September 2014 - An Enzyme In Various Fungi Helps Combat Gram-Negative Bacteria
A group of hybrid strains of fungi shows that it has the enzyme that is an efficient inhibitor of several types of gram-negative bacteria called: metallo - β -lactamases, including New Delhi metallo-B-lactamase-1 (NDM-1). Several patients at research hospitals with severe infectious were not able to fight against an infection with antibiotics. They had very little hope of survival. Various antibiotics could not stop the infection from a few negative gram bacteria. When the patients were fed 20 grams a day of a blend of fungi that showed a success from anecdotal tests with other patients, the patients were able to combat the infections and survive. This fungi blend is made from seven different hybrid strains from four different species that came from university research labs. . The assumption is that this enzyme, is in this fungi product that inhibits the resistance. We are now testing this to determine that this is the mechanism or is there another molecule that is causing the inhibiting of the gram-negative bacteria resistance to antibiotics.
The research team from McMaster University seemed to have found the enzyme that comes from fungi to support antibiotics by blocking the chemistry in negative gram bacteria so that it is resistant to antibiotics.
Various fungi from soil proved to be an efficient inhibitor of several types of metallo - β -lactamases, including New Delhi metallor- β -lactamase-1 (NDM-1), according to Gerard Wright at McMaster University in Hamilton, Ontario, Canada, and his collaborators. Because those β -Lactamases render bacterial pathogens resistant to carbapenem antibiotics, he and his collaborators are continuing to study this natural product as a candidate to use with carbapenems to overcome that resistance and restore their clinical usefulness, he says.
“Metallo-B-lactamases destroy our best β -lactam antibiotics and help life threatening pathogens to spread,” says microbiologist Kim Lewis at Northeastern University in Boston, Massachusetts.
Beta-lactamases are enzymes produced by some bacteria that provide resistance to β-Lacta antibiotics like penicillins, cephamycins, and carbapenems (ertapenem), although carbapenems are relatively resistant to beta-lactamase. Beta-lactamase provides antibiotic resistance by breaking the antibiotics’ structure. These antibiotics all have a common element in their molecular structure: a four-atom ring known as a β-Lactam. Through hydrolysis, the lactamase enzyme breaks the β-Lactam ring open, deactivating the molecule's antibacterial properties.
An advantage of this enzyme is its low molecular weight, which enables it to cross the outer membrane of gram-negative pathogens.
Beta-lactam antibiotics are typically used to treat a broad spectrum of Gram-positive and Gram-negative bacteria.
Beta-lactamases produced by Gram-negative organisms are usually secreted, especially when antibiotics are present in the environment.
Aspergillomarasmine A is an polyamino acid naturally produced by the mold Aspergillus versicolor. The substance has been reported to inhibit two antibiotic resistance carbapenemase proteins in bacteria, New Delhi metallo-beta-lactamase 1 (NDM-1) and Verona integron-encoded metallo-beta-lactamase (VIM-2), and make those antibiotic-resistant bacteria susceptible to antibiotics. Aspergillomarasmine A is toxic to leaves of barley and other plants, being termed as "Toxin C" when produced by Pyrenophora teres.
Aspergillomarasmine A can be made from a culture of molds that produce it. The liquid culture is filtered. Then from the filtrate, a precipitate is formed using calcium chloride, tricalcium phosphate and acetone. From the precipitate, the substance is redissolved at pH 9 in water. Then chromatographic separation in Amberlite IRC 50 with ammonia in water, and finally crystallisation at pH 3.0. At pH 2.5 aspergillomarasmine B crystalises.
Gram negative bacteria have thin cell walls with an outer layer composed of proteins and lypopolysaccharide. This outer layer sometimes reacts with the immune system, causing inflammation and infection. In addition to preventing the bacteria from staining, the outer membrane of the cell also helps the bacteria resist an assortment of drugs, making treatment of infections with Gram-negative bacteria rather challenging.
Some examples of Gram-negative bacteria include Legionella, Salmonella, and E. Coli. Numerous other pathogens are also Gram-negative, including some forms of meningitis, a number of bacterial sources of gastrointestinal distress, and spirochetes. Gram-negative bacteria can be stubborn infectious agents, and many sources of lethal infection are Gram-negative, including the bacteria which contribute to secondary infections in hospitals and clinics.
1. Potera, Carol. “Natural Product from Soil Fungus Blocks Metallo-B-Lactamases”. Microbe (Microbe – Vol, 9, Number 10,2014): 398-399.
3. Neu HC (June 1969). "Effect of beta-lactamase location in Escherichia coli on penicillin synergy". Appl Microbiol 17 (6): 783–6. PMC 377810. PMID 4894721
4. King, Andrew M.; Sarah A. Reid-Yu; Wenliang Wang; Dustin T. King; Gianfranco De Pascale; Natalie C. Strynadka; Timothy R. Walsh; Brian K. Coombes; Gerard D. Wright (2014). "Aspergillomarasmine A overcomes metallo-β-lactamase antibiotic resistance". Nature 510 (7506): 503–506. doi:10.1038/nature13445. ISSN 0028-0836
5. Weiergang, I.; H.J. Lyngs Jørgensen, I.M. Møller, P. Friis, V. Smedegaard-Petersen (2002). "Optimization of in vitro growth conditions of Pyrenophora teres for production of the phytotoxin aspergillomarasmine A". Physiological and Molecular Plant Pathology 60 (3): 131–140. doi:10.1006/pmpp.2002.0383. ISSN 0885-5765
6. Wagman, G.H.; Cooper, R. (1988-12-01). Natural Products Isolation: Separation Methods for Antimicrobials, Antivirals and Enzyme Inhibitors. Elsevier. p. 499. ISBN 9780080858487. Retrieved 27 June 2014
Update on Research
We have completed in vitro and in vivo studies with various strains of medicinal mushrooms, deciphered the chemical analyses and took what we found to be the best cases. Ongoing research is being done with various rare strains of rare species, many of them not commercially available. This involved growing these mushrooms on various substrates to determine the best substrate for the best strain with the desired outcome that would benefit mammalian chemistry. We then conducted experiments to determine the best results by giving certain blends of these rare strains to several cancer patients with remarkable results. The protocol included having individuals refrain from consuming foods with tryalene and pryalene proteins and foods that promoted inflammation. The research papers analyzed showed that the higher consumption of some of these rare strains provided better results. Clinical tests are being planned with other researchers at University Medical Centers.
The rare strains used came from the following species of medicinal mushrooms. The blends and individual strains were developed to target certain diseases and cancers: Ganoderma Lucidum (Reishi), Lentinus edodes (Shiitake), Hericium erinaceus (Lion's Mane), Inonotus obliquus (Chaga), Antrodia camphorata (Antrodia), Agaricus muscarius and californicus (Agarius Blazie), Pleruotus ostreatus (Oyster), Flammulina velutipes and ononidis (Enoki), Cordycep sinensis, unilateralis and bassiana (Cordycep), Grifola frondusa (Maitake), Schizophyilum communa (Suehirotake) and Poria cocos (Hoelen). The whole mushroom was used in our studies, not extracts. We are researching the chemistry in these rare strains to determine the mechanisms that cause these positive outcomes with mammals.
Lyme Disease and Tick-borne Diseases
Other tick-borne pathogens to test for are: Babesia, Human Granulocytic Anaplasma (HGA), Human Monocy Ahrlichia (HME), Bartonelli and Rickettsia. These other tick-borne diseases are seen in approximately 20% of the patients with Lyme Disease. The typical first tests to order for the tick-borne diseases are IFA antibody tests, dependent on the region you live or have traveled in.
Abnormal Response to Wheat Protein May Provide Clue To Cause of Type 1 DiabetesRussell Phillips, PhD
Aug 31, 2009
In a Canadian study involving 42 patients with type 1 diabetes, nearly half of the subjects had an abnormal response to wheat proteins. Scientists at the Ottawa Hospital Research Institute and the University of Ottawa, who conducted the study, found that the patients' over-reaction to wheat is linked to genes that are associated with type 1 diabetes.
The findings have two implications. First, testing for sensitivity to wheat could be a way of establishing whether a person is predisposed to acquiring type 1 diabetes. Second, people at risk for type 1 might forestall its onset by eliminating wheat from their diet.
The presence of wheat generates a response by the body's immune system in the form of attacks by T cell defenders. The Canadians believe that this constant over-reaction puts a strain on the immune system, eventually unbalancing it to the point that it attacks other parts of the body, including the pancreas.
Given the small number of patients in the study, lead researcher Dr. Fraser Scott said that more research would be needed to confirm the link between sensitivity to wheat and the predisposition toward type 1. He noted, however, that previous research with lab animals has shown that a wheat-free diet reduced the risk of developing diabetes.
The study was published in the August 2009 issue of Diabetes.
NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
The National Institute of Neurological Disorders and Stroke (NINDS) supports basic and clinical research on brain and nervous system disorders. There is a growing awareness of the importance of diseases of the brain in our society. In part this arises because our population is aging, and diseases of the brain become more prevalent as one gets older. It is also due to the growing awareness of the importance of a healthy nervous system in early childhood and the brain's role in many problems that have not traditionally been considered as biologically based diseases, conditions such as autism or addiction or Tourette's syndrome. NINDS shares with a number of other Institutes and Centers at NIH responsibility for research on the brain, and cooperates with them in areas of mutual research interest.
NINDS has responsibility for more than 600 neurological disorders that affect every age of the life span, ranging from well-known disorders such as stroke, Alzheimer's disease, and epilepsy, which affect millions of Americans, to a host of less well-known disorders that may affect a only few hundred Americans, but are nevertheless devastating to those with the disease and to their families. Most NINDS-funded research is conducted by extramural scientists in public and private institutions, such as universities, medical schools, and hospitals. They compete for grants and contracts that account for more than 80 percent of the Institute's annual budget. NINDS intramural scientists, working in 22 Institute branches and laboratories, also conduct a wide array of neurological research, ranging from studies uncovering structure and function in single brain cells to tests of new diagnostic tools and treatments for those with neurological disease. By supporting and conducting neurological research, the NINDS seeks better understanding, diagnosis, treatment, and prevention of these disorders.
To achieve this goal, the Institute relies on both clinical and basic investigations. Clinical research applies directly to disease detection, prevention, and treatment, as in studies of brain imaging techniques and in trials to test new drugs or surgeries for stroke. Although scientists studying the brain have made astounding progress in recent years, a great deal is still not known about its complex functions. Basic research pursues an understanding of the structure and activities of the human nervous system. The answers gained through this research can create the foundation for diagnosing and treating brain disease in the future. Learning how the brain stores memory, for example, may help scientists determine what happens when memory fails and may even suggest possible ways to treat certain dementias.
NINDS sponsors a rich portfolio of research focusing on disease and disability associated with the aging brain, including Parkinson's disease and stroke, two major areas of need and opportunity.
Parkinson's Disease. Parkinson's disease (PD) usually strikes in late middle age and affects more than a half million Americans. It impairs control of movement, progressing from symptoms such as tremor and muscular rigidity to total disability and death. Parkinson's disease, like Alzheimer's disease, amyotrophic lateral sclerosis (ALS), and Huntington's disease, is a neurodegenerative disease, the causes of which remain largely unknown. Clinical trials are underway to evaluate several pharmaceutical and surgical interventions to treat PD. Promising studies of new drug therapies for PD are continuing. Scientists conducting basic research studies are investigating the genetic and cellular origins of the disease, and have discovered the gene for one form of PD. These genetic findings open up the possibility for new understanding of the disease and development of new therapies.
Stroke. For several years, NINDS has been reporting significant new findings in the prevention of stroke. In 1995 NINDS-supported research led to the identification of the first emergency treatment for stroke in which a clot blocks a major brain artery. Clinical trial results showed that the drug, tissue plasminogen activator (t-PA), increases chances for recovery by at least 30 percent if given within three hours. The trial findings will guide future attempts to develop additional treatments for stroke. Moreover, the trial demonstrated how community health care systems can organize to provide swift high quality care. To insure such prompt treatment, NINDS is working with patient and professional organizations to publicize the research results, helping public and health care professionals organize acute stroke treatment in a variety of settings.
NINDS also supports research on many neurological disorders that affect the entire lifespan. Examples include:
Epilepsy. Epilepsy refers to a group of disorders which have in common recurrent seizures that are usually unprovoked and unpredictable. Seizures are caused by abnormal activity in the brain and take many forms depending on what parts of the brain are involved. In about half the cases no cause can be found, but head injuries, brain tumors, lead poisoning, problems in brain development before birth, and certain genetic and infectious illnesses can all cause epilepsy. Medication controls seizures for the majority of patients, who are otherwise healthy and able to live full and productive lives. On the other hand, at least 200,000 Americans have seizures more than once a month. Their lives are devastated by frequent, uncontrollable seizures or associated disabilities. As part of an international coalition including the Human Genome Project and scientists from Finland, NINDS-supported investigations discovered a gene for one form of epilepsy. Understanding the processes involved in this form of epilepsy opens up an entirely new area of research that may provide insights about the cause of many forms of epilepsy. This discovery should lead to a screening test, and perhaps to a better understanding of what causes epilepsy and to new treatment approaches. In addition, NINDS is conducting and supporting many ongoing research efforts to identify and test new therapies.
Brain and Spinal Cord Injury. One reason trauma to the central nervous system has such severe consequences is that neurons in the brain and spinal cord fail to regenerate after damage. Now we know they make unsuccessful attempts to regenerate, and in some circumstances can be coaxed to regrow. NINDS is encouraging research in several areas with potential for success:
- High dose methylprednisolone, the first therapy to improve the outcome of spinal cord injury, is now regularly used in emergency rooms. The effects of longer methylprednisolone treatment and of a new class of drugs are now being studied.
- Efforts to understand and repair trauma of the brain and spinal cord are continuing, using grafts, nerve bridges, cell implants, cell survival factors, antibodies, and genetic engineering. The potential use of newly-discovered neural progenitor cells, nerve cells that may have the capacity to replace cells lost because of trauma, is also under investigation.
- Neuroprosthetic devices connect with the nervous system via electrodes to stimulate muscles or provide sensory input. For example, a neural prosthesis developed with NINDS support and recently recommended for approval by the FDA restores hand function to quadriplegics. Future research goals include a splint-free system to allow a paraplegic person to rise, stand and sit again without assistance, and technologies to control muscles using direct brain signals instead of a functional neuromuscular stimulation implant.
Disorders of the Developing Nervous System. More than a third of all genetic disorders affect the nervous system, and hundreds of these first show symptoms in children. In the past several years, research has rapidly progressed in identifying genes for a number of these disorders. Approximately 50 genes have been identified. Finding the defective gene that causes a disease is only a beginning towards developing a therapy, but it allows scientists to develop diagnostic tests, create animal models, learn how the gene and its protein function to promote health or disease, and pursue a reasoned strategy towards counteracting the defect. Another very exciting area of research addresses the development of the brain in early life.
The National Academy of Science and reports published by the US Department of Health showed that a variety of proteins in wheat and rye grains may be the cause of neurological and immune malfunctions and the US department of Mental Health in the 1960's and 1970's completed clinical studies. The results showed that about 50% of all neurological and immune malfunctions were eliminated when the patient was on a wheat and rye grain free diet which included refraining from foods with dirivatives of these grains. This should not be confused with celiac disease which is caused by a reaction to the gluten protein found in wheat, rye, barley and oats. These studies are being shared with researchers to hypothosize the best approaches for further study as to the etiology of disease.
University of Maryland School of Medicine Scientists Pinpoint Critical Molecule to Celiac, Possibly Other Autoimmune DisordersMonday, September 07, 2009
Director of Operations
• See local news coverage of Dr. Fasano's discovery.
• Read Dr. Fasano's PNAS Paper
• Supporting Information for the PNAS Paper
Alessio Fasano, M.D.
Findings Reveal Further Detail About Protein Linked to Inflammatory Disorders
It was nine years ago that University of Maryland School of Medicine researchers discovered that a mysterious human protein called zonulin played a critical role in celiac disease and other autoimmune disorders, such as multiple sclerosis and diabetes. Now, scientists have solved the mystery of zonulin’s identity, putting a face to the name, in a sense. Scientists led by Alessio Fasano, M.D., have identified zonulin as a molecule in the human body called haptoglobin 2 precursor.
Pinpointing the precise molecule that makes up the mysterious protein will enable a more detailed and thorough study of zonulin and its relationship to a series of inflammatory disorders. The discovery was reported in a new study by Dr. Fasano, published September 8 in the online version of the Proceedings of the National Academy of Sciences. Dr. Fasano is a professor of pediatrics, medicine and physiology and director of the Mucosal Biology Research Center and the Center for Celiac Research at the University of Maryland School of Medicine.
Haptoglobin is a molecule that has been known to scientists for many years. It was
identified as a marker of inflammation in the body. Haptoglobin 1 is the original form of the haptoglobin molecule, and scientists believe it evolved 800 million years ago. Haptoglobin 2 is a permutation found only in humans. It’s believed the mutation occurred in India about 2 million years ago, spreading gradually among increasing numbers of people throughout the world.
Dr. Fasano’s study revealed that zonulin is the precursor molecule for haptoglobin 2 — that is, it is an immature molecule that matures into haptoglobin 2. It was previously believed that such precursor molecules served no purpose in the body other than to mature into the molecules they were destined to become. But Dr. Fasano’s study identifies precursor haptoglobin 2 as the first precursor molecule that serves another function entirely — opening a gateway in the gut, or intestines, to let gluten in. People with celiac disease suffer from a sensitivity to gluten.
“While apes, monkeys and chimpanzees do not have haptoglobin 2, 80 percent of human beings have it,” says Dr. Fasano. “Apes, monkeys and chimpanzees rarely develop autoimmune disorders. Human beings suffer from more than 70 different kinds of such conditions. We believe the presence of this pre-haptoglobin 2 is responsible for this difference between species.”
“This molecule could be a critical missing piece of the puzzle to lead to a treatment for celiac disease, other autoimmune disorders and allergies and even cancer, all of which are related to an exaggerated production of zonulin/pre-haptoglobin 2 and to the loss of the protective barrier of cells lining the gut and other areas of the body, like the blood brain barrier,” says Dr. Fasano.
“The only current treatment for celiac disease is cutting gluten from the diet, but we have confidence Dr. Fasano’s work will someday bring further relief to these patients. Zonulin, with its functions in health and disease as outlined in Dr. Fasano’s paper, could be the molecule of the century,” says E. Albert Reece, M.D., Ph.D., M.B.A., dean of the School of Medicine, vice president for medical affairs of the University of Maryland and John Z. and Akiko K. Bowers Distinguished Professor. Dr. Fasano, as a physician scientist, fulfills two of the core missions of the University of Maryland School of Medicine: making basic science discoveries that can impact human health, and finding ways to translate those discoveries into treatments and diagnostic tools.”
People who suffer from celiac disease have a sensitivity to gluten, a protein found in wheat, and suffer gastrointestinal distress and other serious symptoms when they eat it. In celiac patients, gluten generates an exaggerated release of zonulin that makes the gut more permeable to large molecules, including gluten. The permeable gut allows these molecules, such as gluten, access to the rest of the body. This triggers an autoimmune response in which a celiac patient’s immune system identifies gluten as an intruder and responds with an attack targeting the intestine instead of the intruder. An inappropriately high level of production of zonulin also seems responsible for the passage through the intestine of intruders other than zonulin, including those related to conditions such as diabetes, multiple sclerosis and even allergies. Recently, other groups have reported elevated production of zonulin affecting the permeability of the blood brain barrier of patients suffering from brain cancer.
“We hope pre-haptoglobin 2 will be a door to a better understanding of not just celiac disease, but of several other devastating conditions that continue to affect the quality of life of millions of individuals,” says Dr. Fasano. “This is quite a remarkable molecule that was just flying under the radar. We would have never have thought it would be the key. Now that we have identified this molecule, we are able to replicate it in the lab to use for research purposes. We hope to learn much more about it and its potential for treating and diagnosing celiac disease and other autoimmune conditions. This molecule has opened innumerable doors for our research.”
For video or audio of an interview with Dr. Fasano discussing his study, please see these links:
To view the interview using Real Player:
http://media.umaryland.edu:8080/ramgen/oea/090507-fasano-intvu.rmStudy to Investigate AHCC and the Swine Flu Virus
AHCC is a nutritional food supplement developed using a patented process from the mycelia of several species of mushrooms including the shiitake mushroom.
Dr. Kenna Stephenson Interview with CBS News YouTube Video
AHCC and the Flu on Fox's Morning Show YouTube Video
Learn more about AHCC from Dr. Fred Pescatore YouTube Video
AHCC- a Powerful Aid in Fighting Viruses and Infections Total Health Magazine
Mon Jun 1, 2009 5:12pm EDT
Prior studies published in peer-reviewed journals have shown the benefits of
AHCC against influenza, avian flu, west nile virus and MRSA.
BEAVERTON, Ore., June 1 /PRNewswire/ -- A study evaluating the effect of AHCC
(Active Hexose Correlated Compound) on supporting the immune system in
response to the swine flu virus will be conducted at the Southern China
Agricultural University, one of the few research centers in China that have
been approved to conduct studies on highly infectious diseases such as the
swine flu. This controlled study will examine the effects of AHCC when
supplemented to a group of mice infected with the virus.
Two years ago, following the avian flu outbreak, the same university conducted
a study in mice on the effect of AHCC on the bird flu (H5N1) virus. The
findings, published in the Japanese Journal of Complementary and Alternative
Medicine (Vol. 4, (2007), No. 1 pp. 37-40), demonstrated that AHCC enhanced
host resistance against bird flu. Other papers published in peer-reviewed
journals include studies on AHCC and the common influenza (H1N1), the West
Nile Virus and hospital-borne infections including MRSA.
"The animal study published by my group in the recent issue of Journal of
Nutrition suggested that dietary supplementation with AHCC may be
immunotherapeutic for the West-Nile Virus potentially susceptible
populations." said Tian Wang, Assistant Professor at the Departments of
Microbiology, Immunology and Pathology at the University of Texas Medical
Branch at Galveston. "So in light of our findings and similar conclusions of
Drexel University's study on influenza, I think that a study on AHCC and the
swine flu is very timely and likely to yield useful information on the value
of AHCC in combating this virus".
Published human clinical studies have shown that AHCC activates NK cells and
DC Cells, which are the body's frontline defense against viruses and
infections. A recent study at the Yale Medical School also showed that AHCC
increases the production of cytokines which act as chemical messengers that
activate the immune system when the body comes under attack.
"AHCC is the only natural compound that I know of which has been studied not
only for influenza but also for several other important virus strains." said
Fred Pescatore, MD, MPH of the Center for Integrative and Complementary
Medicine. "The combination of the animal research on those specific strains
and the significant clinical data on the efficacy of AHCC in humans makes this
compound a very compelling target for further research specifically against
the swine flu."
What is AHCC (Active Hexose Correlated Compound)?
AHCC is a natural compound derived from the hybridization of several
subspecies of medicinal mushroom. AHCC is used by hundreds of healthcare
facilities around the world and by hospitals as a part of a standard regimen
for incoming patients to reduce the risk of hospital infections. AHCC is
supported by more than 20 studies published in major peer-reviewed medical
journals (key term "active hexose correlated compound" at www.PubMed.org).