Podcasts about tnfa

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.03.29.534819v1?rss=1 Authors: Kim, T. W., Koo, S. Y., Riessland, M., Cho, H., Chaudhry, F., Kolisnyk, B., Russo, M. V., Saurat, N., Mehta, S., Garippa, R., Betel, D., Studer, L. Abstract: Ongoing, first-in-human clinical trials illustrate the feasibility and translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinson's disease (PD). However, a major unresolved challenge in the field is the extensive cell death following transplantation with less than 10% of grafted dopamine neurons surviving. Here, we performed a pooled CRISPR/Cas9 screen to enhance the survival of postmitotic dopamine neurons in vivo. We identified p53-mediated apoptotic cell death as a major contributor to dopamine neuron loss and uncovered a causal link of TNFa-NFkB signaling in limiting cell survival. As a translationally applicable strategy to purify postmitotic dopamine neurons, we performed a cell surface marker screen that enabled purification without the need for genetic reporters. Combining cell sorting with adalimumab pretreatment, a clinically approved and widely used TNFa inhibitor, enabled efficient engraftment of postmitotic dopamine neurons leading to extensive re-innervation and functional recovery in a preclinical PD mouse model. Thus, transient TNFa inhibition presents a clinically relevant strategy to enhance survival and enable the engraftment of postmitotic human PSC-derived dopamine neurons in PD. 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/2023.02.08.527788v1?rss=1 Authors: Schafer, C., Martin-Almedina, S., Kurylowicz, K., Dufton, N. P., Osuna-Almagro, L., Wu, M.-L., Johnson, C., Shah, A., Haskard, D. O., Buxton, A., Willis, E., Wheeler, K., Turner, S., Chlebicz, M., Scott, R., Kovats, S., Cleuren, A., Birdsey, G. M., Randi, A. M., Griffin, C. T. Abstract: Background: During infectious diseases, pro-inflammatory cytokines transiently destabilize interactions between adjacent vascular endothelial cells (ECs) to facilitate the passage of immune molecules and cells into tissues. However, in the lung the resulting vascular hyperpermeability can lead to organ dysfunction. Previous work identified the transcription factor ERG as a master regulator of endothelial homeostasis. Here we investigate whether the sensitivity of pulmonary blood vessels to cytokine-induced destabilization is due to organotypic mechanisms affecting the ability of endothelial ERG to protect lung ECs from inflammatory injury. Methods: Cytokine-dependent ubiquitination and proteasomal degradation of ERG was analyzed in cultured Human Umbilical Vein ECs (HUVECs). Systemic administration of TNFa or the bacterial cell wall component lipopolysaccharide (LPS) was used to cause a widespread inflammatory challenge in mice; ERG protein levels were assessed by immunoprecipitation, immunoblot, and immunofluorescence. Murine Erg deletion was genetically induced in ECs (Ergfl/fl;Cdh5(PAC)-CreERT2), and multiple organs were analyzed by histology, immunostaining, and electron microscopy. Results: In vitro, TNFa promoted the ubiquitination and degradation of ERG in HUVECs, which was blocked by the proteasomal inhibitor MG132. In vivo, systemic administration of TNFa or LPS resulted in a rapid and substantial degradation of ERG within lung ECs, but not ECs of the retina, heart, liver, or kidney. Pulmonary ERG was also downregulated in a murine model of influenza infection. Ergfl/fl;Cdh5(PAC)-CreERT2 mice spontaneously recapitulated aspects of inflammatory challenges, including lung-predominant vascular hyperpermeability, immune cell recruitment, and fibrosis. These phenotypes were associated with a lung-specific decrease in the expression of Tek, a gene target of ERG previously implicated in maintaining pulmonary vascular stability during inflammation. Conclusions: Collectively, our data highlight a unique role for ERG in pulmonary vascular function. We propose that cytokine-induced ERG degradation and subsequent transcriptional changes in lung ECs play critical roles in the destabilization of pulmonary blood vessels during infectious diseases. 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 Aging (listed as "Aging (Albany NY)" by MEDLINE/PubMed and "Aging-US" by Web of Science) Volume 15, Issue 1, entitled, “Genetic deficiency and pharmacological modulation of RORα regulate laser-induced choroidal neovascularization.” Choroidal neovascularization (CNV) causes acute vision loss in neovascular age-related macular degeneration (AMD). Genetic variations of the nuclear receptor RAR-related orphan receptor alpha (RORα) have been linked with neovascular AMD, yet its specific role in pathological CNV development is not entirely clear. In this new study, researchers Chi-Hsiu Liu, Felix Yemanyi, Kiran Bora, Neetu Kushwah, Alexandra K. Blomfield, Theodore M. Kamenecka, John Paul SanGiovanni, Ye Sun, Laura A. Solt, and Jing Chen from Harvard Medical School, UF Scripps Biomedical Research and University of Arizona showed that Rora was highly expressed in the mouse choroid compared with the retina, and genetic loss of RORα in Staggerer mice (Rorasg/sg) led to increased expression levels of Vegfr2 and Tnfa in the choroid and retinal pigment epithelium (RPE) complex. “Here, we investigated whether RORα regulates CNV using a mouse model of laser-induced CNV, mimicking the neovascular features of wet AMD. We found that expression of RORα was enriched in the mouse choroid/RPE complex and upregulated in laser-induced CNV.” In a mouse model of laser-induced CNV, RORα expression was highly increased in the choroidal/RPE complex post-laser, and loss of RORα in Rorasg/sg eyes significantly worsened CNV with increased lesion size and vascular leakage, associated with increased levels of VEGFR2 and TNFα proteins. Pharmacological inhibition of RORα also worsened CNV. In addition, both genetic deficiency and inhibition of RORα substantially increased vascular growth in isolated mouse choroidal explants ex vivo. RORα inhibition also promoted angiogenic function of human choroidal endothelial cell culture. “Together, our results suggest that RORα negatively regulates pathological CNV development in part by modulating angiogenic response of the choroidal endothelium and inflammatory environment in the choroid/RPE complex.” DOI: https://doi.org/10.18632/aging.204480 Corresponding Author: Jing Chen - jing.chen@childrens.harvard.edu Keywords: age-related macular degeneration, angiogenesis, choroidal neovascularization, inflammation, nuclear receptors, RORα, VEGFR2, TNFα Sign up for free Altmetric alerts about this article: https://aging.altmetric.com/details/email_updates?id=10.18632%2Faging.204480 About Aging-US: Launched in 2009, Aging (Aging-US) publishes papers of general interest and biological significance in all fields of aging research and age-related diseases, including cancer—and now, with a special focus on COVID-19 vulnerability as an age-dependent syndrome. Topics in Aging go beyond traditional gerontology, including, but not limited to, cellular and molecular biology, human age-related diseases, pathology in model organisms, signal transduction pathways (e.g., p53, sirtuins, and PI-3K/AKT/mTOR, among others), and approaches to modulating these signaling pathways. Please visit our website at www.Aging-US.com​​ and connect with us: SoundCloud – https://soundcloud.com/Aging-Us Facebook – https://www.facebook.com/AgingUS/ Twitter – https://twitter.com/AgingJrnl Instagram – https://www.instagram.com/agingjrnl/ YouTube – https://www.youtube.com/agingus​ LinkedIn – https://www.linkedin.com/company/aging/ Reddit – https://www.reddit.com/user/AgingUS Pinterest – https://www.pinterest.com/AgingUS/ For media inquiries, please contact media@impactjournals.com.

Gut Immune Body Brain Axis.Dr Gundry:Leaky Gut, gut microbiome and dietRenowned Cardiovascular Surgeon who realised that all he was doing was treating thesymptoms so he studied the underlying causes.The lining of the gut is one cell thickAs bacteria break down the gut that is when ageing startsIbuprofen or roundup disastrous the bacteria populationSkin is a mirror of the lining of the gutJoints do not naturally wear out.Animal model C Elegans as bacteria begin to break down the wall of the gut that is whenageing starts105 year old people have a diverse set of bugs identical to a healthy 30 year old. It is notattacking the wall of the gut.Ecermansia musinophilia. Lives in the mucous layer whose job is to trap lectins plantproteins looking for sugar molecules and to protect the wall of the gut from harmful bacteria.Ecermansia musinophilia eats mucus which in turn makes more mucus.Metformin works by increasing mucous and this change in bacteria makes some peoplehave mild diarrhoea as the bacteria change.If we damage this lining eg ibuprofen or food with roundup destroys the bacteria populationand gut lining.Glyocosade an antibacterial damages Ecermansia Musinophilia even though it does notdirectly affect human cells.Antibiotics in food or direct prescription eg ladies who take low dose for UTI have a higherincidence of heart disease.Heart disease is an autoimmune disease starting in the gut.Cholesterol is an innocent bystander which gets sucked into the inflamed wall of a bloodvessel.Infants with heart transplants have coronary artery disease with pathology identical withtypical coronary artery disease.Lectins which are a foreign protein which can stick to sugar molecules on the surface ofblood vessels are the cause of atherosclerosis and removing lectins reduces those markers.Lectins are one of the plant defence systems. Sticky proteins that look for specific sugarmolecules to stick to which insights an inflammtory response.Joints do not normally wear out. Usually you can find bacterial particles in the joint fluid ofarthritisBecauseLectins broke down the wall of the gut. 65% of the immune system is behind the wall of thegut because the gut is where the outside word gets through. A reason why we store fat inthe gut is to provide energy to the immune system. Similarly fat on the outside ofatherosclerotcic blood vessles correlates with the severity of inflammation.Fat is not the cause . It is there because of the inflammation and the inflammation is theredue to the leaky gut.The immune system responds to antigens on bacteria of viruses. Lectins have antigens withcross reactivity with other proteins in the body. Eg thyroid.Nightshade vegetables or peanutsLectins disrupt the microbiome and break up the lining of the gut allowing entry by lectinsand by bacteria or bacterial particles.Hence if you inject a bacterial lipopolysaccharide into a person you can induce septic shock.Alzheimers Parkinsons is neuroinflammation.Most amyloid is produced by bacteria in the gut. Therefore 40 billion dollars invested inantiamyloid drugs has been a waste because amyloid is produced by the amyloid producingbacteria inthe gut fet by western diet. Then the amyloid has to get through the wall of the gut.Once they get through the gut wall and goto the brain it will produce more amyloid.Cholesterol and amyloid coexist in dementia in those with the apoE gene.The apo E gene codes for a carrier molecule because it is less efficient at transportingcholesterol. It cannot get out of the cell after it has been attracted by inflammation.Faecal microbial transplant:1970s broad spectrum antibiotics came out which made it much quicker to treat infectionsbut it also wiped out the gut bacteria. Normally 10000 species of bacteria.Pseudomembranous colitis was caused by Clostridium Difficile over growing. Initial studydone from the faeces of medical students.Faecal enemas treated the pseudomembranous colitis.Meat with animals treated by antibiotics can also cause problems.60% of faeces is bacteriaOral microbiome and cloud of bacteria around us –Holobiome . This defines our personalspace.Kissing is a human and ape characteristic. Exchanging oral microbiome. Bacteria decidewhether the other person's bacteria are compatible with them.Women have a gut feeling because they are more capable of listening to their microbiome. We inherit our microbiome from our mother. All of the mitochondria are involved with bacteriainherited from our mother. Bacteria communicate to their ‘sisters 'ie the body's mitochondria.Autism: kids have a different microbiome than ‘normal'The placental microbiome is important in educating the foetal immune system.Oral faecal transplants for 6 weeks in autistic kids. Almost immediately 50% autismsymptoms reduced.Ecermansia like tubers, mushrooms, -study in Asia find 90% reduction in Alzheimers withtwo cups of mushrooms a week.Inulin containing compounds eg chicory, radicchio, jerusalem artichoke.Exercise women who exercise routinely from midlife have a 90% reduction in Alzheimers. Inthose who get AD it happens 11 years later. Housework can be important part of exercise.Meditation and yoga also changes the gut microbiome.Lymph system in the brain in deep sleep -early in the sleep cycle-shrinks by 20% and thesebad proteins are squeezed out. You need a 3-4 hour window between sleep and dinnerbecause blood flow diverts to the gut.Olive oil /walnuts / mediterranean low fat diet: first two groups improved memory after 5years. 3rd group lost memoryThose with CVD had a 30% reduction in events, the low fat group continued CVS events.Polyphenos in olive oil grow proteinsTMAO is made by gut bacteria primarily from animal protein especially choline eg egg yolkand carnitine . TMAO damages blood vessels. Polyphenols in certain olive oil and red winebalsamic vinegar that paralyse enzyme systems in the bacteria so they do not make TMAO.However the logical error here is that eggs which are high in choline are not associated withincreased morbidity.Vitamin D at least 5000 units a day . Almost all cancer patietns and autoimmune pateitnshave low vitamin D. HIgher your VItamin D the longer your telomere. Stem cells in the gutare simulated by vitamin D.VItamin CLectins are present in most plant foods but especially high in:legumes, such as beans, lentils, peas, soybeans, and peanutsnightshade vegetables, such as tomatoes and eggplantdairy products, including milkgrains, such as barley, quinoa, and riceThe Roll of Inflammation in Depression and FatigueFrontiers In Immunology:CH Lee 2019:Immune system link to depression first noticed with immunotherapy eg INFa (which activates an inflammatory antiviral response) for Hepatitis C : associated with raised proinflammatory cytokines and depression and fatigue.20% of patients treated with INFa developed depression which resolved on discontinuationbut also increased the risk of depression in future.Also people with higher IL6 aged 9 were more likely to have depression aged 18 in a dosedependent manner.Innate immune system seems to be lower in depression eg NK cells and also less antiinflammatory regulatory T cells whereas inflammatory monocytes are activated.There is commonality in immune activation from autoimmune disorder such as multiplesclerosis or immune reactions in sepsis.Antidepressants reduce inflammation while a higher baseline level of inflammation predicts apoorer treatment response.People with depression have been shown to have higher inflammatory markers which canbe used to predict treatment efficacy and future recurrences of depression.Elevated inflammatory markets eg TNFa after an MI disrupt the blood brain barrier causingdepression.Inflammatory changes in the brain with raised TNFa in the hippocampus and striatumprecede development of depressive symptoms.Neurogenesis is inhibited by the kynurenine pathway which is rescued by both inhibitors ofthis pathway and traditional antidepressants.TNFa also increases glutamate release causing exocytotic damage to surroundingsneurones.Conditions associated with chronic immune activation such as asthma, atopy, diabetes mMS, RhA, SLE are all associated with raised levels of depression eg 36% of asthma havedepression who also had higher TNFa than those who were not depressed. 75% in RhAMS up to 50% risk of depression.Acute inflammation with sepsis also causes depression and raises the risk of depression infuture which in animal models can be reduced by using steroid during the acute sepsis.Antidepressants reduce inflammatory markers perhaps SNRI more effective than SSRI andalso ECT adds in return to normal of NK activity.Directly reducing the immune response eg anti TNF a or Caspase Inhibitors have beenshown to reduce depression. Rituximab which is an antibody that targets and depletes Bcells in the treatment of RhA also reduces depression.Aspirin can reduce depression but can also reduce the effect of an SSRI.

Getting to the Heart of Cannabis Health Risks Cell Press Cannabis has deleterious effects on cardiovascular physiology. Wei et al. (Cell 185, May 12, 2022) confirm that important inflammatory markers increase transiently after a single marijuana joint and prove a mechanistic link between THC induced vascular inflammation, endothelial dysfunction, cellular oxidative stress, and atherosclerosis using cell-based and mouse models of atherosclerosis. They provide a pharmacological model in which THC-mediated activation of CB1 receptor signaling pathways converge on MAP kinase, TNFa, and NF-kB outputs to create a proinflammatory and atherogenic environment in endothelial cells.   That genistein can antagonize the negative effects of THC with minimal central effects is exciting because genistein is a common component of soy and is already a widely consumed dietary product.   Electronic Cigarettes Versus Nicotine Patches for Smoking Cessation in Pregnancy: A Randomized Controlled Trial Nature Medicine Pregnant smokers were randomized to use either nicotine replacement therapy with patches (NRT, n=571) or e-cigarettes (n=569) for smoking cessation. For the primary outcome, validated prolonged quit rates at the end of pregnancy, the results were 4.4% for NRT and 6.8% for e-cigarettes (P=0.08). However, 25 participants in the NRT arm who reported abstinence also used e-cigarettes. When these participants were excluded from data analysis the quit rates were 3.6% for NRT and 6.8% for e-cigarettes (P=0.02). Low birthweight was less common in the e-cigarette arm, 9.6% versus 14.8% for NRT (P=0.01). The authors conclude that “e-cigarettes were markedly more effective than patches” and do not pose more risk.   Daily Cannabis Use, Cannabis Use Disorder, and Any Medical Cannabis Use Among US Adults: Associations Within Racial, Ethnic, and Sexual Minoritized Identities in a Changing Policy Context Preventive Medicine Reports Cannabis use has steadily increased in the United States, with daily and medical use associated with cannabis use disorder (CUD) and the negative consequences more frequent among marginalized groups. In this study, the authors use the National Survey on Drug Use and Health (NSDUH) to examine medical and daily use and CUD across the intersections of racial, ethnic, and sexual minorities. They found that sexual minorities were more likely to have medical and daily use and CUD than their heterosexual counterparts within each racial and ethnic group. However, when examining the intersection of race, ethnicity and sexual identity, there was more heterogeneity across these groups. In addition, they found that in states with medical cannabis laws (MCL) daily cannabis use was higher across all intersectional groups.     Effect of AXS-05 (Dextromethorphan-Bupropion) in Major Depressive Disorder: A Randomized Double-Blind Controlled Trial AJP Psychiatry Altered glutamatergic neurotransmission is implicated in the pathogenesis of major depressive disorder. AXS-05 (dextromethorphan-bupropion) is an oral NMDA receptor antagonist and sigma-1 receptor agonist, which utilizes inhibition of CYP2D6 to increase its bioavailability. This phase 2 trial assessed the efficacy and safety of dextromethorphan-bupropion in the treatment of major depressive disorder. In patients with major depression, dextromethorphan-bupropion (AXS-05) significantly improved depressive symptoms compared with bupropion and was generally well tolerated.  The most common adverse events were dizziness, nausea, dry mouth, decreased appetite, and anxiety. Dextromethorphan-bupropion was not associated with psychotomimetic effects, weight gain, or sexual dysfunction.   Mental Health and Substance Use Among Homeless Adolescents in the US JAMA Network This study evaluated mental health and substance use outcomes among homeless and non-homeless adolescents in 2019. Alcohol, cigarette, marijuana, and binge drinking during the prior 30 days was assessed, along with lifetime use of cocaine, methamphetamine, heroin, ecstasy, and injection drugs or prescription opioid misuse. Results found current substance use ranging from cigarettes to alcohol were higher among homeless adolescents. Lifetime cocaine use was significantly higher among homeless adolescents, as were methamphetamine, heroin, ecstasy, and injection drug use. Homeless adolescents experience worse mental health outcomes, including depression and suicidality, and struggle with more SUDs than their counterparts.   The Importance of Federal Action Supporting Overdose-Prevention Centers NEJM In this prospective piece, the authors discuss the need for new approaches to harm reduction and substance use disorder treatment in the face of substantially increasing overdose deaths, particularly since the start of the COVID-19 pandemic. One such strategy they discuss is overdose-prevention centers, which operate in other countries and are associated with significant reductions in opioid-overdose morbidity and mortality. However, under Section 856 of the Controlled Substances Act, such facilities may be subject to federal legal sanctions. The authors recommend that the Biden administration declare they will not interfere with such public health interventions or declare that section 856 does not apply to legally sanctioned centers. Further, Congress should modify the Controlled Substance Act to exempt overdose-prevention centers.    Complex Persistent Benzodiazepine Dependence—When Benzodiazepine Deprescribing Goes Awry JAMA Psychiatry Benzodiazepines remain popular medications among patients due to rapid symptom relief and reinforcing effects. As clinicians and patients become more aware of potential risks, and clinical guidelines increasingly urge caution in prescribing, guidance for benzodiazepine deprescribing is needed. The authors propose a new clinical concept for patients experiencing significant psychological or functional decline during or after a benzodiazepine taper—complex persistent benzodiazepine dependence (CPBD). CPBD can be described as symptomatic or functional decompensation with or without the development of aberrant medication behaviors in the setting of benzodiazepine deprescribing–in the absence of a benzodiazepine use disorder. Further research is needed to validate this concept.    What is Success in Treatment for Opioid Use Disorder? Perspectives of Physicians and Patients in Primary Care Settings JSAT Opioid abstinence and treatment retention are typically used as measures of success of MOUD treatment. This study sought to identify other important treatment outcomes and patient-centered measures of success. Qualitative, structured interviews were conducted with physicians (n=14)  and patients (n=18) in 2 family medicine residency programs. The physicians (7 faculty and 7 residents) were experienced buprenorphine prescribers. Both patients and physicians identified 5 themes: staying sober, and improvement in physical health, mental health, relationships, and role functioning. Patients, but not physicians, identified 2 additional themes: tapering off buprenorphine, and decreased stigma and shame. The authors conclude that “clinicians and researchers need to consider a broader scope of success indicators.”  

These tests are required if you want to test for inflammation in your body. These tests check for systemic or whole body inflammation. Inflammation is something that you don't want going on in your body. High levels of inflammation in the body increase your risk of developing almost every major disease including autoimmune disease, cancer, heart disease, stroke, and much more. Inflammation has also been associated with accelerated aging! No matter how you look at it, it's not something you want in your body. The good news is that you can test for inflammation in your body by looking at some simple blood tests. These blood tests help identify the presence of both acute and chronic inflammation but they don't tell you WHY the inflammation is present. These tests are so important that I order them in just about every patient that I see. They are especially helpful if you have thyroid problems or just want to stay in optimal health. These tests include: - ESR - CRP - Ferritin - Antibody testing (optional) - IL6, TNFa, and fibrinogen (optional) Learn why these tests are so important and what they mean for you in this video. Download my free thyroid resources here (including hypothyroid symptoms checklist, the complete list of thyroid lab tests + optimal ranges, foods you should avoid if you have thyroid disease, and more): https://www.restartmed.com/start-here/ Recommended thyroid supplements to enhance thyroid function: - Supplements that everyone with hypothyroidism needs: https://www.restartmed.com/product/hy... - Supplement bundle to help reverse Hashimoto's: https://www.restartmed.com/product/ha... - Supplements for those without a thyroid and for those after RAI: https://www.restartmed.com/product/th... - Supplements for active hyperthyroidism: https://www.restartmed.com/product/hy... See ALL of my specialized supplements including protein powders, thyroid supplements, and weight loss products here: https://www.restartmed.com/shop/ Want more from my blog? I have more than 400+ well researched blog posts on thyroid management, hormone balancing, weight loss, and more. See all blog posts here: https://www.restartmed.com/blog/ Prefer to listen via podcast? Download all of my podcast episodes here: https://podcasts.apple.com/us/podcast... Disclaimer: Dr. Westin Childs received his Doctor of Osteopathic Medicine from Rocky Vista University College of Osteopathic medicine in 2013. His use of “doctor” or “Dr.” in relation to himself solely refers to that degree. Dr. Childs is no longer practicing medicine and does not hold an active medical license so he can focus on helping people through videos, blog posts, research, and supplement formulation. To read more about why he is no longer licensed please see this page: https://www.restartmed.com/what-happe... This video is for general informational, educational, and entertainment purposes only. It should not be used to self-diagnose and it is not a substitute for a medical exam, treatment, diagnosis, prescription, or recommendation. It does not create a doctor-patient relationship between Dr. Childs and you. You should not make any changes to your medications or health regimens without first consulting a physician. If you have any questions please consult with your current primary care provider. Restart Medical LLC and Dr. Westin Childs are not liable or responsible for any advice, course of treatment, diagnosis, or any other information, services, or product you obtain through this website or video.

There are  a ton of "functional" labs out there today. In this podcast, I cover the most important labs for managing autoimmunity. If there is one takeaway from this episode, it is: FOCUS ON THE BASICS FIRST! So often someone brings me in a fancy methylation panel, hormone panel, or stool sample and they don't know their basics.  I divide these labs into 3 categories:1. Basics - Basic blood markers are the foundation, and often people have them from multiple practitioners and you can compare apples to apples. These include CBC and CMP (which I discuss how important these are!), iron/TIBC, ferritin, homocysteine, vitamin D, thyroid markers, LDH, HbA1C, inflammatory markers like CRP, TGFbeta, GlycA, TNFa, and more. 2. Autoimmune-Specific Tests - antibodies including ANA, thyroid, rheumatoid factor, gut tissue, neurological tissue; barrier integrity such as gut barrier and blood-brain-barrier; advanced immune tests such as T & B Lymphocytes.  In this section I give a special focus to Cyrex labs, who is in my opinion the leader in autoimmune reactivity testing. 3. Drivers - Foods, Toxins, Stress, Hormones, and Hidden Infections. This includes food sensitivities, stool testing, toxin testing, and more. I discuss some of my favorites, which include Cyrex again, Great Plains, GI Map, Dried Urine Hormones from Meridian Valley, and more. 

Rheumatology describes 'aches & pains' which really includes over 200 autoimmune (and auto-inflammation- what is this?! Listen in!) diseases and, since HIV-AIDs blew open the doors on research into re-discovering the immune system, now includes science ranging from tolerance to the trillions that live in and on us as our microbiome, cancer, autoimmune disease, auto-inflammation (like with chronic inflammatory response syndrome), allergies and mast cell activation, and pathogenic and/or sub-occult infections. What a 'specialty’! In today’s Podcast for Healing Neurology episode, classically-trained and triple-board certified (internal medicine, pediatrics and rheumatology) physician, Anthony Padula, brings his experience to bear on this complicated topic. Gain insight into some of the pharmacology history, like how TNFa medications were 'designed' to treat sepsis (severe infection) but resulted in elevated mortality and then were found useful for inflammation in other realms. It's similar for methotrexate designed to treat cancer can be useful as a once weekly treatment for autoimmune disease. This is foundational information we need to better understand how to address our immune system, with all of the current-day 'new' triggers like light, noise and heavy metal pollution, like microplastics in our air and water (and baby bottles-ugh!) and so many others to which we haven't evolved. It's these 'vague' irritants that can light up our system and persist chronic disease. Listen in to this fun and science-y conversation with Dr Anthony Padula, fellow Jetson Health Gut Council Member . Find more about him at his website: http://www.drpadula.com/ and more about the Jetson Health Gut Council at https://wearejetson.com/pages/jetsons-gut-health-experts

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.22.216572v1?rss=1 Authors: Huie, J. R., Ferguson, A. R., Kyritsis, N., Pan, J., Irvine, K.-A., Nielson, J. L., Schupp, P., Oldham, M. C., Gensel, J. C., Lin, A., Segal, M. R., Ratan, R., Bresnahan, J. C., Beattie, M. Abstract: Traumatic spinal cord injury (SCI) produces a complex syndrome that is expressed across multiple endpoints ranging from molecular and cellular changes to functional behavioral deficits. Effective therapeutic strategies for CNS injury are therefore likely to manifest multi-factorial effects across a broad range of biological and functional outcome measures. Thus, multivariate analytic approaches are needed to capture the linkage between biological and neurobehavioral outcomes. Injury-induced neuroinflammation (NI) presents a particularly challenging therapeutic target, since NI is involved in both degeneration and repair. Here, we used big-data integration and large-scale analytics to examine a large dataset of preclinical efficacy tests combining 5 different blinded, fully counter-balanced treatment trials for different acute anti-inflammatory treatments for cervical spinal cord injury in rats. Multi-dimensional discovery, using topological data analysis (TDA) and principal components analysis (PCA) revealed that only one showed consistent multidimensional syndromic benefit: intrathecal application of recombinant soluble TNFa receptor 1 (sTNFR1), which showed an inverse-U dose response efficacy. Using the optimal acute dose, we showed that clinically-relevant 90 min delayed treatment profoundly affected multiple biological indices of NI in the first 48 hrs after injury, including reduction in pro-inflammatory cytokines and gene expression of a coherent complex of acute inflammatory mediators and receptors. Further, a 90 min delayed bolus dose of sTNFR1 reduced the expression of NI markers in the chronic perilesional spinal cord, and consistently improved neurological function over 6 weeks post SCI. These results provide validation of a novel strategy for precision preclinical drug discovery that is likely to improve translation in the difficult landscape of CNS trauma, and confirm the importance of TNFa signaling as a therapeutic target. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.04.28.064758v1?rss=1 Authors: Wiemann, S., Reinhard-Recht, J., Reinehr, S., Cibir, Z., Joachim, S., Faissner, A. Abstract: Previous studies demonstrated that retinal damage correlates with a massive remodeling of extracellular matrix (ECM) molecules and reactive gliosis. However, the functional significance of the ECM in retinal neurodegeneration is still unknown. In the present study, we used an intraocular pressure (IOP) independent experimental autoimmune glaucoma (EAG) mouse model to examine the role of the ECM glycoprotein tenascin-C (Tnc). Wild type (WT ONA) and Tnc knockout (KO ONA) mice were immunized with an optic nerve antigen (ONA) homogenate and control groups (CO) obtained sodium chloride (WT CO, KO CO). IOP was measured weekly and electroretinographies were recorded at the end of the study. 10 weeks after immunization, we analyzed retinal ganglion cells (RGCs), glial cells and the expression of different cytokines in retina and optic nerve tissue in all four groups. IOP and retinal function was comparable in all groups. Although less severe in KO ONA, WT and KO mice displayed a significant loss of RGCs after immunization. Compared to KO ONA, a significant reduction of {beta}III-tubulin stained axons and oligodendrocyte markers was noted in the optic nerve of WT ONA. In retinal and optic nerve slices, we found an enhanced GFAP+ staining area of astrocytes in immunized WT. In retinal flat-mounts, a significantly higher number of Iba1+ microglia was found in WT ONA, while a lower number of Iba1+ cells was observed in KO ONA. Furthermore, an increased expression of the glial markers Gfap, Iba1, Nos2 and Cd68 was detected in retinal and optic nerve tissue of WT ONA, whereas comparable levels were observed in KO ONA post immunization. In addition, pro-inflammatory Tnfa expression was upregulated in WT ONA, but downregulated in KO ONA. Vice versa, a significantly increased anti-inflammatory Tgfb expression was measured in KO ONA animals. Collectively, this study revealed that Tnc plays an important role in glial and inflammatory response during retinal neurodegeneration. Our results provide evidence that Tnc is involved in glaucomatous damage by regulating retinal glial activation and cytokine release. Thus, this transgenic EAG mouse model offers for the first time the possibility to investigate IOP-independent glaucomatous damage in direct relation to ECM remodeling. Copy rights belong to original authors. Visit the link for more info

Activation of immune cells is the all-important first step in mounting an immune response. Immune cell activation is a popular area of research because so much happens that is key to the downstream goal of fighting infection, cancer, and disease. There are many ways to measure immune cell activation, and they all have utility. Methods can be grouped into four main categories: Proliferation Assays, Cytokine Measurement, Surface Antigen Expression, and Cytotoxicity. In this webinar, we'll discuss specific assays in each of these categories, the joys and pitfalls of each assay, and recommendations on how to choose the best method. You will learn tips and strategies for successful assay development using the following methods: Proliferation: - 3H-Thymidine Uptake - Bromodeoxyuridine Uptake (BrdU) - ATP Luminescence - Fluorescent Dye Reduction (CFSE) Cytokine Measurement: - Multiplex vs. Single Cytokine - Choice of Cytokine (IFNg, TNFa, IL-6, IL-1?, etc.) - Kinetics of Cytokine Release Surface Antigen Expression: - CD69, CD25, PD-1, etc. - Combine with CFSE, Ki67 or BrdU - Kinetics are Important Cytotoxicity: - Two-Label Flow Cytometry - Calcein AM Dye Release - Luciferase Transduced Targets - Annexin V

Gallensäuren stellen potente Signalmoleküle dar, die schon in geringen mikromolaren Konzentrationen, wie sie beim Menschen im Serum beobachtet werden, zentrale Leberzellfunktionen auf transkriptioneller und posttranskriptioneller Ebene beeinflussen. Die hydrophoben und potentiell toxischen Gallensäuren Lithocholsäure (LCA) und Chenodeoxycholsäure (CDCA) induzieren Apoptose und Cholestase, während hydrophile Gallensäuren hepatoprotektiv wirken können. Unter ihnen ist die antiapoptotisch und anticholestatisch wirksame Ursodeoxycholsäure (UDCA) von besonderer Bedeutung. UDCA stellt derzeit das einzige wirksame Therapeutikum bei chronischen cholestatischen Leberkrankheiten dar. Die vorliegende Arbeit untersuchte die Bedeutung intrazellulärer Signaltransduktionswege für die choleretischen, (anti-)cholestatischen und (anti-)apoptotischen Wirkungen physiologischer Gallensäuren in verschiedenen experimentellen Modellen. Hauptziel der Arbeit war die genauere Charakterisierung (i) der für den klinisch bedeutenden anticholestatischen Effekt des Taurinkonjugats der Ursodeoxycholsäure (TUDCA) verantwortlichen Signaltransduktionswege, und (ii) der zentralen Stellung von PI3-Kinasen in der intrazellulären Signalvermittlung der biologischen Effekte hydrophiler und hydrophober Gallensäuren. Im Modell der isoliert perfundierten Rattenleber untersuchten wir die anticholestatische und hepatoprotektive Wirkung der TUDCA in der intakten Leber unter Einsatz pharmakologischer Enzyminhibitoren. Als Leberfunktionsparameter dienten quantitativer Gallenfluß, Sekretion des Modellsubstrats der Konjugatexportpumpe Mrp2, GS-DNP, in die Galle und als Marker der Leberzellschädigung die hepatovenöse LDH-Freisetzung. Simultane Hemmung der cPKCa und der PKA, nicht aber Hemmung von cPKCa oder PKA allein antagonisierte bei Taurolithocholsäure (TLCA)-induzierter Cholestase die protektive Wirkung der TUDCA. Gallenfluß und GS-DNP-Sekretion waren unter gleichzeitiger Hemmung beider Signalwege signifikant reduziert, wohingegen die LDH-Freisetzung deutlich erhöht war. Die Ergebnisse zeigen, dass der posttranskriptionell vermittelte anticholestatische Effekt der TUDCA im etablierten Modell TLCA-induzierter Cholestase durch einen kooperativen cPKC- und PKA-abhängigen Signalweg vermittelt wird. Mitogenaktivierte Proteinkinasen Erk1/2- und p38-abhängige Signalwege hingegen, die als Vermittler von TUDCA-induzierter Cholerese unter nicht-cholestatischen Bedingungen beschrieben wurden, waren im untersuchten Modell ohne Bedeutung für die anticholestatische Wirkung der TUDCA. Mit Hilfe der neu etablierten Biotinylierung von Membranproteinen konnten wir in Ntcp-transfizierten humanen Hepatomzellen (HepG2-Ntcp) zeigen, dass TUDCA unter Cholestase die Insertion von MRP2 in die Hepatozytenmembran anregt. Dieser für die klinische Wirksamkeit der (T)UDCA potentiell bedeutende und im Tiermodell von uns vorbeschriebene Wirkmechanismus konnte damit erstmals in einem humanen Modell nachvollzogen werden. Ein weiterer in vitro Ansatz untersuchte die Phosphorylierung von aus HepG2-Ntcp immunopräzipitiertem MRP2 durch die als Gallensäureneffektoren diskutierten Proteinkinasen cPKCa, nPKCe und PKA. Alle drei Proteinkinasen phosphorylierten, durch den PKC/PKA-Inhibitor Staurosporin hemmbar, MRP2. Diese Phosphorylierung könnte, wie für die Gallensäurentransporter BSEP und NTCP bereits gezeigt, Einfluss auf Aktivität und Membraninsertion von MRP2 haben. Der funktionellen Bedeutung der PI3-Kinasen, welchen in den bisher entschlüsselten Signalwegen sowohl hydrophober/toxischer wie auch hydrophiler/protektiver Gallensäuren eine zentrale Rolle zugesprochen worden war („PI3-Kinasen-Paradoxon“), galten unsere in vitro Untersuchungen zur Aktivität der Isoformen der Klasse I PI3-Kinasen p110a, p110b und p110g nach Stimulation von primären Rattenhepatozyten mit TLCA, GCDCA, TCA und TUDCA in einem neu etablierten isoformspezifischen Kinaseassay. Dabei zeigte sich für jede Gallensäure ein für sie spezifisches Aktivierungsmuster unterschiedlicher PI3-Kinase-Isoformen. PI3-Kinase p110g wurde dabei spezifisch durch die cholestatisch und apoptotisch wirkenden Gallensäuren TLCA und GCDCA aktiviert. In HepG2-Ntcp-Zellen untersuchten wir daher die Bedeutung von p110g für Gallensäuren-induzierte Apoptose nach deren pharmakologischer Hemmung bzw. nach Transfektion mit siRNA gegen p110g. Die apoptotische Wirkung u.a. der Gallensäuren TLCA und GCDCA war unter beiden Methoden der p110g-Antagonisierung deutlich reduziert, wie sowohl in einem Caspase3/7-Assay als auch morphologisch evaluiert. Gallensäuren-unabhängige Apoptose, durch Etoposid bzw. TNFa ausgelöst, war p110g-unabhänig. Die Bedeutung der Aktivierung der PI3-Kinase-Isoform p110a durch TUDCA ist durch weitere experimentelle Untersuchungen zu klären. Die Erkenntnisse der vorliegenden Arbeit tragen zum Verständnis der komplexen Signalgebung im Rahmen cholestatischer Leberschädigung und der therapeutischen Wirkung der (T)UDCA bei und sind damit für die Entwicklung neuer Therapiestrategien bei cholestatischen Leberkrankheiten potentiell von Bedeutung.

Objectives: To study the clinical outcome, treatment response, T-cell subsets and functional consequences of a novel tumour necrosis factor (TNF) receptor type 1 (TNFRSF1A) mutation affecting the receptor cleavage site. Methods: Patients with symptoms suggestive of tumour necrosis factor receptor-associated periodic syndrome (TRAPS) and 22 healthy controls (HC) were screened for mutations in the TNFRSF1A gene. Soluble TNFRSF1A and inflammatory cytokines were measured by ELISAs. TNFRSF1A shedding was examined by stimulation of peripheral blood mononuclear cells (PBMCs) with phorbol 12-myristate 13-acetate followed by flow cytometric analysis (FACS). Apoptosis of PBMCs was studied by stimulation with TNFa in the presence of cycloheximide and annexin V staining. T cell phenotypes were monitored by FACS. Results: TNFRSF1A sequencing disclosed a novel V173D/ p.Val202Asp substitution encoded by exon 6 in one family, the c.194–14G.A splice variant in another and the R92Q/p.Arg121Gln substitution in two families. Cardiovascular complications (lethal heart attack and peripheral arterial thrombosis) developed in two V173D patients. Subsequent etanercept treatment of the V173D carriers was highly effective over an 18-month follow-up period. Serum TNFRSF1A levels did not differ between TRAPS patients and HC, while TNFRSF1A cleavage from monocytes was significantly reduced in V173D and R92Q patients. TNFa-induced apoptosis of PBMCs and T-cell senescence were comparable between V173D patients and HC. Conclusions: The TNFRSF1A V173D cleavage site mutation may be associated with an increased risk for cardiovascular complications and shows a strong response to etanercept. T-cell senescence does not seem to have a pathogenetic role in affected patients.

The cardiovascular hormone ANP is known to exert anti-inflammatory properties in macrophages and endothelial cells. This work provides new insight into the inflammatory signalling pathways influenced by the ANP in the lung. For these purposes, the effects of ANP on both alveolar epithelial cells and a model of LPS-induced lung inflammation were characterized. In alveolar epithelial cells, ANP was shown to inhibit the activation of two major transcription factors, NF-kB and AP-1, in response to TNFa. Astonishingly, this did not result in a reduced expression of the adhesion molecule ICAM-1. ANP was also capable to diminish the activation of AP-1 and NF-kB in lung tissue in vivo using a mouse model of LPS-induced septic shock. The inhibition of NF-kB activation was caused by a delayed phosphorylation and subsequent degradation of IkBa. In addition, ANP treatment elevated total protein levels of IkBa. p38 MAPK and Akt are important mediators in LPS-induced signalling. We demonstrated an activation of these kinases in lung tissue in response to i.p. LPS challenge. ANP treatment was able to lessen this activation. Furthermore, exclusive ANP treatment resulted in an increased p38 MAPK activation, which might contribute to the observed impact on other pathways. ICAM-1 expression was not impaired in whole lung tissue. ANP strongly decreased TNFa serum levels dose-dependently, but had only a slight effect on TNFa tissue levels. Interestingly, TNFa mRNA expression was not significantly reduced. Taken together this work demonstrates that ANP is able to diminish several important inflammatory pathways which are involved in the development of acute respiratory distress syndrome in LPS-induced sepsis.

Diese Dissertation handelt von der TNFa Freisetzung von isoliert, reperfundierten Mäuseherzen nach 15 min Ischämie und 90 min Reperfusion im Vergleich zu normoxisch perfundierten Mäuseherzen. Zudem sollte mit Hilfe von k.o.-Mäusenherzen und pharmakologischen Interventionen versucht werden, Rückschlüsse auf den zellulären Ursprungsort des TNFa zu bekommen und über dessen Freisetzungsmechanismus. TNFa wird nach 15 min Ischämie in zwei Phasen während der Reperfusion freigesetzt: direkt nach der Ischämie und nach 60-90 min. Normoxisch perfundierte Herzen zeigten dagegen einen Basaltonus des TNFa von 1,5pg/min über den gesamten Versuchsablauf. Der erste Gipfel der TNFa Freisetzung konnte als rein kinetisches Phänomen -wash-out- identifiziert werden, während es sich beim zweiten Gipfel um eine de-novo Synthese des TNFa handelt. Interleukin-6 und die Matrix-Metalloprotease-7 sind essentielle Faktoren für die TNFa Freisetung nach 60-90 min. Mastzellen können als Ursprungszelle des zweiten Gipfels ausgeschlossen werden.Sie sind neben Makrophagen und Endothelzellen am Basaltonus der TNFa Freisetzung beteiligt.

During my PhD thesis I was working on the identification of novel apoptosis-inducing genes by using a novel genetic expression screen (Grimm and Leder, 1997). One of the identified genes turned out to be a evolutionary conserved cDNA that codes for a novel BH3-only protein of 219 amino acid residues which was named Spike, for Small protein with inherent killing effect. Spike was then in the course of my PhD thesis extensively characterised, molecularly and biochemically. In summary, upon overexpression in mammalian cells Spike efficiently leads to all features of apoptosis, such as phenotypic alterations, cytochrome c release, caspase activation and DNA degradation. It was shown that Spike localised to the endoplasmic reticulum, where it interacts with a recently identified apoptosis regulating protein complex, consisting of Bap31, Bcl-2, Bcl-XL and an ER-specific isoform of caspase-8: pro-caspase 8L (Breckenridge et al. 2002). Although no direct interaction with anti-apoptotic members of the Bcl2-family could be observed, the importance of the BH3-like sequence for the apoptosis-inducing activity of Spike was demonstrated by using point mutations of conserved amino acid residues of this motif (Mund et al. 2003). Instead of directly interacting with anti-apoptotic members of the Bcl-2 family Spike is able to interfere with the complex formation between Bap31 and Bcl-XL. Based on these data I proposed a model according to which the complex on Bap31 is controlled by Bcl-2/Bcl-XL as long as they are associated. Displacement of Bcl-2/Bcl-XL from Bap31 by Spike leads to the formation of a pro-apoptotic complex. In addition, Spike appears to be implicated in the cell death signal of the Fas receptor. I observed that a dominant-negative version of Spike and a highly effective anti-sense oligonucleotide significantly reduced Fas-mediated DNA fragmentation whereas no reduction was detected in TNFa-induced cell death (Mund et al. 2003).

Primary biliary cirrhosis (PBC) is considered an autoimmune disease characterized by destruction of small intrahepatic bile ducts by lymphocytes. Altered functions of these lymphocytes might reflect an abnormal immune response leading to tissue damage. We investigated lymphokine secretion by mitogen-stimulated T lymphocytes from the liver biopsies of patients with PBC and for comparison also peripheral blood. In PBC, diminished synthesis of lymphotoxin (TNFP), tumor necrosis factor (TNFa) and interferon-y (IFIVy) was found both in T-cell lines from liver tissue and in peripheral blood. The reduction was most prominent for TNFP in early histological stages of PBC, and appeared to be a stable phenomenon when T cells were tested after long-term tissue culture. Analysis of mRNA levels indicates a possible link between reduced TNFP production and a defect in interleukin-2 transcription. The data suggest that diminished lymphokine production in patients with PBC may play ;In important role in the immanopathogenesis of this disease.