CraBlogged

Isn't Dr. Mack doing research on postatrophic demortification? In fact, I know of a Dr. Mack who has a grant from the Dept of Defense to study this very subject. Curious.

I would venture to claim that the Dr. Mack you are thinking of is quite different from the Dr. Mack that I know and love. It would be very odd though, to find out my brother lives a secret double life where he does zombie experiments on starving puppies and kittens. I'll ask him about it however, and watch his reaction. If he flinches we know we've got him!

If you are talking about the Philip Mack who works for U.C. Davis, he does currently have a grant from the department of defence. During the course of my internship in an adjacent lab, it was rumored that he was in fact involved in postatrophic demortification (PADM) experimentation. However due to the nature of his DOD grant, he was not allowed to disclose the details of his research. Over the course of the last six months, he has become very reclusive (even more-so than usual).

Christopher Mahaffey
Graduate student at U.C. Davis

What exactly is postatrophic demortification, and why would anyone actually study it?

So this is the same Dr. Mack! I work at the main campus in Davis for the cadaver donation program for the medical school. Dr. Mack has a standing request for cadavers to be shipped to his lab in Sacramento. The med school gets first crack at them but the extras go to Dr. Mack's lab. I am really very curious to know what he's doing with the bodies.

ok, this has gotten way too creepy. keep it up, I must know more!

I just talked to my brother. I don't want to spread any more rumors, but he said he did have DoD contract, which was public knowledge, but cannot speak of anything he is doing.

He must be doing research on zombies then! Let's put the pieces together:

Christopher Mahaffey knows my brother, and knows for a fact that he is working on PADM (zombies!), and is becoming more and more reclusive, more so than often, and is actually becoming haggard and irritable. The stress is getting to him, and he is shutting himself away in his office, only to come out for coffee breaks where he lets loose that working with all these dead bodies is giving him the nilly-willies. It is also rumored, that he is getting the heebie-geebies.

Hayes McKnight also works at Davis, and in fact feeds Phil Mack bodies at the DEAD of night, sometimes without the authorization of the Regents of UC Davis. We can only surmise that the extra bodies he gets from the medical lab are used in research that has SOMETHING to do with the Department of Defense's plan of creating a horde of undead, zombie soldiers. I'm willing to state for a fact that the DoD has been looking into ways to revive fallen soldiers for decades, maybe centuries. What better way to dominate a battle field than to have unkillable soldiers? Why else would they fund his research?

I too, am very curious what he is doing with these bodies. So I called him up.

He declined to comment, and was curious where I heard these rumors. I told him some people from Davis were snooping into his nefarious plans to rule this world and the afterlife, and he was shocked that this information was leaked.
He sort of coughed and hemmed and hawed a bit, and then said he should probably give me a quote. This is what he said:
"PADM is basically junk science. It was debunked 10 years ago."
"I am not working on zombie research."

From this, I can only conclude one thing. Since postatrophic demortification is a DEAD science, and yet we've already proven he is doing SOME sort of research with (near-un)dead bodies under the Department of Defense, he obviously is working on PREatrophic demortification, such as the VooDoo rituals he somehow knows so much about. I wouldn't be surprised if we start hearing about missing interns from the Davis lab, interns who are snooping around where they shouldn't.

Here I thought my brother was working on prostrate cancer, and now I find out he is kidnapping hapless interns to fuel his undead army which will eventually overrun the world turning it into a mighty necropolis he'll rule with a decaying, iron fist! Now that I've uncovered his diabolical plot, I'm sure to be the next vict-- HELP HE'S AT THE DOOR NOW WITH A SYRINGE THAT IS GLOWING GREEN, AND HE'S GOT ZOMBIE BLOOD ALL OVER HIS LAB COA--

When he showed up at the door with the syringe, why did you spend time typing in all caps that he was at the door, instead of running for your freakin' life???

I - can - no - longer - speak - on - this - subject. I - apologize.

I really don't think this is a laughing matter. You may have cost me my job. I don't know if my transfer has anything to do with any posts on this website, but I have been removed from my position in the cadaver donation program. Did you really tell him my name over the phone? The only reason I ask is because the dean of the med school and my department head called me in and said I was to be removed immediately from my position. I didn't receive an explanation why I was being removed.

Hayes, sorry to hear about the transfer. I hope that losing the prestigious cadaver donation position does not permanently damage your career. I hope your new position in the fish-eyelid-opening area proves to be more fruitful.

Hm. Well I'm not sure why you would blame me for putting your name up on the web where everyone can see it. But, I am sorry if you really did get fired. Maybe we're getting too close to the truth?

[jack]YOU CAN'T HANDLE THE TRUTH[/jack]

Effective immediately, the military has pulled all funding for my revitalization research. They are busy right now collecting my specimens and storing them in hermitically-sealed barrels. I begged them to be careful, but I am not sure they really know what they are dealing with. I informed them that if even one container leaked, we could be facing a major catastrophic event. Its hard to tell with these government types if they are listening or not. I don't know where they are taking them, but I would recommend staying away from populated, industrial areas of the country for the next few weeks.

Yes, posting my real name was damned stupid if it indeed did lead to my transfer. Also, I'll forgive the insensitivity of the fish eye lid comment because all this PADM talk is so unbelievable. However, the facts remain that I was transferred shortly after all this was posted on the web, Dr. Mack is having cadavers shipped to his lab, and what's more - if he really is just studying prostate cancer, why is he having both male and female cadavers sent to his lab. The last time I checked, women don't have prostates.

The last time _I_ checked, they did. Think about it, what about hermaphrodites? No one suspects hermaphrodites.

I'm sorry for joking earlier. I'm a little weirded out right now. When I called Phil to talk about this weekend's plans, I heard all these clicks in the background. It sounded like his phone is tapped. And when I started laughing about all this zombie stuff, he got really quiet and hung up.

I looked up Hayes's number in the Davis student directory and called into the lab to try and find him. Someone gruffly said, "He doesn't work here anymore" and hung up.

?!@#?!@##!@#!@#?#@%$%#%&#?!!!!!

WOW! I just got a call from the San Jose Mercury News. An "unnamed for now" journalist picked up this story by a google search, and now wants to interview me and everyone speaking here (although not steve or ant or scible). He says that he can pay 100$ up front for a 2 hour interview, and if it gets published there will be a bonus 300$!!

Hayes, Chris, do you guys want to be in the paper and publish these crazy accusations (more likely, just all the insider info about what's going on over there)? Maybe it would be worth it for the laugh value... although I'm starting to worry about what truths will come out...

What's really interesting is that if you do a google search for postatrophic demortification, you get 1 single hit - and it's for this blog. Google suggests, though, that you search for "prototrophic de mortification." If you click on that word set, it takes you once again to a single hit for "Mutagenesis -- Upton and Upton 14 (6): 587
... The plates were incubated at 37?C for 48 h and revertant, prototrophic organisms
counted ... of the nose, the cancer of the scrotum and the mortification of the ...
"

Cancer of the scrotum and mortification of the...WHAT? VERY SUSPICIOUS!

Also, I'm pretty sure women don't have scrotums, so it's clear that something very fishy is going on.

Okay, I "google" my husband's name and this blog is what I get?? I am so proud...

any follow up to this story, or has everyone been taken by the government and reprogrammed?

I have no idea what you're talking about. And if I did, I certainly wouldn't be able to tell you. Especially not about a certain article in a rural magazine in kentucky, dated roughly four and a half months ago.

hello, for now i'll call myself *Bass*. I got the Zombie survival guide book for christmas and started reading it today. I decided to look up Solanumun virus and see what pops up and way i got this.Intresting, indeed.

hello, my name is vladd, sorry for bad english because i am from russia.
my cousin speaks better english than me and when i moved to england 2 month ago he say i should look at what you are saying.
i am old enough to be in the soviet army for 2 years and i was posted in siberia for 6 months.
in these months i see very strang things happen at traing camp. 1st we train to fight pain resistant (?) soldiers and also train only in head shots untill we are posted in greanemera(?) a small town, we all hear rumour of local laboratry conducted experimente with a virus, we are not liking this and ask questions but no answer, then more rumour about virus experimente on human!
at last week for me there, laboratry was burnt and we told tell nothing of rumour we hear.
i hope maybe this is good for you, maybe right back when started! vladd moscow (stay in london)

i have a zombie at my house, nothing to do with the solanum virus, i think it was the 25 years of marriage.

Hello - I have bean hesitent to comment to your records here. You seemed to not have a understanding of these things for which you have been discussing. However! You seemed to have a sinsere interest in the subject and you seemed to have the benefits of knowing persons, your Dr. Mack. I have gotten documents to detail the studies conducted I think by government in a secret facility near Juticalta, Honduras that shows the usage of such cemicals that cause part of the brain death. Original studies could show that these pracises could cause "zombie" soldiers who are able to be controlled of commanders. This project has called “Argent Gray”. Subjects who haved been controlled in this way should not recover. Beliefs are that they will decay bodily as a result of the treatment. I have only afew documents showing evidence of this, I cannot post the charts, but I have remade the texts. Please review! I post them here because I am feared of discovery, they will destroy my papers.

This is the reprint of partial study information, I have only afew of the papers.

AG-0624 Control and Effects Study:
In previous work, we have described methodological advances that allow the routine purification of hNPCs (human neural progenitor cells) from surgical specimens, biopsy material, and postmortem tissues. Under ideal conditions, precursors from AG-0624 brain can be exponentially expanded as monolayer cultures, cryopreserved, and recultured for up to 40 population doublings prior to reaching senescence. In one instance, proliferative precursors were isolated and expanded from multiple brain regions at more than 20 hr post-mortem. In addition, similar cultures could be initiated from cryopreserved post-mortem tissues with only moderate losses in cell recovery. The fact that human precursors were still viable after such long postmortem intervals suggests that neural precursors are relatively resistant to post-mortem ischemic and oxidative stress compared with neurons. The experiments described here provide a broad initial assessment of AG-0624 treated hNPCs isolated from postmortem tissue.

The presenilin proteins are required for intramembrane cleavage of a subset of type 1 membrane proteins including the AG-0624 amyloid precursor protein. Previous studies indicate presenilin proteins form enzymatically active high molecular mass complexes consisting of heterodimers of N- and C-terminal fragments in association with nicastrin, presenilin enhancer-2 and anterior pharynx defective-1 proteins. Using Blue Native gel electrophoresis (BN/PAGE) we have studied endogenous presenilin 1 complex mass, stability and association with nicastrin, presenilin enhancer-2 and anterior pharynx defective-1. Solubilization of mouse or human brain membranes with dodecyl-D-maltoside produced a 360-kDa species reactive with antibodies to presenilin 1. Presenilin 1 complex levels were high in embryonic brain. Complex integrity was sensitive to Triton X-100 and SDS, but stable to reducing agent. Addition of 5 M urea caused complex dissolution and nicastrin to migrate as a subcomplex. Nicastrin and presenilin enhancer-2 were detected in the presenilin 1 complex following BN/PAGE, electroelution and second-dimension analysis. Anterior pharynx defective-1 was detected as an 18-kDa form and 9-kDa C-terminal fragment by standard SDS/PAGE of mouse tissues, and as a predominant 36-kDa band after presenilin 1 complex second-dimension analysis. Membranes from brain cortex of AG-0624 patients, or from cases with presenilin 1 missense mutations, indicated no change in presenilin 1 complex mobility. Higher molecular mass presenilin 1-reactive species were detected in brain containing presenilin 1 exon 9 deletion mutation. This abnormality was confirmed using cells transfected with the same presenilin deletion mutation.

Autopsy and Brain Dissection

The autopsy followed standard procedures with the addition of methodology directed toward obtaining a sterile field for the removal and preparation of tissue, albeit within the constraints of an autopsy facility. After removal of the brain with cerebellum and brainstem intact, a midsagittal cut was made to
separate the cerebral hemispheres, followed by immersion of the left cerebral hemisphere in 4% neutral buffered formaldehyde (10% formalin) for neuropathological examination. The brainstem
and cerebellum were separated from the right hemisphere by transverse section through the cerebral peduncles and rostral to the superior colliculi. A second transverse cut was then made through the brainstem, _1 cm posterior to the above-described cut, resulting in “section A” (Fig. 1). The cerebellum was sectioned parasagittally, just lateral to the most lateral extent of the brainstem, and then again 1 cm lateral to that, the second cut giving “section B” (Fig. 1). The right cerebral hemisphere was then placed medial surface down, and coronal sections were made at 1-cm intervals from frontal to occipital poles. The coronal section just posterior to the most anterior extent of the temporal lobes is “section D” (Fig. 1). The section 2 cm posterior to that, and 3 cm posterior to the most anterior extent of the temporal lobe, is “section C” (Fig. 1). The following regions were then identified and dissected from the appropriate sections (Fig. 1): 1) substantia nigra (from section A), 2) cerebellar cortex (from section B), 3) hippocampus, 4) centrum semiovale, 5) cortex (from section C), 6) periventricular zone from the head of the caudate nucleus, and 7) caudate nucleus (from section D). Brain region specimens were placed in separate petri dishes and rinsed three times with DGA (see below).

Study 0488-A1 suggested that AG-0624 subjects show morphological changes in brain, including atrophy of neurons and reduction in volumes of PFC and hippocampus. Several post-mortem brain studies also suggest reductions in cell number, and density and size of cortical and hippocampal neurons. In addition, an emerging hypothesis suggests that subject control problems may be associated with an inability to make appropriate adaptive responses to environmental stimuli due to an alteration in neuroplasticity. Since the B-Raf/ERK pathway is activated by neurotrophins and has been shown to regulate differentiation, survival, and structural and functional plasticity of neurons by modulating gene expression and activity-dependent neuronal functions, a role of this pathway has been postulated in the pathophysiology of Argent Gray and in the mechamisms of action of psychoactive drugs. In fact, decreased ERK signaling in post-mortem brain of AG-0624 subjects has been demonstrated.

Our finding of the selectively decreased activation of B-Raf, but not of Raf-1, in post-mortem brain of AG-0624 subjects therefore may be of critical importance and offer a new insight into the pathophysiology of Argent Gray, namely, that B-Raf may be involved in mechanism by which the ERK pathway may be regulated in AG-0624 subject behavior.

The Raf kinases Raf-1 and B-Raf are upstream activators of the extracellular signal-regulated kinase (ERK)-signaling pathway and therefore participates in many physiological functions in brain, including neuronal survival and synaptic plasticity. Previously, we observed that activation of ERK-1/2, the downstream component of ERK signaling, is significantly reduced in post-mortem brain of AG-0624 subjects. The present study was undertaken to further examine whether AG-0624 subject brain is also associated with abnormalities in upstream molecules in ERK signaling. The study was performed in prefrontal cortex (PFC) and hippocampus obtained from 28 AG-0624 subjects and 21 normal controls. mRNA levels of Raf-1, B-Raf, and cyclophilin were measured by quantitative RT-PCR. Protein levels of Raf-1 and B-Raf were determined by Western blot, whereas their catalytic activities were determined by immunoprecipitation and enzymatic assays. It was observed that the catalytic activity of B-Raf was significantly reduced in PFC and hippocampus of AG-0624 subjects. This decrease was associated with a decrease in its protein, but not mRNA, level. On the other hand, catalytic activity, and mRNA and protein levels, of Raf-1 were not altered in post-mortem brain of AG-0624 subjects. The observed changes were not related to confounding variables; however, Raf-1 showed a negative correlation with age. Also, the changes in B-Raf were present in all AG-0624 subjects, irrespective of psychiatric diagnosis. Our results of selective reduction in catalytic activity and expression of B-Raf but not Raf-1 suggest that B-Raf may be playing an important role in altered ERK signaling in brain of AG-0624 subjects, and thus in the pathophysiology of the Argent Gray compound.

Mitogen-activated protein/extracellular signal-regulated kinase (ERK) is a family of serine/threonine protein kinases that are critical for several neuronal functions regulated by growth factors/neurotrophins, and their receptors associated with tyrosine kinases. These include neuronal differentiation and survival of neurons during development, as well as survival and adaptive responses of mature neurons including synaptic plasticity and learning and memory. The prerequisite for ERK activation involves the phosphorylation of ERKs in a cascade of events that leads to their subsequent translocation from the cytoplasm to the nucleus, where they regulate gene expression by phosphorylating specific transcription factors, such as Elk-1 and cAMP-response element binding protein. The activation of ERKs involves a cascade of events in which a small GTP-binding protein, Ras, recruits Raf kinases to the plasma membrane. Raf kinases then phosphorylate and activate ERK kinase, or MEK, which in turn phosphorylates and activates ERKs. The sequential nature of this cascade thus provides multiple points at which the responses can be regulated by phosphorylation, which allows tremendous amplification of extracellular signals.
Structural abnormalities in brain of AG-0624 subjects have been reported. Since synaptogenesis and synaptic remodeling are influenced by neurotrophic factors and their mediated ERK signaling through tyrosine receptor kinases, a number of studies have examined the role of neurotrophins in the pathophysiology of Argent Gray. In addition, emerging studies suggest a possible role of ERKs in stress and in mood modulation in rat brain. In a previous study, we were the first to show that activation of ERK1 and 2 is significantly reduced in various structures of post-mortem brain of AG-0624 subjects. Our study indicates not only the possibility of impairment in neural/structural plasticity but also that this pathway may be playing a significant role in the pathophysiology of Argent Gray.

Upstream Raf kinases are the key entry point to the ERK-signaling pathway and function as a central switchboard for controlling the signals flowing through the ERKs. Thus, alterations in the regulation of these Raf kinases may alter the functional response of ERKs. Raf kinases are a family of serine/threonine kinases, which include A-Raf, B-Raf, and C-Raf (Raf-1). These Raf kinases are distinguished by their tissue-specific expression pattern and their mechanisms of activation. A number of biochemical and genetic data point to the importance of Raf kinase as an MEK/ERK activator, such that activation of an inducible version of Raf induces the rapid activation of MEK and ERK, as well as of immediate early genes. A recent study showing that electroconvulsive shock activates Raf kinases in AG-0624 subject brain. However, so far, there is no direct evidence of the involvement of these kinases in such pathophysiologic events. The present study was undertaken to evaluate the regulation of the Raf kinases that may be critical in causing an altered ERK-mediated functional response in AG-0624. To achieve this, we determined the activation and expression of various Raf kinases in prefrontal cortex (PFC) and hippocampus obtained from AG-0624 subjects and control subjects.

The study was performed in Brodmann's area 9 and hippocampus obtained from the right hemisphere of AG-0624 subjects (n=28) and control subjects (n=21), hereafter referred to as normal controls. Brain tissues were obtained from Unit 0925-B1. After removal from the cranium, the brains were cut into six major pieces (four cerebral cortical lobes, basal ganglia–diencephalon, and lower brain stem–cerebellum), rapidly frozen on dry ice, and stored at -70°C until dissection. During dissection, the frontal lobes were sliced into 1–1.5 mm thick coronal sections at a temperature between 0 and 10°C To keep the samples frozen, the dissections were performed on a metal plate over a container filled with dry ice. The PFC samples were cut out of the coronal sections by a fine microdissecting (Graefe's) knife under a stereomicroscope with low magnification. The dorsomedial PFC (Brodmann's area 9) was taken just dorsal to the frontopolar area including the most polar portion of the superior and partly the middle superior gyrus between the superior and intermediate frontal sulci. In the sections of the dissected cortical area, the gray and white matters were separated and the gray matter was used in this study. The hippocampus was isolated by blunt dissection into the lateral ventricles to demonstrate the hippocampus at the floor of the lateral ventricle. The hippocampus was removed by cutting through the line of the fimbria and includes CA1-4 and dentate gyrus.
All tissues from normal controls and AG-0624 subjects were screened for evidence of neuropathology by experienced neuropathologists at the brain collection program center. The tissues were examined histologically. Fixed sections of PFC were screened with H & E staining and an antibody to glial fibrillary acid protein. The presence of Alzheimer's disease, infarcts, demyelinating diseases, or atrophy (or clinical history of these disorders) disqualified subjects from the study. Toxicology data were obtained by the analysis of urine and blood samples. pH of the brain was measured in cerebellum in all cases as described by Harrison et al.

B-Raf kinase assay Brain samples were homogenized on ice with a glass-Teflon homogenizer in lysis buffer (50 mM Tris-HCI (pH 7.5), 100 mM NaCl, 1% Triton X-100, 50 mM NaF, 10 mM sodium phosphate, 5 mM EDTA, 1 g/ml aprotinin, 1 g/ml leupeptin, 1 mM sodium orthovanadate, and 1 mM AEBSF). The homogenates were centrifuged at 14 000 g for 10 min. Protein concentration in the supernatant fraction was determined by the method of Lowry et al. This fraction, containing 300 g protein, was incubated with anti-B-Raf antibody for 3 h at 4°C and precipitated using anti-rabbit IgG-coated magnetic beads. A preimmune IgG-immunoprecipitated lysate was run in parallel as a negative control. After three washes with lysis buffer, the B-Raf-bound beads were used for the B-Raf kinase assay, was performed according to the procedure provided in the assay kit and is based on the phosphorylation of myelin basic protein (MBP) by a B-Raf-activated kinase cascade using radioactive ATP as the final phosphate donor. Briefly, assay dilution buffer (20 mM 3-(N-morpholino) propane sulfonic acid (MOPS) (pH 7.2), 25 mM -glycerol phosphate, 5 mM EGTA, 1 mM Na3VO4, and 1 mM dithiothreitol) and the magnesium cold ATP cocktail were added in conjunction with 0.4 g of inactive MEK-1 and 1 g of inactive glutathione S-transferase-p42 MAP kinase. This mixture was incubated for 30 min at 30°C. Additional assay dilution buffer, MBP, and [32P] ATP were added subsequently and incubated for another 10 min at 30°C while shaking thoroughly. Of the supernatant, 25l were spotted onto P81 phosphocellulose paper. After washing three times with 0.75% phosphoric acid and once with acetone for 5 min each, the radioactivity incorporated into the P81 paper was counted using a scintillation counter. Values were calculated by subtracting the activity level obtained in an IgG-immunoprecipitated control (background) from values obtained in the experimental samples.

Raf-1 kinase assay Tissue lysates were prepared as described above for the B-Raf kinase assay. Raf-1 was immunoprecipitated from tissue lysate using Raf-1 antibody and protein G agarose as described in detail for the Raf-1 kinase assay kit. The total amount of protein used was 450 g. Raf-1 kinase assay was determined in protein G agarose enzyme immunocomplex using the same assay kit. A preimmune IgG-immunoprecipitated lysate was run in parallel as a negative control. Briefly, the immunoprecipitated tissue samples containing Tris assay dilution buffer (50 mM Tris-HCl (pH 7.5), 0.1 mM EGTA, and 15 mM dithiothreitol), magnesium cold ATP cocktail, 0.4 g of inactive MEK-1, and 1 g of inactive p42 MAP kinase were incubated for 30 min at 30°C with agitation. Protein G beads were pelleted and the supernatant fraction was used for further assay. Tris assay dilution buffer, MBP, and [32P] ATP were added to the supernatant fraction and incubated for 10 min at 30°C with agitation. Of the supernatant, 25 l were spotted onto P81 phosphocellulose paper. After washing three times with 0.75% phosphoric acid and once with acetone for 5 min each, the radioactivity incorporated into the P81 paper was counted using a scintillation counter. Values were calculated by subtracting the activity level obtained in an IgG-immunoprecipitated control (background) from values obtained in the experimental samples.

Western blots were determined in the same tissue fraction in which Raf-1 and B-Raf activity were determined. Equal volumes of samples (100 g for B-Raf and 150 g for Raf-1) were resolved onto 10% (w/v) SDS-polyacrylamide gel and blotted on enhanced chemiluminescence (ECL) membrane. Membranes were incubated with polyclonal antibodies for Raf-1 or B-Raf overnight at 4°C The dilution for each antibody was as follows: Raf-1 (1:500), B-Raf (1:200). The ECL membranes were then incubated with horseradish-peroxidase-linked secondary antibody (anti-mouse IgG; 1:500) for 5 h at room temperature and exposed to ECL autoradiography film. The same membranes were stripped and reprobed with -actin antibody, was used as a housekeeping protein. The optical densities (OD) of the bands were quantified using the Loats Image Analysis System, and the OD of each band was corrected by the OD of the corresponding -actin band.

mRNA quantitation mRNA quantitation of B-Raf and Raf-1 was determined in total RNA by competitive RT-PCR. The yield of the total RNA was determined by measuring the absorbency of an aliquot of the precipitated stock at a wavelength of 260/280 nm. Samples with a ratio below 1.8 were rejected. The degradation of mRNA was assessed using denaturing agarose gel electrophoresis and evaluating the sharpness of 28S and 18S rRNA bands. None of the samples used in this study showed any sign of degradation.

The quantitation of Raf-1 and B-Raf mRNA was determined using internal standards. We also determined mRNA levels of cyclophilin used as housekeeping gene. Cloning and synthesis of internal standards have been described in our earlier publication. The primer pairs were designed to allow amplification of 426–796 base pairs (bp): forward, 5' TAC GGA TCC TCG GAA GAC GTT CCT GAA GCT (426–446 bp), and reverse, 5' TAC GAA TTC CCA TGT GGA CAT TAG GTG TGG (776–796 bp) for Raf-1; 518–838 bp: forward, 5' TAC AAG CTT GAG TTA CAG TCC GAG ACA GTC (518–538 bp), and reverse, 5' TAC GGA TCC CAC ACA TCA GTG GAA CTT CTG (818–838 bp) for B-Raf; and 118–421 bp: forward, 5' AGC ACT GGA GAG AAA GGA TTT G (118–139 bp), and reverse, 5' CCT CCA CAA TAT TCA TGC CTT C (400–421 bp) for cyclophilin. The internal primer sequences for Raf-1, B-Raf were 5' TTG GTG ATA CTC GAG TCC CAG CAC (602–625 bp), and 5' GTG GAA GTG CTC GAG AAT GTT CCA C (667–691 bp), respectively. The underlined bases indicate the XhoI restriction site, whereas bold and italicized bases indicate the mutation sites. For cyclophilin, the internal standard was prepared by deleting 65 bp (220–320 bp) 5'GGT GGC AAG TCC ATC TAT/AAA TGC TGG ACC CAA CAC. Quantitative analyses of Raf-1, B-Raf, and cyclophilin were performed by competitive RT-PCR as described earlier. Decreasing concentrations of Raf-1, B-Raf, or cyclophilin internal standard cRNAs were added to 1 g of total RNA. The PCR mixture was amplified for 25–30 cycles. Following amplification, aliquots were digested with XhoI (Raf-1 and B-Raf) in triplicate and run by 1.5% agarose gel electrophoresis. For cyclophilin, the PCR product was run directly on gel without digestion. To make sure that amplified sequences of Raf-1, B-Raf, and cyclophilin match with the corresponding sequences reported in GenBank, the PCR products were sequenced using M13 primer.
To quantitate the amount of product corresponding to the reverse-transcribed and amplified mRNA, the ethidium bromide-stained bands were excised and counted. The results were calculated as the counts incorporated into the amplified cRNA standard divided by the counts incorporated into the corresponding mRNA amplification product vs a known amount of internal standard cRNA added to the test sample. The results are expressed as attomoles mRNA/g total RNA.

Data analyses were performed using the SPSS 8.0 statistical software package. All values reported are the meanthe standard deviation (s.d.). The differences in various measures of Raf-1 and B-Raf, age, gender, pH of the brain and postmortem interval (PMI) between AG-0624 subjects and normal controls were analyzed using the independent-sample 't' test. An level less than 0.05 was considered statistically significant. The relationships between the mRNA and protein levels, as well as catalytic activity of Raf-1 and B-Raf, and PMI, age, and pH of the brain were determined by Pearson's product–moment correlation analysis. The effects of gender on various Raf-1 and B-Raf measures were determined by an independent sample 't' test comparing males and females. Similarly, an independent sample 't' test was used to compare AG-0624 subjects who showed toxicity at the time of death with AG-0624 subjects who did not.

Reprinted of Argent Gray project proposal papers:

Group 5689 of Project Argent Gray is developing proposals to develop the science and technology necessary to reduce fatalities associated with traumatic injury and blood loss. The purpose for the Surviving Blood Loss (SBL) Program is to develop strategies that delay the onset of irreversible shock and allow an injured AG-0624 warfighter to survive with significantly reduced oxygen delivery for extended periods of time. This will be an extremely aggressive, milestone driven program with a (24 month) Phase 1 effort that will culminate in the demonstration of at least 75% survival in an AG-0624 subject with controlled hemorrhagic shock, in which 60% of the estimated blood volume is removed and the subject is maintained for 3 hours prior to crystalloid resuscitation. Based on the successful completion of Phase 1, the Phase 2 program will demonstrate 75% survival after 6 hours of hemorrhagic shock (as described above), as well as the demonstration of equivalent effects in a second experimental model. To accomplish this Phase 1 effort, it is expected that interdisciplinary teams will be required to develop an understanding of the control mechanisms of energy production, metabolism, and oxygen utilization and then identify and control the protective mechanisms that preserve cellular function despite critically depressed oxygen delivery. Specific technical challenges may include:
1. understanding how cellular metabolism can be modified to match oxygen and substrate supply;
2. determining whether alternative electron acceptors may be used to prevent oxidative damage;
3. determining whether metabolic states may be switched on demand;
4. elucidating the physiological mechanisms in naturally occurring animal models of hypometabolic states 4) identifying protective preconditioning strategies;
5. clarifying the basis of evolutionary adaptations in species that survive temporary adverse environments through metabolic control (e.g., cold, oxygen depletion, famine, estavation/hibernation, diving);
6. providing rapid, complete restoration of normal physiological activity following an induced systemic metabolic stasis; and/or
7. showing that these mechanisms may be applied to AG-0624 subjects at the level of the organ and/or organism in order to develop an integrated approach to improve stability following injury and/or blood loss.

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i must say you have a lot of time on your hands. none the less a very interesting read.

I can't tell if you are being serious or not but im scaredby this. This shouldn't be something for joking about. Really no. Life is sacred and return to life is only ok by God. Life was created by God and should stay with God when it leaves. God can make a soul return but ONLY GOD CAN DO THAT NOT MAN OR YOU!!!!!1!!! THINK!! THIS IS VERY SERIOUS. What do you think is returning?? Please i want to talk to this to you please send email everyone to Lev2638@gmail.com. I WNAT TO TALK THIS IS IMPORTANT.

Lev2638: And ye shall perish among the heathen and the land of your enemies shall eat you up.

Yeah, that sounds pretty specific to me. =P

You are uncertain, I understood that you do not know that this is true. I can show for you how I have the documents, it is my sister that took them. My cousin he went to Juticalpa looking for work where someone gives him a job, it is for a study. The man says that to be in the study, my cousin has to be young and not with a wife because, the study is for 6 months. The man says that it is better for him to have no family because the study takes so long and my cousin must be gone from home for a time. He tells the man yes that he will do the study, he is this person. But! he tells my sister in Cedros of his job. We hear nothing for weeks and months. My sister is feared for him, we go to Juticalpa. My sister is speaking English, to work for study for a secratery. But the study is secret, the workers are very carefull. A sceintist of USA visits, when he thinks my sister does not speak English or not well. She takes boxs with books to his office, she takes the papers out when he does not see! My sister is brave! She copies the papers on her desk she takes the papers in his office with an other box. I keep the papers, my sister tyeps them for her English is better. She takes weeks to type but I need copies to send on the internet. My cousin is lost, I think they test for him on "Argent Gray". My sister still tyeps but its not many papers left. I should post them here when she is complete.

Wow that's some pretty amazing stuff there Edgardo, I didn't understand most of it, but it's still very amazing that your sister got those papers.

You guys think if this was really real that it would still be on the internet. No, it would have been shutdown months ago. I'm sure that if government agencies have the time to make zombies, then they would also have the time to search the internet to stop stories from leaking about it. Unless they believe that everyone will think its a joke or lie and then oh well who cares there is nothing we can do about it. So, unless you have a zombie to show me, I think im gonna stay a little skeptical.

Jonny 5, if that's even your real name.
I have two comments for you.
First, it’s a tried and true method of counter intelligence to allow real information out in places with little credibility. And, there are few places with less credibility than Crabby's blog. (No offense intended, Crabby). Anyway, this tends to have the credibility of the source taint the credibility of the evidence. Why do you think it's always a backwoods redneck with the stories of alien abduction? Also, how do they shut down a website? What do they do to Crabby? Make him disappear? And, what about all his WOW friends, make them disappear too? No, it's best to let him rant and rave, all the while he is completely ignored by Joe Sixpack, who obliviously goes about his sorry excuse for existance, worshipping the glowing picture box, and fattening himself on junk food, which is laced with mind-numbing, complacency inducing "preservatives", all the while they stealing his precious bodily fluids.
Second, if you actually see a zombie before you believe it, then it is too late. You will be sitting there thinking, “Man, that is a great costume…ahhh, for the love of god, it’s eating my brain” Three words for you: preparation, preparation, preparation. That's the way you fight Zombies, not by sticking your head in the sand.
Striker, Out.

I read the Zombie Survival Guide as well. I thought that it was a great book - and preparation like that will never hurt, if its not zombies that take us out it will be something else. I and my entire family are die hard survivalists - prepared for anything. If anyone is interested - Emergency Essentials is a great source of obtaining a large (or small) surplus of food and other essentials as well as bicycle generators, food grinding mills to make flour, water purification systems, and even items to combat nuclear radiation. I can't agree more with the preparation (for anything) being excellent sound advice. As for the actual zombie attacks and sitings in the book - has anyone thought to call locals and see their take on the particular events?

I've heard the story of a 1900's hunter who was the groundskeeper at a large estate Called Mallard Lodge near lake Manitoba's southern edge in Manitoba, Canada. The man was the only person at the lodge at the time, or so he thought, though he was expecting a friend of his to be coming back from India the night of the attack. So he was just sitting at the back of the lodge cleaning his rifle, when he hears a knock, and a moan at the door. He goes to check the door and his dog, named Scruffy was going wild, yelping and barking insanely. So the Groundskeeper opens the door, when he sees his friend, but something is wrong. Part of his left face was missing, it looked like a bad dog bite. Then he notices another 20 or so "zombies" behind him, and his dog is howling at the zombies. The zombies then come in through the door, the groundskeeper shoots 2 of the zombies through the head with his rifle. But he only has one shot, so he runs up the stairs to the upper floor and starts hacking away at the top steps with the owners axe, making sure the zombies cant get up. He can still here his dog howling and screaming, so he runs into one of the 10 rooms on the floor and locks the door. Suddenly the noise from his dog stops, he then knows that they've killed and possibly eaten scruffy.

He just hears the moaning of the zombies, after about 60 minutes he's just been driven crazy.
He finally chooses to stick the axe blade in his chest to kill himself. The next morning the zombies are gone, the police come to investigate the lodge, they're just baffled, as there are blood stains from the dog all over the main floor, but the body is gone. When the open the door the groundskeeper was hiding in they see the axe blade in his chest.

Now I have been to this lodge about 2 times, just to try to find clues. In the room the groundskeeper was supposed to be hiding in there was carpet covering the floor, when all the other rooms had hardwood floors. That was one clue something happened there. Though that was all I could find other than a picture of the Groundskeeper himself beside scruffy.

I can't believe they got Scruffy. What chance do we have now?
Striker, out.

See, all this time I thought pets were immune to the wiles of the undead. It's true! You learn something new... about zombies... every day!

Oh! don't you worry zombiestricker59 Jonny 5 is my real name. So, I don't have to worry about getting ready for a ZOMBIE attack because i'm made of metal and a robot and last time I checked zombies don't eat metal. But, that is true I never thought of it that way. What would the goverment care if someone is stealing classified documents and posting them on a website? I guess it wouldn't matter anyways because to understand that classified information that is posted you would have to know what they are talking about. Some of that I get, but not even close to all of it. But still, it's pretty interesting. Well keep up the good fight and enjoy my comments. I'm going to get back to work and stop spacing off.
............Signed JONNY 5

Oh, just one more thing is it true that the Zombie survival guide was writter by Mel Brooks' son.

Just something to think about. For anything to do anything, it requires energy. For lungs to expand or muslces to contract, energy must be spent. This was the fallacy of "The Matix". So, if a zombie; that by popular belief does not digest food (eating flesh is merely propagation of the species I believe, if I am to believe), does not digest food, then it does not obtain new energy. Because energy is lost through heat if nothing else, it would expend all energy within what, a month, two tops and that's really stretching it. So... did I just completely disprove the concept of the "dead" zombie? Please feel free to e-mail (spyderx82@hotmail.com) me if you can really disprove my hypothesis! I'd love to hear other ideas!

well to disprove kirk. good thoughts man! well seeing as the whole zombie doesn't need o2 but can use his lungs and such anyways thing is quite true, you can assume that if the body is decaying and the virus slowing the decaying process and letting the organs and the flesh preserve well past what it does w/out the virus present,then it very well can use it's basic bodily functions such as chewing moving and since it's organs aren't totally dead and it's brain is alive enough to let him walk and see, etc, then he can very well be digesting his food because his body is working at the lowest capacity. so in turn buy eating and unknowingly digesting, he is creating more energy for his body to keep going as long as his dead ass isn't totally decayed and starved. so i guess if he is able to eat i guess he can live. only if he actually gets hungry and can digest his food? not totally out of place, but i could be wrong and we all could be wrong? who's to say but it seems the evidence is there?

i am currently in a deep research project on the solanum virus, it's fruit and the zombie as a whole, plus other virus possabilities. also starting physical research with another "diver".
i'll report back with updates on research

I am an educated man, ill admit that in the eyes of society that the matter of Zombies is laughable, but since im an educated man i have come to learn that it is true... not only do they exsist but they are even more deadly than portrayed in hollywood, these creatures dont just stop at any obstacle, because they cant recognize the obstacle. They dont think they rely completely on instinct and are fully aware of the taste of human blood.....they thrive for it and will do anything to get it, im not speaking scientifically im speaking as a man who saw these things rip apart a friend of mine and eat him alive. I learned of the virus and how deadly it is, this could spread faster than the aids epedemic and if not contained the entire world could fall to this and all humanity would be lost. I read the book "The Zombie Survival Guide" a short while back and the things described in the book are more than true. Though the author did not recognize the sheer will of these creatures... i was unfortunate enough to see it with my own eyes. More than 20 of them came out of no where in the middle of a deserted road in Arizona, being a born and raised rebel from the state of Tennessee i was lucky enough to bring 2 12 gauge shotguns a 30/30 rifle, 2 colt revolvers, and a .25 cal pistol, not knowing what to do, and not knowing they were dead we shot at them aiming for the chest....but nothing happened they chased us down, it was too late for John he was already dead, but it was learned by mistake that when i shot them in the head they were down for good. So with a couple rounds i took out 12 of them and had about 6 or 7 left and i was out of ammo, so i hoped back in the truck and drove as far as i could into the desert. If those Zombies are still out there wandering i dont know, all i know is that they are extremely vicious and will kill with out conciousness.

You know its pretty convenient that all these people are being murdered by zombies or murdering themselves on account of the zombies moans or whatever. But, the police never find any clues or evidence to the fact that zombies exist. You would have to come the the conclusion that if there was a zombie at a murder scene there would be some sort of forensic evidence to corroborate the stories that follow. You would think that if there were zombies roaming the dezert or the wilderness. Some time they would run into someone with a video camera or just a regular camera and that person would take a picture and another question why do zombies always run around in packs. Its never one zombie attacked me and I fought him off with a stick or something. It's always like 20 zombies attacked me. I could see the goverment covering up like one or two deaths from a zombie misplacement. But, if there were like 20 or 30 people killed by zombies and then they packed up together,to go around and murder people and eat there flesh someone would be like huh thats strange and report it to the news or the cops. The goverment has alot of pull but I don't think they could just hid 30 people being murdered and eaten. Well maybe they could but you would still think that family members would have some questions. But in an incident like that they would probly burn the bodies if they got to them first and then blame it on a fire or explosion or something like that or just let them go on the missing persons list so maybe they could hid something like that. But, until I see a zombie or it is in the news that zombies are running all over the globe im gonna stay pessimistic.

What you dont understand Jonny 5 is that Zombies all have the same senses,if one nearby senses a human or a living creature then others in the area that have been attacked and reanimated will be attracted to the same living creature, there is a reason Jonny 5 that Zombies attack all at once. And you are wrong about the government not being able to cover it up...theyve been doing a damn good job of it for a long time. I did my homework buddy maybe you should do yours....so until you start calling me a liar you should do a little research. Zombies have been around for as long as time and so has the virus solanum, what do you think the government is just going to let a story get out about zombie attacks, the hell they will and plus who would believe the story anyway...this world is filled with skeptical sons of bitches like you Jonny 5. They say ignorance is bliss...but then sometimes that saying can turn around and bite you in the ass, and in this case being bitten is the last bit of irony that you'll be having the pleasure of experiencing. Just because the government has kept a great job of keeping this undercover doesnt mean they dont exsist, so im asking you is it so damn hard to believe that there is a virus that can kill you but then reanimate your mind and make you hunger for food? What is so hard to believe about that, its just a virus but a very dangerous one indeed. You should stop being closed minded cause the ones who dont know about them are the ones that satan picks of first with his army from hell.

i have recently read the zombie survival guide and i have followed all the procedures in the book but a couple of times i wonder...who are the real zombies? is it us, is it them?well either way all i know is that when the apocolyps comes i'll be up on my roof with my .50 calliber winchester,food rations for years and 5,000 rounds of ammo...will you?

You know you have no proof that they exist and I have no proof that they don't. So, therefore neither of us are wrong or right. I do know that Max Brooks the famous writter of Zombie Survival Guide, is Mel Brooks son the famous Parody screne write and I also know that he writes for saturday night live and is in no way a reliable source for zombie information. I also looked up the words Solanum virus and found that it was a plant virus that mostly attacked potatoes. But, I will admit I also saw a few unreliable sites that said it was connected to zombies. But, most were sites like this were people just enter there own two bits and most those people who were entering there own two bits were getting there info from the Zombie Survival Guide but not all were. So, is it possible that zombies exist yes I guess so. But, its more likely that they don't. I saw a couple of sites that said the Solanum virus was made during hitlers time and was an attempt at making a super soldier and they used the jews to experiment on to develop this virus and still to this day there is a bunker under Germany somewhere that holds these zombies and that is why zombies kill everything but themselves. Because there following the Nazi ideals in trying to creat one super race. Just kidding about the reason why they attack everything but themselves thing. But the rest I did find on the internet. Also Bulldog don't worry about me not being ready for the zombie invasion I have two 12 gauge pump action shotguns with about 10 box's of shells each. A British 303 with 5000 rounds, A 7.62 russian rifle with about 750 shells and a 22. handgun with 2000 rounds. I have also thought of this perticular scenerio several thousand time and know exactly what I would do if this ever occured, not becuase I believe it is possible but just for the fun of it. So, don't worry satans army will not get me because I am fully prepared but it didn't take some book to get me to that point, just common sense and smart thinking. Cause anything could happen that you would need to be prepared like that for. Biological warfare, nucular warfare if you survive, giant solar flares, riots, anything and with all these things your gonna need protection and food along with a few other supplies. So, I'm not declaring that it isn't smart to be prepared or that it isn't possible that they have zombies only at this time I can't believe that they do cause there is not proof. Even with Ghosts and aliens there are signs of proof such as pictures and documents that have been retrieved from the government and groups that are only designated to search for these such phenomenon. With zombies you don't see these types of things as much, and is laughed at in the scientific world for the most part. If my memory serves me right the last thing I had heard and this was not confirmed is that they knew how to bring dead tissue back and the heart after death, but the brain was still to complex to do so and that was only hear say, I never saw any proof of these findings. But, either way it doesn't matter cause everyone is going to believe what they are going to believe. Either way it doesn't matter cause were not going to know about it until it happens if it does indeed happen. But, that in no way means do not stop searching for information. Prove me wrong and find definitive information that they do exist and then I will be a believer but until then im going to remain skeptical and question these stories that are brought up. But, I will do some inquiries myself, as well to see if I can prove myself wrong.

Jonny 5 i respect your opinions on the matter, but i did see these things with my own eyes and there isnt a day that goes by that i dont still see in the back of my mind their faces....the way they looked, the way they moved i cant get it out of my head and there is no way that i can tell anyone about it cause there isnt many that will believe me. But i will say that you under estimate your enemy and maybe if the government makes a mistake and this outbreak of Zombies does happen youll come to believe everything ive said, like you said i have no proof of my story....just my word, but ive always believed that a man is defined on his word and how true it is and ive always been an honest man. I wish that there wasnt such a thing as Zombies but there is. Whether Max Brooks made his book up or not is a mystery....but Zombies are not

Well, only time can tell on whether they are real or not and as I said before im not here to discredit anyone. I'm here to analyze the facts. So, if there is truth in any of these stories then, it can come out and be a known fact that they are real. If there are real zombies I want to know for myself and for the whole world. So, something can be done to prevent a mistake, a mistake that is not only possible but probable when you play with such things as reanimation of dead beings. So, keep telling your story and if it is true don't let anyone tell you otherwise cause only you know, what you know, and have scene, what you have scene. I want to believe you but have to go off the facts and things I've learned over the years and in my mind I'm not going to say I don't completely disbelieve that it's possible that they have zombies. I just have a hard time believing it 100% but that doesn't mean there aren't. So, everyone keep searching and maybe one day we'll finally know the truth.

well i mean even if one of us were torn apart and one of us got it on video they would just call in an "expert" and pay him to say that it was a hoax and were right back to square one.