It was posted by an actual nut—an unfortunate woman whose delusions and paranoia led health authorities to lock her up for a while. (See: http://www.elenacook.org/statejun06.html) Of course, not everything can be cured so she periodically pops up with some new conspiracy fantasy. The latest one—perhaps encouraged by MacDonald—is a wacky attack on Linda Bockenstedt, MD at Yale. Here’s some of it:
Lyme borrelia biofilms expert Dr. Alan Macdonald writes to the Journal of Clinical Investigation to protest as... IDSA Guidelines Author Bockenstedt Contravenes Regulations to Evade Verification of her Findings
by Elena Cook
Dr Linda Bockenstedt is a Yale-based author of the deeply-flawed Lyme Disease guidelines issued by IDSA in 2006. Recently, she published a study purporting to show that signs of persisting Borrelia burgdorferi in antibiotic-treated mice (and, by implication, in humans with chronic Lyme Disease), were nothing more than the remnants of dead bacteria (1). She did not explain why this "debris" should, exceptionally, resist all normal immune mechanisms for clearing dead microbe remnants in a mammalian host. Images accompanying her publication attracted the attention of Dr Alan B. Macdonald, who is currently researching Biofilms of Borrelia along with Dr Eva Sapi and her team at University of New Haven, Connecticut. This collaboration recently resulted in the first-ever study demonstrating Biofilm formation by Borrelia burgdorferi in vitro.
Because her images so strikingly reproduce the known features of a Borrelia biofilm, Dr. Macdonald wrote to her privately, requesting that she carry out a very simple, dye-based test on the material. The test is inexpensive and easily distinguishes between dead and live micro-organisms. She refused. Dr. Macdonald then asked her to share some of the mouse tissue from her experiment so that he could perform this test in his own lab. Once again, Dr. Bockenstedt refused - in flagrant denial of US public health agency regulations, which require taxpayer-funded scientists to make their material available to other researchers for independent verification. Dr. Bockenstedt's study was funded by the National Institutes for Health (NIH) and the Centre for Disease Control (CDC), but also by the non-profit National Research Fund for Tick-Borne Diseases. Her refusal to share tissue or to perform the simple dye test calls into question her integrity as a scientist.
Below is the e-letter submitted to the Journal of Clinical Investigation by Dr. Alan Macdonald.
It’s not clear if MacDonald actually wrote to L.B. or to JCI. (Why he’d write to a journal about getting samples from the author of an article is a complete mystery.) It’s not clear how poor crazy Elena Cook got hold of the letter either. But I suppose the point of posting this stuff was to somehow embarrass L.B. or pressure her into giving MacDonald that to which he is not entitled.Letter Submitted to Journal of Clinical Invesigations by Dr. Alan B. Macdonald
Dear Dr Bockenstedt,
I remark with great interest your inclusion of the brief discussion of Biofilms of Borrelia burgdorferi in your Discussion section, in connection with possible etiologies for the GFP-emitting "amorphous blobs" of Borrelia burgdorferi in deep dermal sites closely apposed to the articular cartilage of the murine arthritic Lyme arthritis mice.
[deleted because MacDonald tends to run on and on and on…]
The assertion that All of the borrelia microbes in the "Amorphous Blobs in murine deep dermis in laboratory induced Chronic Lyme Arthritis" are ALL DEAD and merely represent "debris"....is just that: an assertion, buttressed by what you say are corroborative mRNA supportive data...( not presented in your paper). I have suggested to you in a personal private communication that the way to establish Live versus Dead borrelia in your "Amorphous Blobs" is to pour some Dye ( Invitrogen ::Live Dead Assay) (Ref 10)(Red=dead, and Green = alive).
You have steadfastly refused to accommodate this quality control procedure, which is fast, cheap, reliable and easily accomplished. You have refused me tissue from your "amorphous Globs" dermal tissue from murine subjects, so that I might perform this simple quality control step in my own laboratory. Refusal to provide tissue to an outside scientist, is in violation of the guidelines of the NIH and other Federal funding agencies. Your Research activities were accomplished with the use of public funds. You have an incumbent obligation to provide tissue to outside scientists upon request,to verify your experimental findings.
[deleted due to excessive yakking]
I therefore challenge you to voluntarily participate in Quality Control microscopic exercise -- to establish with the LIVE DEAD Invitrogen Kit that ALL of the Borrelia inside of your "Amorphous Blobs" stain Red(=Dead) and that none of the borrelia within your "Amorphous Globs" stains Green in the Invitrogen Live Dead assay.(Ref 10) If you are correct, there is nothing to lose in this quality control exercise.
Just because the NIH funded some of L.B.’s research does not mean she can be forced to collaborate with whoever sends her a letter. Nor does it mean she has to provide them with anything more than what is provided in the M&M section of her paper. Moreover, she might actually get into legal trouble if she were to ship off biological samples and reagents to anyone who asked for them. A retired guy tinkering in his basement is not likely to be viewed as a qualified scientist or in the employ of a legitimate research center.
L.B.’s colleagues and the journal editors found her methods and conclusions sufficiently credible to have them published. If anyone questions them, they can certainly repeat those experiments on their own.
As for MacDonald’s colorful Live-Dead assertions about Borrelia: it isn’t that simple. Davey recently noted in his minireview:
Whether we are discussing microorganisms or macroorganisms, it is usually easier to distinguish between live and killed individuals than between organisms that are alive and those which have recently died of natural causes. In the case of staining, while control live and dead samples may be clearly separable, environmental stresses often give rise to heterogeneous populations, with some cells showing an intermediate uptake of viability stains. When a population of cells is exposed to stress, depending on the magnitude of the stress, there may be a heterogeneous response in which some cells are killed, others are damaged, and yet others may show no observable phenotypic change. It has recently been shown, even for well-characterized, eukaryotic laboratory organisms when cells are under stress, that propidium iodide (PI) may enter cells during or immediately after application of the stress but that a short period of recovery will allow membrane damage to be repaired such that PI cannot enter the cells. (Hazel M. Davey HM. Life, Death, and In-Between: Meanings and Methods in Microbiology. Appl Environ Micro, 2011;77(16):5571-76.)
The commercially available LIVE/DEAD BacLight kit (Invitrogen) has enjoyed increasing popularity among researchers in various fields since it was released about 10 years ago. The kit consists of two stains, propidium iodide (PI) and SYTO9, which both stain nucleic acids. Green fluorescing SYTO9 is able to enter all cells and is used for assessing total cell counts, whereas red fluorescing PI enters only cells with damaged cytoplasmic membranes. The emission properties of the stain mixture bound to DNA change due to the displacement of one stain by the other and quenching by fluorescence resonance energy transfer. LIVE/DEAD staining was shown to work not only with (eu)bacteria but also with archaea or eukaryotic cells, such as yeast (Saccharomyces cerevisiae). Although this kit enables differentiation only between bacteria with intact and damaged cytoplasmic membranes, it is often used to differentiate between active and dead cells. While it seems accurate to assume that membrane-compromised bacterial cells can be considered dead, the reverse (that intact cells are active cells) is not necessarily true. Microscopic assessment of LIVE/DEAD-stained bacterial cells is usually simplified to either “green”-labeled (live) or “red”-labeled (dead) cells. However, experience with this dye combination and flow cytometry during the last few years by our group and others has shown that the staining of bacterial cells with SYTO9 and PI does not always produce distinct “live” and “dead” populations; intermediate states are also observed. In the kit manufacturer’s manual, the region of intermediate states is referred to as “unknown.” This can lead to difficulties in the interpretation of results and can be critical when, for example, decisions have to be made about the effectiveness of disinfection methods or the amount of viable bacteria in water distribution systems. (Berney M, et al. Appl Environ Micro, 2007;73(10):3283-90)
For MacDonald, it’s back to the basement with a pocketful of mice. As for Cook, maybe it’s time to see the nice mental health people again.