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Archive for the ‘Quorum Sensing’ Category

Biofilm And Bacterial Resistance To Antibiotics When Starved

Saturday, November 19th, 2011

 

Dr. Dao Nguyen, now at McGill University, trained in the University of Washington lab of Dr. Pradeep Singh, a lung specialist who studies bacterial biofilm infections.

“A chief cause of the resistance of biofilms is that bacteria on the outside of the clusters have the first shot at the nutrients that diffuse in,” said Pradeep Singh, associate professor of medicine and microbiology at the University of Washington.

“This produces starvation of the bacteria inside clusters, and severe resistance to (their) killing,” added the senior study author, the journal Science reports.

“Bacteria become starved when they exhaust nutrient supplies in the (infected) body, or if they live clustered together in groups known as biofilms,” said study co-author Dao Nguyen, assistant professor of medicine at Montreal’s McGill University.

Preventing pathogenic bacteria from sensing nutrient starvation may present a new therapeutic approach to increasing antibiotic efficacy and preventing drug resistance, researchers claim. A team led by McGill University investigators has found that blocking an active mechanism used by bacteria to respond to starvation by slowing their growth significantly reduces the natural tolerance to antibiotics that infectious organisms develop when nutrient supplies become low.

The investigators work is reported in Science in a paper titled “Active Starvation Responses Mediate Antibiotic Tolerance in Biofilms and Nutrient-Limited Bacteria.” Pradeep K. Singh, Ph.D., Dao Nguyen, Ph.D., and colleagues

Abstract:

Bacteria become highly tolerant to antibiotics when nutrients are limited. The inactivity of antibiotic targets caused by starvation-induced growth arrest is thought to be a key mechanism producing tolerance. Here we show that the antibiotic tolerance of nutrient-limited and biofilm Pseudomonas aeruginosa is mediated by active responses to starvation, rather than by the passive effects of growth arrest. The protective mechanism is controlled by the starvation-signaling stringent response (SR), and our experiments link SR-mediated tolerance to reduced levels of oxidant stress in bacterial cells. Furthermore, inactivating this protective mechanism sensitized biofilms by several orders of magnitude to four different classes of antibiotics and markedly enhanced the efficacy of antibiotic treatment in experimental infections.

You can read all of my biofilm posts here

Stress, Biofilm and a Predisposition for GI Infections in Type O Blood Individuals

Wednesday, May 19th, 2010

If after reading this post you have questions regarding alternative medicine, integrative medicine, chiropractic, weight-loss, diabetes or pre-diabetes prevention, nutritional supplementation or how to become a new patient, please feel free to contact our office. Advanced Healing Center of Orange County, the practice of Dr. Marcus Ettinger BSc, DC. Phone: 714-639-4360, E-mail: info@advancedhealing.com, Mail: 630 South Glassell Street #103. Orange, CA 92866.

Stress made me solid and less human

Stress Made Me Solid and Less Human

When we are under acute or chronic episodes of physical or emotional stress, our body protects itself by shifting the relative balance of our nervous system, to  sympathetic dominance (self-control), thereby rapidly releasing specific stress hormones such as cortisol, adrenaline aka epinephrine and noradrenaline aka norepinephrine.   The long-term effects of the continual release of these hormones is not good at all and will eventually lead to significant degenerative changes within the body.

The stress hormone norepinephrine affects parts of the brain where attention and responding actions are controlled.   Along with epinephrine, norepinephrine also underlies the so-called fight-or-flight response.  During this stress response, heart rate increases, glucose is triggered to be released from energy stores, and blood flow is increased to skeletal muscle.  At the same time blood and energy is drawn away from the gastrointestinal tract and other internal organs.

1.  Norepinephrine is synthesized from dopamine by utilizing the enzyme dopamine β-hydroxylase.

2.  The gene for dopamine β-hydroxylase has shown some association linkages with the gene that controls the ABO blood types.(1)

3.  Norepinephrine and epinephrine possess a synergistic relationship with AI-3, an autoinducer* and may even substitute itself for the AI-3 auto-inducer, resulting in biofilm growth.

4.  The common denominator between type O blood and dopamine appears to be via the null allele (A null allele is a mutant copy of a gene  that completely lacks that gene’s normal function).  In this case the null allele is the type O blood allele in the human A, B and O blood type system – A, B and AB blood don’t possess it.

A hypothesis  could then be made, based on the above data, that type O blood individuals who are highly stressed (over-activated adrenal glands and sympathetic nervous system dominant) may possess a predisposition to infections or overgrowth of yeasts, bacterias and biofilm in the GI tract, since both epinephrine and norepinephrine are present throughout the gastrointestinal tract, and are involved in the stress response.  This may be especially relevant for those type O individuals who possess the ‘Hunter’ epigenotype.(2)

*Bacteria communicate via signaling molecules called auto-inducers, a type of bacteria pheromone.  These autoinducers can initiate or interfere with Quorum Sensing.  One of the two series of auto-inducer molecules are the Auto-Inducers AI-1, AI-2 and AI-3.

These autoinducers are one of the very few biologically active family of molecules that contain the element boron.  Some evidence indicates that grapefruit juice and its furocoumarins inhibits autoinducer signaling and biofilm formation in bacteria.  The most abundant source of furocoumarins in our diet would be grapefruit juice.  The average levels of furocoumarins were lower in the juice from red grapefruit than the white variety, with the highest level of  this component found in the meat of the grapefruit.

(1) AF Wilson, RC Elston, R M Siervogel, and LD Tran. Linkage of a gene regulating dopamine-beta-hydroxylase activity and the ABO blood group locus. Am J Hum Genet. 1988 January; 42(1): 160-166.

Quorum Sensing and Bonnie Bassler

Monday, April 12th, 2010

Quorum – a quorum is the minimum number of members of a deliberative body, such as a legislature or bacterial colony, necessary to conduct the business of that “group”.

How Bacteria “Talk” An eight minute video explaining, in simple English, how bacteria communicate through Quorum Sensing (QS)

Bonnie Bassler is a Howard Hughes Medical Institute Investigator and professor in the Department of Molecular Biology at Princeton University

Bonnie Bassler is a Howard Hughes Medical Institute Investigator and professor in the Department of Molecular Biology at Princeton University

Bonnie Bassler responded to viewer questions and comments about “talking” bacteria.

Q: Is it possible that even when you make a drug to keep the bacteria from “talking” that the bacteria will work their way around the drug that’s stopping them from communicating, just like they’ve worked around current antibiotics?
Alexandra, 7th grade, Manchester, New Hampshire

A: Hi Alexandra,

Absolutely. We know that bacteria will evolve mechanisms of resistance to anti-quorum-sensing therapies. The hope is that because anti-quorum-sensing strategies do not kill bacteria, only keep them from communicating, this is a less harsh treatment and a less stringent selection for resistance. Because of that, the hope is that resistance will develop more slowly than it does to traditional antibiotics. Thus anti-quorum-sensing therapies may have a “longer shelf life” than traditional antibiotics. Of course, we won’t know if that is indeed the case until we make the drug and monitor its effectiveness.

Q: Do bacteria use quorum sensing for a variety of purposes, e.g., fish form schools for protection, lions form prides to hunt? Also, do the same types of bacteria from different colonies communicate differently than the same types of bacteria from different colonies, i.e., do genetically related bacteria recognize each other aside from their being the same species?
Paula, Washington, D.C.

A: Dear Paula,

Quorum sensing is used to control hundreds of different group processes. In the cases we understand best, the processes controlled are ineffective when carried out by individual bacteria acting alone but become effective when carried out in synchrony by the group. Virulence is a good example of a quorum-sensing-controlled behavior. A few bacteria can’t make us sick. Even if they release toxins or other harmful agents, we are HUGE compared to them, so each bacterium’s measly few molecules of a toxin can’t harm us. But, if the bacteria wait, count themselves with quorum-sensing molecules, and then all of the bacteria release their toxins simultaneously, they can overcome an enormous host. There are all kinds of examples of quorum-sensing-controlled group behaviors: bioluminescence, virulence, exchange of DNA, biofilm formation, symbiosis….

Bacteria have multiple chemical languages. We think each bacterial species has a molecule that is unique and represents its own species-specific language. Each unique molecule allows a particular species to know and communicate with its relatives (i.e., have a private conversation). We know there is a universal molecule, a sort of bacterial trade language that bacteria use to converse between species (i.e., a Bacterial Esperanto). There is mounting evidence that many additional molecules remain to be discovered, such as species non-specific molecules that say “who” the neighbor is. This field is only about 10 years old. So far we have only managed to identify a few molecules. We know that bacteria interpret a rich and complex chemical world, and we are working hard to define the complexity of the lexicon.

Q: Can you provide some thoughts on individuals with knee or hip implants and the biofilm some unfortunately develop and if there is hope for probiotics in this issue?
[first name not given] Raunaque, LaCrosse, Wisconsin

A: Dear Mr. or Ms. Raunaque,

Biofilms are communities of bacteria attached to surfaces (for example, as described in the NOVA piece, your teeth in the morning). Biofilms are a cause of constant concern for people with medical implants, heart valves, etc., because these medical devices provide a niche for bacteria to adhere to and thus cause infection. We know that quorum sensing is required for biofilm formation: If the bacteria can’t talk, they can’t make these large, interacting, adherent communities. The goal of many researchers’ studies is to find methods to interfere with biofilm formation, and, of course, a key focus of study is on interrupting quorum sensing. One strategy is to make molecules that antagonize the natural quorum-sensing molecules to impede biofilm formation. Another strategy is the one you suggest, to develop probiotic quorum-sensing therapies that enhance the conversation among our commensal bacteria at the expense of the biofilm formers. Both these avenues are currently being explored.

Quorum Sensing and Biofilm

Sunday, December 13th, 2009

What is Quorum Sensing and how do bacteria talk to each other?

The discovery that bacteria are able to communicate with each other changed our general perception of many single, simple organisms inhabiting our world. Instead of language, bacteria use signaling molecules which are released into the environment. As well as releasing the signaling molecules, bacteria are also able to measure the number (concentration) of the molecules within a population. Nowadays we use the term ‘Quorum Sensing’ (QS) to describe the phenomenon whereby the accumulation of signaling molecules enable a single cell to sense the number of bacteria (cell density). In the natural environment, there are many different bacteria living together which use various classes of signaling molecules. As they employ different languages they cannot necessarily talk to all other bacteria. Today, several quorum sensing systems are intensively studied in various organisms such as marine bacteria and several pathogenic bacteria.

Quorum Sensing & Biofilm Formation

Quorum Sensing & Biofilm Formation

Why do bacteria talk to each other?

(QS) enables bacteria to co-ordinate their behavior. As environmental conditions often change rapidly, bacteria need to respond quickly in order to survive. These responses include adaptation to availability of nutrients, defense against other microorganisms (biofilm formation) which may compete for the same nutrients and the avoidance of toxic compounds (biofilm formation) potentially dangerous for the bacteria. It is very important for pathogenic bacteria during infection of a host (e.g. humans, other animals or plants) to co-ordinate their virulence in order to escape the immune response of the host in order to be able to establish a successful infection. The University of Nottingham Quorum Sensing Research Group

From Dr. Ettinger’s Biofilm Protocol for Lyme and Gut Pathogens: Pathogenic bacteria known to reside in biofilms include: Borrelia burgdorferi, Escherichia coli, Candida albicans, Clostridium difficile, Clostridium perfringens, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Vibrio cholerae. The number of human diseases shown to be associated with biofilms is expanding and includes chronic bacterial prostatitis, chronic rhinosinusitis, cystic fibrosis pneumonia, infective endocarditis, periodontitis, recurrent otitis media, and virtually all device and implant related infections. Strong evidence is also beginning to emerge for an etiologic role of pathogenic mucosal biofilms in gastrointestinal diseases, such as Irritable Bowel Disorders: Crohn’s disease and ulcerative colitis.

Dr. Ettinger’s Biofilm Protocol for Lyme and Gut Pathogens

Friday, September 25th, 2009

A specific question has been asked a lot lately, as to what is my protocol for handling Biofilm.  Most of these questions have been directed to me by those diagnosed with or think they may have, Lyme disease or H. pylori bacteria.  The reason that I’ve put this “biofilm protocol” post together is because of this fact: the day I discovered how to handle biofilm in the body, was the day that chronic conditions were no longer a ‘project’, so to speck, to handle. I hope this information is helpful to you.

First a little background on biofilm:

biofilm

Fig. 1: The biofilm life cycle. 1: individual cells populate the surface. 2: extracellular polymeric substance (EPS) is produced and attachment becomes irreversible. 3 & 4: biofilm architecture develops and matures. 5: single cells are released from the biofilm. Related PostBiofilm Basics and Quorum Sensing and Biofilm

This is an excerpt from a Klaire Labs product monograph which is a basic primer on the topic (My additions are in RED) The National Institutes of Health estimates that 60% of all human infections and 80% of refractory infections (def. unresponsive to medical treatment) are attributable to biofilm colonies.  I have seen this, most commonly, in cases I’ve worked-up, where the pathogen is: Chlamydia pneumoniae, Pseudomonas aeruginosa, Helicobacter pylori, [Lyme disease - Borrelia burgdorferi] and Candida albicans.

  • The protection conferred upon microorganisms by biofilm allows them to achieve a high level of antibiotic resistance, stealth and invisibility.
  • Biofilm not only provide a physical barrier to antimicrobial agents (pharmaceutical antibiotics) and host antibodies, but facilitate the exchange of antibiotic-resistant genetic material between organisms and may contain antibiotic-degrading (hydrolysing) enzymes such as b-lactamase, effectively neutralizing incoming antibiotic (b-lactam antibiotics) molecules.
  • In fact, biofilm communities can be 1000 times more resistant to antibiotics than free-floating bacteria.
  • The decreased growth rate of sessile microorganisms (def. Permanently attached to a substrate; not free to move about; “an attached oyster”) also reduces their antibiotic susceptibility as most antimicrobial agents require rapid cell growth in order to effectively kill or inhibit the microbes.  Biofilm thus render pathogenic microorganisms enormously difficult to eradicate, and can almost single-handedly contribute to localized or systemic inflammatory reactions and delayed wound healing.
  • Depending on the type of biofilm, one or more species of pathogens may be found embedded in the extracellular polymeric substance (def. Composed primarily of polysaccharides and can either stay attached to the cell’s outer surface, or be secreted into its growth medium).  Bacterial extracellular polymeric substance (EPS) maybe a carrier of, or may have heavy metals embedded in them, thus the indication for chelation w/EDTA. EDTA, ethylenediaminetetraacetic acid, is a chelating agent used to lower one’s body burden of heavy metals).

Pathogenic bacterial known to reside in biofilms include, but are not limited to: Borrelia burgdorferi (Lyme bacteria), Escherichia coli, Candida albicans (yeast and fungal mutation), Clostridium difficile, Clostridium perfringens, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, and Vibrio cholerae. The number of human diseases shown to be associated with biofilms is ever expanding and includes: chronic bacterial prostatitis, chronic rhinosinusitis (chronic sinus infections), cystic fibrosis pneumonia, infective endocarditis, periodontitis, recurrent otitis media, and virtually all device and implant related infections.  Strong evidence is also beginning to emerge for an etiologic (causative) role of pathogenic mucosal biofilm in gastrointestinal diseases, such as Irritable Bowel Disorders (IBS): Crohn’s disease and ulcerative colitis.

S. aureus biofilm

S. aureus biofilm

Dr. Marcus Ettinger’s Biofilm Protocol – Only the eradication phase is presented here.  There is a pre,  post and toxin reduction step as well.  You can get help with any of these steps – HERE

A. Products (mandatory products in red). These are ONLY the basics. Additional nutraceuticals may be needed, based on each individuals unique situation.

  1. Monolaurin [AKA Glyceryl laurate or glycerol monolaurate] (monolaurin information) or Lauricidin
  2. Nutiva Extra-Virgin Coconut Oil (42-52% Medium Chain Fatty Acids [MCFA], lauric acid, by volume)
  3. Nattokinase (a potent fibrinolytic enzyme) I like this better than Lumbrokinase.
  4. InterFase Plus™ (broad-spectrum enzyme formula w/EDTA)
  5. Serrapeptase (a potent fibrinolytic enzyme)
  6. Vitamin C (ascorbic acid – Not buffered, as most of these contain metals)
  7. NAC (N-Acetyl-Cysteine)
  8. Lactoferrin (specifically Nutricillin by Ecological Formulas) Dr. Anju Usman of Illinois states, “Our bodies make proteins, transferrin and lactoferrin, which mop up iron and block the ability of biofilm to form,” she said. “But pathogenic bacteria secrete iron chelators to snatch up iron and thus compete with the transferrin and lactoferrin for what they need to survive.”

B. Avoid supplemental forms of: magnesium, iron and calcium during the biofilm protocol, as they may contribute to  biofilm formation or decrease the effectiveness of the biofilm protocol.

C. Take a broad-spectrum probiotic and prebiotic.  I like the combination of Now Foods brand Probiotic-10 and their Probiotic Defense Powder (contains gluten).  These products will help to crowd out the bad bacteria, and also help disrupt and replace biofilm colonies along the mucus membrane.

D. Specific additions based on condition (not a complete list):

  1. Candida albicansSF722* (10-Undecenoic Acid  50 mg) Thorne Research. This is as close as you can get to a medication and still be a natural substance.  There are a few chat rooms blasting this product, based on who knows what – can’t make everyone happy.  I’ve used SF722 for over 15 years and it is amazing – never a problem!  *Do not take SF722 if you are allergic to fish. There are many other amazing products that can be added to complement the SF722. It’s really a matter of how many pills someone wants/doesn’t want to take per day or the severity of one’s condition, that will determine, if or which, additional products will be added. If the Candida albicans overgrowth is severe, has not responded to holistic methods or has mutated into its more virulent hyphal form/fungal infection (nails, underarms, groin or skin); Diflucan (fluconazole), a prescription medication, is my personal preference, but Nizarol (ketoconazol) can also be used. In Azole-resistant Candida albicans, lactoferrin must be added to either medication in order to increase their effectiveness. There is a certain B vitamin, mineral and amino acid that possesses synergistic qualities and I find them indispensable when taking Diflucan (fluconazole), Nizarol (ketoconazole) or for supporting candida die-off symptoms.
  2. Chlamydia pneumonia, Klebsiella pneumoniae or Pseudomonas aeruginosa Pneumotrophin PMG by Standard Process, Inc. How it works.  I use this because it helps direct the body’s attention and healing efforts to the lung, where it’s needed most. Apex Energetics, H-PLR is also a mandatory addition. I also like to use OOrganik-15™ and Pneuma-Zyme™ by Biotics Research with some of my patients who manifest asthma, a chronic cough and/or emphysema like symptoms.     
  3. H. pyloriComplete write-up on another post.
  4. Chronic bacterial prostatitis – Quercitin (600mg’s) and Bromelain (200mg’s) combination by Now Foods. Decreases inflammation and oxidant stress in the prostate while increasing local concentrations of beta-endorphins. Apex Energetics, H-PLR is also a mandatory addition.

E. Certain dietary restrictions and additions will need to be taken. These are determined on a case by case basis.

Important Note:

All dosages will be provided if you purchase some or all of your “biofilm protocol” products through my office. I truly do want to help all who are interested, but it’s finally gotten to the point where too many people want free advice and an increasing amount of my time, and then buy all of their products elsewhere. I am a firm believer in fair exchange and I feel I have done that by providing the information in this post.

I also offer tailor made protocols for your individual situation, please contact our office for product prices and distance patient information (714) 639-4360. 

 

 

 

Biofilm testing is also available through Fry Laboratories. Fry Laboratories, L.L.C. is an independent clinical diagnostic and research laboratory located in Scottsdale, Arizona. We are committed to understanding chronic diseases and contributing to their cure through advancements in diagnostics and basic science research with emphasis on chronic inflammatory diseases, vector-borne diseases, and their intersection. Our clinical diagnostic laboratory offers general and targeted immunology services in conjunction with standard and cutting edge infectious disease detection and identification technologies. Our signature services include microscopy for visual identification and quantification of a wide range of blood-borne pathogens, co-infection serology, biofilm detection, and genus wide molecular detection technology with sequencing for individualized species and/or strain identification. We participate in both CAP and API quality control programs and provide worldwide testing service.

Diseases of Interest: Chronic Fatigue Syndrome, Fibromyalgia, Gulf War Veterans Illness, Chronic Lyme Disease, ALS (Lou Gehrig’s Disease), Parkinson’s Disease, Multiple Sclerosis, Autism, Lupus, Ulcerative Colitis, Scleroderma, Rheumatoid Arthritis, Osteoarthritis, Crohn’s Disease.

Infections of Interest: Borrelia (Lyme), Babesia, Bartonella, Anaplasma, Ehrlichia, Q-Fever (Coxiella), Toxoplasma, Rickettsia, Plasmodium, XMRV

Important: This post is not a substitute for medical advise or treatment and is for informational purposes only. Please consult with a physician before starting any nutritional or biofilm protocol on your own.


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