Podcasts about hepg2

Being AuDHD + gifted is being an outlier of outliers. I'd guess most of us feel like aliens. And even when we start to understand the shape of our experiences, it can feel overwhelming to even begin to order our understanding in a satisfying way. (Autoethnography, anyone?)Our depth and intensity needs can be quite challenging to meet, especially if we have lower physiological capacity due to trauma/CPTSD, and other intersectional identities that are often marginalized/mistreated.This became part one of two!Mentioned in episode: 1. Some giftedness models: Rainforest mindRuf's 5 levelsInterGifted & HEPG2. Neurocomplexity & PDA as existential intelligence3. Autism/ADHD/Gifted Venn Diagram4. Overexcitabilities & being "too much"5. Ember Green's quick IQ doesn't exist overview + Aspie Supremacy deep dive (p.s. this video is 2 hrs long and made me cry a LOT bc of historical horrors... still worth it if/when you have the spoons)Resources:Transcript DocEmail Newsletter: Nothing Wrong With UsLike Your Brain community space (Patreon) Hosted on Acast. See acast.com/privacy for more information.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.06.24.546395v1?rss=1 Authors: Tao, H., Dar, H. Y., Tian, C., Banerjee, S., Glazer, E. S., Srinivasan, S., Zhu, L., Pacifici, R., He, P. Abstract: Males show higher incidence and severity than females in hepatic injury and many liver diseases, but the mechanisms are not well understood. Ferroptosis, an iron-mediated lipid peroxidation-dependent death, plays an important role in the pathogenesis of liver diseases. We determined whether hepatocyte ferroptosis displays gender difference, accounting for sexual dimorphism in liver diseases. Compared to female hepatocytes, male hepatocytes were much more vulnerable to ferroptosis by iron and pharmacological inducers including RSL3 and iFSP1. Male but not female hepatocytes exhibited significant increases in mitochondrial Fe2+ and mitochondrial ROS (mtROS) contents. Female hepatocytes showed a lower expression of iron importer transferrin receptor 1 (TfR1) and mitochondrial iron importer mitoferrin 1 (Mfrn1), but a higher expression of iron storage protein ferritin heavy chain 1 (FTH1). It is well known that TfR1 expression is positively correlated with ferroptosis. Herein, we showed that silencing FTH1 enhanced while knockdown of Mfrn1 decreased ferroptosis in HepG2 cells. Removing female hormones by ovariectomy (OVX) did not dampen but rather enhanced hepatocyte resistance to ferroptosis. Mechanistically, OVX potentiated the decrease in TfR1 and increase in FTH1 expression. OVX also increased FSP1 expression in ERK-dependent manner. Elevation in FSP1 suppressed mitochondrial Fe2+ accumulation and mtROS production, constituting a novel mechanism of FSP1-mediated inhibition of ferroptosis. In conclusion, differences in hepatocellular iron handling between male and female account, at least in part, for sexual dimorphism in induced ferroptosis of the hepatocytes. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

A new research paper was published in Oncotarget's Volume 14 on March 21, 2023, entitled, “Attenuation of cancer proliferation by suppression of glypican-1 and its pleiotropic effects in neoplastic behavior.” Glypicans (GPC1-6) are associated with tumorigenic processes and their involvement in neoplastic behavior has been discussed in different cancer types. In this recent cancer-wide GPC expression study, researchers Fang Cheng, Victor Chérouvrier Hansson, Grigorios Georgolopoulos, and Katrin Mani from Lund University and Genevia Technologies used clinical cancer patient data in The Cancer Genome Atlas to reveal net upregulation of GPC1 and GPC2 in primary solid tumors. On the other hand, GPC3, GPC5 and GPC6 displayed lowered expression patterns compared to normal tissues. “[...] we identify and propose a mechanism where GPC1 interacts with extracellular matrix mediating signal transduction by mitogenic molecules involving TGF-β and p38 MAPK.” Focusing on GPC1, the researchers conducted survival analyses of the clinical cancer patient data that revealed a statistically significant correlation between high expression of GPC1 and poor prognosis in 10 particular cancer types: bladder urothelial carcinoma, brain lower grade glioma, liver hepatocellular carcinoma, colon adenocarcinoma, kidney renal clear cell carcinoma, lung adenocarcinoma, mesothelioma, ovarian serous cystadenocarcinoma, uterine corpus endometrial carcinoma, and uveal melanoma. In vitro studies targeting GPC1 expression by CRISPR/Cas9 or siRNA or treatment with an anti-GPC1 antibody resulted in attenuation of proliferation of cancer cells from bladder carcinoma, glioma and hepatocellular carcinoma patients (T24, U87 and HepG2 cells). Further, GPC1 overexpression exhibited a significant and negative correlation between GPC1 expression and proliferation of T24 cells. Their attempt to reveal the mechanism through which downregulation of GPC1 leads to attenuation of tumor growth using systematic Ingenuity Pathway Analysis indicated that suppression of GPC1 results in ECM-mediated inhibition of specific pro-cancer signaling pathways involving TGF-β and p38 MAPK. The team also identified differential expression and pleiotropic effects of GPCs in specific cancer types. This emphasizes their potential as novel diagnostic tools and prognostic factors, and open doors for future GPC targeted therapy. “It is plausible to measure circulating GPCs in serum, plasma or urine using a variety of methods including ELISA, urine cell sediments or exosome isolation [13, 24]. Further, detection and quantification of GPC1 by histopathological and immunohistochemical methods in tumor biopsies could be a new way to predict the biological outcome. The results of this investigation would also emphasize the potential of GPCs as novel tumor antigens, and open for GPC targeted immunotherapy. GPC targeted immunotherapy would be of high value, especially as we move into an era of precision and personalized cancer therapy.” DOI: https://doi.org/10.18632/oncotarget.28388 Correspondence to: Katrin Mani - katrin.mani@med.lu.se Keywords: Glypican-1, TCGA, bladder carcinoma, hepatocellular carcinoma, glioma About Oncotarget Oncotarget is a primarily oncology-focused, peer-reviewed, open access journal. Papers are published continuously within yearly volumes in their final and complete form, and then quickly released to Pubmed. On September 15, 2022, Oncotarget was accepted again for indexing by MEDLINE. Oncotarget is now indexed by Medline/PubMed and PMC/PubMed. To learn more about Oncotarget, visit https://www.oncotarget.com and connect with us: SoundCloud - https://soundcloud.com/oncotarget Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/@OncotargetJournal LinkedIn - https://www.linkedin.com/company/oncotarget MEDIA@IMPACTJOURNALS.COM

VIDEOS: Video Emerges Where Fauci and Others Planned for a “Universal mRNA Flu Vaccine” Which Became the “COVID-19 mRNA Vaccine” Because People were not Afraid Enough of the Flu Virus (1:51) You're Not Going To Believe This! | Mark Steyn & Eva Vlaardingerbroek (3:03) Neil Oliver – ‘…it's a toxic hell…' (START @ 9:00) Gravitas: Who helped Taliban repair the abandoned American aircraft? (7:25)   Healthy plant-based diets associated with lower colorectal cancer risk in men Kyung Hee University, South Korea, November 28, 2022 Eating a plant-based diet rich in healthy plant foods—such as whole grains, vegetables, and legumes—and low in unhealthy plant foods—including refined grains, fruit juices, and added sugars—is associated with a lower risk of colorectal cancer in men. The findings are published in the open access journal BMC Medicine.Jihye Kim, the corresponding author, said, “Colorectal cancer is the third-most common cancer worldwide, and the risk of developing colorectal cancer over a lifetime is one in 23 for men and one in 25 for women. Although previous research has suggested that plant-based diets may play a role in preventing colorectal cancer, the impact of plant foods' nutritional quality on this association has been unclear. Our findings suggest that eating a healthy plant-based diet is associated with a reduced risk of colorectal cancer.” Researchers from Kyung Hee University, South Korea found that among a population of 79,952 American men, those who ate the highest average daily amounts of healthy plant-based foods had a 22% lower risk of colorectal cancer, compared to those who ate the lowest amounts of healthy plant foods. However, the authors did not identify any significant associations between the nutritional quality of plant-based diets and colorectal cancer risk among a population of 93,475 American women. Jihye Kim said, “We speculate that the antioxidants found in foods such as fruits, vegetables, and whole grains could contribute to lowering colorectal cancer risk by suppressing chronic inflammation, which can lead to cancer. As men tend to have a higher risk of colorectal cancer than women, we propose that this could help explain why eating greater amounts of healthy plant-based foods was associated with reduced colorectal cancer risk in men but not women.” The authors found that the association between the nutritional quality of plant-based diets and colorectal cancer risk among men varied by race and ethnicity. Among Japanese American men, colorectal cancer risk was 20% lower for those who ate the highest amount of healthy plant foods per day than for those who ate the lowest amount. Among white men, those who ate the highest amount of highest amount of healthy plant foods had a 24% lower colorectal cancer risk than those who ate the lowest amount. The authors did not identify any significant associations between plant-based diets and colorectal cancer risk among African American, Latino or Native Hawaiian men. (next) Green Mediterranean diet reduces twice as much visceral fat as traditional Mediterranean diet Ben-Gurion University of the Negev (Israel), November 28, 2022 Following the green Mediterranean diet significantly reduces visceral adipose tissue, a type of fat around internal organs that is much more dangerous than the extra “tire” around your waist. Recently, researchers compared the green Mediterranean diet to the traditional Mediterranean diet and a non-Mediterranean healthy diet in a large-scale clinical interventional trial—the DIRECT PLUS. Subsequent analysis found that the green Mediterranean diet reduced visceral fat by 14%, the Mediterranean diet by 7% and the non-Mediterranean healthy diet by 4.5%. The study was published in BMC Medicine. Reducing visceral fat is considered the true goal of weight loss, as it is a more important indicator than a person's weight or the circumference of their waist. Visceral fat aggregates over time between organs, and produces hormones and poisons linked to heart disease, diabetes, dementia and premature death. The DIRECT-PLUS trial research team was the first to introduce the concept of the green Mediterranean diet. This modified Mediterranean diet is further enriched with dietary polyphenols and is lower in red/processed meat than the traditional Mediterranean diet. On top of a daily intake of walnuts (28 grams), the participants consumed 3-4 cups of green tea/day and 100 grams (frozen cubes) of duckweed green shake/day. The aquatic green plant duckweed is high in bioavailable protein, iron, B12, vitamins, minerals, and polyphenols and substituted meat intake. The team has shown in previous studies that the green Mediterranean diet has a variety of salutary effects ranging from the microbiome to age-related degenerative diseases. A group of 294 participants took part in the 18-month long trial. “A 14% reduction in visceral fat is a dramatic achievement for making simple changes to your diet and lifestyle. Weight loss is an important goal only if it is accompanied by impressive results in reducing adipose tissue,” notes Dr. Hila Zelicha. (next) Are older women being over-screened for cervical cancer? University of Illinois at Chicago, November 28, 2022 A new study published in JAMA Internal Medicine suggests that women over the age of 65 may be undergoing unnecessary cervical cancer screenings and that more public health data is needed on the utilization of cervical cancer screening-associated services among older women to prevent potential harm and unnecessary costs. The study, which is authored by experts from the University of Illinois Chicago, the University of California San Francisco and the U.S. Centers for Disease Control and Prevention, looked at Medicare claims data from 1999 to 2019 for fee-for-service care for women over the age of 65. The analysis showed that in 2019 more than 1.3 million women received cervical cancer screening-associated services, such as a Pap test, colposcopy, and other cervical procedures after age 65. While these services cost more than $83 million, the researchers concluded they were of “unclear clinical appropriateness.” “Cervical cancer screening and other preventive services are among our most important tools for keeping people healthy throughout life, but screenings should also follow evidence-based guidelines to prevent overspending, potential complications and patient discomfort,” said study co-author Dr. Hunter Holt, assistant professor of family and community medicine at the University of Illinois Chicago. According to recommendations and guidelines from the U.S. Preventive Services Task Force, the American Cancer Society and the American College of Obstetrics and Gynecology, women considered to be of average risk can stop undergoing routine cervical cancer screening once they reach the age of 65 if they have had adequate prior screening. “The decision to end cervical cancer screening for women after age 65 requires review of past screening results and related medical history. This process can promote cervical cancer prevention and prevent harms and costs from unnecessary tests and procedures,” said Jin Qin, study co-author and epidemiologist in CDC's Division of Cancer Prevention and Control. The researchers say that the high rates of screening among older women is potentially concerning. “It could be that women are getting screened when they do not need to, or that these women are considered to be at higher-than-average risk, for example, because they have not been adequately screened prior to 65. We do not want to see either of these things and unfortunately, there is not enough public health data to shed light on the causes,” said Holt, who is also affiliated with the University of Illinois Cancer Center at UIC. (next) Chemotherapy could increase disease susceptibility in future generations Washington State University, November 28, 2022 A common chemotherapy drug could carry a toxic inheritance for children and grandchildren of adolescent cancer survivors, Washington State University-led research indicates. The study, published online in iScience, found that male rats who received the drug ifosfamide during adolescence had offspring and grand-offspring with increased incidence of disease. While other research has shown that cancer treatments can increase patients' chance of developing disease later in life, this is one of the first-known studies showing that susceptibility can be passed down to a third generation of unexposed offspring. “The findings suggest that if a patient receives chemotherapy, and then later has children, that their grandchildren, and even great-grandchildren, may have an increased disease susceptibility due to their ancestors' chemotherapy exposure,” said Michael Skinner, a WSU biologist and corresponding author on the study. Given this study's implications, the researchers recommend that cancer patients who plan to have children later take precautions, such as using cryopreservation to freeze sperm or ova before having chemotherapy. In the study, researchers exposed a set of young male rats to ifosfamide over three days, mimicking a course of treatment an adolescent human cancer patient might receive. Those rats were later bred with female rats who had not been exposed to the drug. The resulting offspring were bred again with another set of unexposed rats. The first-generation offspring had some exposure to the chemotherapy drug since their fathers' sperm was exposed, but researchers found greater incidence of disease in not only the first- but also the second-generation, who had no direct exposure to the drug. While there were some differences by generation and sex, the associated problems included greater incidence of kidney and testis diseases as well as delayed onset of puberty and abnormally low anxiety, indicating a lowered ability to assess risk. The results of the researchers' analysis showed epigenetic changes in two generations linked to the chemotherapy exposure of the originally exposed rats. The fact that these changes could be seen in the grand-offspring, who had no direct exposure to the chemotherapy drug, indicates that the negative effects were passed down through epigenetic inheritance. (next) Saffron can fight liver cancer, reveal UAE researchers United Arab Emirates University, November 20, 2022 It may be an expensive spice but you cannot put a label or price on health, said Professor Amr Amin who has researched a breakthrough in the properties of saffron in fighting liver cancer. Professor Amin from Cellular & Molecular Biology at United Arab Emirates University said that researchers have investigated and found saffron to have anti-liver cancer properties. “Safranal, a major biomolecule of the golden spice saffron arrests and stops the cancer cell division at two different stages,” he said. The UAE researchers have been working on this project since 2011 when they first published the research in the Hepatology Journal. The study suggests a novel mechanism of anti-proliferative activity of safranal against human liver cancer cells. “This molecule could serve as a novel and/or adjuvant drug to treat liver cancer,” said Dr Amin. The findings are now also published in a Nature journal Scientific Reports. “The ingredient works in two ways; it stops cell division and promotes cell death,” he explained. Prof Amin and colleagues concluded that safranal exerts its anticancer effect in HepG2 cells by inhibiting DNA repair, resulting in increased DNA damage. (next) Japanese researchers say that ultrasound therapy can be used to treat patients with dementia Tohoku University (Japan), November 20, 2022 A new therapy based on ultrasound waves might be able to improve the cognitive powers of patients suffering from Alzheimer's disease and other forms of dementia. According to an article on the Tohoku University news page, the approach improved the condition of mice with symptoms similar to human dementia. In their experiment, the Tohoku University research team sent low-intensity pulsed ultrasound (LIPUS) waves through the brain of the mice. They found that the waves improved the creation of blood vessels and the rate of regeneration of nerve cells. Furthermore, the treatment did not cause any notable side effects on the mice. The results led the researchers to believe that they can replicate their experimental success in actual human patients one day. “The LIPUS therapy is a non-invasive physiotherapy that could apply to high-risk elderly patients without the need for surgery or anaesthesia, and could be used repeatedly,” explained TU researcher Hiroaki Shimokawa. The Tohoku researchers applied LIPUS therapy to the whole brain of mice with symptoms that resemble those of Alzheimer's disease or vascular dementia. They did this three times a day, with each session lasting for 20 minutes. The mice that simulated vascular dementia underwent surgery that reduced the amount of blood that reached the brain. These animals underwent LIPUS treatment on the first, third, and fifth days after that surgery. Meanwhile, the mice that modeled Alzheimer's disease got 11 LIPUS treatments over a three-month-long trial period. By the end of the experiment, the researchers found that LIPUS activated genes involved with the cells that made up the inner lining of blood vessels. Furthermore, an enzyme that promoted blood vessel formation displayed increased activity, as did a protein which helped nerve cells grow. Based on their findings, whole-brain LIPUS therapy can help alleviate the symptoms of certain forms of dementia by encouraging the development of cells that are normally affected by the condition. The technique is currently undergoing initial clinical trials that will determine its efficacy and safety.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.22.517223v1?rss=1 Authors: Calla, J., Mittal, N., LaMonte, G., Liffner, B., Godinez-Macias, K., Carolino, K., Walker, G. T., Zou, B. Y., Paytas, E., Guerra, L., Tong-Rios, C., Campo, B., Vinetz, J. M., Gamboa, D., Raffatellu, M., Absalon, S., Winzeler, E. Abstract: Changes in host cell morphology and transcription after apicomplexan parasite infection have long been noted, but there have been few studies of the functional consequences of host cell remodeling. Here we show, using time-dependent immunofluorescence microscopy of multiple human cell lines (HFF, HepG2, HC-04, Huh7.5.1 and primary human hepatocytes), infected with multiple Plasmodium species (Plasmodium berghei, P. falciparum and P. vivax (hypnozoites and schizonts)), and antibodies to multiple human proteins (HsNR4A3, HsMUC13, HsGOLGA8A, HsCGA, HsBiP, HsCXCL2), that human protein trafficking is extensively modified in Plasmodium infected cells. Using conventional as well as ultrastructure expansion immunofluorescence microscopy we show that newly-synthesized human proteins are trafficked to the parasitophorous vacuole instead of the infected-cell plasma membrane, nucleus or extracellular space. Universal redirection of human signaling proteins cells the parasitophorous vacuole may provide a mechanistic explanation for how apicomplexan parasites can block host cells response to infection. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.05.515282v1?rss=1 Authors: Lujan, A. L., Foresti, O., Brouwers, N., Mateo Farre, A., Vignoli, A., Wojnacki, J., Malhotra, V. Abstract: We show that TANGO2, which lacks a transmembrane domain localizes predominantly to mitochondria and transiently to endoplasmic reticulum (ER) and lipid droplets (LDs). Evaluation of lipids in HepG2 cells lacking TANGO2 revealed an increase in the size of lipid droplets and reactive oxygen species production. There is also a marked increase lysophosphatidic acid (LPA) and a concomitant decrease in its biosynthetic precursor phosphatidic acid (PA). These changes are exacerbated in nutrient starved cells. Based on our data, we suggest that the principle function of TANGO2 is in acyl-CoA metabolism, which is necessary for the acylation of LPA to generate PA. This defect subsequently affects metabolism of many other fatty acids. These data help explain the physiological consequence of TANGO2 that induce acute metabolic crisis including rhabdomyolysis, cardiomyopathy and cardiac arrhythmias often leading to fatality upon starvation and stress. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Red and processed meat linked to increased risk of heart disease, study shows Oxford University, July 21, 2021 Globally, coronary heart diseases (caused by narrowed arteries that supply the heart with blood) claim nearly nine million lives each year1, the largest of any disease, and present a huge burden to health systems. Until now, it has been unclear whether eating meat increases the risk of heart disease, and if this varies for different kinds of meat. Researchers at the University of Oxford's Nuffield Department of Population Health have conducted the largest systematic review of the prospective evidence to date, including thirteen cohort studies involving over 1.4 million people. The study participants completed detailed dietary assessments, and their health was tracked for up to 30 years. The results are published today in Critical Reviews in Food Science and Nutrition. Overall, the evidence from the analysis indicated that: Each 50 g/day higher intake of processed meat (e.g. bacon, ham, and sausages) increased the risk of coronary heart disease by 18%. Each 50 g/day higher intake of unprocessed red meat (such as beef, lamb and pork) increased the risk of coronary heart disease by 9%. There was no clear link between eating poultry (such as chicken and turkey) and an increased risk of coronary heart disease. The findings may be because of the high content of saturated fat in red meat, and of sodium (salt) in processed meat. High intakes of saturated fat increase levels of harmful low-density lipoprotein (LDL) cholesterol, whilst excess salt consumption raises blood pressure. Both LDL cholesterol and high blood pressure are well-established risk factors for coronary heart disease. Previous work from the same research team has also indicated that even moderate intakes of red and processed meat are associated with increased risk of bowel cancer2. Dr. Keren Papier (Nuffield Department of Population Health), co-lead author of the study, said: "Red and processed meat have been consistently linked with bowel cancer and our findings suggest an additional role in heart disease. Therefore, current recommendations to limit red and processed meat consumption may also assist with the prevention of coronary heart disease." Dr. Anika Knüppel, from the Nuffield Department of Population Health and the other co-lead author of the study, added: "We know that meat production is a major contributor to greenhouse gas emissions and we need to reduce meat production and thereby consumption to benefit the environment. Our study shows that a reduction in red and processed meat intake would bring personal health benefits too." Currently in the UK, about 10 in 100 people would be expected to eventually die from coronary heart disease. Based on the findings from the present study and current red and processed meat intakes in the UK,4 if all these 100 people reduced their unprocessed red meat intake by three-quarters (for example from four times a week to one time a week), or if they stopped consuming processed meat altogether, deaths from coronary heart disease would decrease from 10 in 100 down to 9 in 100. The studies involved in this analysis were mostly based on white adults living in Europe or the U.S.. The research team say more data are needed to examine these associations in other populations, including East Asia and Africa.   C is for Vitamin C -- a key ingredient for immune cell function Harnessing the combined power of Vitamin C and TET proteins may give scientists a leg up in treating autoimmune diseases   La Jolla Institute for Immunology and Emory University, July 22, 2021 You can't make a banana split without bananas. And you can't generate stable regulatory T cells without Vitamin C or enzymes called TET proteins, it appears.  Regulatory T cells (Tregs) help control inflammation and autoimmunity in the body. Tregs are so important, in fact, that scientists are working to generate stable induced Tregs (iTregs) in vitro for use as treatments for autoimmune diseases as well as rejection to transplanted organs. Unfortunately, it has proven difficult to find the right molecular ingredients to induce stable iTregs. Now scientists at La Jolla Institute for Immunology and Emory University School of Medicine report that Vitamin C and TET proteins can work together to give Tregs their life-saving power.  "Vitamin C can be used to stabilize iTregs generated in vitro," says LJI Instructor Xiaojing Yue, Ph.D., who served as co-first author for the EMBO Reports study. "We hope that these kinds of induced Tregs can be used in the future for treatment of autoimmune diseases and organ transplantation." The recent study, led by LJI Professor Anjana Rao, Ph.D., and Emory Instructor Benjamin G Barwick, Ph.D., builds on the previous discovery that Vitamin C can enhance the enzymatic activity of TET proteins and prompt the generation of stable iTregs under lab conditions. This finding was encouraging, but the scientists did not want to work toward new autoimmune therapies without first analyzing the gene expression patterns and other key epigenetic features of the induced Tregs.  "We wanted to study the entire system at a whole genome level using next generation sequencing technology to better understand the molecular features of these cells," says Yue. Study co-first author Daniela Samaniego-Castruita, a graduate student at LJI, spearheaded the analysis of gene expression and epigenetic changes in the iTregs. A major type of epigenetic modification involves the DNA itself through the addition or removal of molecules called methyl groups from cytosines, one of the four DNA bases. The methyl groups can be further oxidized by TET enzymes. All of these interactions can eventually change how cells "read" the DNA code.  Another type of epigenetic change involves the alteration of DNA accessibility: whether DNA is loosely or tightly coiled. As the DNA coils unwind, regulatory regions become exposed which subsequently influence gene expression. In their analysis, the researchers found TET proteins are absolutely required for maintaining the gene expression and epigenetic features that make Tregs as what they are; and adding Vitamin C led to iTregs with similar similar gene expression and epigenetic features as normal "wild type" Tregs found in the body. The study also reveals an intriguing connection between TET enzymatic activity, Vitamin C and IL-2/STAT5 signaling. "In mice that are deficient for components of IL-2/STAT5 signaling, such as IL-2, IL-2 receptors or STAT5, the Tregs cannot develop properly or they can have impaired function," Yue says. The researchers demonstrate that on one hand, TET-deficiency in Treg cells leads to impaired IL-2/STAT5 signaling; on the other hand, Vitamin C confers iTregs enhanced IL-2/STAT5 signaling by increasing the expression level of IL-2 receptor and the functional form of STAT5, and STAT5 binding to essential regions in the genome, rendering these cells survive better in tough environments with low IL-2 supplementation. "We are looking for more small molecules to stabilize TET activity and generate induced Tregs that are even more stable," says Yue. "These induced Tregs could eventually be used to treat patients." "This research gives us a new way to think about treating autoimmune diseases," says Samaniego-Castruita.       Resveratrol ameliorates high-fat-diet-induced abnormalities in liver glucose metabolism in mice via the AMPK pathway Hebei Medical Institute (China), July 19, 2021 A new study on high fat diet is now available. According to news originating from the Department of Internal Medicine by NewsRx correspondents, research stated, “Diabetes mellitus is highly prevalent worldwide.” Our news reporters obtained a quote from the research from Department of Internal Medicine: “High-fat-diet (HFD) consumption can lead to liver fat accumulation, impair hepatic glycometabolism, and cause insulin resistance and the development of diabetes. Resveratrol has been shown to improve the blood glucose concentration of diabetic mice, but its effect on the abnormal hepatic glycometabolism induced by HFD-feeding and the mechanism involved are unknown. In this study, we determined the effects of resveratrol on the insulin resistance of high-fat-diet-fed mice and a hepatocyte model by measuring serum biochemical indexes, key indicators of glycometabolism, glucose uptake, and glycogen synthesis in hepatocytes. We found that resveratrol treatment significantly ameliorated the HFD-induced abnormalities in glucose metabolism in mice, increased glucose absorption and glycogen synthesis, downregulated protein phosphatase 2A (PP2A) and activated Ca2+/CaM-dependent protein kinase kinase b (CaMKKb), and increased the phosphorylation of AMP-activated protein kinase (AMPK). In insulin-resistant HepG2 cells, the administration of a PP2A activator or CaMKKb inhibitor attenuated the effects of resveratrol, but the administration of an AMPK inhibitor abolished the effects of resveratrol. Resveratrol significantly ameliorates abnormalities in glycometabolism induced by HFD-feeding and increases glucose uptake and glycogen synthesis in hepatocytes.” According to the news editors, the research concluded: “These effects are mediated through the activation of AMPK by PP2A and CaMKKb.”     Hundreds of chemicals, many in consumer products, could increase breast cancer risk List includes potential carcinogens that act by stimulating production of hormones that fuel breast tumors Silent Spring Institute, July 22, 2021 Every day, people are exposed to a variety of synthetic chemicals through the products they use or the food they eat. For many of these chemicals, the health effects are unknown. Now a new study shows that several hundred common chemicals, including pesticides, ingredients in consumer products, food additives, and drinking water contaminants, could increase the risk of breast cancer by causing cells in breast tissue to produce more of the hormones estrogen or progesterone. "The connection between estrogen and progesterone and breast cancer is well established," says co-author Ruthann Rudel, a toxicologist and research director at Silent Spring Institute. "So, we should be extremely cautious about chemicals in products that increase levels of these hormones in the body." For instance, in 2002, when the Women's Health Initiative study found combination hormone replacement therapy to be associated with an increased risk of breast cancer, women stopped taking the drugs and incidence rates went down. "Not surprisingly, one of the most common therapies for treating breast cancer is a class of drugs called aromatase inhibitors that lower levels of estrogen in the body, depriving breast cancer cells of the hormones they need to grow," adds Rudel. To identify these chemical risk factors, Rudel and Silent Spring scientist Bethsaida Cardona combed through data on more than 2000 chemicals generated by the U.S. Environmental Protection Agency (EPA)'s ToxCast program. The goal of ToxCast is to improve the ability of scientists to predict whether a chemical will be harmful or not. The program uses automated chemical screening technologies to expose living cells to chemicals and then examine the different biological changes they cause. Reporting in the journal Environmental Health Perspectives, Rudel and Cardona identified 296 chemicals that were found to increase estradiol (a form of estrogen) or progesterone in cells in the laboratory. Seventy-one chemicals were found to increase levels of both hormones. The chemicals included ingredients in personal care products such as hair dye, chemical flame retardants in building materials and furnishings, and a number of pesticides. The researchers don't yet know how these chemicals are causing cells to produce more hormones. It could be the chemicals are acting as aromatase activators, for instance, which would lead to higher levels of estrogen, says Cardona. "What we do know is that women are exposed to multiple chemicals from multiple sources on a daily basis, and that these exposures add up." The Silent Spring researchers hope this study will be a wakeup call for regulators and manufacturers in how they test chemicals for safety. For instance, current safety tests in animals fail to look at changes in hormone levels in the animal's mammary glands in response to a chemical exposure. And, although high throughput testing in cells has been used to identify chemicals that activate the estrogen receptor, mimicking estrogen, the testing has not been used to identify chemicals that increase estrogen or progesterone synthesis. "This study shows that a number of chemicals currently in use have the ability to manipulate hormones known to adversely affect breast cancer risk," says Dr. Sue Fenton, associate editor for the study and an expert in mammary gland development at the National Institute of Environmental Health Sciences. "Especially concerning is the number of chemicals that alter progesterone, the potential bad actor in hormone replacement therapy. Chemicals that elevate progesterone levels in the breast should be minimized." The researchers outlined a number of recommendations in their study for improving chemical safety testing to help identify potential breast carcinogens before they end up in products, and suggest finding ways to reduce people's exposures, particularly during critical periods of development, such as during puberty or pregnancy when the breast undergoes important changes. The project is part of Silent Spring Institute's Safer Chemicals Program which is developing new cost-effective ways of screening chemicals for their effects on the breast. Knowledge generated by this effort will help government agencies regulate chemicals more effectively and assist companies in developing safer products.     Antioxidant activity of limonene counteracts neurotoxicity triggered by amyloid beta 1-42 oligomers in cortical neurons University of Naples (Italy), July 19, 2021 According to news reporting from Naples, Italy, by NewsRx journalists, research stated, “Many natural-derived compounds, including the essential oils from plants, are investigated to find new potential protective agents in several neurodegenerative disorders such as Alzheimer's disease (AD).” The news editors obtained a quote from the research from School of Medicine: “In the present study, we tested the neuroprotective effect of limonene, one of the main components of the genus * * Citrus* * , against the neurotoxicity elicited by Ab [ [1-42] ] oligomers, currently considered a triggering factor in AD. To this aim, we assessed the acetylcholinesterase activity by Ellman's colorimetric method, the mitochondrial dehydrogenase activity by MTT assay, the nuclear morphology by Hoechst 33258, the generation of reactive oxygen species (ROS) by DCFH-DA fluorescent dye, and the electrophysiological activity of K [ [V] ] 3.4 potassium channel subunits by patch-clamp electrophysiology. Interestingly, the monoterpene limonene showed a specific activity against acetylcholinesterase with an IC [ [50] ] almost comparable to that of galantamine, used as positive control. Moreover, at the concentration of 10 g/mL, limonene counteracted the increase of ROS production triggered by Ab [ [1-42] ] oligomers, thus preventing the upregulation of K [ [V] ] 3.4 activity. This, in turn, prevented cell death in primary cortical neurons, showing an interesting neuroprotective profile against Ab [ [1-42] ] -induced toxicity.” According to the news editors, the research concluded: “Collectively, the present results showed that the antioxidant properties of the main component of the genus * * Citrus* * , limonene, may be useful to prevent neuronal suffering induced by Ab [ [1-42] ] oligomers preventing the hyperactivity of K [ [V] ] 3.4.”     Meditation And Yoga Change Your DNA To Reverse Effects Of Stress, Study Shows Coventry University (UK), July 22, 2021 Many people participate in practices such as meditation and yoga because they help us relax. At least those are the immediate effects we feel. But much more is happening on a molecular level, reveal researchers out of Coventry University in England. Published in the journal Frontiers in Immunology, this new research examined 18 studies on mind-body interventions (MBIs). These include practices such as mindfulness meditation and yoga. Comprehensively, these studies encompassed 846 participants over 11 years. The new analysis reveals that MBIs result in molecular changes in the human body. Furthermore, researchers claim that these changes are beneficial to our mental and physical health. Body's Response to Stress Causes Damage To elaborate, consider the effect that stress has on the body. When we are under stress, the body increases the production of proteins that cause cell inflammation. This is the natural effect of the body's fight-or-flight response. It is widely believed that inflammation in the body leads to numerous illnesses, including cancer. Moreover, scientists also deduct that a persistent inflammation is more likely to cause psychiatric problems. Unfortunately, many people suffer from persistent stress, therefore they suffer from pro-inflammatory gene expression. But there is good news! According to this new analysis out of Coventry, people that practice MBIs such as meditation and yoga can reverse pro-inflammatory gene expression. This results in a reduced risk of inflammation-related diseases and mental conditions. Lead investigator Ivana Buric from Coventry University's Centre for Psychology, Behaviour and Achievement stated: Millions of people around the world already enjoy the health benefits of mind-body interventions like yoga or meditation, but what they perhaps don't realise is that these benefits begin at a molecular level and can change the way our genetic code goes about its business. These activities are leaving what we call a molecular signature in our cells, which reverses the effect that stress or anxiety would have on the body by changing how our genes are expressed. Put simply, MBIs cause the brain to steer our DNA processes along a path which improves our wellbeing. More needs to be done to understand these effects in greater depth, for example how they compare with other healthy interventions like exercise or nutrition. But this is an important foundation to build on to help future researchers explore the benefits of increasingly popular mind-body activities.               Large-scale study finds greater sedentary hours increases risk of obstructive sleep apnea Study finds that maintaining an active lifestyle can reduce the risk of OSA, encourages physicians to recommend exercise-based interventions for those at risk Brigham and Women's Hospital, July 22, 2021 A new study by investigators from Brigham and Women's Hospital examined the relationship between active lifestyles and the risk of obstructive sleep apnea (OSA). The study followed around 130,000 men and women in the United States over a follow-up period of 10-to-18 years and found that higher levels of physical activity and lower levels of sedentary behavior were associated with a lower risk of OSA. Their results are published in the European Respiratory Journal.  "In our study, higher levels of physical activity and fewer hours of TV watching, and sitting either at work or away from home were associated with lower OSA incidence after accounting for potential confounders," said Tianyi Huang, MSc, ScD, an Associate Epidemiologist at the Brigham. "Our results suggest that promoting an active lifestyle may have substantial benefits for both prevention and treatment of OSA."  OSA is a type of sleep apnea in which some muscles relax during sleep, causing an airflow blockage. Severe OSA increases the risk of various heart issues, including abnormal heart rhythms and heart failure.  Using the Nurses' Health Study (NHS), Nurses' Health Study II (NHSII) and Health Professionals Follow-Up Study (HPFS), the research team used statistical modeling to compare physical activity and sedentary hours with diagnoses of OSA. Both moderate and vigorous physical activity were examined separately and both were strongly correlated with lower risk of OSA, showing no appreciable differences in the intensity of activity. Moreover, stronger associations were found for women, adults over the age of 65 and those with a BMI greater than or equal to 25 kg/m2.  "Most prior observational studies on the associations of physical activity and sedentary behavior with OSA were cross-sectional, with incomplete exposure assessment and inadequate control for confounding," said Huang. "This is the first prospective study that simultaneously evaluates physical activity and sedentary behavior in relation to OSA risk."  This study also differs from others because of its large sample size and detailed assessment pf physical activity and sedentary behaviors. The research team was able to take many associated factors into account, making the findings more credible.  The authors note that all collected data, both of OSA diagnosis and physical activity or sedentary behavior, were self-reported. While all study participants were health professionals, mild OSA is often difficult to detect and can remain clinically unrecognized. Furthermore, only recreational physical activity was taken into consideration, leaving out any physical activity in occupational settings. Sedentary behavior was only counted as sitting while watching TV and sitting away from home or at work.  According to Huang, the next research steps would be to collect data using actigraphy, home sleep apnea tests and polysomnography, rather than self-reports.  In light of the findings, investigators encourage physicians to highlight the benefits of physical activity to lower OSA risk.  "We found that physical activity and sedentary behavior are independently associated with OSA risk," said Huang. "That is, for people who spend long hours sitting every day, increasing physical activity in their leisure time can equally lower OSA risk. Similarly, for those who are not able to participate in a lot of physical activity due to physical restrictions, reducing sedentary hours by standing or doing some mild activities could also lower OSA risk. However, those who can lower sedentary time and increase physical activity would have the lowest risk."

Greater adherence to Mediterranean diet associated with better cognitive performance in older individuals University of South Australia, July 13, 2021 According to news originating from Adelaide, Australia, by NewsRx correspondents, research stated, “Adherence to a Mediterranean diet is associated with higher cognitive function and reduced risk of dementia in Mediterranean populations. However, few studies have investigated the association between Mediterranean diet adherence and cognition in populations outside of the Mediterranean basin.” Our news journalists obtained a quote from the research from the University of South Australia, “Furthermore, it is currently unknown whether the association between Mediterranean diet adherence and cognitive function differs between middle-aged and older individuals. Cross-sectional (n = 894) and longitudinal (n = 530) multivariable analyses were undertaken using data from community-dwelling adults from the Maine-Syracuse Longitudinal Study (MSLS). Mediterranean diet adherence was measured by applying a literature-based Mediterranean diet score to food frequency questionnaire data. Cognitive function was assessed with a battery of tests and composites scores were computed for global cognitive function, Visual-Spatial Organisation and Memory, verbal memory, working memory, scanning and tracking and abstract reasoning. No cross-sectional associations between Mediterranean diet adherence and cognitive function were detected. Over a period of five years, higher adherence to a Mediterranean diet was associated with improvements in Global Cognitive Function, Visual-Spatial Organisation and Memory and scanning and tracking in participants >= 70 years.” According to the news editors, the research concluded: “No significant longitudinal associations were observed for participants Conclusion: Our findings suggest that higher adherence to a Mediterranean diet is associated with better cognitive performance, and therefore less cognitive decline, in older but not middle-aged individuals.” This research has been peer-reviewed.     Coffee and veggies may protect against COVID-19 Northwestern University, July 20, 2021 Sip a venti dark roast and eat a salad. A new Northwestern Medicine study shows coffee consumption and eating lots of vegetables may offer some protection against COVID-19. The authors believe this is the first study using population data to examine the role of specific dietary intake in prevention of COVID-19. "A person's nutrition impacts immunity," said senior author Marilyn Cornelis, associate professor of preventive medicine at Northwestern University Feinberg School of Medicine. "And the immune system plays a key role in an individual's susceptibility and response to infectious diseases, including COVID-19." Being breastfed may also offer protection as well as eating less processed meats, the study found. "Besides following guidelines currently in place to slow the spread of the virus, we provide support for other relatively simple ways in which individuals can reduce their risk and that is through diet and nutrition," Cornelis said.  The paper on nutrition and COVID-19 protection was published recently in the journal Nutrients. One or more cups of coffee per day was associated with about a 10% decrease in risk of COVID-19 compared to less than one cup per day. Consumption of at least 0.67 servings per day of vegetables (cooked or raw, excluding potatoes) was associated with a lower risk of COVID-19 infection. Processed meat consumption of as little as 0.43 servings per day was associated with a higher risk of COVID-19. Having been breastfed as a baby reduced the risk 10% compared to not having been breastfed.  While the study shows diet appears to modestly reduce disease risk, the Centers for Disease Control and Prevention recommends vaccines as the most effective way to prevent COVID-19 disease, especially severe illness and death. COVID-19 vaccines also reduce the risk of people spreading the virus that causes COVID-19. Thus far, most COVID-19 research has focused on individual factors assessed after a positive COVID-19 test. Individuals with suppressed immune systems such as the elderly and those with existing comorbidities including cardiovascular diseases, hypertension, diabetes and obesity, are more likely to experience severe outcomes of COVID-19.  But other than weight management, less attention has focused on other modifiable risk factors preceding COVID-19 infection, said Cornelis, who studies how diet and nutrition contribute to chronic disease.  Dr. Thanh-Huyen Vu, the study's first author and a research associate professor of medicine at Northwestern, is now leading analyses to determine whether these protective diet behaviors are specific to COVID or respiratory infections more broadly.  Exact mechanisms linking these diet factors to COVID are unknown.  "Coffee is a major source of caffeine, but there are also dozens of other compounds that may potentially underlie the protective associations we observed," Cornelius said. "Associations with processed meat, but not red meat, point to non-meat factors." Using data from the UK Biobank, researchers examined the associations between dietary behaviors measured in 2006-2010 and COVID-19 infections in March to December 2020, before vaccines were available. They focused on 1) diet factors for which data were available and previously implicated in immunity based on human and animal studies; 2) self-reported intakes of coffee, tea, vegetables, fruit, fatty fish, processed meat and red meat. An early-life exposure to breastmilk also was analyzed.  Among the 37,988 participants tested for COVID-19 and included in the study, 17% tested positive.  The observational nature of the UK Biobank research limits the extent to which mechanisms of protection can be tested, Cornelis said. However, much of her nutrition research uses genetics, and with all UK Biobank participants currently genotyped, she hopes to use this information to gain better insight into how diet and nutrition offer protection from the disease.   Biologic age reversed with lifestyle improvement plus supplements Institute for Functional Medicine (Seattle), July 14 2021.  The April 15, 2021 issue of Aging published the results of an eight-week randomized trial which resulted in a reduction in biologic age among men who participated in lifestyle changes and consumed nutritional supplements. "The combined intervention program was designed to target a specific biological mechanism called DNA methylation, and in particular the DNA methylation patterns that have been identified as highly predictive of biological age,” explained lead author Kara Fitzgerald, ND. “We suspect that this focus was the reason for its remarkable impact.”  The trial included 38 men between the ages of 50 and 72 years. Eighteen participants consumed a plant-based, low carbohydrate diet that included limited nutrient-dense animal proteins. The diet was supplemented with a vegetable and fruit powder and the probiotic Lactobacillus plantarum 299v. The group was advised to participate in a minimum of 30 minutes of exercise daily and to perform breathing exercises twice per day for stress reduction.  According to the Horvath DNAmAge clock, which evaluates DNA methylation patterns as a marker of biologic age, men who participated in the lifestyle program had scores that averaged 1.96 younger at the end of the program in comparison with the beginning, while control participants scored 1.27 years older. Additionally, triglycerides were reduced in the lifestyle program group. “To our knowledge, this is the first randomized controlled study to suggest that specific diet and lifestyle interventions may reverse Horvath DNAmAge epigenetic aging in healthy adult males,” the authors announced.  “These early results appear to be consistent with, and greatly extend, the very few existing studies that have so far examined the potential for biological age reversal,” Dr Fitzgerald commented. “And it is unique in its use of a safe, non-pharmaceutical dietary and lifestyle program, control group, and the extent of the age reduction."       Research suggests L-carnitine could aid burn recovery Anhui Medical University (China), July 12, 2021 According to news reporting from First Affiliated Hospital of Anhui Medical University research stated, “Impaired hepatic fatty acid metabolism and persistent mitochondrial dysfunction are phenomena commonly associated with liver failure. Decreased serum levels of L-carnitine, a amino acid derivative involved in fatty-acid and energy metabolism, have been reported in severe burn patients.” Our news editors obtained a quote from the research from First Affiliated Hospital of Anhui Medical University: “The current study aimed to evaluate the effects of L-carnitine supplementation on mitochondrial damage and other hepatocyte injuries following severe burns and the related mechanisms. Serum carnitine and other indicators of hepatocytic injury, including AST, ALT, LDH, TG, and OCT, were analyzed in severe burn patients and healthy controls. A burn model was established on the back skin of rats; thereafter, carnitine was administered, and serum levels of the above indicators were evaluated along with Oil Red O and TUNEL staining, transmission electron microscopy, and assessment of mitochondrial membrane potential and carnitine palmitoyltransferase 1 (CPT1) activity and expression levels in the liver. HepG2 cells pretreated with the CPT1 inhibitor etomoxir were treated with or without carnitine for 24 h. Next, the above indicators were examined, and apoptotic cells were analyzed via flow cytometry. High-throughput sequencing of rat liver tissues identified several differentially expressed genes (Fabp4, Acacb, Acsm5, and Pnpla3) were confirmed using RT-qPCR. Substantially decreased serum levels of carnitine and increased levels of AST, ALT, LDH, and OCT were detected in severe burn patients and the burn model rats. Accumulation of TG, evident mitochondrial shrinkage, altered mitochondrial membrane potential, decreased ketogenesis, and reduced CPT1 activity were detected in the liver tissue of the burned rats. Carnitine administration recovered CPT1 activity and improved all indicators related to cellular and fatty acid metabolism and mitochondrial injury. Inhibition of CPT1 activity with etomoxir induced hepatocyte injuries similar to those in burn patients and burned rats; carnitine supplementation restored CPT1 activity and ameliorated these injuries. The expression levels of the differentially expressed genes Fabp4, Acacb, Acsm5, and Pnpla3 in the liver tissue from burned rats and etomoxir-treated hepatocytes were also restored by treatment with exogenous carnitine.” According to the news editors, the research concluded: “Exogenous carnitine exerts protective effects against severe burn-induced cellular, fatty-acid metabolism, and mitochondrial dysfunction of hepatocytes by restoring CPT1 activity.”     Red blood cell ‘traffic' contributes to changes in brain oxygenation   Penn State University, July 19, 2021 Adequate blood flow supplies the brain with oxygen and nutrients, but the oxygenation tends to fluctuate in a distinct, consistent manner. The root of this varied activity, though, is poorly understood. Now, Penn State researchers have identified one cause of the fluctuations: inherent randomness in the flow rate of red blood cells through tiny blood vessels called capillaries. According to the researchers, this randomness could have potential implications for understanding the biological build-up mechanisms underlying neurodegenerative diseases, such as Alzheimer's disease. They published their findings in PLOS Biology today. “These oxygenation fluctuations also occur in other tissues, like muscle,” said Patrick Drew, Huck Distinguished Associate Professor of Engineering Science and Mechanics, Neurosurgery and Biomedical Engineering. “The question we had was: Are these fluctuations caused by neural activity or something else?” The fluctuations resemble 1/f-like noise, a statistical pattern showing large fluctuations made up of many small fluctuations and naturally occurring in a variety of phenomena, from stock-market prices to river heights. The researchers investigated the fluctuations in mice due to their brains' similarities to those of humans, according to Drew, who also serves as associate director of the Penn State Neuroscience Institute. First, the researchers monitored the blood flow, oxygenation and electrical signals produced by brain activity—the first time the latter two had been tracked simultaneously, according to Drew—in awake mice. They collected the data as mice moved on a spherical treadmill for up to 40 minutes at a time. Next, to investigate the relationship between brain activity and oxygenation fluctuations, the researchers used pharmacological compounds to temporarily and reversibly silence neural signals in the mice's brains. Despite the silencing, the fluctuations continued, showing little correlation between neural activity and oxygenation. The passage of red blood cells, however, told a different story. Using two-photon laser scanning microscopy, an imaging technique used to visualize cells deep inside living tissue, the researchers could visualize the passage of individual red blood cells through capillaries. “It's like traffic,” Drew said. “Sometimes there are a lot of cars going by, and the traffic gets plugged up, and sometimes there aren't. And red blood cells go either way when they approach a junction, so this random flow can lead to bottlenecks and stalls in the vessel.” Importing experimental data into a statistical model allowed the researchers to run further simulations and make inferences based on massive amounts of data produced by the model. The researchers discovered that these random red blood cell stoppages contributed to the fluctuations in oxygenation, further supporting a relationship between the flow of red blood cells through capillaries and the tiny changes in oxygenation that formed larger trends. Better understanding the regulation of blood flow and subsequent transport of oxygen can help researchers improve medical technology and explore causes of diseases such as Alzheimer's, according to Drew. While the researchers identified the link between red blood cell transport and oxygenation, further research is needed to investigate additional contributors to oxygenation fluctuations that could play a role in neurodegenerative diseases. Kyle Gheres, a graduate student in the intercollege Graduate Program in Molecular Cellular and Integrative Biosciences, also contributed to this paper. Qingguang Zhang, assistant research professor of engineering science and mechanics, served as first author on the paper. This work was supported by the National Institutes of Health.     EGCG in Green Tea inhibits the growth of breast cancer cells Chonbuk National University School of Medicine (S Korea), July 21, 2021   Findings on Breast Cancer Reported by Researchers at Chonbuk National University School of Medicine(Epigallocatechin gallate inhibits the growth of MDA-MB-231 breast cancer cells via inactivation of the b-catenin signaling pathway) According to news reporting originating in Chonbuk, South Korea, research stated, "Epigallocatechin gallate (EGCG), a major constituent of green tea, has potential as a treatment for a variety of diseases, including cancer. EGCG induces apoptosis and inhibits tumorigenesis through multiple signaling pathways in breast cancer cells. b-catenin signaling modulators could be useful in the prevention and therapy of breast cancer." The news reporters obtained a quote from the research from the Chonbuk National University School of Medicine, "However, the precise anticancer effect of EGCG through the b-catenin signaling pathway in breast cancer is unclear. The present study investigated the association between b-catenin expression and clinicopathological factors of breast cancer patients, and the effect of EGCG on b-catenin expression in breast cancer cells. b-catenin expression was analyzed according to the clinicopathological factors of 74 patients with breast cancer. All patients were females diagnosed with invasive ductal carcinoma. Western blot analysis revealed that b-catenin was expressed at higher levels in breast cancer tissue than in normal tissue. b-catenin expression was associated with lymph node metastasis (p=0.04), tumor-node-metastasis stage (p=0.03) and estrogen receptor status (p

In this episode, Dr. Donohoe and Dr. Panno discuss two of the most commonly used interventions, both in mainstream medical and functional medicine fields, Metformin and Berberine. Dr. Panno recently wrote a paper on this topic where he read through numerous research articles in order to gain more clarity and understanding of these two products (how they work and how effective they are). We get into the science behind them but also the clinical takeaways and what it all means. Below we reference all the articles used.    PLEASE SUBSCRIBE TO THE PODCAST For more Diabuddies content follow us on The Diabuddies Podcast Facebook page. Twitter: @TheDiabuddies Instagram: @thediabuddiespodcast You can email us at TheDiabuddiesPodcast@gmail.com  Time Stamps 1:10 - Start/Introduction 6:25 - Berberine: What Does It Do And How Effective Is It? 16:30 - Where Does Berberine Come From? 22:45 - Where Does Metformin Come From? 31:30 - How Does Metformin Work? (MOA) 39:50 - How Does Berberine Work? (MOA) 44:10 - Can you take Metformin and Berberine Together? 57:30 - Burst My Beta Cells   Resources/Links Discussed in the episode: References Arayne, M., Sultana, N., & Bahadur, S., (2007). The berberis story: berberis vulgaris in therapeutics. Park J Pharm Sci; 20(1): 83-92. PMID: 17337435 Bailey, C., & Day, C., (2004). Metformin: its botanical background. Pract Diabetes Int; 21: 115-117. https://doi.org/10.1002/pdi.606 Bailey, C., & Day, C., (1989). Traditional plant medicines as treatments for diabetes. Diabetes Care; 12(8): 553-564. PMID: 2673695 Center of Disease Control (2020). National Diabetes Statistic Report 2020, Estimates of Diabetes and its burden in the united states. Retrieved at: https://www.cdc.gov/diabetes/pdfs/data/statistics/national-diabetes-statistics-report.pdf Chang, W., Zhang, M., Li, J., Meng, Z., Wei, S., Du, H., Chen, L., & Hatch, G., (2013). Berberine improves insulin resistance in cardiomyocytes via activation of 5'adenosine monophosphate-activated protein kinase. Metabolism; 62(8): 1159-1167. PMID: 23537779 Chang, W., Chen, L., & Hatch, G., (2015). Berberine as a therapy for type 2 diabetes and its complication: from mechanism of action to clinical studies. Biochem Cell Biol; 93(5): 479-486. PMID: 19800084 Cheuh, W., & Lin, J., (2012). Protective effect of berberine on serum glucose levels in non-obese diabetic mice. International Immunopharmacology; 12(3): 534-538. PMID: 22266065 Choi, YH., & Lee, MG., (2006). Effects of enzyme inducers and inhibitors on the pharmacokinetics of metformin in rats: involvement of CYP2C11, 2D1, 3A1/2 for the metabolism of metformin. B J Pharmacol; 149(4): 424-30. PMID: 16940989 ClinCal (2020). The Top 200 Drugs of 2021. Medical Expenditure Panel Survey (MEPS). Retrieved at: https://clincalc.com/DrugStats/Top200Drugs.aspx Dong, H., Wang, H., Zhao, L., & Lu, F., (2012). Berberine in the treatment of type 2 diabetes mellitus: a systemic review and meta-analysis. Evidence Based complement alternative med; 2012: 591654. PMID: 23118793 Elbere, I., Kalnina, I., Silamikelis, I., Konrade, I., Zaharenko, L., Sekace, K., et al., Klovins, J., (2018). Association of metformin administration with gut microbiome dysbiosis in healthy volunteers. PLoS One; 13(9): e0204317. PMID: 30261008 Foretz, M., Herbrad, S., Leclerc, J., Zarrinpashneh, E., Soty, M., Mithieux, G., et al., & Viollet, B., (2010). Metformin inhibits hepatic gluconeogenesis in mice independently of LKB1/AMPK pathway via a decrease in hepatic energy state. The Journal of Clinical investigation; 120(7): 2355-2369. PMID: 20577053 Fullerton, M., Galic, S., Marcinko, K., Sikkema, S., Pulinikunnil, T., Chen, Z., et al., & Steinberg, G., (2013). Single phosphorylation sites in ACC1 and ACC2 regulate lipid homeostasis in the insulin-sensitizing effects of metformin. Nat Med; 19(12): 1649-1654. PMID: 24185692 Lan, J., Zhao, Y., Dong, F., Yan, Z., Zheng, W., Fan, J., & Sun, G., (2015). Meta-analysis of the effect and safety of berberine in the treatment of type 2 diabetes mellitus, hyperlipemia, and hypertension. Journal of Ethnopharmacology; 16H: 69-81. PMID: 25498346 Le Bastard, Q., Grégoire, M., Chapelet, G., Javaudin, F., Dailly, E., Batard, E., Montassier, E., Al, G. G. A., & Knights, D. (2018). Systematic review: human gut dysbiosis induced by non‐antibiotic prescription medications. Alimentary Pharmacology & Therapeutics, 47(3), 332. PMID: 29205415  Li, Y., Wang, Y., Kong, W., Yang, P., Wang, Y., Li, Y., Yi, H., et al., & Jiang, J., (2011). Bioactivates of berberine metabolites after transformation throughout CYP450 isoenzymes. Journal of translational medicine; 9(62): 62. PMID: 21569619 Liang, Y., Xu, X., Yin, M., Zhang, Y., Haung, L., Chen, R., & Ni, J., (2019). Effects of berberine on blood glucose in patients with type 2 diabetes mellitus: a systematic literature review and meta-analysis. Endocrine Journal; 66(1): 51-63. PMID: 30393248 Lyu, Y., Zhang, Y., Yang, M., Lin, L., Yang, X., Cheung, S., et al., & Zuo, Z., (2019). Pharmacokinetic interactions between metformin and berberine in rats: Role of oral administration sequences and microbiota. Life Sciences; 235: 116818. PMID: 31473193 Mao, L., Chen, Q., Gong, K., Xu, X., Xie, Y., Zhang W., et al., & Zhang, Y., (2018). Berberine decelerates glucose metabolism via suppression of mTOR-dependent HIF-1α protein synthesis in colon cancer cells. Oncol Rep; 39(5). 2436-2442. PMID: 29565467 Miller, R., Chu, Q., Xie, J., Foretz, M., Viollet, B., & Birnbaum, M., (2013). Biguanides suppress hepatic glucagon signaling by decreasing production of cyclic AMP. Nature; 494(7436):256-260. PMID: 23292513 Neag, M., Mocan, A., Echeverria, J., Pop, R., Bocsan, C., Crisan, G., & Buzoianu, A., (2018). Berberine: Botanical Occurrence, Traditional Uses, Extraction Methods, and Relevance in Cardiovascular, Metabolic, Hepatic, and Renal Disorders. Front Pharmacology; 9: 557. PMID: 30186157 Owen, M., Doran, E., & Halestrap, A., (2000). Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondria respiratory chain. Biochem J; 348; 607-614. PMID: 10839993 Petersen, M., American Diabetes Association. Economic Costs of Diabetes in the U.S. in 2017. Diabetes care; 41(5): 917-928. PMID: 29567642 Pryor R., & Cabreiro F., (2015). Repurposing metformin: an old drug with new tricks in its binding pockets. Biochemical Journal; 471(3): 307-322. PMID: 26475449 Ren, G., Guo, J., Quian, Y., Kong, W., & Jiang, J., (2020). Berberine improves glucose and lipid metabolism in HepG2 cells through AMPKα1 activation. Front Pharmacology; 11: 647. PMID: 32457629 Yin, J., Xing, H., & Ye, J., (2008). Efficacy of berberine in patients with type 2 diabetes mellitus. 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ENCODEプロジェクトにおけるサイレンサー探索の論文紹介と現代生物学の複雑性と多様性について話しました。Shownotes ENCODE…ENCODEとはEncyclopedia of DNA Elementsの略！ ENCODE (Wikipedia) ヒトゲノムの機能解明に向けたENCODEの試み (pdf) Initial sequencing and analysis of the human genome. Nature 2001…ヒトゲノム解読の論文。 The Sequence of the Human Genome. Science 2001…ヒトゲノム解読の論文。 ENCODE 4…ENCODEのPhase 4についてはここにaimなどが書いてある。 ENCODE3勉強会…Sohも論文読みをしました。 ENCODE 3 paper collection…ENCODE 3の論文リストはここにあります。 DNase I hypersensitive site (Wikipedia) Systematic identification of silencers in human cells. Nature Genetics, 2020…今回Sohが解説する論文はこれです。　 転写制御因子 (脳科学辞典) … プロモーターやエンハンサー、サイレンサーまわりについて。遺伝子発現とは、遺伝子というDNA上にコードされた情報がいかにRNA、タンパク質として実体を伴って機能するかということであるのだが、この量を調節する機能、とくにDNA->RNAの段階(転写)で調節することを転写制御という。ここで重要となるのがプロモーターやエンハンサー、サイレンサー、さらにはインスレーターなどである。 遺伝子発現制御機構: クロマチン, 転写制御, エピジェネティクス (Amazon) … tadasuはこの本で一度転写周りを勉強しなおしたが、自分が思っていたプロモーターやエンハンサーと実際の定義はかなりズレがあったことを思い知らされた。 STARR-seq (Wikipedia)…ゲノム中に含まれるエンハンサーやプロモーター配列をバーコードと次世代シーケンサーを用いて大量にアッセイする方法の一つ。オリジナルの論文はここ: Genome-Wide Quantitative Enhancer Activity Maps Identified by STARR-seq, Science 2013 Caspase-9 (Wikipedia)…Caspase9はアポトーシスを担う重要なタンパク質の一つ。 EF1aプロモーター…ヒトやマウスの細胞の中で高い活性を持つ恒常発現プロモーターのうちの一つ。CMVプロモーターも使われることが多い。 サイレンサー…哺乳動物細胞における転写を制御する配列のうち、転写活性に対して抑制的に働く配列を指しています。恒常発現プロモーターの下流にあるとそのさらに下流にある遺伝子発現を強く抑制する配列（200bp程度）を指しています。 ハウスキーピング遺伝子…細胞機能を維持するために常に発現している遺伝子。これを「恒常的な遺伝子発現」と呼んだりします。 K562…ヒト慢性骨髄性白血病由来の培養細胞株。 293T…ヒト胎児腎由来の培養細胞株。 HepG2…ヒト肝癌由来の細胞株。 ルシフェラーゼアッセイ pGL4.51[luc2/CMV/Neo] Vector…スクリーニングで見つかったサイレンサーはこのベクターを用いてルシフェラーゼアッセイによってその活性が調べられた。 ヒストン修飾 (Abcam) ヒストン (Wikipedia) Purification of Proteins Associated with Specific Genomic Loci. Cell, 2009.…PICh法を用いるとゲノムに結合したタンパク質を同定することができる。 Hi-C…Hi-Cについてはエピソード9などで話しました。 ChiA-PET (Wikipedia) FANTOM…“FANTOMは、理化学研究所のマウスゲノム百科事典プロジェクトで収集された完全長cDNAのアノテーション（機能注釈）を行うことを目的に、林崎良英博士が中心となり2000年に結成された国際研究コンソーシアムです” NIH Roadmap Epigenomics Mapping Consortium…ヒトのエピジェネティクスに関連する大量のデータを計測しアトラスを作ることを目的とした国際コンソーシアム。 Illumina Body Map 2.0 Human Cell Atras…ヒトの体を構成する全ての細胞の分類とマッピングを目指す国際共同プロジェクト。 4D Nucleome Project…染色体構造のマッピングのためのコンソーシアム。 The 4D nucleome project. Nature, Dekker at al 2017 The Cancer Genome Atlas Program (TCGA)…がんゲノムの大量のデータを収集している国際プロジェクト。 ゲノム編集とはなにか (ブルーバックス) (Amazon)…山本先生が書かれたゲノム編集について非常にわかりやすい本が出ております。ぜひ。 ep47, Researchat.fm…エピソード47ではCas13を用いた微量の核酸検出SHERLOCKやDETECTRについて紹介しています。 Advances in Chromatin and Chromosome Research: Perspectives from Multiple Fields. Molecular Cell, 2020 … tadasuが参加しているBoston Chromatin Clubで書いた筆頭著者17人のレビュー。 ep37, Biological Enigma, Researchat.fm…セントラルドグマを含む分子生物学の基礎についても話しました。 ep7, In the golden age of molecular biology … 遺伝暗号解明周りの研究について話しました。 ep9, One-shot beautiful experiment … 細胞系譜について ep2, An emerging technology is always not perfect … CRISPRとゲノム編集周辺について話しました。 Gene drive (Wikipedia) … 遺伝子ドライブ ワトソンの遺伝子の分子生物学 (Amazon) 後成説 (Wikipedia) … エピジェネシス。アリストテレスから始まる発生の理論。 前成説 (Wikipedia) 分子進化のほぼ中立説 偶然と淘汰の進化モデル (ブルーバックス) (Amazon) ep36, DNA-of-things … DNAを練りこんだ3Dプリンティング技術について話しました。 情報量 … 情報量、エントロピー シャノンの情報理論入門 (ブルーバックス) (Amazon) ファインマンの計算機科学 (Amazon) … tadasuおすすめの情報理論の教科書。ゲノムやセントラルドグマの話についての情報理論についても記述がある(あった気がする)。 Developmental Biology (Amazon) … 発生のGilbert本 Principles of Development (Amazon) … 発生のWolpert本 Molecular Biology of the Cell (NCBI) Shownotes 正直、プロモーターとかエンハンサーとかサイレンサーって言っていますが、実際にプロモーターとエンハンサーの違いなどよく考えるとわからなくなってきます。とても難しい。雰囲気でやっています。(soh) データは山のようにある。あとは君たち次第だ！(いいのかそれで？)(tadasu) 作業が楽しくても3日以上寝ないと空から奴らが襲ってくるので要注意です(coela)

Speaker 1:           Hi, everyone. Welcome to episode 21 of Getting Personal, Omics of the Heart from October 2018. I'm Jane Ferguson, an Assistant Professor at Vanderbilt University Medical Center and an Associate Editor at Circulation: Genomic and Precision Medicine. We have a great issue this month. So, let's dive straight in. First up, an article on "Loss-of-Function ABCC8 Mutations in Pulmonary Arterial Hypertension" from Michael Bohnen, Wendy Chung and colleagues from Columbia University. In pulmonary arterial hypertension, or PAH, compromised pulmonary arterial function can raise pressure in the pulmonary artery which leads to increased pulmonary vascular resistance. This ultimately results in right heart failure. While PAH is relatively rare, it has a high rate of mortality. Some genetic underpinnings have been identified, notably the KCNK3 gene identified by the same research group where they find that mutations result in potassium channelopathy. However, here the authors hypothesized that other genetic contributors also exist and that identification of these could highlight new therapeutic targets to improve treatment and outcomes in PAH. In their study, the authors performed exome sequencing for discovery of novel disease variants in 233 PAH patients, 99 of whom had pediatric-onset and 134 with adult-onset. They sequenced a replication sample of 680 individuals with adult-onset PAH. They found a de novo missense variant in the ABCC8 gene in one patient and then found 10 more ABCC8 variants in other unrelated patients in the discovery and replication samples. Half of these were novel mutations and all were located in conserved regions and predicted to be deleterious. They screened over 33,000 subjects from the Exome Aggregation Consortium and over 49,000 from the Regeneron-Geisinger DiscovEHR study and found significant overrepresentation on rare ABCC8 variants in the PAH cases compared with population controls. ABCC8 encodes sulfonylurea receptor ... part of the potassium ATP channel. The authors determined that it is expressed in lungs in both PAH and healthy individuals and is particularly localized to alveolar macrophages and proximal pulmonary arteries. They expressed eight of the newly discovered ABCC8 mutations in COS cells, which are a monkey-derived, fiberglass-like cell line and they assessed the effects on function. They used patch-clamp experiments to assess potassium ATP channel activity and recorded efflux rates of Rubidium-86. Every mutation was associated with impairments in one or both functional assays, suggesting that mutations in ABCC8 are responsible for PAH by a modulating potassium channel function and flux. An existing drug, Diazoxide, targets ABCC8 and has anti-hypertensive and insulin-lowering effects. The authors find that all mutants were pharmacologically activated by Diazoxide in the functional assays. Now, whether this drug would be safe or effective in PAH remains unknown, but these findings open up targeting of ABCC8 as a possible treatment in PaH and highlight the importance of potassium channels in PAH. Our next paper also used whole-exome sequencing for novel discovery. Marzia de Bortoli, Alessandra Rampazza and colleagues from the University of Padua in Italy published "Whole-Exome Sequencing Identifies Pathogenic Variants in TJP1 Gene Associated With Arrhythmogenic Cardiomyopathy". Arrhythmogenic Cardiomyopathy, or ACM, is one of the most common causes of sudden unexpected death in athletes and young people. It is known to be frequently caused by mutations in genes encoding mechanical junction proteins of the intercalated disks within the cardiac muscle. However, some individuals with ACM do not have any mutations in known genes. This research group was interested in finding novel causal gene mutation and they use whole-exome sequencing to identify mutations from a single patient in Italy. They used InSilica tools to screen for potentially damaging mutations which brought their list of candidate mutations down to 52 and this was topped by a novel mutation in the TJP1 gene which was predicted to be highly deleterious using various algorithms. Using Sanger sequencing, they found that this mutation was also present in several family members. A second mutation in TJP1, also predicted to be damaging, was identified in a second Italian family. They then screened a sample of 43 Dutch and German subjects diagnosed with ACM and found that, once again, mutations in TJP1 topped the list as predicted to be damaging. The TJP1, or tight junction protein 1, encodes the intercalated disk proteins ZO1. The identified mutations may affect folding and local interactions within the protein, affecting protein-protein interactions and gap junction organization. Well, within this paper, they were not able to fully disentangle the mechanisms linking these mutations to disease, given that the prevalence of TJP1 mutations in their ACM samples was almost 5%. Screening for TJP1 mutations in ACM cohorts may identify many additional affected subjects. Further research into TJP1 is needed to identify how these variants may cause ACM. If you want to read more about this paper, you can check out the accompanying editorial from Jason Roberts ... Western University, Ontario ... in this same issue. Next up is a paper from Natsuko Tamura, Yasuhiro Maejima, Mitsuaki Isobe and colleagues from Tokyo Medical and Dental University entitled "Single-nucleotide Polymorphism of the MLX Gene Is Associated With Takayasu Arteritis". Takayasu Arteritis, or TAK, is an autoimmune disease causing aortic vasculitis that is poorly understood and disproportionately affects young Asian women. In previous genome-wide associations, study of TAK in Japanese individuals conducted by this group, indicated SNPs in the MLX gene. In this paper, the authors aim to identify mechanisms linking MLX mutations with TAK. The top GWAS SNP rs665268 is a missense mutation causing L-Glutamine Arginine substitution in the DNA binding site of MLX. They found that this SNP was associated with severity in disease in TAK. With additional copies of the risk alleles associated with more severe aortic regurgitation and greater number arterial lesions. In mice, the highest expression of MLX was found in the aortic valves. Using crystallography, they found that the missense mutation likely stabilizes a complex formed between MLX and MondoA. Immunoprecipitation experiments confirmed that the missense mutation was associated with enhanced MLX MondoA heterodimer formation and MLX transcriptional activity. This resulted in upregulation of TXNIP and higher TXNIP expression is associated with increased intracellular oxidative stress and the authors found for increased oxidative stress in cells carrying the MLX mutation. Further, additional cell experiments showed evidence of this MLX mutation reduces autophagy and stimulates inflammasome activation. Overall, through a series of really elegant experiments, the authors demonstrate that a missense mutation in MLX leads to inflammasome activation and accumulation of cells within the aorta, potentially underlying the pathophysiology seen in TAK patients and highlighting novel causal pathways that may be probed therapeutically.regular Our next paper from Danxin Wang, Wolfgang Sadee and colleagues from the University of Florida and The Ohio State University, also delves into the functional impact of disease-associated SNPs. In their paper, "Interactions Between Regulatory Variants in CYP7A1 Promoter and Enhancer Regions Regulate CYP7A1 Expression", they used a series of experiments to demonstrate how SNPs in CYP7A1 ... which have been associated with cholesterol and cardiovascular disease ... are related to gene function. CYP7A1 is a gene which coordinates a key pathway for cholesterol removal from the body because it encodes an enzyme which is rate-limiting for bioassay synthesis from cholesterol. Although several SNPs in the gene have been associated with cardiovascular phenotypes, the reported effects on gene function have been inconsistent and/or unclear. Because of the linkage disequilibrium between SNPs, it has been hard to understand which SNP or SNPs are actually functional. What this team set out to do was to systematically screen functionality of individual CYP7A1 SNPs to understand the independent effects of each functional variant. First, they used chromatin conformation capture, or 4C assay, to identify regions that associated with a CYP7A1 promoter. They found three distinct regions with evidence of enhancer function and [phonetic 00:09:05] active A>G regulation. They, next, used CRISPR Cas9 to delete each of the three regions in HepG2 cells and assess effects on CYP7A1 expression. One region had no effect, while one led to increased expression and one led to decreased expression ... thus, identifying the presence of both enhancer and repressor regions. Using reporter gene assays, they confirmed the effects seen in CRISPR experiments. Based on reported SNP associations, they narrowed down candidate functional SNPs within the regions and constructed reporter assays containing haplotypes of potential functional SNPs. They were able to identify two SNPs acting together to determine differences in CYP7A1 gene expression. Because these SNPs are in LD, but the minor alleles have effects in opposite directions, considering genotype at both SNPs is required to understand the effects on gene expression. This explains why previous studies found inconsistent results. Both during the functional experiments, they went to human samples and they assessed the combined effect of the two SNPs on clinical phenotypes. Designating people as high or low activity based on the two SNPs, they found significant differences in cholesterol and in the likelihood to reach cholesterol targets on statin, as well as in the risk of MI. This paper is a lovely example of how careful functional interrogation can tease out a complex problem and I think it highlights how much more of this type of work needs to be done for the many other genomic regions with confusing or discord in associations. The last full-length article concerns the "Effect of Ascertainment Bias on Estimates of Patient Mortality in Inherited Cardiac Diseases" and comes from Eline Nannenberg, Imke Christiaans and colleagues at the Academic Medical Center, Amsterdam. They were interested in how much ascertainment bias and the tendency to publish findings from more severe disease cases affects the mortality estimates that are used to guide clinicians and genetic counselors when helping patients understand their disease prognosis. They revisited three inherited cardiac diseases including idiopathic ventricular fibrillation associated with a mutation in DPP6, SCN5A overlap syndrome associated with SCN5A mutations, and Arrhythmogenic Cardiomyopathy caused by a founder PLN mutation. They analyzed mortality over 2-10 years of clinical screening and cascade screening and found that the median age of survival quickly increased in all three conditions. In many cases, the reason that a mutation was identified was because of severe disease in that patient or family, but as the authors highlight here, this can bias publications towards associating the variant with more severe phenotypes and higher mortality. Following up the initial findings with additional screening and tracking of affected individuals is important to subsequently give a more accurate estimation of the effect of the mutation which can be used to inform treatment plans. Moving on to this month's research letters, Catherine Hajek, Jerome Rotter and colleagues from LA BioMed and the University of South Dakota, published the results of their study, "A Coronary Heart Disease Genetic Risk Score Predicts Cardiovascular Disease Risk in Men, Not Women: The Multi-Ethnic Study of Atherosclerosis". The genetic risk scores are being increasingly applied to estimate disease risk in individuals. However, these scores are based on the GWAS discovery from specific populations which have often been disproportionately male and with individuals of European ancestry. In this letter, the authors wanted to understand whether coronary heart disease genetic risk scores performed the same in men and women of European ancestry. Using data from the MESA Study, they applied a 46 locus genetic risk score to over 2500 individuals. In men, this risk score was strongly associated with event rates. However, in women, there was no association. Given the known differences in disease pathophysiology and manifestation between men and women, this finding additionally highlights the need to conduct genetic studies in underrepresented groups so that we can design scores that accurately predict risk within specific groups. Our next letter comes from Xiao Wang and Kiran Musunru at the University of Pennsylvania ... "Confirmation of Causal rs9349379- PHACTR1 Expression Quantitative Trait Locus in iPSC Endothelial Cells". They were interested in understanding the affect of a coronary disease SNP in the PHACTR1 gene on gene expression. Previous efforts to investigate this had yielded conflicting results showing either a significant eQTL effect for PHACTR1 and vascular tissue or no effect on PHACTR1, but an effect on a distal gene EDN1 in endothelial cells. For this study, the authors used CRISPR Cas9 to introduce the SNP to iPS cells and then expanded isogenic lines at the major and minor allele homozygous and differentiated these into endothelial cells. They find that the major allele was associated with significantly higher factorial expression, but no difference in EDN1 expression. Thus, based on these experiments, it appears that PHACTR1 may indeed be the causal gene in that region underlying the GWAS signal and whether or not EDN1 is involved remains unclear. Our next letter is a clinical letter from Nosheen Raza, Anjali Owens and co-authors at the University of Pennsylvania. They report on "ACTA1 Novel Likely Pathogenic Variant in a Family With Dilated Cardiomyopathy". In this case report, they describe that the discovery of a mutation in ACTA1 in a family with dilated cardiomyopathy, but no skeletal muscle symptoms. As a gene that is predominantly expressed in skeletal muscle, ACTA1 mutations have previously been associated with skeletal muscle myopathies and would not have been expected to cause cardiac symptoms in the absence of skeletal muscle dysfunction. However, sequencing suggests that this variant is a causal mutation in this family, highlighting the need to consider potential mechanisms for cardiac muscle specifics of highly expressed skeletal muscle genes. Our second clinical letter comes from Laura Zahavich, Seema Mital and co-authors from the Hospital for Sick Children in Ontario. They report a "Novel Association of a De Novo CALM2 Mutation With Long QT Syndrome and Hypertrophic Cardiomyopathy". They report finding mutation in the calcium transporter CALM2 gene in the child who presented with hypertrophic cardiomyopathy and ultimately died from sudden cardiac death. While this patient also had some variants of un-insignificance, the CALM2 gene is highly conserved and mutations are likely to be pathogenic. The CALM2 is not on all of the clinical genetic testing panels and in this case, whole-exome sequencing was required to identify a mutation. CALM2 have been described in other individuals and together with the findings reported here, there's compelling evidence for inclusion of CALM2 on cardiomyopathy in clinical testing panels. This issue also contains a perspective article from Michael Mackley, Elizabeth Ormondroyd and colleagues from the University of Oxford entitled "From Genotype to Phenotype: Clinical Assessment and Participant Perspective of a Secondary Genomic Finding Associated with Long QT Syndrome". They describe some of the challenges arising from more widespread genetic testing including how to deal with incidental findings. A larger number of people including apparently healthy individuals are receiving sequencing results that highlight potential disease-related mutations, but with varying penetrance and uncertain effects. This perspective paper highlights the issues through case study and discusses future directions and challenges in this rapidly growing area. Finally, we ride out this issue with an AHA scientific statement on "Cardiovascular Health in Turner Syndrome: A Scientific Statement From the American Heart Association" led by Michael Silberbach and Jolien Roos-Hesselink and a group of co-authors representing the American Heart Association Council on Cardiovascular Disease in the Young; Council on Genomic and Precision Medicine; and Council on Peripheral Vascular Disease. In this statement, they discuss the cardiovascular complications that commonly occur in girls and women Turner syndrome. Cardiovascular disease contributes significantly to premature death in individuals with Turner syndrome. Because of the unique nature of the cardiac presentations in Turner syndrome, better clinical guidelines are needed to improve diagnosis and treatment of [phonetic 00:17:26] ischemia in these individuals. This statement takes a first step to outline suggestions to improve clinical practice and highlights the work that still remains to be done to inform disease management. That rounds out the October issue of Circulation: Genomic and Precision Medicine. Thanks for listening! You can go online to ahajournals.org/journal/circgen to access the latest issue and browse previous issues. As a last reminder, AHA Sessions is approaching fast and I hope to see many of you in Chicago, November 10-12. This podcast was brought to you by Circulation: Genomic and Precision Medicine and the American Heart Association Council on Genomic and Precision Medicine. This program is Copyright American Heart Association, 2018.  

In this work, we were able to take advantage of a deregulated wnt signaling pathway – a condition which is found in most gastrointestinal cancers, in particular in colorectal carcinomas. In order to restrict reporter gene expression to the desired cell type, we utilized the β-catenin dependent CTP4-promoter to restrict the expression of Firefly Luciferase and enhanced green fluorescent fusion protein (EGFPLuc) to cell lines with deregulated wnt signaling including SW480, LS174T, HepG2, Coga2 and Coga12. Stable cell lines containing this CTP4-driven EGFPLuc construct were established with the help of a lentiviral vector to monitor wnt activity by transgene expression. With these stably transduced cell lines, we performed a therapeutic target screen via siRNA-mediated knock-down of a number of potentially therapeutic targets within the wnt pathway – osteoprotegerin (OPG), Traf2 and Nck-interacting kinase (TNIK), SRY-related HMG-box (Sox2), protease-activated receptor 1 (PAR-1), β-catenin and transcription factor 4 (TCF4). The in vitro screening system was utilized as a prevalidation tool for therapeutically relevant targets. The degree of interference of our novel targets was determined and the search for a suitable siRNA target in colorectal cancer cells was narrowed down to β-catenin, PAR-1 and TNIK. As proof of principle the siRNA-mediated knock down of β-catenin was verified on mRNA and protein level in LS174T cells. After the initial read-out of various cell lines with different siRNAs has been established via the reduction of Luciferase expression levels, the biological effect of these targets were validated. For this purpose colony formation and cell motility/invasion assays were conducted for all relevant target cell lines. Furthermore in the in vitro experiments, the tumor-selectivtiy of the CTP4-promoter was employed in the delivery of the cytotoxic protein diphteria toxin A (DTA) in colorectal cancer target cells. Data evaluation of all in vitro assays pointed at reduced levels of proliferation, invasive behavior and aggressiveness, which yielded three candidates (PAR-1, TNIK and β-catenin) considered as viable for a treatment attempt in vivo. In the in vivo experiments, systemic delivery of siRNA against β-catenin, sticky siRNA targeting PAR-1 and plasmid DNA encoding for CTP4 controlled DTA were evaluated in a disseminated liver metastasis model of LS174T colorectal cancer. Specific knock-downs of β-catenin and PAR-1 were achieved which was confirmed via mRNA analysis. As for CTP4-DTA pDNA delivery the overall tumor load of the liver was reduced without any significant systemic toxicity, indicating specific DTA expression in tumor tissue. Also knock down of PAR1 using sticky siRNA significantly reduced tumor growth. All in all, the therapeutic effect of PAR-1 and β-catenin knock-down could be verified in various in vitro assays analyzing invasive behavior and anchorage independent growth and ultimately also in vivo. The tumor-specific expression of DTA pDNA could also be confirmed in vitro and was further investigated in an orthotopic liver dissemination model in NMRI nude mice.

Anticancer effects of aloe-emodin on HepG2 cells, Murine plasma protein response to sepsis, Biomarkers for type 2 diabetic nephropathy.

Anticancer effects of aloe-emodin on HepG2 cells, Murine plasma protein response to sepsis, Biomarkers for type 2 diabetic nephropathy.

The nucleotide and complete amino acid sequence for the human ß2-glycoprotein I (ß,I) was derived by sequencing the cDNA clone pB2I- 1. In addition to the 326 amino acid residues of the mature protein this clone codes for a putative leader peptide and contains sequence representing 5' and 3' untranslated regions. When this amino acid sequence was compared with the previously published primary sequence, three major amino acid substitutions were found, two involving cysteine residues. These substitutions lead to a new alignment of the complement control protein (CCP) repeats present in fl2I and a prediction of the complete disulphide bond organization. Northern-blot analysis indicates that hepatocytes are a major site of biosynthesis for this protein. A transcription signal of about 1.5 kb was detected by using RNA from HepG2 cells