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[b]Fiber-Famished Gut Microbes Linked to Poor Health
While probiotics receive more attention, key fibers remain the workhorses in maintaining a healthy gut microbiom[/b]e
March 23, 2015 |By Katherine Harmon Courage
Another recent study shows that when microbes are starved of fiber, they can start to feed on the protective mucus lining of the gut, possibly triggering inflammation and disease.
Diet is one of the most powerful tools we have for changing the microbiota," Justin Sonnenburg, a biologist at Stanford University, said earlier this month at a Keystone Symposia conference on the gut microbiome. "Dietary fiber and diversity of the microbiota complement each other for better health outcomes." In particular, beneficial microbes feast on fermentable fibers—which can come from various vegetables, whole grains and other foods—that resist digestion by human-made enzymes as they travel down the digestive tract. These fibers arrive in the large intestine relatively intact, ready to be devoured by our microbial multitudes. Microbes can extract the fiber's extra energy, nutrients, vitamins and other compounds for us. Short-chain fatty acids obtained from fiber are of particular interest, as they have been linked to improved immune function, decreased inflammation and protection against obesity.
Today's Western diet, however, is exceedingly fiber-poor by historical standards. It contains roughly 15 grams of fiber daily, Sonnenburg noted. For most of our early history as hunter-gatherers, we were likely eating close to 10 times that amount of fiber each day. "Imagine the effect that has on our microbiota over the course of our evolution," he said.
What does that mean?As fiber consumption increased, the activity of genes associated with protein metabolism declined, a finding that researchers hope will help them understand the complicated puzzle of diet and weight loss. "We're getting closer to what is actually cause and effect," Swanson says.
A third group of mice received high-fiber chow and fiber-free chow on alternating days—"like what we would do if we were being bad and eating McDonald's one day and eating our whole grains the next," Martens joked. Even the part-time high-fiber diet was not enough to keep guts healthy: these mice had a mucus layer about half the thickness of mice on the consistently high-fiber diet. If we can extend these results to humans, he said, it "tells us that even eating your whole fiber foods every other day is still not enough to protect you. You need to eat a high-fiber diet every day to keep a healthy gut."
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More disturbing news was revealed this week on new sources of antibiotic resistance in the environment. First, in a troublesome report in mBio, the journal of the American Society for Microbiology, researchers showed that three commercial herbicides—Monsanto’s dicamba (Kamba) and glyphosate (Roundup), and 2,4-dichlorophenoxyacetic acid (2,4-D)—could make strains of Escherichia coli and Salmonella enterica serovar Typhimurium less sensitive to antibiotics. (The response varied with different combinations of antibiotic, herbicide, and bacterial strain).
This is hugely important for several reasons: Herbicides are fairly ubiquitous in the environment. Glyphosate (Roundup) has been found in the milk and meat of cows, and in human urine. According to German researchers, “Glyphosate residues cannot be removed by washing and they are not broken down by cooking. Glyphosate residues can remain stable in foods for a year or more, even if the foods are frozen, dried or processed.” Thus, there is great chance for interaction of herbicides with antibiotics. Interestingly, Roundup alone had once been considered as an antibiotic, but resistance was found to develop rapidly. Dr. Jack Heinemann, the study’s lead author and professor at the University of Canterbury in New Zealand explains that while a bacteria alone might have been killed by an antibiotic, when exposed to an herbicide, a resistance gene is turned on, in effect “‘immunizing’ the bacteria to the antibiotic.”
n additional problem is that some weeds have become resistant to Roundup. New genetically modified soybeans and corn have been engineered by Dow to resist the “Enlist Duo” herbicide, which is a combination of glyphosate and 2, 4-D. Monsanto also is looking to sell soybean and cotton GMO varieties resistant to both herbicides.
Another study showed that heavy metals, added to feed as growth promoters, can also select for antibiotic resistance. Pollution with heavy metals—even at very low levels— can promote bacteria with multiresistance plasmids (small bits of extrachromosomal DNA). This combination can fuel resistance, as the antibiotics can act synergistically with the heavy metals in selecting for resistance. In this Swedish study, investigators examined an extended-spectrum beta-lactamase (ESBL) plasmid from a hospital outbreak of Klebsiella pneumoniae and Escherichia coli. This plasmid carried resistance not only to beta-lactam and other antibiotics, but also to quaternary ammonium compounds, copper, silver, and arsenic. This is important as copper and silver are being explored for their properties that might treat infections without antibiotics and “quats” are a commonly used hospital disinfectant. Resistance to these metals has not yet been a problem.
Intuition Explained
by John Douillard on May 5, 2015 | Leave a Comment
Average Reading Time: 3 minutes and 24 seconds
A recent study has linked gut feelings and intuitive decision-making to newly-discovered neurobiological gut-brain pathways. (1) A gut-brain pathway carries gut-based, emotionally-charged signals carried by microbes from your gut to the brain where they can affect brain function.
While “gut feelings” and “intuition” seem like terms better suited for the metaphysical rather than neurobiology, the new science linking our microbiome to higher brain functions like intuition is emerging.
According to research, intuition and gut feelings are built on a series of both positive and negative gut-brain signals beginning at birth. These include gut-brain emotions such as reward hormones (dopamine), and hunger and craving hormones (ghrelin) which train the central nervous system. (1)
Years of repetitive gut-based brain signals form memories of emotional states that, according to the research, are used for complex social emotions, including predictions about the future and intuitive decision-making. (1) In Ayurveda, predictions and intuition are similar concepts; both based on the extension of logic.
According to Ayurveda, Intuition Is Not Magic.
In 1986, during my studies in India, I would follow my Ayurvedic teacher around each day as he performed his medical rounds. Dr. Raju was a pulse diagnosis and herbal expert, and I would be wowed on a regular basis by some of his pulse readings.
Once, after taking the pulse of a middle-aged man from England, Dr. Raju asked the Englishman, “Did you fall off a ladder when you were young and break a few vertebrae in your back?” The man confirmed that he had, in fact, fallen off a ladder and broken his back. Dr. Raju then contended that it was his back injury some 30 years earlier that was causing his current condition of anxiety and insomnia.
After hearing Dr. Raju make similar diagnoses on a regular basis, I asked him how he could make such predictions. I asked him if he was psychic. He gave me a strange look as if he didn’t know what psychic was. I asked him if he used intuition. He said, “No.”
He told me that intuition is nothing more than the extension of logic. He said, “When I take the pulse, it is logic, not magic. When you are a child, adding 2 + 2 is a challenge but soon, with practice, the answer comes without having to think about it. Soon, with the extension of logic, more difficult problems also come easy without having to think about them. It is just a ‘knowing.’ Taking the pulse is the same. Over time and with practice, it becomes natural to know things that seem unknowable by simply extending logic.”
Ayurvedic Intuition
According to Ayurveda, there are many explanations for intuition, higher cognitive function, and higher states of consciousness. Intuition starts with the emotional state of mind during the process of eating food. One of my favorite concepts is how food becomes ‘emotionally-charged’ as a result of the state of mind during the process of eating. Eating while angry or stressed is considered a no-no in Ayurveda. Of course, now we know that it is in fact the microbes carried by food that can be emotionally-charged that carry these emotional messages. (1) Additionally, lifestyle, stress levels, and emotions can alter the gut-to-brain signals carried by the microbes. (2,3)
Other research in support of the intuitive process has shown that the gut feelings of one person (as measured by electrogastrogram activity) increase in response to the emotions of a distant person beyond the influence of ordinary sensory interactions. (4) Interestingly, research has shown that intuition seems to be enhanced in conjunction with positive emotions, as opposed to negative emotions. (5-7)
Ayurveda suggests living a positive, loving, giving, caring, lifestyle, called a sattvic lifestyle. A sattvic lifestyle is a prerequisite for the development of intuition and higher states of consciousness. A positive, loving lifestyle also supports the proliferation of beneficial microbes, inhibition of harmful bacteria and the production of a health promoting hormone called oxytocin. (2,3,8)
According to Ayurveda, the subtle digestive nutrient fluid is called rasa and it carries the emotional charge, possibly via the microbiology, into every cell of the body. If the charge is positive or sattvic, then the nutrient fluid positively affects all the tissues of the body. The final product of digesting this nutrient fluid, which can take up to 30 days to complete, is a substance called ojas. Ojas is not only responsible for immunity and vitality; it is an essential substance required for intuition, gut feelings and the development of higher states of consciousness.
While much more research is needed to confirm the relationship between lifestyle, stress, emotionally-charged microbes, the production of ojas through digestion, and higher cognitive functions like intuition and gut feelings, there is emerging evidence that suggests that there is an intimate relationship between digestion, gut microbes, the brain, emotions and intuition. (1)
References
http://www.ncbi.nlm.nih.gov/pubmed/21750565
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC414848/
http://www.ncbi.nlm.nih.gov/pubmed/18227772
http://www.ncbi.nlm.nih.gov/pubmed/15750366
http://www.ncbi.nlm.nih.gov/pubmed/20515252
http://www.ncbi.nlm.nih.gov/pubmed/12930470
http://www.ncbi.nlm.nih.gov/pubmed/19203169
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3537144/
New Self, New World explores the implications of the little-known fact that we have two brains: in addition to the familiar cranial brain in the head, there is a “second brain” in the gut. This is not a metaphor. Scientists recognize the web of neurons lining the gastrointestinal tract as an independent brain, and a new field of medicine — neurogastroenterology — has been created to study it.
According to Shepherd, there is a good reason that we talk about “gut instinct.” If cranial thinking sets us apart from the world, the thinking in the belly joins us to it. If the cranial brain believes itself surrounded by a knowable world that can be controlled, the brain in our belly is in touch with the world’s mystery. The fact that the second brain has been discovered, forgotten, and rediscovered by medicine three times in the past century suggests how complicated our relationship with our bodily intelligence is.
Tim Spector, professor of genetic epidemiology at King's College London, said: "Another dimension that could be as important are our gut microbes, which also change between seasons and could be driving these changes because of seasonal changes in diet."
New York (April 23, 2015) — An international research team led by Weill Cornell Medical College investigators has discovered an answer to why the human immune system ignores roughly 100 trillion beneficial bacteria that populate the gastrointestinal tract. The findings, published April 23 in the journal Science, advance investigators' understanding of how humans maintain a healthy gastrointestinal tract, and may provoke new ways to treat inflammatory bowel disease — including Crohn's disease and ulcerative colitis — whose origins have been mysterious and treatment difficult.
The investigators studied T cells — critical components of the adaptive immune system — which have the capacity to recognize, eliminate and remember foreign microbes that invade our bodies. T cells are named after the thymus, an organ where they develop and are taught not to attack normal human tissues and organs, leaving them free to target and eradicate disease-causing foreign invaders. One question that had puzzled scientists until now is how these cells learn to ignore beneficial bacteria in the intestine that are also foreign, but not harmful.
In the study, the research team discovered that once they leave the thymus, T cells are again educated in the gastrointestinal tract, or gut, to leave beneficial bacteria alone. This dual education strategy is vital to supporting healthy immune function, the investigators say. Disruption in the pathway that facilitates this education, they add, causes the immune system to attack beneficial bacteria in the intestine, which is often linked to the development and progression of diseases like inflammatory bowel disease, HIV, viral hepatitis, cardiovascular disease, obesity, diabetes and cancer. Therapeutic strategies to promote and boost the activity of this education pathway may be beneficial in treating patients with these chronic inflammatory disorders, the investigators say.
"In many chronic human diseases, the immune system attacks bacteria in the intestine that are normally beneficial.
Be good to your gut — do more exercise
Exercise alone doesn’t lead to significant weight loss, argues Professor Spector. Research also shows it won’t help keep it off.
However, it is good for your heart and brain — and your gut bacteria.
Results from the professor’s studies of 3,000 twins show that the amount of exercise they took is the strongest factor in promoting the richness of their gut microbes.
The findings are supported by a study of elite athletes in the national Irish rugby squad, published last year in the journal Gut.
Nutritionists at University College Cork found that the athletes had much more diverse stomach bacteria than normal, as well as lower levels of inflammation.
So, how does exercise have this effect? One way in which it does this is by stimulating the immune system, which, in turn, sends stimulating chemical signals to the microbes in our guts, according to a 2011 study in the journal Immunology Investigations.
Exercise also benefits our balance of gut bugs directly, according to a 2008 report in the journal Bioscience, Biotechnology and Biochemistry.
The study of lab rats showed that those exercising on a wheel produced twice as much of the fatty acid butyrate in their guts compared with sedentary rats.
Butyrate is produced by our gut microbes and has a broad range of beneficial effects on the immune system, says the report. Exercise stimulates microbes to produce more of it.
Read more: http://www.dailymail.co.uk/health/artic ... z3Zy57nsKR
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[/color]The Stanford study relied on data from the Women's Health Initiative, a long-term federally funded project that tracked the health of nearly 162,000 women 50 to 79 years of age at 40 U.S. hospitals.
Of those women, the researchers focused on about 132,000 postmenopausal women to see if their exercise levels had any effect on lung cancer risk or death. During nearly 12 years of follow-up, just over 2,200 women developed lung cancer and 1,400 women died from the disease.
But women who spent more minutes per week on the move were less likely to either develop lung cancer or die from it, according to the findings.
The study looked only at minutes spent moving around, not at intensity of movement, Wang said. A minute of walking or mowing the lawn weighed the same as a minute jogging or lifting weights.
"It seems to indicate that you don't have to kill yourself," she said. "It doesn't need to be strenuous. You just have to put the time in."
What's more, Wang and her colleagues found that physical activity seemed to help even smokers. Former heavy smokers and current smokers developed lung cancer and died from the disease less often if they were active, compared with sedentary women who smoked.
The benefits of physical activity were most prominent among women who were not obese, with a BMI -- body mass index, a ratio of weight to height -- under 30, according to findings. A BMI of 30 or greater is considered obese.
However, "it would be the wrong conclusion that women with higher BMI won't benefit from physical activity," Wang added.
It should be noted that the study only revealed an association between exercise and lung cancer risk. It did not prove cause-and-effect.
The French study found that physical activity appeared to reduce breast cancer risk.
In that study, researchers reviewed 38 previous studies published between 1987 and 2014 that involved 4.18 million women and more than 116,000 cases of breast cancer.
Women with the highest levels of physical activity experienced an 11 percent to 20 percent reduction in breast cancers, compared to women with the lowest levels of activity, researchers reported.
Overall, a sedentary woman who began engaging in four to seven hours a week of mainly vigorous physical activity seemed to reduce her risk of breast cancer by 31 percent, according to lead author Cecile Pizot, a biostatistician with the International Prevention Research Institute in Lyon, France.
"This reduction occurred irrespective of the type of physical activity, the place of residence, obesity and menopausal status," Pizot said. "Also, breast cancer risk seems to decline with increasing physical activity, and we observed no threshold."
However, that benefit only applied to women who had never used hormone replacement therapy. Taking replacement hormones appeared to wipe out whatever protective benefit that exercise conferred.
Dr. Ruth Heidrich healed stage 4 breast cancer with nutrition in 1982, then became a world class athlete winning more than 900 trophies, 6 Ironman Triathlons, 8 Senior Olympics Gold Medals and 67 marathons!
The mammalian gut microbiome is involved in controlling the circadian rhythm of its host, according to a mouse study published today (April 16) in Cell Host & Microbe. In both mice and humans, timing of feeding and diet type can impact the bacterial populations of the gut. Now, Eugene Chang of the University of Chicago Medical Center and his colleagues have found that mouse gut microbiota produce metabolites in diurnal patterns, and these can influence the expression of circadian clock genes in the liver.
nviting us to consider what a scaled-up version of this “molecular chatter” might mean, Turney wants us to see our microbiome as engaged in a series of overlapping conversations involving the body’s four “information and control” systems – the genome, endocrine, brain/nervous systems and especially the immune system.
Researchers found that prolonged exposure to a toxin produced by Staphylococcus aureus (staph) bacteria causes rabbits to develop the hallmark symptoms of Type 2 diabetes, including insulin resistance, glucose intolerance and systemic inflammation.
Obesity is a known risk factor for developing Type 2 diabetes, but obesity also alters a person's microbiome - the ecosystem of bacteria that colonise our bodies and affect our health.
"What we are finding is that as people gain weight, they are increasingly likely to be colonised by staph bacteria - to have large numbers of these bacteria living on the surface of their skin," Schlievert said.
"People who are colonised by staph bacteria are being chronically exposed to the superantigens the bacteria are producing," Schlievert said.
The study shows that superantigens - toxins produced by all strains of staph bacteria - interact with fat cells and the immune system to cause chronic systemic inflammation, and this inflammation leads to insulin resistance and other symptoms characteristic of Type 2 diabetes.
Among their long list of tasks, gut bacteria may help train the immune system to distinguish between human and microbe so that it can confront what’s bad, tolerate what’s not, and recruit a diverse army of cellular foot soldiers to stop invading germs. Writing in Nutrition Journal in June 2014, one scientist likened gut microbiota to a sparring partner, providing a regular workout that strengthens the contender for a true opponent.
To function at their best, though, gut microbes, like most living things, need to be well fed. And many of the species responsible for immune equilibrium don’t seem to care for junk food. In a study reported in Nature Communications in April, African-American volunteers who shifted from an American diet to high-fiber, low-fat African cuisine experienced a drop in inflammation in just two weeks.
These are the kinds of compounds that TIM digests all day, and the hope is that more of them will reach our supermarkets and kitchens, in products better targeted to help our bodies and even our minds. Fantasy foods like juices that can ward off autoimmune diseases or cereals that can encourage fat reduction now seem like attainable goals to some scientists. Other researchers think the microbiota could launch an era of personalized nutrition, in which a thorough analysis of your unique gut ecosystem could guide the way to a customized diet, optimized for your health.
The plummeting cost of microbial genome sequencing has triggered an avalanche of research that’s prompting optimism about potential benefits—more than 6,000 studies of the microbiome have been published since 2006. The attendant publicity hasn’t gone unnoticed by consumers: In a recent survey by market-research firm NPD Group, 31% of Americans said they wanted to ingest more probiotics. Seeing an opportunity, foodmakers, from corporate giants like Nestlé and General Mills to fledgling startups, are pouring money into research. “Drug companies, food companies, academics, federal scientists, food scientists, neuroscientists, institutes,” says George Fahey, professor emeritus of nutritional sciences at the University of Illinois. “Everybody wants a piece of it.”
In this crowded race, Danone, the world’s leading dairy producer, has a pronounced headstart. Yogurt, one of the best-known gut-friendly foods, is Danone’s bestselling product globally (including under the Dannon brand in the U.S.). Danone was one of the first food companies to fund microbiota research; it’s currently staging some 100 clinical trials and collaborating with more than 40 academic or commercial partners in the field. Gérard Denariaz, director of strategic R&D partnerships at Danone, describing the company’s scatter-the-seeds philosophy, says: “Who knows who will come up with the next breakthrough?”
If companies do crack the microbiome, the rewards could be huge: Global sales of “fortified/functional foods” reached $275 billion in 2014, according to Euromonitor, and foods that could tout themselves with scientific precision as suppressing obesity or fostering childhood development could be enormously lucrative.
nol’s observation reflects a reality that makes food executives queasy—the chance that big food won’t be the industry that profits first, or most, from microbiome research. A number of pharmaceutical companies, including Johnson & Johnson JNJ 0.11% , Pfizer PFE -0.23% , and Sanofi, have research initiatives focusing on the gut microbiome. The tech sector could also capitalize on the trend: Just about everyone in microbiome research describes a not-far-off future when some kind of diagnostic tool (most have their bets on smart toilets) will analyze your gut bacteria and steer you to the foods you need most—a point of connection between the Internet of things and a new normal of personalized nutrition.
Having a healthy gut may well depend on maintaining a complex signaling dance between immune cells and the stem cells that line the intestine. Scientists at the Buck Institute are now reporting significant new insight into how these complex interactions control intestinal regeneration after a bacterial infection. It's a dance that ensures repair after a challenge, but that also goes awry in aging fruit flies—the work thus offers important new clues into the potential causes of age-related human maladies, such as irritable bowel syndrome, leaky gut and colorectal cancer.
Read more at: http://phys.org/news/2015-05-complex-bl ... n.html#jCp
Jasper says aging makes it harder for the stem cells to switch gears between proliferation and quiescence and that flies suffer from age-related intestinal dysfunctions similar to those experienced by humans. Jasper says when the flies are young they are able to fend off infection and repair tissues, but that the cumulative effect of damage over a lifetime takes a toll - signaling goes awry, and stem cells get chronically activated, causing inflammation and dysplasia, which makes the animal more prone to infection and dysfunction. "This is another classic example of 'what is good for us in youth, turns against us with age'," said Jasper. "When we think of interventions, we need to find the sweet spot. We want to promote stem cell repair and regeneration without having those responses become chronically activated."
Fat, sugar cause bacterial changes that may relate to loss of cognitive function
Date:
June 22, 2015
Source:
Oregon State University
Summary:
A study indicates that both a high-fat and a high-sugar diet, compared to a normal diet, cause changes in gut bacteria that appear related to a significant loss of 'cognitive flexibility,' or the power to adapt and adjust to changing situations. This effect was most serious on the high-sugar diet, which also showed an impairment of early learning for both long-term and short-term memory.
Severe Burns May Let 'Bad Bacteria' Take Over the Gut
Gillian Mohney, Live Science Contributor | July 08, 2015 05:21pm ET
People who have gotten severe burns are known to be at risk for a host of complications, but there may be other consequences lurking deeper within the body: A new study finds that a burn may change the community of bacteria within a person's gut, and possibly lead to an increased risk of infection.
In the study, researchers analyzed fecal bacteria from four patients with severe burns over at least 30 percent of their body, and compared these bacteria with fecal bacteria of people with minor burns. They found that the severely burned patients had higher levels of a potentially dangerous group of bacteria called Enterobacteriaceae, which includes E. coli and Salmonella.
In fact, Enterobacteriaceae made up 31.9 percent of the gut bacteria in the people with severe burns, compared with just 0.5 percent in those with minor burns, according to the study, published July 8 in the journal PLOS ONE.
This increase in "bad bacteria" may explain why burn patients are at increased risk for sepsis, a dangerous inflammatory response that can lead to organ failure, Choudhry said. It's possible that the severe burns compromise the defense mechanism in the gut, thus allowing the harmful bacteria to leave the gut and trigger inflammation in the body, Choudhry noted.
A new animal study lends support to the idea that high fat diets make it harder for our brains to tell us we're full by changing the bacterial environment of the
The authors treated the rats in their study to one of two meal plans: a regular diet with a daily fat content of about six percent and a sinful smorgasbord fit for a whiskered king that clocked in at a whopping 35 percent fat. They then, apparently without a hint of guilt, dissected the rats and examined their guts and brain, finding that there were distinctive differences between the two groups.
They also found that there was noticeable changes along certain neural pathways in the rats made obese by the high fat diet, changes that may explain why obesity is notoriously difficult to combat. "When we switch the rats to a high fat diet, it reorganizes brain circuits," Czaja said. "The brain is changed by eating unbalanced foods. It induces inflammation in the brain regions responsible for feeding behavior. Those reorganized circuits and inflammation may alter satiety signaling."
ALT LAKE CITY — The easiest way to stop germs from getting in your house is to leave your shoes at the door. A new study found 40 percent of shoe soles contain the bacteria C. difficile.
The study, published in the New England Journal of Medicine, was conducted by researchers from the University of Houston. They looked at samples from 30 houses in Houston and found C. diff on 40 percent of shoe soles and 33 percent of bathrooms and toilets.
Companies are also mining the microbiome for uses beyond therapeutic ones. Cambridge, Massachusetts-based AOBiome is pursuing a consumer route, having developed a live bacterial spray it says can help users cut back on deodorant and soap. The product, AO+ Mist, contains ammonia oxidizing bacteria that the company calls AOBs.
"They convert ammonia to nitrate and nitric oxide," CEO Spiros Jamas said in an interview at AOBiome's offices, which are decked out in gear touting the benefits of bacteria, including shirts that say "bacteria is the new black; wear it every day."
"The company's hypothesis is that humans evolved with AOBs as a natural component of our skin," Jamas said. "And really only in the last 50 years, with the advent of cleaning and hygiene routines and antibacterial products, we've lost AOBs from our skin and also lost a key regulatory element from our skin microbiome."
http://mag.uchicago.edu/science-medicine/microbial-me
Microbial me
BY LYDIALYLE GIBSON | THE UNIVERSITY OF CHICAGO MAGAZINE—JULY–AUG/15
Scientists are discovering how microbes not only make us sick but also keep our bodies working.
Bacteria use antibiotics as a weapon and even produce more antibiotics if there are competing strains nearby. This is a fundamental insight that can help find new antibiotics. Leiden scientists Daniel Rozen and Gilles van Wezel published their research results in the authoritative Proceedings of the National Academy of Sciences USA on 28 July 2015.
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