Herpes killing herbs xbox,hsv oncolytic virus immunotherapy,natural remedies for pediatric asthma - .

admin 19.09.2014

Pete Sartin says he's been watching over his horses carefully since the Equine Herpes Virus [EHV-1] broke out after a recent cutting horse show in Utah. Washington state veterinarian Leonard Eldridge says this particular strain of the virus can't be contracted by humans, but it's very deadly for horses.
Morgellons Disease Awareness -  Artist and Morgellons sufferer, Ayla, journals her experience with the disease   I'd like to introduce myself, I'm Ayla, a 59year old artist and graphic designer who contracted Morgellons in Jan.
The natural antiseptic, antibacterial, anti-viral, and anti-fungal tea tree oil derived from the plant that is native to Australia has established itself as one of the best remedies for herpes and other sores.
If you are using tea tree oil on your skin for the first time, use a carrier oil like olive or almond oil. If you are used to using tea tree oil, you may omit mixing it with other oil and apply it directly over your sores. Traditionally the gel taken out of the aloe vera leaf has been used for various medical purposes, one being skin conditions. Know using the knife again, cut open the aloe leaf slice and take out its sticky thick juice. Goldenseal has this dependable reputation in traditional medicine as a healing agent due to its components alkaloids berberine and hydrastine.
To use it in undiluted form, apply goldenseal tincture on your herpes affected area 3 times a day. If using on sensitive areas like genital areas, dilute the goldenseal tincture by adding 3-5 drops of it into a cup of water. Even Hippocrates, the father of western medicine, used garlic to heal a variety of infections. After putting the mixture in your mouth, hold the garlic there as long as you can and then swallow it. As a preventive measure, you may have 1 tsp of raw garlic-honey mix everyday to keep outbreaks at bay. After having raw garlic, follow it with a cup of echinacea tea which is also our next herbal remedy for herpes.
Echinacea herb has been shown to be antibacterial, antiviral and antifungal along with having anti-inflammatory and immune strengthening properties. Warning: Avoid having echinacea if suffering from autoimmune diseases or if if you are pregnant. Prunella vulgaris herb also known as “self-heal” or “all heal” herb, is high on flavonoids due to which it gets its anti-inflammatory, anti-microbial, and anti-viral properties. Once the syrup reduces to half, put off the flame and strain the liquid with the help of cheesecloth, sieve or your coffee filter.
While you can always have 1 tsp of this syrup daily to boost your immune system, increase the amount to 2-3 tsp daily once you see the onset of herpes outbreak. There are at least two components in this sweet tasting herb licorice root that help in healing herpes.
These herbal remedies for herpes have used commonly available herbs so that you may be at ease while selecting and applying your choice of herbs to cure herpes. This isn't an herb, but I guarantee if you start taking the amino acid L-Lysine, your sores will vanish within a week.
Up until the late ‘90s, cytotoxins were as deadly to the healthy cells as they were to the cancer cells, but scientists have managed to find a way to tag them, so that they only entered the cells with specific molecules.
The modified stem cells were put inside a biodegradable gel capsule and put where the tumor was located before it was surgically removed.
Enter your email address to subscribe to this blog and receive notifications of new posts by email. A method for killing malignant brain tumor cells in vivo entails providing replication competent herpes simplex virus vectors to tumor cells. BACKGROUND OF THE INVENTIONThe present invention relates to the use of an altered herpes simplex virus that is capable of killing tumor cells.
If you have sores around your mouth and lips, it can be herpes caused by Herpes type 1 (HSV-1). If you apply tea tree oil at the first sign of an outbreak, you may prevent the outbreak altogether. Now many clinical researches too have proved that aloe gel can heal the herpes lesions effectively and fast. Modern researches have shown berberine to have antibiotic, immune strengthening and anticancer properties.
Garlic, the humble herb readily found in any kitchen, is one of the nature’s best anti-microbial that kills a host of bacteria, viruses, and parasites, not to name herpes simplex 2 virus too.
Hence, this herb can help you in treating as well as preventing infections due to herpes viruses.
As ginger is an excellent anti-inflammatory, it will increase the effectiveness of this herbal tea. If you are taking immunosuppressive drug corticosteroids, then also this herb should be avoided. Among its various properties, tulsi can be mentioned here for herpes for its adaptogen, anti-microbial, antioxidant, immunomodulating, and analgesic properties.
Traditionally, prunella vulgaris has been used to treat sore throats and gum diseases in the form of a mouthwash. Also use the back of your spoon or spatula to squeeze the elderberries so their juice is fully extracted into the syrup. You may want to keep the elderberries syrup in your fridge where it will stay for about 2 months.
One is glycyrrhizin which is an anti-inflammatory compound and the other is flavonoids that combats the growth of the herpes simplex virus. A ray of light can kill cancer cells without leaving terrible scars - so why are so few offered it? A ray of light: Beams of light can now be used to diagnose AND treat cancer - so why aren't patients being told? A new study has proven that stem cells release cancer-killing toxins if genetically altered. Shah had proven that stem cells loaded with herpes could kill brain tumors and now the group of scientists managed to alter the cells so that they would kill brain cancer cells without any damage done to themselves. They work by entering the cancer cells’ walls and slowly stopping them from making proteins, which prevents them from reproducing, dividing and growing. A replication competent herpes simplex virus vector, with defective expression of the ?34.5 gene and the ribonucleotide reductase gene, specifically destroys tumor cells, is hypersensitive to anti-viral agents, and yet is not neurovirulent.
A method according to claim 1, wherein said virus is targeted to a tumor cell of non-nervous tissue origin. A method according to claim 1, wherein said virus is targeted to a specific tumor type with a tumor cell-specific promoter. A method according to claim 1, wherein said promoter is basic fibroblast growth factor promoter.
A method according to claim 1, wherein an essential viral gene product of said virus is under the control of a tumor cell-specific promoter rather than its own viral promoter. More specifically, the present invention relates to a mutated, replication-competent Herpes Simplex Virus-1 (HSV-1) which contains mutations in two genes, is hypersensitive to antiviral agents such as acyclovir, is not neurovirulent and does not replicate in non-dividing cells, yet can kill nervous system tumor cells.Malignant tumors of the nervous system usually are fatal, despite many recent advances in neurosurgical techniques, chemotherapy and radiotherapy. State agriculture officials are asking stables to practice biosecurity for at least two more weeks due to an outbreak of Equine Herpes Virus Type 1 (EVH-1).
NPR reserves the right to use the comments we receive, in whole or in part, and to use the commenter's name and location, in any medium. I've been moved to create awareness tools that help educate healthcare practitioners and the public about Morgellons Disease (see Awareness Poster) as well as share what I've learned from this experience.Many of the skin photos in the photo galleries are mine. Aloe vera is composed of various antioxidants and is particularly helpful in reducing inflammation.
Goldenseal is particularly beneficial in treating viral skin disease such as genital herpes and even shingles.
Researches indicate that allicin- ajoene is the plant chemical found in raw garlic which is effective in killing viruses. If taken after garlic and along with goldenseal, echinacea works faster to increase immunity, reduce inflammation and heal sores. It is one of the very effective herbs used by Chinese medicine and has come to be known as soothing remedy for herpes by modern herbalists too. While this herb has been used since ages for preventing and curing inflammation, cough, cold, and flu, modern herbalists have been using it also for curing herpes. You need to experiment with one or multiple herbs for herpes and find out the best natural remedies that work for your herpes! The results of this study have been published in the journal Stem Cells and it was funded by the National Institutes of Health and the James S. He and his fellow researchers have found that after they had genetically modified stem cells, they released a toxin that eliminated the cancer cells in the brains of mice after the brain tumor had been surgically removed.
We tested these stem cells in a clinically relevant mouse model of brain cancer, where you resect the tumors and then implant the stem cells encapsulated in a gel into the resection cavity. In particular, there is no standard therapeutic modality that has substantially changed the prognosis for patients diagnosed with malignant brain tumors. It has changed and become a lot more virulent, and showing a lot more neurological signs and a lot more contagious than the older stains that we saw." Pete Sartin has a new regimen for his horses. It is not that herpes type 1 cannot cause genital herpes but most commonly, it is caused by the type 2 virus. Many researches have shown that echinacea may alleviate pain and discomfort due to by virus.
Stress triggers outbreak of herpes and as an adaptogen, tulsi helps your body adapt to stress along with promoting energy and endurance.
This herbal remedy will get you rid of itching, pain and discomfort within minutes to hours depending upon how your body reacts to it.
Prunella vulgaris has tannins in it which helps in reducing the moisture thus making it difficult for virus to stay alive.
While the licorice root extract is easily available which can be applied to herpes lesions and sores, you can make your own licorice root infusion and tea to wash the affected area.
For example, high mortality rates persist in malignant medulloblastomas, malignant meningiomas and neurofibrosarcomas, as well as in malignant gliomas.Gliomas are the most common primary tumors arising in the human brain. For the last few weeks he's been watching his horses' behavior closely for any signs of paralysis and takes their temperatures twice a day. However, it is always advisable to use fresh aloe vera gel, from leaf itself if possible instead of the products that claim using aloe vera gel. Today, medical herbalists use Oregon grape root but its dried leaves too had been used traditionally to treat various ailments including herpes. A topical microbicide which contained phytochemicals obtained from Echinacea proved to be a good antiviral against strains of both HSV-1 and HSV-2 viruses. Due to its anti-microbial properties, holy basil herb is used topically as well as internally to cure bacterial, viral and fungal infections including genital herpes.
If you want to cure your herpes naturally, you may make Prunella vulgaris oil for topical application.
Modern researches have shown elderberry components smooth out the minute spikes that are on the outside of viruses.
The most malignant glioma, the glioblastoma, represents 29% of all primary brain tumors, some 5,000 new cases per year in the United States alone. Like most, I panicked bought a lot of unnecessary products—some toxic, then managed to pull myself back from the brink of actually hurting myself. When you apply lemon balm topically, you get instant relief from swelling, itching, and pain.
While you can have St John’s Wort tea internally to build your immune system, for immediate relief you can apply herbal oil made from this herb. This stops them from entering the body cells where they reproduce and grow just to burst the cell and replicate themselves. Glioblastomas are almost always fatal, with a median survival of less than a year and a 5-year survival of 5.5% or less.
What’s more important is that lemon balm herb combats the outbreak or reactivation of herpes viruses. Thus, if you want to prevent viruses from penetrating and infecting your healthy cells, you may like to make this elderberry syrup which also includes other herbs that fight off herpes. Always wary of doctors, I tried to stay out of that loop but finally caved when my beloved ones pressured me to seek help. Just understand that if you have sores in any part of your body, they may be Herpes caused either by HSV-1 or HSV-2. A dermatologist gave me Ivermectin which caused a Herxheimer (die-off) reaction and many of the black particles I'd been seeing with a scope surfaced. This gave me great relief from the discomfort of what felt like fiberglass embedded in my skin. The difference between herpes and genital warts is that while herpes is caused by HSV, genital warts are caused by HPV (Human Papilloma Virus). Also, herpes show in form of blisters and sores while genital warts look like skin tags or cauliflower tops. There were NO fibers visible in my skin in the previous months before taking the medication. They duck and dodge against their opponents a€“ cattle a€“ in massive arenas filled with cheering crowds.
After glioblastomas are treated with radiotherapy, recurrent disease usually occurs locally; systemic metastases are rare. Within two days of taking the Ivermectin, I watched in horror as fibers started to rise to the surface of my skin. Neurologic dysfunction and death in an individual with glioblastoma is due to the local growth of the tumor.In the past, viruses have been tested for their ability to treat various types of tumors in animals or humans. That plus coming off the tail-end of the pharmaceutical seemed to stop all the symptoms of itching, biting and crawling for approximately the next 4 months. The proposed therapeutic mechanisms of viral cancer therapy in the prior art includes: (i) producing new antigens on the tumor cell surface to induce immunologic rejection, a phenomenon called “xenogenization”, and (ii) direct cell killing by the virus, called oncolysis. The only thing that hadn't cleared up was the hyper-sensitivity to my skin which I thought I was stuck with for the rest of my life, an unfortunate consequence of having Morgellons. Had I not been actively using a microscope to look at my skin during this period, I might have presumed I was well.
The skin however, doesn't lie and told a whole other story for under it's surface, fibers and particles were clearly still present—multitudes of them. Early on, I'd heard about the testing that Pam Crane of Morgellons Focus On Health (site no longer active) was championing and felt that it was probably the way to go.
My only problem with moving forward was a lack of practitioner, as I felt having the best testing in the world wouldn't mean anything unless someone was up to the task of decoding it's contents into a meaningful protocol, sort of like buying a Cadillac that arrives without the key. I wanted to have a conversation with whatever doctor I was going to see and to hear them speak about their philosophy of healthcare. We don't want to raise their body temperature until this fear goes away." Anna King: "Are these guys kind of lonely in here? Just try to get a physician on the phone before actually making an appointment and you will find it is an impossible task (especially the high-end specialists). They like to get out and play a bit and they really like to work cattle, but right now we're not doing any of that." Before he starts back on his chores, Sartin flips on the radio for the horses.
She talked about The Great Plains Labs and why the testing used in Functional Medicine is so important for our community.
She sounded like she knew how to do her job well and was more than capable, indeed she had impressed me as understanding how to approach the disease in a holistic manner that aligned with my own healthcare philosophies. I believe my first words to her were, "Where do I sign the dotted line?" (While focusonhealth has been taken down, here is another radio show with Nancy speaking with Pam. Treatments for tumors in both animals and in humans have been based on wild-type virus, passage attenuated virus, or infected cell preparations. Let me remind you that I felt fine when I first contacted Nancy, but the emerging fibers showed me I was not.
Nancy forewarned me that treatment would kick up all the initial symptoms I had when first ill with Morgellons, such as itching and stinging and that I might experience more fatigue with die-off, possibly even get lesions although I hadn't had many lesions previously. Within a week and a half of beginning this new protocol all the discomforts she mentioned came back.
Black particles began coming out from the skin, sand-like crystals were suddenly pushing from the scalp, I was itchy and uncomfortable but this time around it was at a much lower volume then at onset. I have experienced some lesions, in the form of 'papercuts' and blood spots appearing on the skin. They were tiny and healed quickly. It took close to 3 months for the symptoms of this detox to disappear, lessening with each week. We added in another few products to my protocol fine-tuning the process. Once all my symptoms had abated I was ready to move on to the next step and do a colon cleanse.
I used supplements for one month that gently cleansed the colon (Colon Cleanse by DaVinci Labs??).
After that, I began with chelation therapy (Chelex by Xymogen) to remove heavy metals. Nancy has taken me through this journey with constant support through email. The fact that she is able to tell me what I can expect and interpret what I am going through is invaluable.
Having gone through the early treatment phases and emerging symptom-free, makes me feel that I did indeed work through something—hopefully clearing biofilm. So where am I right now and what are the changes that happened over these few months of treatment?My skin cleared of many of black particles, although I still have them (my great lament was that I didn't have a camera back when I first got sick, because my skin was so saturated it was alarming!).
At least nine viruses have been shown to be capable of inducing some degree of tumor regression in a variety of tumors in mice, rats, rabbits, and guinea pigs.
A major drawback seen in these early animal studies, however, was systemic infection by the virus.To avoid systemic infection, the genetic engineering of viruses for use as antineoplastic agents has focused on generating altered viruses that are not capable of replication in non-dividing cells. The replication-defective herpes simplex virus vector system consists of an amplicon plasmid which, in herpes simplex virus infected cells, is replicated and packaged into viral particles. Defective herpes simplex virus vectors require helper virus to generate a herpes simplex virus vector.The use of replication-defective retroviruses for treating nervous system tumors requires producer cells and has been shown to be limited because each replication-defective retrovirus particle can enter only a single cell and cannot productively infect others thereafter. Because these replication-defective retroviruses cannot spread to other tumor cells, they would be unable to completely penetrate a deep, multilayered tumor in vivo. Retroviral vector-containing cells have been implanted into brain tumors growing in human patients. Actually, I feel spry and 10 years younger than my 55 years of age (at the time this was written), an unexpected and very welcome gift.
Still, they are here and I am monitoring them carefully. I am convinced of the value of the Great Plains and Metametrix testing. These retroviral vectors carried the HSV-1 thymidine kinase (HS-tk) gene into the surrounding brain tumor cells, which conferred sensitivity of the tumor cells to the anti-herpes drug ganciclovir.


It just makes sense that one would want to know the particular weaknesses of their body before embarking on a journey of healing. Of eight patients with recurrent glioblastoma multiforme or metastatic tumors treated by stereotactic implantation of murine fibroblast cells producing retroviral vectors, five patients demonstrated some evidence of anti-tumor efficacy but none were cured. Getting the labwork done and seeing Nancy are the two smartest things I did throughout my ordeal. Morgellons for me is like that impossible teacher I had in my youth. Some of the limitations of current retroviral based therapy as described by Oldfield are (1) the low titer of virus produced, (2) virus spread limited to the region surrounding the producer cell implant, (3) possible immune response to the producer cell line, (4) possible insertional mutagenesis and transformation of retroviral infected cells, (5) single treatment regimen of pro-drug, ganciclovir, because the “suicide” product kills retrovirally infected cells and producer cells and (6) the bystander effect limited to cells in direct contact with retrovirally transformed cells. But it has also left me savoring hard-won battles and set me straight on how to live more in balance. Therefore, I try not to rail against the unfairness of the experience, but to look at how much I learned and the ways in which it has stretched me as a human being. Yes it has been a miserable teacher, but by it's hand I have been become an attentive student.I wish you all full recovery.
A replication-competent virus has the advantage of being able to enter one tumor cell, make multiple copies, lyse the cell and spread to additional tumor cells. AylaWhile I am not comfortable sharing my protocol as it is specific to my lab results, I will outline what I feel are the bare-bones minimum I'd look to take if I were building a protocol. A thymidine kinase-deficient (TK?) mutant, dlsptk, was able to destroy human malignant glioma cells in an animal brain tumor model.
This would be a good probiotic (I use 3 different kinds throughout the day as was indicated by the results of my testing). Unfortunately, the dlsptk mutants were only moderately attenuated for neurovirulence and produce encephalitis at the doses required to kill the tumor cells adequately. FOS, according to Nancy can promote yeast growth so I'd stay away from a probiotic that includes it.
Residual neurovirulence, as evidenced by a 50% lethality of intracranially-administered, replication-deficient herpes simplex virus viral vectors at 106 plaque forming units (pfu) limits the use of such vectors for tumor therapy. Furthermore, known TK? HSV-1 mutants are insensitive to acyclovir and ganciclovir, the most commonly used and efficacious anti-herpetic agents.Therefore, it remains of utmost importance to develop a safe and effective viral vector for killing tumor cells. Even though various attempts have been made to engineer a viral vector able to kill human tumor cells in vivo, no viral vector has provided attenuated neurovirulence at the dose required to kill tumor cells while exhibiting hypersensitivity to antiviral agents and an inability to revert to wild-type virus. I'd include a biofilm buster which breaks down the cellular matrix in the gut that inhibits absorption of necessary nutrients. I used Interfase Plus and Wobenzym at different times (there are different kinds of Wobenzyme. The tumor cells can be of a nervous-system type selected from the group consisting of astrocytoma, oligodendroglioma, meningioma, neurofibroma, glioblastoma, ependymoma, Schwannoma, neurofibrosarcoma, and medulloblastoma.
Other kinds of tumor cells which can be killed, pursuant to the present invention, include those selected from the group consisting of melanoma cells, pancreatic cancer cells, prostate carcinoma cells, breast cancer cells, lung cancer cells, colon cancer cells, lymphoma cells, hepatoma cells and mesothelioma and epidermoid carcinoma cells.In accordance with still another aspect of the present invention, a method is provided for killing tumor cells in a subject, comprising the steps of administering to the subject a herpes simplex virus vector, wherein the vector comprises a tumor cell-specific promoter wherein the promoter controls expression of at least one viral protein necessary for viral replication and wherein the promoter is induced selectively or at a higher level in tumor cells than in normal cells.
I've used Carlson's liquid D3 which is lipid based, but recently have switched because of assimilation issues with fats to a dry encapsulated D3 (Prothera brand 5,000 I.Us a day). Certain of the B's can become depleted when the body is fighting infection, so a good B-complex seems necessary. Also, Mr. 2 shows the sequence arrangement of a mutant herpes simplex virus, G207-1, compared to its parental wild-type background (strain F). Common Sense and now Clifford Carnicom have written about NAC and it's benefits along with vitamin C in helping quell the symptoms of our condition.
I'd look into this as a close friend from the community began taking both high amounts of Vitamin C and NAC and her symptoms disappeared.
She was taking 3,000-5,000 mg of C spread throughout the day and 600 mg 2x's a day of NAC both in the morning and at night.
4 is a graph illustrating the ganciclovir (GCV) sensitivity of R3616, G207-1, and G207-2 which reveals that G207-1 and G207-2 are ten-times more sensitive to ganciclovir than R3616.
She eventually reduced it to 2,000 mg of vitamin C and 600 mg of NAC because the NAC was giving her headaches.
The lesser dosage has been fine for keeping her symptoms of crawling, biting and stinging away. 5 is a graph illustrating the ability of G207-1 and G207-2 to inhibit the growth of human brain tumor cells (U-87MG) in the subcutaneous human brain tumor model in athymic mice.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTSThe present invention exploits the ability of mutant, replication-competent HSV-1 to enter a tumor cell in situ, make multiple copies, lyse the tumor cell and spread to additional tumor cells with relatively minor effects on the surrounding normal cells. The mutant herpes simplex virus of the present invention has each of the following characteristics: (1) efficacy in killing human brain tumor cells, (2) marked attenuation of generalized neurovirulence to protect the normal brain, (3) multiple deletions so that a single mutation cannot cause reversion to the wild-type viral phenotype, and (4) hypersensitivity to ganciclovir so that undesired spread of the virus can be prevented.
When she went off the protocol symptoms came back and when she again resumed, they went away. I might mention, that my protocol has always called for 1000 mg of vitamin C, three times a day.
Alterations in this regard include any that disrupt the expression of the product of both of the ?34.5 gene and the ribonucleotide reductase gene.
While I was not on NAC as a separate supplement, it is in another product I take daily ?(Liver Protect by Xymogen). The presence of such multiple mutations further reduces the possibility of reversion to wild-type pathogenicity.
The present invention provides methods for sequentially constructing and testing viruses for the ability to effectively kill brain tumor cells without harming surrounding normal brain.
Nancy has stated that the clients she sees that do this first make themselves much worse and take longer to get better. If I am going to do this right, I want to know what my baseline toxic load is so I can measure my progress over time.
The hrR3 mutant contains an Escherichia coli lacZ gene insertion in the ICP6 gene, which encodes the large subunit of ribonucleotide reductase.
The results arrived right after I finished my third week of chelation. My total toxic representation was OFF the chart with high levels of antimony, silver, cadmium, tin and magnesium.
Other ribonucleotide reductase herpes simplex virus mutants are suitable for constructing the mutant viral vector of the invention.
I have a host of other metals as well, they just were at lower levels than those mentioned above. HSV-1 encodes its own RR (UL39 and UL40 genes), which is composed of two non-identical subunits. Once Nancy reviewed my test results and knew how I was responding to the treatment she modified my protocol to include a liver support, and the addition of 3-4 new products that would also help balance me out. I keep an ongoing visual journal using photos which I share with her so she can see any new developments. During chelation I was more tired, needing to take naps and sleep longer on most days. I also had some tiny red spots appear on the skin without breaking through.Red spots appear on the skin.
The large subunit (140 k molecular weight), designated ICP6, is tightly associated with the small subunit (38 k molecular weight). Based on what was going, on Nancy pulled back on the chelation so that the process is even slower and gentler until my body can adapt. Herpes simplex virus RR is required for efficient viral growth in non-dividing cells but not in many dividing cells. Using shea butter and coconut oil mixture as a moisturizer seems to draw fibers and black particles from my skin if left on for 3-4 hours. Most are not readily apparent unless you really look for them. Hair The Morgellons has moved into my hair. An acupuncturist, pointed out after looking at my hair photos, that there were multiple "hairs" emerging from one follicle. This is probably why my hair feels thicker even though it has been brittle, weak and falling out. Helicobacter pylori, the bacteria I had with high levels the first time I was tested, is completely gone. Because TK? HSV-1 mutants known in the art are resistant to these anti-viral agents, such mutants could be difficult to eliminate in the event of systemic infection or encephalitis.
There was a new bacteria that showed up on this round of testing that wasn't on the previous. Nancy feels that this is something that could have morphed, or, could have been present all along, but like peeling an onion back, was hidden under layers. Therefore, these mutants are attenuated for neurovirulence and less likely to propagate in the event of a fever in the infected host. She is adding seven new supplements to my protocol to address adrenal support as sleep has become fitful and I feel somehow too 'plugged in.' Fight or flight mode. I'll be taking a prebiotic to address the new bacteria as well as some new products from Xymogen to open detox pathways. I'm excited to be moving in a new direction. Every time we do a new test, I see the value of this approach. The decreased neurovirulence of R3616 is putatively associated with the cessation of neuronal protein synthesis, which is preempted in wild-type herpes simplex virus infection. Randomly throwing supplements at the body without being tested, with the hope that something might work, seems a dangerous practice. The ?34.5 gene product can be detected by Western blot or ELISA analysis of infected cell proteins with antibodies or lack of replication in confluent primary cells.
Not terrible full-blown itchy, just enough to let me know that I was reinfected or maybe this is an allergic flare-up. This episode might have lasted longer than necessary had I immediately washed all my clothes but I got lazy and didn't wash them for a few days. Was not constantly itchy, but found I'd get itchy at odd intervals, sometimes once a day, sometimes not for days at a time and sometimes for a mere few seconds or so. I am putting a garden in upstate and often have my hands in the soil (without gloves) and have found that I am fine. The ?34.5 gene has been sequenced in four strains of HSV-1, namely F, 17, MGH-10 and CVG-2. The only itching I've got is from the occasional mosquito bite (and yes, these are real mosquitoes). Now, instead of direct application of GSE I brush my teeth twice, once with toothpaste and once with a clean brush that has two drops of GSE on it. Such alterations render the product of both genes non-functional or reduce their expression such that the mutant herpes simplex virus vector has the properties of the instant invention. The mutated herpes simplex virus vector of the instant invention is a replication competent herpes simplex virus whose genome is altered in the ?34.5 gene and the ribonucleotide reductase gene. Alterations in the ?34.5 gene and the ribonucleotide reductase gene include modifications in either the structural or regulatory sequences of these genes. Genetic alterations can be determined by standard methods such as Southern blot hybridization of restriction endonuclease digested viral DNA, sequencing of mutated regions of viral DNA, presence of reporter gene (for insertions), new restriction endonuclease site, enzymatic assay for ribonucleotide reductase activity [Huszar and Bacchetti, J. Note that I have been letting my supplements run out in the last few months because I needed a break from taking approx.
The high rate of recombination in herpes simplex virus and the fact that transfected viral DNA is infectious renders genetic manipulation very straightforward. These genetically-altered, replication-competent viruses can be used in the safety and efficacy assays described below.HSV-1 contains a double-stranded, linear DNA genome, 153 kilobases in length, which has been completely sequenced by McGeoch.
I wonder if the return to itchiness had something to do with my letting down my guard and not watching my pH.
After the first day of swimming, I noticed small circular red blotches appear under the skin of my calves and thought, uh-oh, here comes trouble. I FINALLY have a microscope again (it was replaced after more than 7 months at the manufacturer!).
Late in infection, concatemeric viral DNA is cleaved into genome length molecules which are packaged into virions. In the CNS, herpes simplex virus spreads transneuronally followed by intraaxonal transport to the nucleus, either retrograde or anterograde, where replication occurs.DNA constructs employing HSV-2 based on those illustrated herein using the HSV-1 genome are encompassed by the present invention. I had a few breakthrough 'papercut lesions' on the hands which I don't know what to make of. I stopped the Ormus because I wanted to document F-6, which has gotten quite a buzz in the Morgellons community. I still have 1.5 bottles left and intend to take it in a few months feeling I didn't give it a proper shot.
For now, I want to concentrate on doing one therapy at a time, so I can really be sure what each does.
October 17, 2011: Began using F-6 and have started a new link documenting that experience including photos showing what is emerging from the skin.
I'm going to take it slow with this product as I don't want to have any major herxheimer episodes. Caroline Carter, A healthcare worker that has Morgellons and works with ozone therapy (and who made her Morgellons worse with ozone saunas) said she cannot even be around it anymore. I have since talked with another person that had strong outbreaks from doing an ozone sauna. The “at” sequence also contains sequences for cleavage of unit length DNA from HSV-1 concatamers, packaging of HSV-1 DNA into capsids and inversion of L and S components.
Ozone therapy is typically great for cancer, AIDS and herpes, but for this disease might make us worse.
There seems to be improvement to the hair which feels stronger and seems to be falling out less.
The promoter region of ICP6 has been mapped to the 5? upstream sequences of the ICP6 structural gene. A lot of "debris" is coming up through the skin, fibers, black particles, granules and many more fibers then ever seen before. The transcription start site for the small subunit of RR, namely UL40, falls within the coding region of ICP6. A week and a half later I ate some quinoa with my lunch and two hours later had 3 oatmeal cookies (oatmeal, walnuts, carob chips, baking soda, vanilla).
This forage into complex carbs was unusual for me.While eating the cookies I started to itch. By the third cookie, I was having full-blown symptoms of itching, stinging and biting sensations (no crawling, thank God!). The fact that it came on with such force and was related to food, makes me think that Morgellons counts among it's causes a hyper-allergic reaction to food. In this case, I don't really know what specific ingredient triggered the reaction.By eliminating carbs again, symptoms quieted down. Because herpes simplex virus genes are regulated differently when present in the cellular genome, the effects of each alteration in the ?34.5 or ribonucleotide reductase regulatory component would be assessed in various mammalian target cells. I am noticing flare-ups of the arthritis in my foot which had so improved at the beginning of my treatment (but I kill my feet dancing one night a week, so that could play into this). The ones I can visually see without a scope are bigger then I've had before but there is no increase in the amount I generally see with my scope in the skin. Will keep you posted of new developments.I've noticed that when it's very hot (93-97 degrees) and my skin is exposed to the air, I have more sensations on my arms.
It's not unusual to feel a single pin prick sensation on my left upper arm, often I'll feel the same type of prick mirrored in the same spot on right arm sometime later, as if they were somehow linked.The other day on my bike I felt as if I hot poker were being pressed momentarily to my arm. When I looked down, there were three distinct tract marks in that spot, about a half, to an inch long that hadn't been there before. Since having Morgellons, it feels like the skin is no longer a reliable friend but is capable of erratic changes suddenly.
They took about 5 days to disappear.?September 27, 2012 • Live blood microscopyJust posted a new link with photos of my recent session with Rick Panson, a microbiologist and healthy body coach who did a live blood and dried blood microscopy and gave me additional guidance for my health.
It's a bird's eye view into the system, giving a fuller, deeper understanding of what's going on  then just looking at lab results—although lab tests have their place! In the next two weeks I get more testing done so will post those results when in. There were many problems with the blood.
We are in a seasonal shift and my fingers have had a few tiny papercut lesions since the change began.Started the protocol Oct. Beginning with one pill a day of capyrilic acid with one pill olive leaf extract and 1 tsp. The L-glutamine is supposed to strengthen the gut lining and prevent leaky gut syndrome caused by the yeast die-off.
Clinical studies indicate that even wild-type HSV-1 viruses generally do not spread far from the site of initial infection or cause serious systemic disease in immunocompetent individuals. Will stay on this for a few days and slowly double the dosage, then determine where to go from there. 16, 2012 Went back to Rick Panson for another live blood analysis and posted the photos under my last blood microscopy.
This is encouraging for all of you who wonder how long it will take to knock back candida and even if our protocols are working. Any mutant replication-competent viral vector that is more sensitive to the anti-viral agent than its wild-type parent is deemed hypersensitive to the anti-viral agent, potentially providing a means to abort an undesired spread of the mutant virus.In constructing herpes simplex virus mutants for use in vivo, the mutants are tested for their sensitivity to current anti-herpetic drug therapies in order to control unforeseen virulent infections.
Rick recommended Candex and Nancy told me she loved the product except now they are adding corn to Candex (and it's unknown if it's GMO corn), so she is no longer recommending it. A number of drugs currently are available to treat herpes infections in humans, the most effective being nucleoside analogs which block herpes simplex virus DNA replication. Three herpes simplex virus genes are known to be involved in sensitivity to nucleoside analogs: herpes simplex virus DNA polymerase (UL30, pol), herpes simplex virus thymidine kinase (UL23,tk), and CMV UL97 which shares homology with, protein kinases and bacterial phosphotransferases.
Adrenal system had a number of markers indicating a lot of stress to the system, very low levels of vitamins C and B's, four different pathogenic bacteria showed up—Helicobacter Pylori was high, Clostridia, E. Kreb cycle needs support. Heavy metals hair test—all toxic metals were down except arsenic which went up with cadmium. Lead which was extremely high on my first test showed a considerable decrease as well as silver but lots more chelation ahead. Unfortunately, intracranial injections of AraAr9 led to premature death and had no effect on subcutaneous tumor growth.
It's been 2 years since I started using supplements to pull the metals out and I think it will take a couple more years of work.
Another mutant herpes simplex virus, the dlsptk virus, is no longer drug sensitive, at least to nucleoside analog drugs, and therefore potentially uncontrollable in vivo.Attenuation for NeurovirulenceAttenuated or decreased generalized neurovirulence means that life-threatening encephalitis does not ensue after infection with the double mutant herpes simplex virus vector of the instant invention. The process is slow but steady which is a good thing because I think if the body released too much metal into the system at once there would be negative impact on the health.


Because herpes simplex virus-induced encephalitis in humans is very difficult to treat and can be fatal, even with adequate pharmacologic measures, decreased generalized neurovirulence is an important feature of the instant invention. The mutant virus of the present invention is capable of replicating in neoplastic cells but spares surrounding non-neoplastic tissue.Different herpes simplex virus strains vary in neurovirulence and more attenuated strains may be employed in the construction of the double mutant to further decrease neurovirulence. Nancy pointed out that lead and cadmium are antagonistic to calcium absorption and bone health, so seeing this rise in cadmium and knowing I still have too high levels of lead makes me think that I might have to give up the painting at some point or switch mediums rather than compromise bone health. The heavy metals test also measures "essential and other elements," (minerals) It seems as a consequence of pulling heavy metals out, the minerals have been compromised. Nancy suggested this organism is a pleomorph moving from bacteria to fungus according to favorable conditions in the body. This is the third time it's changed. Feb 3, 2013 Just an acknowledgment that I just passed my 3rd anniversary since getting sick with Morgellons.
Herpes simplex virus encephalitis is usually localized to the temporal lobe and the limbic system and histological examination of autopsy cases demonstrates viral antigen at these sites. A number of drugs are available to control infection, including acyclovir 9-92-hydroxyethoxy-methyl)guanine, Zovirax®, adenine arabindside (Vidarabine®), foscarnet (phosphonoformic acid, PFA) and ganciclovir 9(1,3-dehydroxy-2-propoxy)methylguanine, DHPG, 2?NDG, Cytovene®. The only time I really have symptoms is when I really mess up on diet (itchiness will start) or when ?very overheated and sweating from physical exertion like dancing, where it sometimes feels like needle pinpricks are sticking my skin from the INSIDE out, accompanied by itchiness. Thieves is a great oil for preventing flu or when one is coming down with illness, but I am unclear if it can damage beneficial bacterias. Stopped taking thieves after a few weeks but do still take lemon and added in lemongrass and grapefruit to my daily capsule. The term “tumor cell-specific promoter” indicates a promoter that is induced selectively or at a higher level in the target tumor cell than in a normal cell.
Itchiness did not return. September 22, 2013 Decided to go on Stephen Buhners herbal protocol for Lyme disease (crafted from his excellent book, Healing Lyme). Tumor cell-specific promoters include promoters that are induced selectively or at a higher level in a particular cell type or a tumor cell.The vectors of the invention also can be designed to selectively replicate in and kill a tumor cell of non-nervous tissue origin.
Although I tested negative for Lyme and feel fine, I don't trust the traditional testing one gets with their regular MD, and even though I don't think I'm dealing with Lyme, I can't discount the fact that so many Morgellons sufferers also have Borrelia and other co-infections that might be related to our disease which seem to improve with Lyme treatment.
The herpes simplex virus vector of the invention is engineered to place at least one viral protein necessary for viral replication under the control of a cell specific or tumor cell-specific promoter. Also, his protocol addresses other issues I feel are pertinent to my health (even before Morgellons). Rendering the essential herpes simplex virus gene non-functional can be achieved by genetic inactivation or replacement of promoter with tumor cell-specific promoter.The instant invention encompasses a host-range conditional herpes simplex virus mutant where an essential viral gene product is under the control of a tumor cell-specific promoter rather than its own viral promoter. Certainly, I see over time that my immune system is weak, and my gut reaction now to coasting is that it could be an eventual time bomb. In permissive cells, containing the proper regulatory proteins for this specific promoter, the essential viral gene product is expressed and the virus is able to replicate and spread to adjacent cells until a non-permissive cell is infected. These studies are applicable to the replication-competent herpes simplex virus of this invention. I'm taking Cats Claw, Sarsaparilla root (Jamaican—which I grind in my coffee grinder), Eleuthero (Siberian Ginseng), Astragalus, Japanese Knotweed and Andrographis. To his formula I've added in Ashwagandha and Sweet Cinnamon (from Sri Lanka) for the brain, as well as nettles (excellent herb for trace minerals and general support of system) and Pau D'arco (fungal).
One can take advantage of this altered expression pattern to construct tumor cell-specific viruses. I take a quarter level teaspoon of each herb (which is roughly the equivalent of one 00 capsule) added to some water and drink it down. In addition, the instant invention employs the replication competent herpes simplex virus vector having decreased neurovirulence as a tumor cell modulator or inducer of an immune response against the tumor cells.
The mutant herpes simplex virus vector of the invention can be further altered to express cytokines in the tumor target cell in order to elicit an immune response against the tumor cells. Surprisingly, I did not feel exhausted, although the protocol was pretty intense and caused a lot of flushing through the digestive system. For example, a mutant herpes simplex virus vector can induce viral-mediated killing of tumor cells, which then is amplified by a cytokine-enhanced immune response, a cytokine having been expressed by the vector itself. The expression of cytokines, or other gene products, from the mutant herpes simplex virus vector would occur within hours of infection so that sufficient gene products would be synthesized prior to cell killing.
This is not a new symptom and usually happens in the first few weeks of starting any new protocol. As these breakouts have always been so minor and short-lived? (an occasional papercut lesion or what looks like a pin prick), I paid them little mind. In this case, the skin continued to break open for the duration of treatment and I had some long "scratch" marks come up as well. It seemed like the last papercut I got was deeper and for the first time I started to worry that perhaps I was giving myself a lesion problem that I might not be able to turn off. For tumors in the brain, MRI, CT, or other imaging guided stereotactic technique will be used to direct viral inoculation or virus will be inoculated at the time of craniotomy.The pharmaceutical compositions of the present invention would be advantageously administered in the form of injectable compositions.
A typical composition for such purpose would comprise a pharmaceutically acceptable vehicle.
This little spot has bothered me on occasion on and off throughout the years of my Morgellons experience, but during the Buhner protocol, it itched almost constantly. For instance, the composition could contain human serum albumin in a phosphate buffer containing NaCl.
Other pharmaceutically acceptable carriers include aqueous solutions, non-toxic excipients, including salts, preservatives, buffers and the like. See REMINGTON'S PHARMACEUTICAL SCIENCES (15th ed.) 1405-1412 & 1461-1487, Mack Publishing Co. On the plus side, I could tell there was a lot of detoxification going on and all of those bitter herbs were really helpful for quelling sugar cravings. I believe I will revisit this protocol modified in the future, but for now, am taking a rest from it. Note that when I ceased treatment, the papercut and pinprick lesions stopped, although I recently had a three-inch long "scratch" appear on my leg weeks after stopping his treatment some weeks back. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oil and injectable organic esters such as ethyloleate.
This was a surprise, since problems usually only show up on my hands and 3 inches is rather a statement!For the first time, the scratch resembled images I've seen for Bartonella. Aqueous carriers include water, aqueous solutions, saline solutions, parenteral vehicles such as sodium chloride, Ringer's dextrose, etc. The pH and exact concentration of the various components the pharmaceutical composition are adjusted according to routine skills in the art.
There is much about it that I think is helpful, and I encourage people to study these herbs and supplements carefully, and decide for themselves if they want to incorporate them. The active immunogenic ingredient is often mixed with an excipient which is pharmaceutically-acceptable and compatible with the active ingredient. This is a very difficult protocol and for any who have detoxification pathway issues, this could impact their health negatively.
Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof. The release of toxins in a system that cannot remove them properly can be deadly, so it is important to not rush carelessly forward with something this strong. In addition, if desired, the vector may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH-buffering agents, adjuvants or immunopotentiators which enhance the effectiveness of the vector vaccine.Additional formulations which are suitable for other modes of administration include oral formulations. I did feel like the protocol helped me on many levels, but I could not tolerate the heart issues with pressure on the chest, burning, and breathing difficulties.
Oral formulations include such typical excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. The creator of the protocol feels that Bartonella is at the core of the Morgellons infection, and Bartonella likes to set up shop in the endothelial tissue of the heart. The quantity to be administered, both according to number of treatments and amount, depends on the subject to be treated, capacity of the subject's immune system to synthesize antibodies, and degree of protection desired. Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner and are peculiar to each individual.
However, suitable dosage ranges are on the order of one to several hundred micrograms of active ingredient per individual. Thinking this was just a severe Herxheimer reaction, I went back on the protocol but was unable to advance more than one drop of each tincture before heart and breathing issues returned with a vengeance. This time, the heart issues did not abate immediately, and it's been some weeks now in which they often come back on a low level.
His original protocol (which he posted on Jenna's blog and which is now removed) did not use MSM, or Silver Sol or various other supplements.
It also did not include baking soda, which is not inherently a problem, but at 2 teaspoons a day, might become one.
R3616, which is derived from HSV-1 strain F, contains a 1-kilobase-pair deletion in both copies of the ?34.5 gene.
The viral DNA is suitable for transfection after precipitation with ethanol and resuspension in water. For the generation of recombinant viruses, the piece of DNA to be recombined into the viral genome is excised from a plasmid. The linear DNA is co-transfected with viral DNA into cells capable of supporting propagation of the recombinant progeny virus. Recombinant viruses are then plated on permissive cells under selectable or screenable conditions. For example, LacZ+ recombinant plaques are stained by adding X-gal and blue plaques (LacZ+) are selected. Both of these genes are non-essential for viral growth in culture and therefore null mutants are viable in culture. When wide spread cytopathic effects were observed, progeny are harvested and titers determined on Vero cells.On day 2 or 3 following infection, plaques were stained with X-gal solution.
Recombinant virus from blue plaques is purified and the DNA analyzed by restriction endonuclease mapping to confirm the DNA structure of the mutant viruses.
Blue plaques were purified three times by passage in Vero cells in a limiting dilution method before stocks were made.The plaque morphology of G207-1 and G207-2 was analyzed as well as the effect of various concentrations of IFCS-containing medium on plaque morphology. The diameters of plaques were measured using a micrometer under an inverted phase-contrast microscope at 40? magnification.
2's top line represent the inverted repeat sequence flanking the long (UL) and short (Us) components of herpes simplex virus genome, which is represented by thin lines. Mutant G207 contains the structural gene of lacZ inserted into the BamHI site in the ICP6 gene. Total viral DNAs (KOS, hrR3, Strain F, R3616, and G207) were digested with restriction endonuclease, separated by agarose gel electrophoresis in a Tris-borate-EDTA buffer and transferred by the method of Southern. Recombinant DNAs used as probes for hybridization were labeled by ECL labeling Kit (Amarsham) as suggested by the supplier. To confirm that the viral mutants contain the lacZ gene at the appropriate position, total DNA was digested with Xho I and subjected Southern blot hybridization in duplicate. Filters were hybridized with labeled pkpX2?, which contains wild-type sequences of ICP6 gene. HSV-1 strain F contains an approximately 6.0 kB Xho I fragment due to a polymorphism between herpes simplex virus wild-type strains.
Viral DNAs of strain F, R3616, G207 were digested with Bam HI and subjected to Southern blot hybridization. Strain F contains the wild-type Bam HI SP and S versions of these fragments, whereas R3616 and G207 contain the deleted versions of these fragments.
Herpes simplex virus ICP? can be constructed by deletions or insertions into the ICP4 coding region. The vector of the invention can also be made ICP4? by a deletion or insertion into the ICP4 coding region. Alternatively, the ICP4 regulatory region of the herpes simplex virus vector is replaced with the tumor cell-specific promoter so that ICP4 is only produced in cells capable of expressing the replaced promoter. To evaluate tumor inhibition in vivo, subcutaneous U-87MG xenografts in nude mice are treated separately with inoculations of each viral mutant or vehicle, and tumor growth rates were analyzed. To investigate the potential effects of the herpes simplex virus mutant treatment on survival, nude mice with intracranial U-87MG xenografts are treated with virus or vehicle inoculations, and overall survival is compared.To evaluate the degree of tumor eradication, as well as the potentially retained neurovirulence of the viruses when used at doses necessary to achieve prolonged survival, the brains of long-term survivors with intracranial xenografts are sectioned, stained, and microscopically examined. For effective in vivo tumor inhibition and survival prolongation, careful choice of mutant employing the assays described herein is essential. The following methods provide clear guidance to those of skill in the art to screen for mutant viral vectors that are effective in vivo in inhibiting tumor growth and prolonging survival.
To establish the relative safety of these viruses as potential anti-glioma agents, their susceptibility to the common antiherpetic agent ganciclovir is investigated. Finally, to establish the safety of intracerebral inoculation of the mutant viral vector, animals receive an intracerebral inoculum of the mutant virus and are subsequently assessed for encephalitis.In vitro Cytopathic KillingThe ability of the herpes simplex virus vectors of the invention to kill tumor cells is first tested in cell culture. To maximize the titer of the viral mutant, the initial viral suspension was centrifuged at 34,500 g for 2 h at 4° C., and the pellet was subsequently suspended in media and again titered. All viral-infected cell cultures were compared with control cultures (DMEM+only, no virus). Cells that had become rounded, losing normal morphology, and those lifting from the plate were considered dead. Monolayers were considered completely destroyed when 99% or more of the cells exhibited such cytopathic effects.Either the mutant or its wild-type parent were applied to a human glioma line (U-87MG) and African green monkey kidney (Vero) cells at multiplicities of infection (MOIs) from 10?4 to 101 in DME+ (Dulbecco's modified Eagle's medium with 1-5% heat-inactivated fetal calf serum ((IFCS) and antibiotics). The malignant human glioma line U-87MG was obtained from American Tissue Cell Collection, Rockville, Md. Additionally, two primary human malignant gliomas were obtained as surgical tumor specimens. All cells were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and antibiotics (DMEM+).Subconfluent monolayers of U-87MG and Vero cells were infected with the mutant viral vectors of the invention at varying MOIs.
The mutant expressing cytopathic effects at 24 hours that is proportional to the MOI and expressing >99% cytopathic effect after 10 days in U-87MG is deemed to possess the ability to kill glioma cells in vitro.
The lowest inoculum of the mutant virus that can sustain a spreading infection to destroy the entire monolayer of U-87MG cells will provide one of the doses at which the mutant is evaluated in vivo.
The mutant viral vector also is tested against a different human glioma line (T98G) at various MOIs and assessed for its ability to produce monolayer destruction within 10 days.Short-term glioma cultures were established by explanting three malignant human gliomas (one anaplastic astrocytoma and two glioblastomas) obtained at surgery in DME+ and were studied at the second passage. The herpes simplex virus mutant and dose that is cytopathic in all three primary malignant gliomas is deemed to be able to kill a wide variety of human brain tumor cells in vitro.In addition to glioma cultures, the viral mutants are tested for their ability to kill 3 human malignant meningiomas, 1 atypical meningioma, 5 neurofibrosarcomas, and 2 medulloblastomas in cell culture, and in the in vivo models.
Significant tumor inhibition by the mutant virus reveals a wide range of nervous system tumors for which the viral mutant is efficacious in killing human brain tumor cells.FIG. The number of surviving cells relative to the number of cells in mock-infected control cultures (100%) was assessed. The cytopathic efficacy of these mutants can also be tested on the human glioma cells lines T98G, U-138MG and A172.The herpes simplex virus vector of the instant invention can be used to mediate the destruction of other human tumors.
Examples of other human tumors that may be amenable to this invention include melanoma, carcinoma of the prostate, breast cancer, pancreatic cancer, lung cancer, colon, cancer, lymphoma, hepatoma, and mesothelioma.
For example, the CMV UL97 gene (ganS; pGMT7-UL97) is inserted into TK? HSV-1 mutants and tested for its ability to complement the inability of TK? HSV-1 to replicate in serum-starved cells and confer ganciclovir sensitivity on this recombinant. After the viral vector containing the suicide gene is tested for ganciclovir sensitivity, a comparison of the ED50 (in vitro) and Mean Survival Time of the suicide containing and suicide absent viral vectors (eg. 4, which reveals that G207-1 and G207-2 are ten times more sensitive to ganciclovir than R3616. The dotted line indicates the ED50.Temperature Sensitivity of Mutant Viral VectorTo provide an additional safety feature that further compromises viral replication in the presence of encephalitis and fever, the sensitivity of the mutant viral vectors to temperatures greater than the basal temperature of the host are ascertained. Table 2 demonstrates the decreased plaquing efficiency of G207-1 and G207-2 at elevated temperatures. The hrR3 mutant showed temperature sensitivity compared to the parental strain KOS as previously reported. The HSV-1 wild-type strain F, which is the parental strain of R3616, G207-1, and G207-2, is also temperature sensitive. The R3616, G207-1, and G207-2 mutants remain as temperature sensitive as their parental strains.TABLE 2Plaquing Efficiencies of KOS, hrR3, R3616, G207-1, and G207-2on Vero Cells at 37° C.
To test the effect of the herpes simplex virus mutants on human glioma in vivo, 1 mm3 minced glioma pieces (obtained from nude mice previously injected subcutaneously with cultured U-87MG cells) are implanted subcutaneously into nude mice.
Animals dying within 48 hours of any procedure are considered perioperative deaths and are excluded from analysis. Deaths in the subcutaneous tumor experiments are excluded from analysis (no significant difference in deaths occurred between virus-treated groups and their corresponding controls).Between weeks 4 and 5, animals growing tumors (?8 mm in diameter) are divided into two groups of 7 to 10 animals per group.
Controls received intraneoplastic injections of 50 or 60 ?l of DMEM+; treated animals received similar intraneoplastic injections of virus suspended in DMEM+. Similar experiments are conducted for each of the virus mutants at various doses.Tumors were measured weekly or twice weekly with Vernier calipers.
For pathological studies, tumor-bearing mice (>10 mm in diameter) were treated with 1?107 pfu of G207 and sacrificed on day 8, 15 postinjection.
Tumors were removed, placed in fixative for 1 hr and submerged in cold phosphate buffered saline. Tumors were measured twice weekly with calipers and the growth ratio calculated by dividing the tumor volume by the tumor volume on the day of initial inoculation. The mean tumor growth rate was significantly inhibited in tumors treated with G207 compared to control tumors treated with medium alone.Subrenal Capsule ModelThe effects of G207 on U-87MG cells grown in the subrenal capsule of the nude mouse also would be tested because the subrenal capsule is a site used for monitoring growth of other nervous system tumors. Ten days later, the tumors are measured and inoculated with varying pfus of G207 in 1 ?l DME+ or 1 ?l DME+ alone. All mice were re-examined at 14 days and 26 days following inoculation to measure tumor size.
Inoculations would be in 2 ?l DME+ at the stereotactic co-ordinates initially used to inject the U-87MG cells.
A mutant viral vector of the instant invention is one that kills intracerebral brain tumors by keeping a significant number of the mice alive by seven weeks post-treatment. The absence of evidence of encephalitis would reveal that the viral vector possesses the characteristic of decreased or attenuated generalized neurovirulence.



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