1983), and calpain-mediated spectrin proteolysis has been associated with marked and lasting effects on the morphology and physiology of synapses (Lynch and Baudry, 1987; Bednarski et al., 1995). Calpain-mediated spectrin degradation results in the formation of 145-150 kDa breakdown products, and represents site-specific and activity-dependent markers of synaptic activity (Lynch and Baudry, 1987; Saido et al., 1993; Dosemeci and Reese, 1995; Vanderklish et al., 2000; Zakharov and Mosevitsky, 2001). It has been proposed that calpain activation and spectrin degradation could reflect rapid and long lasting reorganizations of postsynaptic structures under physiological or pathological conditions (Masliah et al., 1990; Lee et al., 1991; Vanderklish and Bahr, 2000; Neumar et al., 2001; Caba et al., 2002).
Glutamate release in the brain elicits a fast postsynaptic response mediated by the opening of AMPA-type glutamate receptor (AMPAr) channels. This response depends on binding of glutamate to the receptor and can be positively modulated by a group of allosteric effectors of these receptors called ampakines. Ampakines have been shown to bind to the AMPA receptors at a site distinct from the glutamate site and to increase AMPAr-mediated current in acute and cultured hippocampal slices and in vivo, to improve performance on several behavioral tasks when administered to animals, and to enhance memory encoding in humans (Ingvar et al., 1997; Hampson et al., 1998a,b; Arai et al., 2000; Suppiramaniam et al., 2001). Furthermore, several ampakines have been shown to facilitate LTP induction in hippocampal slices and in vivo, and this effect has been linked to their cognitive enhancing properties. In view of the proposed role of calpain in LTP induction, it was of interest to determine the effect of ampakines on calpain activation and spectrin degradation in hippocampus. Our results indicate that treatment with some, but not all, positive modulators of AMPA receptors results in calpain activation and spectrin degradation.
2. Materials and methods
2.1. Hippocampal slice culture. Ten to 13 day-old rat pups were obtained from Jackson Laboratories (Bar Harbor, Maine) and used to prepare hippocampal slice cultures as previously described (Stoppini et al., 1991). Briefly, animals were decapitated and the brains were dissected, trimmed, and 400 mm thick hippocampal slices were cut using a McIllwain tissue slicer (Fotodyne Inc., New Berlin, WI). Slices were plated onto Millipore insert membranes, placed in six-well culture plates, and fed with regular slice culture medium (50% basal Eagle medium, 25% Earl’s balanced salt solution, 136 mM sodium chloride, 2 mM calcium chloride, 2.5 mM magnesium sulfate, 5 mM sodium bicarbonate, 3 mM glutamine, 40 mM glucose, 0.5 mM ascorbic acid, 20 mM HEPES buffer, 1 mg/l insulin, 5 U/ml penicillin and 5 mg/l streptomycin; pH 7.3) supplemented with 5% fetal bovine serum and 5% horse serum. Slices were kept in culture for 2 weeks and fed with 1 ml of fresh medium every other day. Slices were then treated with ampakines, glutamate receptor antagonists, calpain inhibitor, or cycloheximide at the indicated concentrations: CX614, 1, 5, 10, 25 and 50 mM; CX546, 100 mM; cyclothiazide, 100 mM; AP-5, 100 mM; CNQX, 50 mM; calpain inhibitor III, 10 mM; cycloheximide, 20 mM. All concentrations were selected on the basis of the maximal effects of these drugs on their specified targets. At the indicated times after treatment initiation, slices were collected and processed for immunoblotting. The AMPAr positive modulators, CX614 and CX546, were generously provided by Cortex Pharmaceuticals (Irvine, CA). Cyclothiazide, AP-5, CNQX, calpain inhibitor III, and cycloheximide were purchased from Sigma (St. Louis, MO).
2.2. Brain membrane preparation and proteolysis by calpain. Crude synaptic/mitochondrial membrane fractions were prepared as previously described (Massicotte et al., 1990). Briefly, adult rat forebrain tissues were homogenized in 10 ml of 0.32 M sucrose solution containing 1 mM EGTA and 0.1 mM leupeptin using a glass-Teflon homogenizer. The homogenate was then centrifuged at 1000 × g for 10 min, and the supernatant was collected and centrifuged at 16,000 × g for 20 min. The membrane pellet was then washed twice by re-suspension in H2O containing 1 mM EGTA and 0.1 mM leupeptin, followed by centrifugation at 48,000 × g for 20 min. Subsequently, the pellet was resuspended twice in 50 mM Tris-acetate buffer (pH 7.4) containing 0.1 mM EGTA and centrifuged at 48,000 × g for 20 min. After the final centrifugation/re-suspension, the membrane fraction was stored in aliquots at 80 °C until use. Membranes were incubated at 37 °C with or without 2 mM CaCl2 for 30 min and then denatured as indicated below. These samples were run in parallel with samples obtained from the ampakine experiments and were used as indicators of calpain activation/spectrin degradation.
2.3. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Slices were collected into cold homogenizing buffer (50 mM Tris pH 7.5; 150 mM NaCl, 5 mM EDTA, 1 mM EGTA, 1 mM PMSF, 10 mM NaF, 1 mM Na3VO4; all from Sigma, St. Louis, MO) supplemented with the protease inhibitor cocktail (Sigma) [4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride, 2.08 mM; aprotinin, 1.6 mM; leupeptin, 40 mM; bestatin, 80 mM; pepstatin A, 30 mM; and trans-3-carboxyoxirane-2-carbonyl-l-leucylagmatine, 28 mM], sonicated five times using a Virsonic Cell Disruptor set at 50% for 4-7 s intervals, and protein concentrations were determined. Equal amounts (20-40 mg) of sample proteins were separated according to their molecular weight on 8-10% sodium dodecyl sulfate (SDS) polyacrylamide gels and transferred onto nylon/PVDF membranes (Bio-Rad Laboratories, Hercules, CA). Following transfer, membranes were blocked with 5% milk in 1 TBS-T (137 mM NaCl, 20 mM Tris base, 0.1% Tween 20; pH 7.4) for 1.5 h. The membranes were probed with the primary antibody diluted with 5% milk and incubated overnight at 4 °C. Membranes were then washed with 1 × TBS-T and incubated with horseradish peroxidase-conjugated second-ary antibodies (Amersham, Piscataway, NJ) in a 1:10,000 dilution with 1 × TBS-T for 1 h followed by washing in TBS-T. The membranes were exposed to ECL Western blotting detection reagent (Amersham) for 2 min, placed in an autoradiography cassette and exposed to Hyperfilm (Amersham) for different lengths of time. For Western blotting, the following primary antibodies were used at the indicated final concentrations: monoclonal mouse anti-actin (0.1 mg/ml; Sigma, St. Louis, MO), and monoclonal mouse anti-spectrin (0.2 mg/ml; Chemicon, Temecula, CA). All blots were re-probed with the anti-actin antibody as a loading control. Bands were scanned and quantified using the NIH Image 1.62 software. Because of the inherent variability in basal levels of the 145-150 kDa band, results from each experiment were expressed as percent of the values found in control samples. P value of < 0.05 indicates results significantly different from control values. All error bars correspond to standard errors of the mean.
3. Results
3.1. Treatment of cultured hippocampal slices with CX614 increases spectrin degradation. Calpain is activated by increased intracellular Ca2+ concentrations, and calpain-mediated spectrin degradation specifically produces a doublet band of 145-150 kDa. We monitored this characteristic band as an indicator of calpain activation. Brain membrane fractions treated with 2 mM CaCl2 (see Section 2) were used as a positive control for calpain-mediated spectrin degradation (Massicotte et al., 1990; Lu et al., 2001). Hippocampal slices were incubated with the ampakine CX614 (50 mM for 48 h) or the DMSO vehicle (0.1%). Under these conditions, previous experiments have shown that ampakines increase the levels of spontaneous electrical activity in slices, presumably as a result of the potentiation of the activity of AMPA receptors stimulated by endogenously released glutamate (Arai et al., 2000, 2002a,2002b; Baumbarger et al., 2001a,b). Equal amounts of protein samples extracted from hippocampal cultures were processed for immunoblotting with spectrin antibody (Fig. 1). Treatment of slices with CX614 resulted in the formation of a doublet band with a 145-150 kDa molecular weight, which was identical in molecular weight to that found in Ca2+-treated membrane fractions, and had previously been shown to be produced by calpain-mediated spectrin proteolysis (Siman et al., 1984; Bahr et al., 1995) Quantification of the data obtained in four independent experiments showed that CX614 treatment resulted in a 90% increase in the amount of the 145-150 kDa spectrin breakdown products (SBDP). CX614 has EC50 values for enhancing AMPAr-mediated currents ranging from 18 to 157 mM depending on the type of AMPAr subunits (Arai et al., 2000). In order to assess the effective concentrations at which CX614 induces spectrin degradation, we incubated hippocampal slices in the presence of increasing concentrations of CX614 for 48 h (Fig. 2). Increasing concentrations of CX614 resulted in increased spectrin degradation; significant increases were observed at a concentration of CX614 as low as 1 mM and the estimated EC50 value was found to be 6.5 mM.
The previous experiments were performed following a 48 h treatment with CX614. We determined the effects of shorter incubation times to evaluate the time course of CX614-mediated spectrin degradation. Cultured hippocampal slices were incubated with or without 50 mM CX614 for 0, 4, 8, 12, 24, and 48 h. Samples were then collected and processed for Western blotting (Fig. 3). CX614 treatment induced an increase in the degradation of spectrin that was significant after 4 h of incubation and reached a maximum after 8 h of incubation. Results from six similar experiments indicated that incubation beyond 8 h did not result in significantly higher spectrin degradation.
3.2. Cycloheximide treatment potentiates CX614-mediated spectrin degradation. The protein synthesis inhibitor cycloheximide (CHX) was used to determine the contribution of protein synthesis in the mechanisms underlying CX614-mediated spectrin degradation. Slices were treated with 20 mM CHX with or without 50 mM CX614 for 48 h, or were left untreated. Treatment with cycloheximide alone resulted in a 40% decrease in the levels of native spectrin (Fig. 4). However, the combined treatment with CX614 and cycloheximide dramatically increased the formation of the 145-150 kDa SBDP. The results from four similar experiments were quantified and plotted. While spectrin proteolysis in slices treated with CHX alone were not significantly different from control values (~20% larger than in control), treatment with CX614 alone resulted in a 55% increase in spectrin degradation as compared to control. Interestingly, in the absence of newly synthesized proteins, the combined treatment with CHX and CX614 produced twice as much spectrin degradation as in the control condition (+110% above control levels).
3.3. Effects of different ampakines on spectrin degradation. Various positive modulators of AMPAr have recently been developed by different groups (Hampson et al., 1998a; Johnson et al., 1999; Arai et al., 2000, 2002a,b; Lockhart et al., 2000; Arai, 2001; Baumbarger et al., 2001a,b; Goff et al., 2001; Knapp et al., 2002; Marenco et al., 2002; Munirathinam et al., 2002; Quirk and Nisenbaum, 2002). They have been proposed to belong to two categories of allosteric effectors of AMPA receptors, with one category increasing the rate of opening of the channel, and the other one reducing the rate of closing of the channel following receptor activation (Arai et al., 2000). It was therefore of interest to determine whether different AMPA receptor modulators could produce differential effects on synaptic cytoskeletal proteins. To this end, we incubated hippocampal slices for 48 h with the following compounds: CX546 (100 mM), CX614 (25 mM), and cyclothiazide (CTZ, 100 mM), which is generally considered a blocker of AMPAr desensitization. These AMPAr modulators produced very different effects on spectrin degradation (Fig. 5). While treatment with CX614 increased spectrin degradation, cyclothiazide treatment did not, and treatment with CX546 resulted in decreased spectrin degradation as compared to control.
3.4. Calpain inhibitor blocks CX614-induced spectrin degradation. The pattern of CX614-induced spectrin degradation (formation of 145-150 kDa breakdown products) was suggestive of calpain activation. To further confirm this possibility, we treated hippocampal slices with or without CX614 (50 mM for 48 h) and in the absence or presence of calpain inhibitor III (10 mM for 48 h) (Fig. 6). Calpain inhibitor III alone did not modify basal levels of spectrin degradation, but completely prevented CX614-induced formation of the 145-150 kDa SBDP.
3.5. AMPAr antagonist, but not NMDAr antagonist, blocks CX614-mediated spectrin degradation. CX614-induced spectrin degradation could reflect activity-dependent calpain activation in postsynaptic structures resulting from stimulation of AMPA and/or NMDA receptors, as it has been repeatedly shown that NMDA receptor stimulation produces calpain activation. Therefore, we incubated hippocampal slices with CX614 in the presence or absence of the NMDA or AMPA receptor antagonists, AP-5 and CNQX, respectively (Fig. 7). Blockade of AMPA receptors by CNQX completely prevented CX614-mediated spectrin degradation. In contrast, NMDA receptor blockade resulted in a small though insignificant decrease in CX614-mediated spectrin degradation.
4. Discussion
The present results indicate that treatment of cultured hippocampal slices with some but not all positive AMPA receptor modulators was associated with relatively rapid and sustained calpain activation, as evidenced by the accumulation of spectrin breakdown products identical to those generated by calpain-mediated spectrin degradation (Siman et al., 1984; Bahr et al., 1995; Bi et al., 1996). In addition, a calpain inhibitor prevented the accumulation of spectrin breakdown products. CX614 is a potent ampakine and has been shown to produce large increase in AMPA receptor-mediated synaptic responses in acute and cultured hippocampal slices (Arai et al., 2000; Lauterborn et al., 2000). The EC50 for CX614-induced calpain activation (6.7 mM) was relatively similar to its EC50 for increasing AMPA receptor-mediated synaptic transmission in hippocampal slices (18-30 mM) (Arai et al., 2000), suggesting that the effect of CX614 on calpain activation was directly the consequence of increased synaptic transmission at glutamatergic synapses. This result is in agreement with electrophysiological data indicating that treatment of cultured hippocampal slices with CX614 results in increased spontaneous firing activity (Lauterborn et al., 2000). The effect of CX614 on calpain activation was maximal after 8 h, and remained constant up to 48 h. It has previously been shown that prolonged (48 h) incubation of cultured hippocampal slices with CX614 resulted in decreased receptor responsiveness (Arai and Lynch, 1998; Arai, 2001; Arai et al., 2000, 2002a;Nagarajan et al., 2001). More recently, we demonstrated that, during this period, the expression of AMPA receptors was down-regulated, and it is thus plausible that further spectrin proteolysis after 8 h becomes less likely with increasing time of incubation with the ampakine (Lauterborn et al., 2003).
Co-treatment of slices with CX614 and cycloheximide potentiated spectrin degradation. This indicates that new protein synthesis is not required for CX614-mediated spectrin degradation. The potentiation could possibly be due to a reduction in the levels of the endogenous inhibitor of calpain, calpastatin, resulting from protein synthesis inhibition, and therefore in increased calpain activity. This interpretation could account for the small effect of treatment with CHX alone, which decreased basal levels of spectrin (presumably because of reduced synthesis) and increased levels of SBDP. Alternatively, this could also be the result of decreased elimination of spectrin breakdown products.
Interestingly, different classes of positive AMPA receptor modulators produced divergent effects on calpain activation. In particular, both cyclothiazide and CX546 did not activate calpain; if anything, CX546 produced a decrease in basal level of activation as evidenced by lower levels of SBDP as compared to control. Positive AMPA receptor modulators have been shown to affect AMPA receptor function through different mechanisms, with some modulators interfering with desensitization kinetics of the receptors, while others affect rates of channel opening or closing (Arai et al., 2002b). For instance, CX614 and CX546 mainly decreased the rate of deactivation of the receptor, thus prolonging the duration of AMPA receptor-mediated synaptic responses. On the other hand, cyclothiazide affects primarily desensitization kinetics, and has been shown to have little effect on AMPA receptor-mediated synaptic transmission. Our results would therefore imply that positive AMPA receptor modulators with similar effects on AMPA receptor kinetics are able to have opposite effects on calpain activation. As an antagonist of AMPA receptors completely blocked CX614-induced calpain activation, our results also indicate that AMPA receptor stimulation is necessary for ampakine-elicited calpain activation. The reasons for the decrease in calpain activation elicited by CX546 are not clear at the moment, but might be related to differential effects of various ampakines on pyramidal neurons versus inhibitory interneurons. Testing the effects of additional ampakines should provide interesting information regarding these interpretations.
Calpain is a calcium-dependent protease, and the origin of calcium required to activate it as a result of increased AMPA receptor function is not clear. In the hippocampus, AMPAr are mostly Na+ permeable channels and largely impermeable to Ca2+. In contrast, NMDA-type glutamate receptor activation does lead to calcium ion influx and to the activation of downstream signaling pathways including kinases and proteases such as calpain. Although the slice medium contained magnesium, it has previously been shown that NMDA receptors can still be tonically activated, as AP-5 decreased excitability in hippocampal slices (Sah et al., 1989). According to our results however, calpain activation does not seem to be due to depolarization-induced activation of NMDAr as AP-5 did not significantly inhibit CX614-induced calpain activation. Alternatively, Ca2+ increase in the cytosol could result from the activation of voltage-dependent calcium channels or from the release of Ca2+ from intracellular stores. In this regard, it is worth mentioning that ryanodine receptors are targets for proteolysis by calpain (Gilchrist et al., 1992; Shoshan-Barmatz et al., 1994). Although the source of Ca2+ needed for the initial activation of calpain remains unknown, it is possible that the truncation of ryanodine receptors could accentuate CX614-induced calpain-dependent spectrin proteolysis by enhancing calcium release from intracellular stores. Experiments are currently underway to determine whether voltage-dependent calcium channels or calpain-mediated proteolysis of ryanodine receptors are implicated in CX614-mediated rise in cytosolic Ca2+ and the subsequent spectrin proteolysis.
Calpain activation and spectrin degradation have been shown to be associated with the induction of long-term potentiation as well as with excitotoxicity (Najm et al., 1992; Bahr et al., 1995; Vanderklish and Bahr, 2000). A previous study from our laboratories has shown that CX614 treatment increased spontaneous synaptic activity (Arai et al., 2000). However, under our experimental conditions we did not detect any sign of excitotoxicity at any time during treatment of cultured hippocampal slices with CX614. As has been reported previously, this treatment is associated with large induction of the survival-promoting brain-derived neurotrophic factor (BDNF) in the absence of neuronal damage or seizure activity (Lauterborn et al., 2000, 2003). Thus, it is logical to propose that CX614-induced increases in calpain activation and spectrin degradation could contribute to structural and functional reorganization of glutamatergic synapses similar to what is achieved by long lasting synaptic phenomena (Bahr et al., 1995; Bednarski et al., 1995; Caba et al., 2002; Del Cerro et al., 1994; Vanderklish et al., 1996). Although we cannot exclude the possibility that part of the ampakine-induced spectrin degradation could take place in glial cells, all previously published data related to glutamate-mediated calpain activation indicate that this activation occurs in neurons (Roberts-Lewis et al., 1994; Vanderklish et al., 1995, 1996, 2000), suggesting that our results also reflect a neuronal activation of calpain and spectrin degradation. Several groups have shown that long-term depression is associated with receptor internalization and that receptor removal from synaptic membranes is correlated with reorganization of the postsynaptic cytoskeleton (Blanpied et al., 2002; Ehlers, 2002; Hanley et al., 2002; Iwakura et al., 2001; Lu et al., 2000, 2001; Marchand and Cartaud, 2002; Murase et al., 2002; Hering et al., 2003). We also recently reported that prolonged incubation of cultured hippocampal slices with CX614 resulted in decreased levels of the AMPAr subunits GluR1 and GluR2/3 in synaptic membranes (Lauterborn et al., 2003). Further studies are currently underway to determine the duration of this new state of organization, the effects of various ampakines, and to evaluate the reversibility of the ampakine effects on spectrin and on postsynaptic anchoring proteins such as the PDZ proteins, and on AMPAr homeostasis.
In conclusion, our data indicate that prolonged treatment with some positive AMPA receptor modulators is accompanied by increased calpain activation and spectrin degradation, and that these are mediated by AMPAr activation. Taken together, our results suggest that this treatment might produce synaptic reorganization similar to what occurs in synaptic plasticity such as during the establishment of long-term potentiation or depression at hippocampal synapses. This might be an added benefit for potential clinical applications of these drugs in various diseases. <<<
Here's a scary new abstract, an Ampakine investor's worst nightmare. If this was happening with CX-717 however, it would be clearly visible in frozen slices, and one wouldn't need an electron microscope to see it. Still, it makes for creepy reading -
>>> : Eur J Pharmacol. 2007 Feb 28;557(2-3):106-14. Epub 2006 Nov 22. Links
Involvement of calpain in AMPA-induced toxicity to rat cerebellar Purkinje neurons.
Mansouri B, Henne WM, Oomman SK, Bliss R, Attridge J, Finckbone V, Zeitouni T, Hoffman T, Bahr BA, Strahlendorf HK, Strahlendorf JC.
Department of Physiology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
AMPA receptor-elicited excitotoxicity is manifested as both a type of programmed cell death termed dark cell degeneration and edematous necrosis, both of which are linked to increased intracellular Ca(2+) concentration. The appearance of marked cytoskeletal changes in response to abusive AMPA receptor activation, coupled with increased intracellular Ca(2+) concentration suggests activation of various destructive enzymes such as calpains, a family of Ca(2+)-dependent cysteine proteases. Since calpains and AMPA have been linked to both necrotic cell death and programmed cell death, we sought to determine the role of calpains in mediating both types of AMPA-mediated toxicity in Purkinje neurons of the cerebellum. These studies employed immunohistochemistry for cytoskeletal breakdown products of calpain activity coupled with confocal microscopy and pharmacological interventions with calpain and AMPA receptor antagonists. The present study identifies an early involvement of calpains in mediating AMPA-induced dark cell degeneration, but not edematous necrosis, based upon the effectiveness of AMPA to generate calpain-derived alpha-spectrin cleavage products in cerebellar Purkinje neurons that express dark cell degeneration, and the effectiveness of calpain antagonists, PD150606 and MDL28170, to attenuate AMPA-induced dark cell degeneration. Moreover, the AMPA receptor antagonist CNQX, a proven inhibitor of AMPA-elicited dark cell degeneration, also blocked AMPA-induced increases in alpha-spectrin, further suggesting interplay between abusive AMPA receptor activation, calpain activation and dark cell degeneration. Since AMPA-induced dark cell degeneration possesses morphological changes that resemble those that occur following brain ischemia in vivo, hypoglycemia, or extended seizure episodes, the involvement of calpains as mediators of cell death is potentially far reaching and has widespread therapeutic implications in numerous CNS disorders.
PMID: 17188264 [PubMed - in process] <<<
AeroVironment (2-07) Gets $46.8M U.S. Army
Tuesday February 20, 5:51 pm ET
AeroVironment Gets $46.8 Million U.S. Army Contract for Small Unmanned Aircraft
WASHINGTON (AP) -- The U.S. Army said Tuesday it awarded a $46.8 million contract boost to unmanned aircraft developer AeroVironment Corp.
The Monrovia, Calif.-based AeroVironment will supply the Army with the RQ-11, a small unmanned aircraft, known as the Raven, and initial spare packages. The Army granted the pact on Feb. 14.
The firm will perform the work in Simi Valley, Calif., by Jan. 11, 2008.
Shares of AeroVironment gained 65 cents to close at $21.72 in afternoon trading on the Nasdaq Stock Market.
I see that Otis Day and the Knights will be the evening's entertainment at the Roth Conference. Otis my man !
>>> 6:00 PM - 10:00 PM - Evening Entertainment
Toga! Toga! Toga! with
Otis Day and the Knights <<<
I'm picturing Dr. Stoll in full toga regalia getting down on the dance floor with Maria Messinger, with Coleman and Varney doing backing vocals with the Knights as they do their rendition of "Shout". Toga, Toga, Toga!
I remember a toga party our fraternity had back in college, well, sort of remember...
Cubist (2-07) Pharma Focuses On Abandoned Antibiotic Market
Tuesday February 20, 3:57 am ET
>>> Faisal Laljee submits: Cubist Pharmaceuticals (NasdaqGS: CBST) is a biopharmaceutical company that engages in the research, development, and commercialization of pharmaceutical products for the acute care and anti-infective markets; specifically, antibiotic drugs that kill drug-resistant bacteria.
In the U.S. alone, almost 2 million people get bacterial infections every year, of which almost 5% succumb to the infections. Bacterial infections are a relatively smaller focus for biotechs and drug companies, who usually look to develop blockbusters that can rake in a few billion rather than the millions that Cubist is looking to bring in with their focus on the abandoned market of antibiotics. Cubicin is the first major antibiotic in over 13 years and is the most successful one of its kind ever, bringing in $350 million in revenue so far.
On the flip side, CBST’s earnings have not always yielded the result investors have hoped for. The last 4 quarters have seen CBST beat estimates once by a whopping 125%, and fall short 3 times. What this means is simply that analysts have not been able to get their arms around Cubist’s earnings.
With drug resistant bacterial infections on the rise, CBST is well poised to address this market with its current offerings and its pipeline of drugs due to be approved over the next couple of years. Technically, the stock has broken out of its last down-trend and looks to be on the way up. I recommend buying small positions at dips under $20, but keep in mind that the stock is going to be volatile, specially around earnings.<<<
Sigma-Aldrich (2-07) - Supelco Introduces Ascentis(R) Express HPLC Columns for High Speed and High Efficiency Separations
Tuesday February 20, 4:00 pm ET
ST. LOUIS, Feb. 20 /PRNewswire-FirstCall/ -- Supelco, a division of Sigma-Aldrich (Nasdaq: SIAL - News), has introduced Ascentis Express (http://sigma-aldrich.com/express), an innovative HPLC column designed to provide the analysis speed and resolving power of sub-2-micron columns with a 50% reduction in backpressures. Ascentis Express columns are designed to help researchers achieve extreme performance from their current HPLC or UPLC(TM) instruments.
"The high efficiencies and low backpressures of Ascentis Express enable the researcher to either double the speed or resolving power above current HPLC methods," said Wayne K. Way, Marketing Manager at Supelco in Bellefonte, PA. "A strategic partnership between Advanced Materials Technology, Inc. (AMT) and Supelco has permitted this product offering to reach the marketplace. Our scientists have worked diligently with AMT and Dr. Jack Kirkland, who developed the Fused-Core particle technology."
Ascentis Express will initially be offered in C18 and C8 stationary phases, with plans to introduce other phases. The enabling technology for Ascentis Express is the Fused-Core(TM) particle. These high purity silica particles measure 2.7 micron overall with a 0.5 micron thick porous shell. Relative to conventional sub-2-micron particles, Ascentis Express columns generate comparable efficiencies but with half the backpressure.
Supelco's offering of HPLC products is one part of Sigma-Aldrich's technological and value-driven products that, combined with our services and support, provide complete analytical applications solutions.
About Sigma-Aldrich: Sigma-Aldrich is a leading Life Science and High Technology company. Its biochemical and organic chemical products and kits are used in scientific and genomic research, biotechnology, pharmaceutical development, the diagnosis of disease and as key components in pharmaceutical and other high technology manufacturing. The Company has customers in life science companies, university and government institutions, hospitals, and in industry. Over one million scientists and technologists use its products. Sigma-Aldrich operates in 35 countries and has 7,300 employees providing excellent service worldwide. Sigma-Aldrich is committed to Accelerating Customer Success through Leadership in Life Science, High Technology and Service. For more information about Sigma-Aldrich, please visit its award-winning Web site at sigma-aldrich.com.
Fearfrost, I think perhaps "Phenoptin" sounded too much like a treatment for the eyes/optical (?) so they changed the name. I don't think they'll have much trouble getting Phenoptin approved.
For the cardio indications however, 6R-BH4 now has an uphill battle ahead. In the conf call, their science guy explained that the nitric oxide mechanism of 6-BH4 should theoretically lend itself more to some of the other cardio areas they're looking at. They're pretty much doing a shotgun approach to find where they see some activity. They may still hit with one of the indications, but they definitely need to round out their pipeline. JJ mentioned that they're starting to get approached by other companies since Biomarin has an established a sales network. I figure they'll probably partner some late stage programs over the next several years. Aldurazyme, Naglazyme, and Phenoptin (Kuvan) give Biomarin a solid foundation for a very successful company over the long haul. Also, JJ seems like an excellent CEO.
First chink in the 6R-BH4 armor. This won't affect Phenoptin/PKU at all, but the outlook for the other cardiovascular indications just got a lot riskier. If I were JJ I'd start looking for an additional compound for the pipeline just in case. Biomarin still looks great with their approved products plus Phenoptin, but they have a lot riding on 6R-BH4 for the cardio indications, probably too much.
>>> BioMarin Announces Results From Phase 2 Clinical Study of 6R-BH4 in Poorly Controlled Hypertension
Tuesday February 20, 9:00 am ET
No Significant Difference Observed Between 6R-BH4 and Placebo
NOVATO, Calif., Feb. 20 /PRNewswire-FirstCall/ -- BioMarin Pharmaceutical Inc. (Nasdaq and SWX: BMRN - News News) today announced results from its Phase 2a placebo-controlled double-blind study of 6R-BH4 in patients with poorly controlled hypertension. Results demonstrate that there was no statistically significant or clinically meaningful effect of 6R-BH4 on any efficacy or safety parameter measured, relative to placebo.
Jean-Jacques Bienaime, Chief Executive Officer of BioMarin stated, "We are surprised and disappointed by these results, especially considering the numerous encouraging pre-clinical and clinical studies of 6R-BH4 in diseases with endothelial dysfunction. We plan to analyze the data in detail to better understand the background therapy and other characteristics of patients in the study. We have no immediate plans to change the course of ongoing or planned clinical studies of 6R-BH4."
Mr. Bienaime continued, "We remain on track to file our new drug application for Phenoptin in PKU next quarter. While 6R-BH4 is the active ingredient in Phenoptin, this development has no effect on our program for Phenoptin in PKU. 6R-BH4 works by an entirely different mechanism of action and metabolic pathway in PKU, and the safety data in the hypertension study did not change the safety profile of the drug."
Study Design
The 8-week multi-center, randomized, double-blind, placebo-controlled study enrolled 116 patients with poorly controlled systemic hypertension, approximately half with type 2 diabetes. Among other eligibility criteria, to participate in the study, patients had elevated blood pressure while on at least two different medications for hypertension. Study patients received oral doses of 5 mg/kg of 6R-BH4 or placebo twice daily for an eight-week period.
Primary Efficacy Endpoint Results
Patients receiving placebo experienced a 6.4 mm Hg drop in systolic blood pressure compared to a drop of 4.8 mm Hg for patients receiving 6R-BH4. The difference was not statistically significant.
Conference Call Information
BioMarin will hold a conference call today, February 20, 2007, at 11:00 a.m. ET to discuss the results of the Phase 2 study of 6R-BH4 in poorly controlled hypertension and fourth quarter and year-end 2006 financial results. This event can be accessed on the investor section of the BioMarin website at http://www.BMRN.com.
Date: February 20, 2007
Time: 11:00 a.m. ET
U.S. and Canada Toll-Free Dial in #: 800.901.5217
International Dial in #: 617.786.2964
Participant Code: 35915200
Replay Toll-Free Dial in #: 888.286.8010
Replay International Dial in #: 617.801.6888
Replay Code: 94899251
About 6R-BH4
6R-BH4, commonly known as BH4 or tetrahydrobiopterin, is a naturally occurring enzyme cofactor that is required for numerous biochemical and physiologic processes, including the synthesis of nitric oxide (NO). NO has been shown to play a key protective role throughout the cardiovascular system and produces multiple positive effects, such as relaxing smooth muscle, reducing blood pressure, controlling inflammation and reducing platelet aggregation. Researchers have demonstrated that a deficiency of BH4 can disrupt NO synthesis, resulting in a loss of normal endothelial NO production. This loss of endothelial NO production, commonly referred to as endothelial dysfunction, has been associated with many cardiovascular diseases, including hypertension, diabetic vascular disease, peripheral arterial disease, coronary arterial disease and pulmonary hypertension, and has been shown to be a strong predictor of cardiovascular adverse events in a number of clinical studies.
6R-BH4 is the same enzyme cofactor currently being evaluated in BioMarin's Phenoptin(TM) (sapropterin dihydrochloride) for phenylketonuria (PKU). In March 2006, BioMarin and Merck Serono (a division of Merck KGaA, Darmstadt, Germany), BioMarin's corporate partner for the Phenoptin and 6R-BH4 programs, announced positive results from the Phase 3 clinical study of Phenoptin for PKU. All primary and secondary endpoints of the study were met. The type and incidence of adverse events was similar in the Phenoptin and placebo groups. Phenoptin was well tolerated and investigators reported that no serious adverse event occurred.
About BioMarin
BioMarin develops and commercializes innovative biopharmaceuticals for serious diseases and medical conditions. The company's product portfolio is comprised of two approved products and multiple clinical and preclinical product candidates. Approved products include Naglazyme® (galsulfase) for mucopolysaccharidosis VI (MPS VI), a product wholly developed and commercialized by BioMarin, and Aldurazyme® (laronidase) for mucopolysaccharidosis I (MPS I), a product which BioMarin developed through a 50/50 joint venture with Genzyme Corporation. Investigational product candidates include Phenoptin(TM) (sapropterin dihydrochloride), a Phase 3 product candidate for the treatment of phenylketonuria (PKU), and 6R-BH4 for cardiovascular indications, which is currently in Phase 2 clinical development for the treatment of peripheral arterial disease. For additional information, please visit http://www.BMRN.com. Information on BioMarin's website is not incorporated by reference into this press release. <<<
Meridian Bioscience (2-07) Receives FDA Clearance for New E. coli Test
Tuesday February 20, 9:47 am ET
CINCINNATI--(BUSINESS WIRE)--Meridian Bioscience, Inc., Cincinnati, Ohio (NASDAQ:VIVO - News) today announced that it has received clearance from the U.S. Food and Drug Administration (FDA) to market ImmunoCard STAT!® EHEC, a revolutionary new test for the diagnosis of E. coli infection. The new product detects all Shiga-toxin producing E. coli and is the first rapid (20 minute) E. coli diagnostic that differentiates between Toxin 1 & Toxin 2.
With the recent national E. coli outbreaks (spinach in September 2006 and lettuce in December 2006), there has been an increased need in the hospital market for a highly accurate, quick, easy-to-use E. coli test. New CDC recommendations published in September 2006 recommend that laboratories use a test that detects all Shiga-toxin producing E. coli, not just a test that detects the O157 E. coli strain. ImmunoCard STAT!® EHEC, with its easy-to-perform procedure, provides a practical solution that will enable laboratories to adopt those new CDC recommendations.
ImmunoCard STAT!® EHEC is the first product resulting from a strategic partnership that was announced in August 2006 with the Performance & Life Science Chemicals Division of Merck KGaA, Darmstadt, Germany and its American company EMD. This new-to-the-world product introduction demonstrates the value of the long-term strategic partnership that has been formed.
John A. Kraeutler, President and Chief Operating Officer, stated, "Disease caused by E. coli can be devastating, particularly with children, and Meridian is proud to introduce an advanced method for rapid E. coli testing that will significantly improve patient care. The launch of ImmunoCard STAT!® EHEC further solidifies Meridian's position as a leader in the field of E. coli testing."
Meridian is a fully integrated life science company that manufactures, markets and distributes a broad range of innovative diagnostic test kits, purified reagents and related products and offers biopharmaceutical enabling technologies. Utilizing a variety of methods, these products and diagnostic tests provide accuracy, simplicity and speed in the early diagnosis and treatment of common medical conditions, such as gastrointestinal, viral and respiratory infections. Meridian's diagnostic products are used outside of the human body and require little or no special equipment. The Company's products are designed to enhance patient well-being while reducing the total outcome costs of healthcare. Meridian has strong market positions in the areas of gastrointestinal and upper respiratory infections, serology, parasitology and fungal disease diagnosis. In addition, Meridian is a supplier of rare reagents, specialty biologicals and related technologies used by biopharmaceutical companies engaged in research for new drugs and vaccines. The Company markets its products and technologies to hospitals, reference laboratories, research centers, veterinary testing centers, physician offices, diagnostics manufacturers and biotech companies in more than 60 countries around the world. The Company's shares are traded through NASDAQ's Global Select Market, symbol VIVO. Meridian's website address is www.meridianbioscience.com.