Sunday, January 29, 2006

RNA isolation: required reading for any student, postdoc, or P.I.

Yes, I know this is a natural products/herbal medicine blog but, believe it or not, even pharmacologists sometimes have cause for isolating RNA in their experiments. I'm also really keen on good science journalism since there are so few writers qualified enough to cover science and medicine.

How many of you have used Chomczynski & Sacchi's RNA isolation method using acid phenol-guanidinium thiocyanate-chloroform extraction? Y'ever wonder why chloroform? Y'ever have a student ask you who ever thought to put isoamyl alcohol in the chloroform? Y'ever wonder why 4M guanidinium and not 1M or 2.7M? Or, do you just use kits and never think about these things?

It always amazes me how techniques develop and how often we take for granted today the technologies we have available to us in the lab.

So, while sneaking into lab this Sunday morning before PharmGirl and PharmToddler wake up, I grabbed the new copy of Nature Methods and got a great history lesson from an emerging science journalism star, Michael Eisenstein.

You've got to read his quick one-pager, "A look back: message in a test tube," that precedes the entire classic protocol. (Apologies if you don't have institutional access to Nature Methods; send a gmail to abelpharmboy and I'll send you my PDF).

I won't spoil it for you, but it makes you appreciate those who came before us and how happy accidents come from the ingenuity of folks working in resource-challenged laboratories.

P.S. to any grad students out there about to take their qualifying exams: if I'm on your committee, this stuff is officially fair game!

Saturday, January 28, 2006

Grant deadline approaching

My apologies, friends. Just as I have started to build a readership that is approaching 'dozens,' I've gotten quite bogged down in writing my parts of several research grant applications that are due on Feb 1.

Something about the letdown after the holidays has frozen my brain and amplified my already excellent aptitude for procrastination. Not to mention this week was the year's 'Day of Gloom', and you've got the formula to explain, but perhaps not excuse, my 8-day hiatus. I wasn't able to get 'on a roll' with my work writing until this past Wednesday but may now actually get some free time to post something useful and interesting.

How do my bloggerific colleagues do it? I guess I'll learn as this blog develops.

Stick with me, devoted reader, and I shall work for your redemption.

Friday, January 20, 2006

Can we use this herb to treat my husband's cancer?

That was the question awaiting me Tuesday morning when I first checked e-mail and my telephone messages. Odd, I thought, because my institution is very protective of our e-mail addresses and direct telephone numbers. Ah-ha, the woman must have read our most recent paper where all of my contact info is in black-and-white in the corresponding author section.

This inquiry highlighted for me the chasm that exists for the general public between in vitro cell culture results and the outcomes of clinical trials. All too often, though, herbal manufacturers report in vitro results as "scientific research proves that our product does X, Y, and Z," when, in fact, the remedy would have no effect in a real person. Or, you'd have to take three bottles of the remedy per day just to achieve plasma concentrations of the active components consistent with the observed in vitro effects. (It kills me to give you this link, but I want you to see an example).

My laboratory works on an herbal medicine that others have shown to shrink human prostate tumors in nude mice and prolong their survival. I don't want to mention the herb by name for fear of misleading cancer patients and/or being quote-mined by companies that sell this herb. But just do a PubMed search for "herbal" and "cancer" and you'll see some candidates that have been publicized in the press far more than our little project.

Anyhoo, my chemistry colleagues have now isolated the individual compounds present in this herb, some for the first time. We recently published a paper showing that most of the compounds inhibit human prostate cancer cells from growing in culture, but one was particularly potent and caused the cells to die rather than simply differentiate and just sit there.

So, we're gearing up with a collaborator to test the pure herbal compounds in human tumor xenografts (the nude mouse thing above - I personally no longer work with lab animals). But even if we were a drug company, we'd be at least 5 years away from getting the pure compound approved as a drug...and only if the pure compound had a clear patent position.

If I were a weasel and were selling this compound as an herbal medicine under the US dietary supplement laws, I suspect I could promote my peer-reviewed and published in vitro results as having direct application to humans and sell tons of this dietary supplement to unsuspecting cancer patients. Instead, I closed my e-mail response as follows:

"I apologize that I have no other concrete recommendations, but please recognize that we are still in the early stages of cell culture and mouse experiments. [These] products have both been shown effective in animals that harbor human prostate cancer implants and we are currently testing each extract to see which compounds are most effective. Your last question may refer to our paper showing that a component of [this herbal extract] is the most effective at suppressing prostate cancer cell growth in culture, but it is only about 3-fold more active than other compounds in the mixture. However, we can't make any real recommendations until the extracts and pure compounds are tested in people.

Please feel free to follow-up with any other questions you may have, but recognize that I am not a physician. Since you are in [her hometown], I would encourage you to also look at the integrative oncology programs at Thomas Jefferson Univ Hospital in Philadelphia and Memorial Sloan-Kettering in New York."

I'm certain that the woman wanted me to tell her the doses and products to recommend to her husband, but I'm not in the legal and authoritative position to do so. And, frankly, we really don't yet know how much of this compound gets into the blood stream of people who take X dose of even the best herbal product off the shelf.

What would you have done?

Wednesday, January 18, 2006

"Immediate danger" with "natural" supplements...

...but not for the reason stated in this warning that I found in my snail-mail box from a large dietary supplement manufacturer .

Late Friday night, the US Food and Drug Administration (FDA) released a warning about two diet supplements imported from Brazil found to contain prescription drugs and an unapproved amphetamine. I wasn't going to write about this since I got tied up with working on a NIH grant this past weekend and, moreover, the story seems to have been beaten to death in the press and blogosphere. However, getting this mailer to contact my Congressional representatives to oppose legislation on adverse event reporting for dietary supplements just screamed for a response.

Without getting into the tedious issues of drug regulation, let it suffice to say that US dietary supplement manufacturers benefit from a 1994 law that allows them to market products without any proof of safety or effectiveness, so long as they don't make direct disease treatment claims on the labels. In fact, the US FDA can only step in and remove a product from the market if reports accumulate that it is unsafe. Currently, most such reports come from poison control centers meaning that there have to be injuries or even deaths before unsafe supplements can be taken off the shelves.

In this case, the reports likely stemmed from a consumer getting an unexpected positive drug screen result for amphetamine. Samples from the products in question, Emagrece Sim Dietary Supplement, also known as the Brazilian Diet Pill, and Herbathin Dietary Supplement, were found to contain chlordiazepoxide HCl (the active component of the original anxiolytic benzodiazepine, Librium) and fluoxetine HCl (the active component of the serotonin-selective reuptake inhibitor antidepressant, Prozac). So where was the amphetamine? Well, some samples were also found to contain Fenoproporex (N-2-cyanoethylamphetamine), that is converted in the body to amphetamine. Incidentally, it is apparently used commonly by Brazilian truck drivers to stay awake. Hell, why take NoDoz when you can get an amphetamine prodrug?

This adulteration of herbal supplements with prescription drugs occurs almost exclusively with supplements imported from Asia and South America. Generally, these are probably accidents of poor manufacturing practices. Many generic drug manufacturers outside the US also make herbal formulations and there is a good chance of carryover of the previous batch into some of the herbal product. That may explain the presence of fluoxetine and chlordiazepoxide, both of which are currently available in generic forms.

Of course, it's tough to say whether the addition of these two compounds could be intentional on the part of the manufacturer. Benzodiazepines like chlordiazepoxide can cause weight loss, perhaps by reducing anxiety associated with hunger when dieting. But they could equally cause weight gain by making one feel too sedated to exercise. Not surprisingly, most benzodiazepine package inserts note "weight gain or loss" as potential side effects. The inclusion of fluoxetine could be accidental or a misinterpretation of reports that other SSRIs like Meridia are used for weight loss; bad idea to put fluoxetine in there because it is more commonly associated with weight gain. Again, package inserts for SSRI antidepressants indicate the risk of either weight loss or gain. However, the presence of the amphetamine compound is the most likely indication of an intentional addition since amphetamines are well-known appetite-suppressants (also known as 'anorectics'), no doubt about it.

But I digress...

Why, as detailed in the mailer above, would a US herbal manufacturer oppose legislation that would improve the safety of botanicals they sell to their patients...I mean, consumers? Saying that Big Pharma is behind this is lame. US sales of individual herbal remedies is only in the hundreds of millions of dollars for big sellers like Echinacea, but those numbers are usually the sum of products from dozens of manufacturers. Big Pharma has gotten involved mildly in herbal products, usually under their consumer products divisions, as with Remifemin brand of black cohosh (GlaxoSmithKline, until recently). Correct me if I am wrong Pharma readers, but big companies won't get interested until herbs start selling several billions of dollars per year.

I'm equally stunned by this mailer since herbal trade organizations and some of the bigger manufacturers are actually in favor of a monitoring system for herb safety. I attended a botanical safety conference at the University of Minnesota almost two years ago where I head Michael McGuffin, president of the American Herbal Products Association (AHPA), welcome an adverse event reporting system so as to show once and for all that herbal supplements are far safer than prescription drugs. (This may be factual and/or due to the fact that many herbal products lack either sufficient levels of pharmacologically-active constituents or adequate bioavailability of these compounds.). Moreover, larger herbal manufacturers have been hurt by the unscrupulous practices of some of the smaller companies who have less means for authoritative botanical quality control and other good manufacturing practices. However, I sense that all might still prefer the current state of self-regulation, creating a medicinal double-standard in this country.

But the herbal industry has done well with scare tactics that depict any proposed regulatory guidelines as the government's attempt to undermine your rights and freedoms to take vitamins and supplements. The watered-down 1994 legislation that allows herbs to be sold in the US under such relaxed quality and safety restrictions was the results of a similar letter-writing campaign.

I'm certainly not a big fan of more government, particularly since our own FDA has taken a good deal of heat as of late for prescription drug regulation. I just think that there is ample precedent for herbs to be regulated in a manner that is not as extensive as that for prescription drugs, but with improved assurances for quality and safety, particularly for warnings of potential interactions with precription drugs the consumer may also be taking Canada's Natural Health Products Directorate is an ideal place to start, but I can tell that I'm already losing you with talking about regulations.

Take home message: Just because a supplement bottle has great graphics and is often sold side-by-side with over-the-counter drugs, the content, effectiveness, and safety of such products is a crapshoot.

Tuesday, January 17, 2006

New Grand Rounds is up

Many thanks to GruntDoc for hosting Grand Rounds, vol. 2, no. 17.

I'm honored to have my explanatory missive, "Why Terra Sigillata?," selected for the Science section of this issue.

I also got a huge kick out of Insider's take at PharmaGossip on Ode's 'homeopathy for flu pandemic' cover story in the Humor in Medicine section. My treatment of the same issue is far too verbose and pales in comparison for overall impact.

Anyway, hearty thanks to the Doc and to those who suggested that I send something in.

Equally, many pleasant welcomes to new readers.

Monday, January 16, 2006

Confessions of the Blogger-Challenged

I've only hung out my e-shingle for a month and already have to confess to being a computer idiot.

Here I was, pissing and moaning that I have yet to get any thread comments, even doing so on someone else's blog. I now realize that I had set up Blogger to moderate comments, partly because others had warned me of some spammer problems.

Well, it would've helped for me to 1) put in an e-mail address to send incoming comments for review and 2) click on the "moderate comments" tab in Blogger. I was so pitiful that two folks kindly came over to comment just to make me feel better.

Even more sad was that Ms.PhD and BotanicalGirl came over within 12-36 hrs of the launch without any acknowledgment on my part. For the record, Ms.PhD of YoungFemaleScientist was my very first commenter, and a very thoughtful one at that. Thanks!

My apologies also to bridgett at Strange Conceit, abbey at Hunting for Mr Right, and a couple of anonymous commentors. You are very kind to give me a read and some feedback.

As a professor and grad student advisor, I was never bashful about sharing my screw-ups with my students to simply to show that 1) no one was infallable and 2) professors and P.I.s still get judged and scored on things (grants, manuscripts, teaching evals, annual performance evaluations, etc.) where they still have the chance to crash and burn and be humiliated.

But, in this case, here I was not completely understanding a simple blog posting program that my daughter could probably already navigate at age three - sheesh! That's part of the trouble of being too young to claim ignorance of computers, but too old to have the comprehensive understanding of html, etc. since all I learned as an undergrad was how to program in Basic. It's amazing that I can even do BLAST searches on my own. Analysis of microarray data - fuggedaboutit! Can you say, "bioinformatics sabbatical?"

So, to my less senior readers, this explains why I keep coming to you for advice.

Thursday, January 12, 2006

Good Things

A favorite song of mine from the 90s is a tune called, "Good Things," done by The BoDeans of Waukesha, Wisconsin, on their 1991 disc called "Black and White." The heart of The BoDeans was the duet of Kurt Neumann and Sammy Llanas. Musically, they couldn't seem more far apart. But together, Kurt and Sammy were magical, vocally and instrumentally. As individuals, they never seemed as successful as when together, as though they were missing part of themselves when performing without the other.

In the time leading up to the launch of this blog and in the month of its existence, I have been the beneficiary of Good Things from both ends of the academic spectrum.

First, hearty thanks go to BotanicalGirl at The World of BotanicalGirl for her blogiversary link-love to this site. I stumbled across her blog when doing Blogger and Technorati searches for anyone writing on botanical or herbal medicines because I wanted to be sure I wasn't adding unnecessary bandwidth. I work primarily on medicinally-active compounds from plants, but my lab is far removed from the actual botany of our specimens. What resulted from my initial contact with her was a prolonged e-correspondence where the student became professor and vice versa. I was fortunate to previously have three direct Ph.D. trainees and serve on almost two dozen Ph.D examining committees, many for folks I considered and still consider friends, despite the cautions of colleagues in The Chronicle of Higher Education. However, my current appointment is not associated with a graduate program and I now realize how much working with graduate students contributed to my persona as an investigator and a person. Hence, it is very refreshing to interact with BG.

BotanicalGirl reminds me of the intellectual and philosophical purity with which one enters this business of doctoral training in the life sciences and the personal tribulations of serving under often-dysfunctional P.I.s. More importantly, she exhibits wisdom far beyond her years and her blog is excellent reading for any P.I. who wonders what folks in the lab are thinking, regardless of your discipline. Yes, we get Ph.D.s and become lab directors, but we don't always have the management skills we think we have. Moreover, she serves as a great mentor to fellow graduate students who often experience isolation in their respective programs, only to realize that their issues are quite universal.

BotanicalGirl has become a dear friend and is a great teacher to both her peers and to any of us who lead laboratories. Yes, it's tough for us to write grants and worry about supporting our lab personnel, but it is refreshing to read her blog and be reminded of the daily stresses that we once faced umpteen years ago. She also just got engaged to a great guy (yeah!) and is about to undergo that rite of passage, the oral qualifying examination. Good luck, my friend.

At the other end of the spectrum of Good Things I have received is the advice of Orac at Respectful Insolence. Orac does what I do professionally, but a ton more. I surmise that we are about the same age (me, 16 yrs out from my Ph.D.), but Orac is also an M.D. who keeps an NIH funded cancer research lab on top of being a surgical oncologist at a major academic medical center and still teaches residents and fellows. That is, while I am bargaining with the Invitrogenics Cell Technologies rep for lower prices on antibodies, Orac is operating on someone like my Mom with breast cancer, all while hoping to keep his lab gainfully funded. To do what he does requires great dedication and selflessness, all while being a prolific blogger and husband (well, maybe a husband and a prolific blogger).

Orac is a very thoughtful critic of alternative medicine, the field where my work on therapeutic natural products is often lumped, incorrectly. Nevertheless, he has provided links and off-line advice on how best to be unique yet be myself.

The bottom line is that I have been very pleasantly surprised at how helpful and thoughtful folks have been in the blogosphere. Even with people who don't always agree with me, I have received useful suggestions, usually offline via e-mail. Although I have yet to receive A SINGLE COMMENT on any of my posts, I have had great dialogues with people at both ends of the life science and medical science training spectrum.

BotanicalGirl and Orac collectively remind me not to forget 1) where I came from and 2) to help the next generation get where they want to go.

Many thanks to you both for promoting what we're trying to do over here in our little piece of the ether.

Tuesday, January 10, 2006

All hail Dr Akira Endo!

The Wall Street Journal has a few great science reporters and hats off today to Peter Landers for his story on the discovery of the cholesterol-lowering statins.

Dr. Akira Endo's work of the early 1970s has been buried in the annals of what ultimately became the first double-digit billion-dollar blockbuster drug class. I'm proud that I at least acknowledged his early work in in my very first post of this little ol' blog.

Mindful of natural history and fungal behavior in the wild, Endo made careful observations, used appropriate animal models and, of course, received almost no financial reward for his discovery of compactin. Merck wisely adopted the technology given to them by Endo's company and discovered lovastatin (Mevacor).

A great story about a great scientist.

Sunday, January 08, 2006

Stetson Kennedy - I'm STILL writing his name in

My profile notes one of my favorite books as Freakonomics, a partnership of NYT writer, Stephen Dubner, and U of Chicago economics Nobelist-to-be, Steven Levitt. A great book that chronicles Dr. Levitt's unique way of using economics theory and actual data to ask and answer difficult societal questions.

One of their chapters compares the KKK to real estate agents, the connection being that information asymmetry provides one side with an economic or philosophical advantage. Just as real estate agents create 'competition' when you are trying to buy or sell a house, the KKK created a more hostile racist environment by making it appear there were more racists than in actuality (although one racist is one too many in my book).

The beauty of this chapter of Freakonomics is the retelling of the story of Stetson Kennedy, a folklorist and civil rights activist who among other things documented his infiltration of the KKK in his 1954 book, The Klan Unmasked. I've not yet met Mr. Kennedy but knew of him during my pharmacology graduate training at a certain Southern university. I won't do adequate justice to him here. Check instead his website and a recent NPR radio interview.

Stetson Kennedy is the stuff of legends: turned down for WWII due to a back injury, he chose to fight fascism at home by signing up for the KKK and reporting their codes and activities to radio shows and ultimately, in a book. Still around at age 89, he continues to stand as one who has dedicated his life, at great risk of personal injury, to letting us each live without fear of persecution. Kennedy was also a good friend and frequent host to Woody Guthrie and immortalized by Billy Bragg and Wilco in their adaptation of Guthrie's lyrics in the song, "Stetson Kennedy."

In 1940s North Florida and Georgia, it would take great courage to pretend to be a Klansman and then reveal their innermost workings to the press. No blogs, no Internet in these days, folks.
Messers. Dubner and Levitt got great mileage, in part, out of Mr. Kennedy's story: a national bestseller for most of this year.

But today, Mr. Dubner has backpedalled saying that some of Kennedy's Klan infiltration was done by a 'John Brown" who was enlisted by Kennedy. The Freakonomics website details a number of documents where Kennedy himself notes that another individual may have provided some of the information that he claimed to have obtained personally.

I don't get it. Why does an accomplished NYT writer feel so motivated to discredit an 89-year-old civil rights leader who had more balls than he could ever hope to have? Wasn't 'John Brown' just a pseudonym for Mr. Kennedy for the official record in case the Klan was ever to discover his identity. After all, the bottom line is the same: the actions of Stetson Kennedy embarrassed the Klan and high-ranking Southern politicians who were linked to the Klan. The Klan's momentum was clearly stunted following Kennedy's revelations.

Freakonomics has done quite well, but Dubner now promises to include the allegedly updated Klan story in a new edition. Dubner has first ridden the coattails of a genius economist (Dr. Levitt) together with a remarkable civil rights advocate (Kennedy). Is there no more original material for Mr. Dubner to generate without making a story out of questioning the reputation of an old man?

I look forward to the response from Kennedy and his associates since he has more than once been challenged by writers lesser than Dubner. I just don't get it. Is it the aftermath of the Jayson Blair thing? Is Dubner starved for new material?

In the meantime, and in the words of Mr. Guthrie, Stetson Kennedy is the man for me.

Friday, January 06, 2006

Lesson 1: Homeopathy is NOT herbal medicine

I love Ode Magazine - much, much more often than not.

Today, I have a bone to pick.

In the aftermath of the 2004 US presidential election, I had to cancel my subscription to The Nation just because I got so depressed about negative news coming into every conduit of my house.

Ode is a Dutch-based mag that offers original and reprinted stories describing where people and ideas are working around the world to create positive change. From their mission statement: "We publish stories that bridge the gap between thinking and doing, between rage and hope, and the painful gap between the rich and the poor. By doing so we build peace and sustainability."

Cool. A breath of fresh air when everyone is screaming at one another. And, largely, Ode succeeds. Until this issue, when I ran screaming from my mailbox like Steve Martin in The Jerk.
In the January 2006 issue, Kim Ridley offers an overview of homeopathy as "a healing idea whose time has come - again?" The article does wisely posit, "Is homeopathy a 200-year-old hoax, or a powerful paradigm for healing?" But the cover statement (above) that homeopathic remedies produced much higher survival rates than conventional medicine during the 1918 influenza pandemic is poorly substantiated in a related article. More disturbingly, an Indian homeopath who uses these remedies to treat cancer is quoted as saying, "The only things I don't approve of are chemo and radiation."

Together with surgery, I know of no two other modalities that HAVE been shown conclusively to produce long-term cancer remissions (I hate to use the word 'cure'). Yet the article irresponsibly provides further details on how to seek this 'healer' who claims to have cured 80 percent of cancers over the last 10 years.

Homeopathy is a late 1700s/early 1800s practice of using extremely dilute preparations, largely of plant extracts and toxic metals, to treat diseases based upon the so-called 'law of similars'. The philosophy that 'like cures like' was first espoused by a German physician named Samuel Hahnemann who took high, conventional doses of plant medicines, observed the symptoms produced, and then used extremely dilute versions of the same plant to treat diseases that produced similar symptoms. For example, the vomit-inducing syrup of ipecac is offered in an extremely dilute form as a homeopathic treatment for any disorder where the patient is experiencing vomiting.

Where homeopathy is most controversial is in the claim that a remedy becomes more potent as it is diluted. Even experts quoted cannot account for how this is scientifically possible, although some invoke a sort of "quantum physics" change in the structure of water. Hmmm.

More erudite fellow bloggers have already commented more concisely on the implausibility of homeopathy. Even the British journal The Lancet, one of the most alternative medicine-friendly among high-impact conventional medical journals, published results last year of a meta-analysis demonstrating that homeopathy is no better than placebo.

But where I object further is when photographs of herbal medicines are placed within an article on homeopathy as is done throughout this issue of Ode. Herbal medicine is NOT homeopathy. Herbal medicine and the use of pure chemical constituents from plants still subscribe to dose-response pharmacology: that the biological response varies in direct proportion to the dose or concentration of the remedy. While some medicinal plants are used as a source for homeopathic treatments, the rationale for dosing in medicine vs. homeopathy are diammetrically opposed. Lumping together herbal medicine with homeopathy gives the former practice the same air of impossibility and detracts from the demonstrated benefits and future promise of using plants as a source of novel therapeutic molecules.

Many would love to see homeopathy proven as an effective medical practice. What's not to like: non-existent doses of a remedy that cure diseases without any side effects.

Anecdotes abound. Show me the data. Until then, I find homeopathy difficult to swallow.