Vitamin and Herb Stores
Health Research Report
156th Issue Date 1 Jun 2013
Compiled By Ralph Turchiano
www.facebook.com/engineeringevil
In this issue:
1. Coffee consumption associated with reduced risk of autoimmune liver disease
2. Ginger compounds may be effective in treating asthma symptoms
3. The compound in the Mediterranean diet that makes cancer cells ‘mortal’
4. Study finds vitamin C can kill drug-resistant TB
5. Common Food Supplement Fights Degenerative Brain Disorders
6. Calcium supplements linked to longer lifespans in women
7. Soda and illegal drugs cause similar damage to teeth
8. Changing gut bacteria through diet affects brain function, UCLA study shows
Coffee consumption associated with reduced risk of autoimmune liver disease
A range of new research studies presented at DDW® 2013
Orlando, FL (May 18, 2013) — Research presented today at Digestive Disease Week® (DDW) explores new discoveries in liver disease research, with findings about the impact of coffee on autoimmune disease and palliative care for cirrhotic patients.
While coffee consumption recently has been associated with reduced risk of fibrosis, a new study found that even a few more cups of java each month also correlate with lower risk for a particular autoimmune liver disease. Researchers at the Mayo Clinic, Rochester, MN, linked coffee consumption with reduced risk of primary sclerosing cholangitis (PSC), a disease of the bile ducts that causes inflammation and subsequent duct obstruction that ultimately can lead to cirrhosis of the liver, liver failure and biliary cancer.
“While rare, PSC has extremely detrimental effects,” said Craig Lammert, MD, instructor of medicine at Mayo Clinic. “We are always looking for ways to mitigate risk, and our first-time finding points to a novel environmental effect that might also help us to determine the cause of this and other devastating autoimmune diseases.”
Funded by grants from the National Institutes of Health and the American Liver Foundation, the study examined the largest cohort of patients with PSC and primary biliary cirrhosis (PBC) in the U.S. as well as a healthy control group. Data showed that coffee consumption was associated with reduced risk of PSC, but not PBC. PSC patients were much more likely to never consume coffee compared with the control group. The control group also spent nearly 20 percent more of their life regularly drinking coffee.
Study highlights need of terminally ill cirrhotic patients
Other DDW research illuminates the need for better palliative care for terminally ill cirrhotic patients who are rejected for a liver transplant. A retrospective cohort review of patients previously assessed or listed for liver transplant by the University of Alberta in Canada found that only 3 percent of patients in the study died while in hospice, a hallmark of palliative care.
“In our study, less than 10 percent of patients had even been referred to palliative care,” said Constantine Karvellas, assistant professor of medicine at the University of Alberta. “We need to be better about ensuring quality of life for these patients.”
Palliative care is specialized medical care for people with serious, often terminal, illnesses. Its goal is to improve patients’ quality of life by concentrating on relief from symptoms, pain and stress.
The patients in Dr. Karvellas’s study had been de-listed or declined for liver transplantation. The most common reason was noncompliance with restrictions on substance use, but other reasons related to cancer and multiple organ dysfunction. Researchers examined patients’ medical trajectory and the symptoms prominent at their end of life and found that more than half had pain and nausea. Others had symptoms of depression, anxiety, breathlessness and anorexia. Eighty percent required repeat hospital admissions and invasive procedures such as paracentesis, in which fluid accumulation is drained from the abdomen.
“Palliative care offers a way to avoid some of these costly procedures and at the same time improve the quality of life for these patients. These data help to start the conversation on how we can make a positive difference in the lives of many patients and families,” Dr. Karvellas said.
Ginger compounds may be effective in treating asthma symptoms
ATS 2013, PHILADELPHIA ─ Gourmands and foodies everywhere have long recognized ginger as a great way to add a little peppery zing to both sweet and savory dishes; now, a study from researchers at Columbia University shows purified components of the spicy root also may have properties that help asthma patients breathe more easily.
The results of the study will be presented at the ATS 2013 International Conference.
Asthma is characterized by bronchoconstriction, a tightening of the bronchial tubes that carry air into and out of the lungs. Bronchodilating medications called beta-agonists (β-agonists) are among the most common types of asthma medications and work by relaxing the airway smooth muscle (ASM) tissues. This study looked at whether specific components of ginger could help enhance the relaxing effects of bronchodilators.
“Asthma has become more prevalent in recent years, but despite an improved understanding of what causes asthma and how it develops, during the past 40 years few new treatment agents have been approved for targeting asthma symptoms,” said lead author Elizabeth Townsend, PhD, post-doctoral research fellow in the Columbia University Department of Anesthesiology. “In our study, we demonstrated that purified components of ginger can work synergistically with β-agonists to relax ASM.”
To conduct their study, the researchers took human ASM tissue samples and caused the samples to contract by exposing them to acetylcholine, a neurotransmitting compound that causes bronchoconstriction. Next, the researchers mixed the β-agonist isoproterenol with three separate components of ginger: 6-gingerol, 8-gingerol or 6-shogaol. Contracted tissue samples were exposed to each of these three mixtures as well as unadulterated isoproterenol and the relaxation responses were recorded and compared.
At the conclusion of their study, the researchers found that tissues treated with the combination of purified ginger components and isoproterenol exhibited significantly greater relaxation than those treated only with isoproterenol; of the three ginger components, 6-shogaol appeared most effective in increasing the relaxing effects of the β-agonist.
Once they were able to demonstrate that the ginger components enhanced the relaxing effects of the β-agonist, they turned their attention to learning why. First, the researchers wanted to determine if the ginger components might work by affecting an enzyme called phosphodiesterase4D (PDE4D). Previous studies have shown that PDE4D, which is found in the lungs, inhibits processes that otherwise help relax ASM and lessen inflammation. Using a technique called fluorescent polarization, they found that all three components significantly inhibited PDE4D.
Next, the study looked at F-actin filaments, a protein structure which previous studies have shown plays a role in the constriction of ASM, and found that 6-shogaol was effective in speedily dissolving these filaments.
“Taken together, these data show that ginger constituents 6-gingerol, 8-gingerol and 6-shogaol act synergistically with the β-agonist in relaxing ASM, indicating that these compounds may provide additional relief of asthma symptoms when used in combination with β-agonists,” Dr. Townsend noted.”By understanding the mechanisms by which these ginger compounds affect the airway, we can explore the use of these therapeutics in alleviating asthma symptoms.”
Dr. Townsend and her colleague, Dr. Charles Emala, hope future studies will enable them to gain a better understanding of the cellular mechanisms that facilitate ASM relaxation and to determine whether aerosol delivery of these purified constituents of ginger may have therapeutic benefit in asthma and other bronchoconstrictive diseases.
The compound in the Mediterranean diet that makes cancer cells ‘mortal’
Scientists design ‘fishing’ technique to show how foods improve health
COLUMBUS, Ohio – New research suggests that a compound abundant in the Mediterranean diet takes away cancer cells’ “superpower” to escape death.
By altering a very specific step in gene regulation, this compound essentially re-educates cancer cells into normal cells that die as scheduled.
One way that cancer cells thrive is by inhibiting a process that would cause them to die on a regular cycle that is subject to strict programming. This study in cells, led by Ohio State University researchers, found that a compound in certain plant-based foods, called apigenin, could stop breast cancer cells from inhibiting their own death.
Much of what is known about the health benefits of nutrients is based on epidemiological studies that show strong positive relationships between eating specific foods and better health outcomes, especially reduced heart disease. But how the actual molecules within these healthful foods work in the body is still a mystery in many cases, and particularly with foods linked to lower risk for cancer.
Parsley, celery and chamomile tea are the most common sources of apigenin, but it is found in many fruits and vegetables.
The researchers also showed in this work that apigenin binds with an estimated 160 proteins in the human body, suggesting that other nutrients linked to health benefits – called “nutraceuticals” – might have similar far-reaching effects. In contrast, most pharmaceutical drugs target a single molecule.
“We know we need to eat healthfully, but in most cases we don’t know the actual mechanistic reasons for why we need to do that,” said Andrea Doseff, associate professor of internal medicine and molecular genetics at Ohio State and a co-lead author of the study. “We see here that the beneficial effect on health is attributed to this dietary nutrient affecting many proteins. In its relationship with a set of specific proteins, apigenin re-establishes the normal profile in cancer cells. We think this can have great value clinically as a potential cancer-prevention strategy.”
Doseff oversaw this work with co-lead author Erich Grotewold, professor of molecular genetics and director of Ohio State’s Center for Applied Plant Sciences (CAPS). The two collaborate on studying the genomics of apigenin and other flavonoids, a family of plant compounds that are believed to prevent disease.
The research appears this week in the online early edition of the journal Proceedings of the National Academy of Sciences.
Though finding that apigenin can influence cancer cell behavior was an important outcome of the work, Grotewold and Doseff point to their new biomedical research technique as a transformative contribution to nutraceutical research.
They likened the technique to “fishing” for the human proteins in cells that interact with small molecules available in the diet.
“You can imagine all the potentially affected proteins as tiny fishes in a big bowl. We introduce this molecule to the bowl and effectively lure only the truly affected proteins based on structural characteristics that form an attraction,” Doseff said. “We know this is a real partnership because we can see that the proteins and apigenin bind to each other.”
Through additional experimentation, the team established that apigenin had relationships with proteins that have three specific functions. Among the most important was a protein called hnRNPA2.
This protein influences the activity of messenger RNA, or mRNA, which contains the instructions needed to produce a specific protein. The production of mRNA results from the splicing, or modification, of RNA that occurs as part of gene activation. The nature of the splice ultimately influences which protein instructions the mRNA contains.
Doseff noted that abnormal splicing is the culprit in an estimated 80 percent of all cancers. In cancer cells, two types of splicing occur when only one would take place in a normal cell – a trick on the cancer cells’ part to keep them alive and reproducing.
In this study, the researchers observed that apigenin’s connection to the hnRNPA2 protein restored this single-splice characteristic to breast cancer cells, suggesting that when splicing is normal, cells die in a programmed way, or become more sensitive to chemotherapeutic drugs.
“So by applying this nutrient, we can activate that killing machinery. The nutrient eliminated the splicing form that inhibited cell death,” said Doseff, also an investigator in Ohio State’s Davis Heart and Lung Research Institute. “Thus, this suggests that when we eat healthfully, we are actually promoting more normal splice forms inside the cells in our bodies.”
The beneficial effects of nutraceuticals are not limited to cancer, as the investigators previously showed that apigenin has anti-inflammatory activities.
The scientists noted that with its multiple cellular targets, apigenin potentially offers a variety of additional benefits that may even occur over time. “The nutrient is targeting many players, and by doing that, you get an overall synergy of the effect,” Grotewold explained.
Doseff is leading a study in mice, testing whether food modified to contain proper doses of this nutrient can change splicing forms in the animals’ cells and produce an anti-cancer effect.
Study finds vitamin C can kill drug-resistant TB
May 21, 2013 — (Bronx, NY) — In a striking, unexpected discovery, researchers at Albert Einstein College of Medicine of Yeshiva University have determined that vitamin C kills drug-resistant tuberculosis (TB) bacteria in laboratory culture. The finding suggests that vitamin C added to existing TB drugs could shorten TB therapy, and it highlights a new area for drug design. The study was published today in the online journal Nature Communications.
TB is caused by infection with the bacterium M. tuberculosis. In 2011, TB sickened some 8.7 million people and took some 1.4 million lives, according to the World Health Organization. Infections that fail to respond to TB drugs are a growing problem: About 650,000 people worldwide now have multi-drug-resistant TB (MDR-TB), 9 percent of whom have extensively drug-resistant TB (XDR-TB).TB is especially acute in low and middle income countries, which account for more than 95 percent of TB-related deaths, according to the World Health Organization.
The Einstein discovery arose during research into how TB bacteria become resistant to isoniazid, a potent first-line TB drug. The lead investigator and senior author of the study was William Jacobs, Jr. Ph.D., professor of microbiology & immunology and of genetics at Einstein. Dr. Jacobs is a Howard Hughes Medical Institute investigator and a recently elected member of the National Academy of Sciences.
Dr. Jacobs and his colleagues observed that isoniazid-resistant TB bacteria were deficient in a molecule called mycothiol. “We hypothesized that TB bacteria that can’t make mycothiol might contain more cysteine, an amino acid,” said Dr. Jacobs. “So, we predicted that if we added isoniazid and cysteine to isoniazid-sensitive M. tuberculosis in culture, the bacteria would develop resistance. Instead, we ended up killing off the culture— something totally unexpected.”
The Einstein team suspected that cysteine was helping to kill TB bacteria by acting as a “reducing agent” that triggers the production of reactive oxygen species (sometimes called free radicals), which can damage DNA.
“To test this hypothesis, we repeated the experiment using isoniazid and a different reducing agent— vitamin C,” said Dr. Jacobs. “The combination of isoniazid and vitamin C sterilized the M. tuberculosis culture. We were then amazed to discover that vitamin C by itself not only sterilized the drug-susceptible TB, but also sterilized MDR-TB and XDR-TB strains.”
To justify testing vitamin C in a clinical trial, Dr. Jacobs needed to find the molecular mechanism by which vitamin C exerted its lethal effect. More research produced the answer: Vitamin C induced what is known as a Fenton reaction, causing iron to react with other molecules to create reactive oxygen species that kill the TB bacteria.
“We don’t know whether vitamin C will work in humans, but we now have a rational basis for doing a clinical trial,” said Dr. Jacobs. “It also helps that we know vitamin C is inexpensive, widely available and very safe to use. At the very least, this work shows us a new mechanism that we can exploit to attack TB.”
Common Food Supplement Fights Degenerative Brain Disorders
Nutritional supplement delays advancement of Parkinson’s and Familial Dysautonomia, TAU researchers discover
Widely available in pharmacies and health stores, phosphatidylserine is a natural food supplement produced from beef, oysters, and soy. Proven to improve cognition and slow memory loss, it’s a popular treatment for older people experiencing memory impairment. Now a team headed by Prof. Gil Ast and Dr. Ron Bochner of Tel Aviv University‘s Department of Human Molecular Genetics has discovered that the same supplement improves the functioning of genes involved in degenerative brain disorders, including Parkinson’s disease and Familial Dysautonomia (FD).
In FD, a rare genetic disorder that impacts the nervous system and appears almost exclusively in the Ashkenazi Jewish population, a genetic mutation prevents the brain from manufacturing healthy IKAP proteins — which likely have a hand in cell migration and aiding connections between nerves — leading to the early degeneration of neurons. When the supplement was applied to cells taken from FD patients, the gene function improved and an elevation in the level of IKAP protein was observed, reports Prof. Ast. These results were replicated in a second experiment which involved administering the supplement orally to mouse populations with FD.
The findings, which have been published in the journal Human Molecular Genetics, are very encouraging, says Prof. Ast. “That we see such an effect on the brain — the most important organ in relation to this disease — shows that the supplement can pass through the blood-brain barrier even when administered orally, and accumulate in sufficient amounts in the brain.”
Slowing the death of nerve cells
Already approved for use as a supplement by the FDA, phosphatidylserine contains a molecule essential for transmitting signals between nerve cells in the brain. Prof. Ast and his fellow researchers decided to test whether the same chemical, which is naturally synthesized in the body and known to boost memory capability, could impact the genetic mutation which leads to FD.
Researchers applied a supplement derived from oysters, provided by the Israeli company Enzymotec, to cells collected from FD patients. Noticing a robust effect on the gene, including a jump in the production of healthy IKAP proteins, they then tested the same supplement on mouse models of FD, engineered with the same genetic mutation that causes the disease in humans.
The mice received the supplement orally, every two days for a period of three months. Researchers then conducted extensive genetic testing to assess the results of the treatment. “We found a significant increase of the protein in all the tissues of the body,” reports Prof. Ast, including an eight-fold increase in the liver and 1.5-fold increase in the brain. “While the food supplement does not manufacture new nerve cells, it probably delays the death of existing ones,” he adds.
Therapeutic potential for Parkinson’s
That the supplement is able to improve conditions in the brain, even when given orally, is a significant finding, notes Prof. Ast. Most medications enter the body through the blood stream, but are incapable of breaking through the barrier between the blood and the brain.
In addition, the researchers say the supplement’s positive effects extend beyond the production of IKAP. Not only did phosphatidylserine impact the gene associated with FD, but it also altered the level of a total of 2400 other genes — hundreds of which have been connected to Parkinson’s disease in previous studies.
The researchers believe that the supplement may have a beneficial impact on a number of degenerative diseases of the brain, concludes Prof. Ast, including a major potential for the development of new medications which would help tens of millions of people worldwide suffering from these devastating diseases
Calcium supplements linked to longer lifespans in women
Study finds calcium-rich diet, supplements provide equal benefits
Chevy Chase, MD—Taking a calcium supplement of up to 1,000 mg per day can help women live longer, according to a recent study accepted for publication in The Endocrine Society’s Journal of Clinical Endocrinology & Metabolism (JCEM).
Calcium, an essential nutrient for bone health, is commonly found in dairy products as well as vitamins. Although calcium is an essential nutrient for bone health, past studies have linked calcium supplements to heart disease risk. Researchers analyzing data from the large-scale Canadian Multicentre Osteoporosis Study (CaMos) sought to clarify this issue and found moderate doses of calcium supplements had a beneficial effect in women.
“Our study found daily use of calcium supplements was associated with a lower risk of death among women,” said the study’s lead author, David Goltzman, MD, of McGill University in Montreal, Canada. “The benefit was seen for women who took doses of up to 1,000 mg per day, regardless of whether the supplement contained vitamin D.”
The longitudinal cohort study monitored the health of 9,033 Canadians between 1995 and 2007. During that period, 1,160 participants died. Although the data showed women who took calcium supplements had a lower mortality risk, there was no statistical benefit for men. The study found no conclusive evidence that vitamin D had an impact on mortality.
“Higher amounts of calcium were potentially linked to longer lifespans in women, regardless of the source of the calcium,” Goltzman said. “That is, the same benefits were seen when the calcium came from dairy foods, non-dairy foods or supplements.”
Soda and illegal drugs cause similar damage to teeth
CHICAGO (May 28, 2013)—Addicted to soda? You may be shocked to learn that drinking large quantities of your favorite carbonated soda could be as damaging to your teeth as methamphetamine and crack cocaine use. The consumption of illegal drugs and abusive intake of soda can cause similar damage to your mouth through the process of tooth erosion, according to a case study published in the March/April 2013 issue of General Dentistry, the peer-reviewed clinical journal of the Academy of General Dentistry (AGD).
Tooth erosion occurs when acid wears away tooth enamel, which is the glossy, protective outside layer of the tooth. Without the protection of enamel, teeth are more susceptible to developing cavities, as well as becoming sensitive, cracked, and discolored.
The General Dentistry case study compared the damage in three individuals’ mouths—an admitted user of methamphetamine, a previous longtime user of cocaine, and an excessive diet soda drinker. Each participant admitted to having poor oral hygiene and not visiting a dentist on a regular basis. Researchers found the same type and severity of damage from tooth erosion in each participant’s mouth.
“Each person experienced severe tooth erosion caused by the high acid levels present in their ‘drug’ of choice—meth, crack, or soda,” says Mohamed A. Bassiouny, DMD, MSc, PhD, lead author of the study.
“The citric acid present in both regular and diet soda is known to have a high potential for causing tooth erosion,” says Dr. Bassiouny.
Similar to citric acid, the ingredients used in preparing methamphetamine can include extremely corrosive materials, such as battery acid, lantern fuel, and drain cleaner. Crack cocaine is highly acidic in nature, as well.
The individual who abused soda consumed 2 liters of diet soda daily for three to five years. Says Dr. Bassiouny, “The striking similarities found in this study should be a wake-up call to consumers who think that soda—even diet soda—is not harmful to their oral health.”
AGD Spokesperson Eugene Antenucci, DDS, FAGD, recommends that his patients minimize their intake of soda and drink more water. Additionally, he advises them to either chew sugar-free gum or rinse the mouth with water following consumption of soda. “Both tactics increase saliva flow, which naturally helps to return the acidity levels in the mouth to normal,” he says.
Changing gut bacteria through diet affects brain function, UCLA study shows
UCLA researchers now have the first evidence that bacteria ingested in food can affect brain function in humans. In an early proof-of-concept study of healthy women, they found that women who regularly consumed beneficial bacteria known as probiotics through yogurt showed altered brain function, both while in a resting state and in response to an emotion-recognition task.
The study, conducted by scientists with UCLA’s Gail and Gerald Oppenheimer Family Center for Neurobiology of Stress and the Ahmanson–Lovelace Brain Mapping Center at UCLA, appears in the June edition of the peer-reviewed journal Gastroenterology.
The discovery that changing the bacterial environment, or microbiota, in the gut can affect the brain carries significant implications for future research that could point the way toward dietary or drug interventions to improve brain function, the researchers said.
“Many of us have a container of yogurt in our refrigerator that we may eat for enjoyment, for calcium or because we think it might help our health in other ways,” said Dr. Kirsten Tillisch, an associate professor of medicine at UCLA’s David Geffen School of Medicine and lead author of the study. “Our findings indicate that some of the contents of yogurt may actually change the way our brain responds to the environment. When we consider the implications of this work, the old sayings ‘you are what you eat’ and ‘gut feelings’ take on new meaning.”
Researchers have known that the brain sends signals to the gut, which is why stress and other emotions can contribute to gastrointestinal symptoms. This study shows what has been suspected but until now had been proved only in animal studies: that signals travel the opposite way as well.
“Time and time again, we hear from patients that they never felt depressed or anxious until they started experiencing problems with their gut,” Tillisch said. “Our study shows that the gut–brain connection is a two-way street.”
The small study involved 36 women between the ages of 18 and 55. Researchers divided the women into three groups: one group ate a specific yogurt containing a mix of several probiotics — bacteria thought to have a positive effect on the intestines — twice a day for four weeks; another group consumed a dairy product that looked and tasted like the yogurt but contained no probiotics; and a third group ate no product at all.
Functional magnetic resonance imaging (fMRI) scans conducted both before and after the four-week study period looked at the women’s brains in a state of rest and in response to an emotion-recognition task in which they viewed a series of pictures of people with angry or frightened faces and matched them to other faces showing the same emotions. This task, designed to measure the engagement of affective and cognitive brain regions in response to a visual stimulus, was chosen because previous research in animals had linked changes in gut flora to changes in affective behaviors.
The researchers found that, compared with the women who didn’t consume the probiotic yogurt, those who did showed a decrease in activity in both the insula — which processes and integrates internal body sensations, like those form the gut — and the somatosensory cortex during the emotional reactivity task.
Further, in response to the task, these women had a decrease in the engagement of a widespread network in the brain that includes emotion-, cognition- and sensory-related areas. The women in the other two groups showed a stable or increased activity in this network.
During the resting brain scan, the women consuming probiotics showed greater connectivity between a key brainstem region known as the periaqueductal grey and cognition-associated areas of the prefrontal cortex. The women who ate no product at all, on the other hand, showed greater connectivity of the periaqueductal grey to emotion- and sensation-related regions, while the group consuming the non-probiotic dairy product showed results in between.
The researchers were surprised to find that the brain effects could be seen in many areas, including those involved in sensory processing and not merely those associated with emotion, Tillisch said.
The knowledge that signals are sent from the intestine to the brain and that they can be modulated by a dietary change is likely to lead to an expansion of research aimed at finding new strategies to prevent or treat digestive, mental and neurological disorders, said Dr. Emeran Mayer, a professor of medicine, physiology and psychiatry at the David Geffen School of Medicine at UCLA and the study’s senior author.
“There are studies showing that what we eat can alter the composition and products of the gut flora — in particular, that people with high-vegetable, fiber-based diets have a different composition of their microbiota, or gut environment, than people who eat the more typical
Western diet that is high in fat and carbohydrates,” Mayer said. “Now we know that this has an effect not only on the metabolism but also affects brain function.”
The UCLA researchers are seeking to pinpoint particular chemicals produced by gut bacteria that may be triggering the signals to the brain. They also plan to study whether people with gastrointestinal symptoms such as bloating, abdominal pain and altered bowel movements have improvements in their digestive symptoms which correlate with changes in brain response.
Meanwhile, Mayer notes that other researchers are studying the potential benefits of certain probiotics in yogurts on mood symptoms such as anxiety. He said that other nutritional strategies may also be found to be beneficial.
By demonstrating the brain effects of probiotics, the study also raises the question of whether repeated courses of antibiotics can affect the brain, as some have speculated. Antibiotics are used extensively in neonatal intensive care units and in childhood respiratory tract infections, and such suppression of the normal microbiota may have longterm consequences on brain development.
Finally, as the complexity of the gut flora and its effect on the brain is better understood, researchers may find ways to manipulate the intestinal contents to treat chronic pain conditions or other brain related diseases, including, potentially, Parkinson’s disease, Alzheimer’s disease and autism.
Answers will be easier to come by in the near future as the declining cost of profiling a person’s microbiota renders such tests more routine, Mayer said.
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