Podcasts about tlr2

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.05.535712v1?rss=1 Authors: Steinberg, N., Galleguillos, D., Zaidi, A., Sipione, S. Abstract: Chronic activation and dysfunction of microglia have been implicated in the pathogenesis and progression of many neurodegenerative disorders, including Huntington Disease (HD). HD is a genetic condition caused by a mutation that affects the folding and function of huntingtin (HTT). Signs of microglia activation have been observed in HD patients even before the onset of symptoms. It is unclear, however, whether pro-inflammatory microglia activation in HD results from cell-autonomous expression of mutant HTT or is the response of microglia to a diseased brain environment, or both. In this study, we used primary microglia isolated from HD knock-in mice (Q140) and wild-type (Q7) mice to investigate their response to inflammatory conditions in vitro in the absence of confounding effects arising from brain pathology. We show that naive Q140 microglia do not undergo spontaneous pro-inflammatory activation and respond to inflammatory triggers, including stimulation of TLR4 and TLR2 and exposure to necrotic cells, with similar kinetics of pro-inflammatory gene expression as wild-type microglia. Upon termination of the inflammatory insult, the transcription of pro-inflammatory cytokines is tapered off in Q140 and wild-type microglia with similar kinetics. However, the ability of Q140 microglia to develop tolerance in response to repeated inflammatory stimulations is partially impaired, potentially contributing to the establishment of chronic neuroinflammation in HD. We further show that ganglioside GM1, a glycosphingolipid with anti-inflammatory effects in wild-type microglia, not only decreases the production of pro-inflammatory cytokines and nitric oxide in activated Q140 microglia, but also dramatically dampen microglia response to re-stimulation with LPS in an experimental model of tolerance. These effects are independent from the expression of interleukin 1 receptor associated kinase 3 (Irak-3), a strong modulator of LPS signaling involved in the development of innate immune tolerance and previously shown to be upregulated by immune cell treatment with gangliosides. Altogether, our data suggest that external triggers are required for HD microglia activation, but a cell-autonomous dysfunction that affects the ability of HD microglia to acquire tolerance might contribute to the establishment of neuroinflammation in HD. Administration of GM1 might be beneficial to attenuate chronic microglia activation and neuroinflammation. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

This month on Episode 45 of Discover CircRes, host Cynthia St. Hilaire highlights four original research articles featured in the February 3rd and February 17th issues of Circulation Research. This episode also features an interview with Dr Hind Lal and Dr Tousif Sultan from the University of Alabama at Birmingham about their study Ponatinib Drives Cardiotoxicity by S100A8/A9-NLRP3-IL-1β Mediated Inflammation.   Article highlights:   Pi, et al. Metabolomic Signatures in PAH   Carnevale, et al. Thrombosis TLR4-Mediated in SARS-CoV-2 Infection   Cai, et al. Macrophage ADAR1 in AAA   Koide, et al. sEVs Accelerate Vascular Calcification in CKD   Cindy St. Hilaire:        Hi, and welcome to Discover CircRes, the podcast of the American Heart Association's journal, Circulation Research. I'm your host, Dr Cynthia St. Hilaire from the Vascular Medicine Institute at the University of Pittsburgh, and today I'm going to be highlighting the articles from our February 3rd and 17th issues of Circulation Research. I'm also going to have a chat with Dr Hind Lal and Dr Tousif Sultan from the University of Alabama at Birmingham about their study, Ponatinib Drives Cardiotoxicity by S100A8/A9-NLRP3-IL-1β Mediated Inflammation. But before I get to the interviews, here are a few article highlights.   Cindy St. Hilaire:        The first article I want to highlight comes from the laboratory of Dr Peter Leary at the University of Washington, and the title is Metabolomic Signatures Associated With Pulmonary Arterial Hypertension Outcomes. Pulmonary Arterial Hypertension or PAH is a rare but life-threatening disease in which progressive thickening of the walls of the lung's blood vessels causes increased blood pressure and that increased blood pressure ultimately damages the heart's right ventricle.   Interestingly, progression to heart failure varies considerably among patients, but the reasons why there is variability are not well understood. To find out, this group turned their attention to patient metabolomes, which differ significantly from those of healthy people and thus may also change with severity. Blood samples from 117 PAH patients were analyzed for more than a thousand metabolites by mass spectrometry and the patient's progress was followed for the next three years. 22 patients died within a three-year period and 27 developed significant right ventricle dilation. Other measures of severity included pulmonary vascular resistance, exercise capacity and levels of BNP, which is a metric of heart health. Two metabolic pathways, those relating to polyamine and histidine metabolism, were found to be linked with all measures of severity suggesting a key role for them in disease pathology. While determining how these pathways influence disease as a subject for further study, the current findings may nevertheless lead to new prognostic indicators to inform patient care.   Cindy St. Hilaire:        The next article I want to discuss is coming from our February 3rd issue of Circulation Research and this is coming from the laboratory of Dr Francisco Violi at the University of Rome and the title is Toll-Like Receptor 4-Dependent Platelet-Related Thrombosis in SARS-CoV-2 Infection. Thrombosis can be a complication of COVID-19 and it is associated with poor outcomes, including death. However, the exact mechanism by which the virus activates platelets, which are the cells that drive thrombosis, is not clear. For one thing, platelets do not appear to express the receptor for SARS-CoV-2. They do however, express the TLR4 receptor and that's a receptor that mediates entry of other viruses as part of the immune response. And TLR4 is ramped up in COVID-19 patient platelets. This group now confirms that, indeed, SARS-CoV-2 interacts with TLR4, which in turn triggers thrombosis.   The team analyzed platelets from 25 patients and 10 healthy controls and they found that the platelet activation and thrombic activity were both boosted in the patient samples and could not be blocked using a TLR4 inhibitor. Additionally, immunoprecipitation and immunofluorescent experiments further revealed colocalization between the virus protein and the TLR4 receptor on patient platelets. The team went on to show that the signaling pathway involved reactive oxygen species producing factors p47phox and Nox2, and that inhibition of phox 47, like that of the TLR4 receptor itsel,f could prevent platelet activation. As such, this study suggests that inhibiting either of these proteins may form the basis of an antithrombotic treatment for COVID-19.   Cindy St. Hilaire:        The third article I want to highlight is coming from the lab of Shi-You Chen at University of Missouri and the title of this article is ADAR1 Non-Editing Function in Macrophage Activation and Abdominal Aortic Aneurysm. Macrophage activation plays a critical role in abdominal aortic aneurysm development, or AAA development. Inflammation is a component of this pathology; however, the mechanisms controlling macrophage activation and vascular inflammation in AAA are largely unknown. The ADAR1 enzyme catalyzes the conversion of adenosine to inosine in RNA molecules and thus this conversion can serve as a rheostat to regulate RNA structure or the gene coding sequence of proteins. Several studies have explored the role of ADAR1 in inflammation, but its precise contribution is not fully understood, so the objective of this group was to study the role of ADAR1 in macrophage activation and AAA formation.   Aortic transplantation was conducted to determine the importance of nonvascular ADAR1 in AAA development and dissection and angiotensin II infusion of ApoE knockout mice combined with a macrophage specific knockout of ADAR1 was used to study the role of ADAR1 macrophage specific contributions to AAA formation and dissection. Allograft transplantation of wild type abdominal aortas to ADAR1 haploinsufficient recipient mice significantly attenuated AAA formation. ADAR1 deficiency in hematopoietic stem cells also decreased the prevalence and the severity of AAA and it also inhibited macrophage infiltration into the aortic wall. ADAR1 deletion blocked the classic macrophage activation pathway. It diminished NF-κB signaling and it enhanced the expression of a number of anti-inflammatory microRNAs. Reconstitution of ADAR1 deficient but not wild type human monocytes to immunodeficient mice blocked the aneurysm formation in transplanted human arteries. Together these results suggest that macrophage ADAR1 promotes aneurysm formation in both mouse and human arteries through a novel mechanism of editing the microRNAs that target NF-κB signaling, which ultimately promotes vascular inflammation in AAA.     Cindy St. Hilaire:        The last article I want to highlight is also from our February 17th issue of Circulation Research and it is coming from the lab of Shintaro Mandai at Tokyo Medical and Dental University and the title of the article is Circulating Extracellular Vesicle Propagated MicroRNA signatures as a Vascular Calcification Factor in Chronic Kidney Disease. Chronic Kidney Disease or CKD accelerates vascular calcification in part by promoting the phenotypic switching of vascular smooth muscle cells to osteoblast like cells. This study investigated the role of circulating small extracellular vesicles or SUVs from the kidneys in promoting this osteogenic switch. CKD was induced in rats and in mice by an adenine induced tubular interstitial fibrosis and serum from these animals induced calcification in in vitro cultures of A-10 embryonic rat smooth muscle cells. Intraperitoneal administration of a compound that prevents SEV biosynthesis and release inhibited thoracic aortic calcification in CKD mice under a high phosphorus diet. In Chronic Kidney Disease, the microRNA transcriptome of SUVs revealed a depletion of four microRNAs and the expression of the microRNAs inversely correlated with kidney function in CKD patients.   In vitro studies found that transected microRNA mimics prevented smooth muscle cell calcification in vitro. In silico analyses revealed that VEGF-A was a convergent target of all four microRNAs and leveraging this, the group used in vitro and in vivo models of calcification to show the inhibition of the VEGF-A, VEGFR-2 signaling pathway mitigated calcification. So in addition to identifying a new potential therapeutic target, these SUV propagated microRNAs are a potential biomarker that can be used for screening patients to determine the severity of CKD and possibly even vascular calcification.   Cindy St. Hilaire:        Today I have with me Dr Hind Lal who's an associate professor of medicine at the University of Alabama Birmingham and his post-doctoral fellow and the lead author of the study Dr Tousif Sultan. And their manuscript is titled Ponatinib Drives Cardiotoxicity by S100A8/A9-NLRP3-IL-1β Mediated Inflammation. And this article is in our February 3rd issue of Circulation Research. So thank you both so much for joining me today.   Tousif Sultan:              Thank you.   Hind Lal:                     Thank you for taking time.   Cindy St. Hilaire:        So ponatinib, it's a tyrosine kinase inhibitor and from my understanding it's the only treatment option for a specific group of patients who have chronic myelogenous leukemia and they have to harbor a specific mutation. And while this drug helps to keep these patients alive essentially, it's extremely cardiotoxic. So cardiotoxicity is somewhat of a new field. So Dr Lal, I was wondering how did you get into this line of research?    Hind Lal:                    So I was fortunate enough to be in the lab of Dr Tom Force and he was kind of father of this new area, now is very developed, it's called cardio-oncology. On those days there were basically everything started in cardio-oncology. So I just recall the first tyrosine kinase approved by FDA was in 2000 and that was... Imagine and our paper came in Nature Medicine 2005 and discovering there is... so to elaborate it a little bit, the cancer therapy broadly divided in two parts. One is called non-targeted therapy like chemotherapy, radiations, et cetera, and then there are cytotoxic drugs. So those cytotoxic drugs because they do not have any targeted name on it so they are, cardiotoxic are toxic to any organ was very obvious and understanding. When these targeted therapy came, which is mainly kinase inhibitor are monoclonal antibodies. So these are targeted to a specific pathway that is activated only in the cancer cells but not in any other cells in the body so they were proposed as like magic bullets that can take off the cancer without any cardiotoxity or minimal side effects. But even in the early phase like 2005 to 2010, these came out, these so-called targeted, they are not very targeted and they are not also the magic bullets and they have serious cardiotoxicity.   Cindy St. Hilaire:        And so what's the mechanism of action of ponatinib in the leukemia and how does that intersect with the cardiovascular system?   Hind Lal:                     Yeah, so this is very good question I must say. So what we believe at this point because, so leukemia if you know is driven by the famous Philadelphia chromosome, which is a translicational gene, one part of human chromosome nine and one part of human chromosome 22 and they translocate make a new gene which is BCR-ABL gene. And because it was discovered in Philadelphia UPENN, is named that Philadelphia chromosome, which is very established mechanism, that's how CML is driven. But what we have discovered that the cardiotoxicity driven by totally, totally different from the ponatinib is one of the inflammatory So it's kind of goodening. So this question is so good. One kind of toxicity is called on-target, when toxicity is mediated by the same mechanism, what is the mechanism of the drug to cure the cancer? So in that case your absolute is minimal because if you manipulate that, the drug's ability to cure the cancer will be affected but if the toxicity and the efficacy is driven by two different mechanism, then as in case of ponatinib seems like it's NLRP3 and inflammasome related mechanism. So this can be managed by manipulating this pathway without hampering the drug efficacy on the cancer.   Cindy St. Hilaire:        So what exactly is cardiotoxicity and how does it present itself in these patients?   Hind Lal:                     So these drugs like ponatinib, they call broader CVD effects. So it's not just cardiac, so they also in hypertensives and atherosclerosis and thrombosis, those kind of thing. But our lab is primarily focused on the heart. So that's why in this paper we have given impresses on the heart. So what we believe at this point that ponatinib lead to this proinflammatory pathway described in this paper, which is just 108A9-NLRP3-IL-1β and this inflammatory pathway lead to a cytokine storm very much like in the COVID-19 and these cytokine storms lead to excessive myocarditis and then finally cardiac dysfunction.   Cindy St. Hilaire:        Is the cytokine storm just local in the cardiac tissue or is it also systemic in the patients? Is cardiotoxicity localized only or is it a more systemic problem?   Tousif Sultan:              I would like to add in this paper we have included that we look this cytokine things and explain blood circulation, bone marrow. So the effect is everywhere, it's not local. So we didn't check other organs, maybe other organs also being affected with the ponatinib treatment.   Cindy St. Hilaire:        And what's the initial phenotype of a patient has when they start to get cardiotoxicity, what's kind of like a telltale symptom?   Hind Lal:                     So good thing that in recent years cardio-oncology developed. So initially the patient that were going for cancer treatment, they were not monitored very closely. So they only end up in cardiology clinic when they are having some cardiac events already. So thanks to the lot of development and growth in the cardio-oncology field, now most patients who going for a long-term cancer treatment, they are closely monitored by cardiology clinics.   Cindy St. Hilaire:        Got it. So they can often catch it before a symptom or an event. That's wonderful.   Hind Lal:                     Yeah, so there's a lot of development in monitoring.   Cindy St. Hilaire:        Wonderful. So you were really interested in figuring out why ponatinib induces cardiotoxicity and you mentioned that really up until now it's been very difficult to study and that's because of the limitation of available murine models. If you just inject a wild type mouse with ponatinib, nothing happens really. So what was your approach to finding relatively good murine models? How did you go about that?   Hind Lal:                     So this is the top scientific question you can ask. So like science, the field is try and try again. So initially this is the first paper with the ponatinib toxicity using the real in vivo models. Any paper before this including ours studies published, they were done on the cellular model in hiPSC, that isolated cardiomyocytes. So you directly putting the ponatinib directly the isolated cells. So this is first case when we were trying to do in vivo, maybe other attempt in vivo but at least not published. So first we also treated the animals with ponatinib and that failed, we don't see any cardiotoxic effect. And then when we going back to the literature, the clinical data is very, very clear from pharmacovigilance that ponatinib is cardiotoxic in humans. So when we're not able to see any phenotype in mouse, we realize that we are not mimicking what's happening in the humans.   So we certainly missing something. Now once again I quote this COVID-19, so many people get infected with COVID-19 but people are having preexisting conditions are on high risk to developing CVD. So there was some literature on that line. So we use this very, very same concept that if there is preexisting conditions, so likely who'd have developing future cardiac event will be more. So we use two model in this paper one atherosclerosis model which is APoE null mice mice, another is tag branding which is pressure overload model for the heart and as soon as we start using what we call comorbidity model like patient is having some preexisting conditions and we very clearly see the robust defect of ponatinib on cardiac dysfunction.   Cindy St. Hilaire:        Yeah, it's really, really well done and I really like that you use kind of two different models of this. Do you think it's also going to be operative in maybe like the diabetic mirroring models? Do you think if we expand to other comorbidities, you might also recapitulate the cardiotoxicity?   Hind Lal:                     So you got all the best questions.   Cindy St. Hilaire:        Thank you. I try.   Hind Lal:                     So because this is CML drug and lot of the risk factor for cardiovascular and cancer are common and even metabolic disease. So most of the time these patients are elderly patients and they're having metabolic conditions and most of the time they have blood pressure or something CVD risk factors. So I agree with you, it'll be very relevant to expand this to the diabetes or metabolic models, but these were the first study, we put all our focus to get this one out so news is there then we can expand the field adding additional models et cetera. But I agree with you that will be very logical next step to do.   Cindy St. Hilaire:        Yeah. And so I guess going back to what you know from the human study or the clinical trials or the human observations, are different populations of patients with CML more predisposed to cardio toxicity than others or is that not known yet?   Hind Lal:                     So one other area called pharmacovigilance. So what pharmacovigilance does patient all over the world taking these drugs. So WHO have their own vigilance system and FDA have their own, so it's called BG-Base for the WHO and it's called the FAERS for the FDA. So one can go back in those data sets and see if X patient taking this Y drug and what kind of symptoms or adverse effect they are seeing and if these symptoms are associated with something else. So there is data that if patients having CVD risk factor, they are more prone to develop ponatinib induced cardiac events. But it needs more polish like you asked the just previous question, diabetes versus maybe blood pressure means hypertension, atherosclerosis, or thrombosis. So it has not been delineated further but in a one big bucket if patients are having CVD risk factor before they are more prone and more likely to develop the cardiac events.   Cindy St. Hilaire:        So after you established that these two murine models could pretty robustly recapitulate the human phenotype, what did you do next? How did you come upon the S100A8/A9-NLRP3-IL-1β signaling circuit? How did you get to that?   Hind Lal:                     So in basic science work, whenever we do mouse is called until we get there is cardiac dysfunction, it's called phenotype, right? So mouse had a cardiac phenotype. So next step is, "Why? What is leading to that phenotype?" That's what we call mechanism. So there the best idea to fit the mechanism is using one of the unbiased approaches like you do unbiased proteomics, unbiased RNC analysis, something like this that will analyze the entire transcript like RNC and say, "Okay, these pathway are," then you can do further analysis that will indicate these pathway are different, are altered. So in this case we used RNC analysis and it came out that this yes A8 and yes A9, 100A8 and nine, they were the most upregulated in this whole set. And thereafter we were very lucky. So we started this study at Vanderbilt, where my lab was and thereafter we very lucky to move here and found Sultan who had a lot of experience with this inflammation and immune system and then Sultan may add something on this so he'll be the better person to say something on this.   Tousif Sultan:              So after our RNC analysis, so we got this S100A8 and nine as top hit with the ponatinib treatment. So then we validated this finding with our flow cytometric, qRT PCR aand then we started which pathway is going to release cytokine and all that. So we found that is NLRP3 inflammasome.   Cindy St. Hilaire:        Yeah and well and I guess maybe step back, what is S100A8/A9? What are those? Tousif Sultan:              Yeah, S10A8/A9 is a calcium binding protein. So that's also called alarmin and they basically binds with the pathogen associated pattern and other TLR2 like receptors and then start inflammatory pathway to release cytokine and all that and it's stable in heterodimer form. So S100A8 heterodimer with A9 and then bind with TLR and a start in this inflammatory pathway.   Cindy St. Hilaire:        And what type of cell is that happening in? Is that happening in the immune cells only or is it also in the cardiomyocyte, or...?   Tousif Sultan:              Yeah, we have included all this data. So from where this alarmin is coming with ponatinib treatment, so literature also suggested that neutrophils and monocytes, those cells are the potential to release the alarmin. So here we also found these two type of cells, neutrophils and monocytes. They release huge alarmin with the treatment of ponatinib.   Cindy St. Hilaire:        And so really taking this really neat mechanism to the next level, you then tried attenuating it by using broad anti-inflammatory steroid dexamethasone but also by targeting these specific components, the NLRP and the S100A specific inhibitors and they worked well. It worked really nicely. Does your data show that any of these therapies work better than the other and then are these viable options to use in humans?   Hind Lal:                     Yeah, we have some data in the paper. Are very broad which help a lot in COVID patients, far very acute infections. So in this case, situation is very different cause most of CML patients will going to take ponatinib for lifelong, there is no remission, right? So in those case, its certainly not a very attractive option. We have shown data in the paper that dexamethasone help with the heart but lead to some metabolic changes. So we have compared those with the NLRP3 inhibitors, those metabolic alterations, dexa versus the NLRP3 inhibitors, CY-09. And we demonstrated that targeting is specifically with paquinimod, our NLRP3 inhibitor CY-09, feel better. It can still rescue the cardiac phenotype without having those adverse effect on metabolic parameters.   Cindy St. Hilaire:        That's wonderful. Do you think though that because you have to take ponatinib for life, that long-term NLRP inhibition would also cause problems or...?   Hind Lal:                     So because not every patient who taking ponatinib would develop the cardiac phenotype, right? Which is like a 10%, 12%, patient developing cardiac dysfunction. So I think someone like I strongly believe paquinimod, which is inhibitor of S100A9, will be really good option or at least we have enough data that make us nail for at least a small clinical trial. And we quickly moving on that. At UAB we have our clinical cardio-oncology program and we are already in touch with the director for the clinical cardio-oncology program. So what we trying to do in that small trial is if one of the standard therapy for heart like beta blocker or ARBs inhibitor, is there any preference like one work better than the other in the standard care? So first we doing that project, then we obviously looking forward if one small clinical trial can be done with paquinimod. I strongly believe it should be helpful.   Cindy St. Hilaire:        That is wonderful. And so do you think... There's other chemotherapeutic agents or probably even other non-cancer drugs that cause cardiotoxicity, do you think this mechanism, this pathway, this S100A-NLRP-IL-1β axis is operative in all cardiotoxicities or do you think it's going to be very specific to the ponatinib?   Hind Lal:                     So it's certainly not all, but it'll be certainly more than ponatinib. So in our lab we are using another kinase inhibitor, which is osimertinib and it's not published yet, but now we know that it's also cardiotoxic because it's taking metabolic root or energetics disruption but not this pro-inflammatory part, but we're doing another project which is strep pneumonia induced cardiac dysfunction, which is called pneumonia. So strep pneumoniae, which leads to the pneumonia ,and lot patient die because of the failing heart we see here in the hospitals and we see these pathways operational over there and we gearing up to do clinical trial on that aspect as well, but it's not generalized like all kind of heart will have the same mechanism.   Cindy St. Hilaire:        It's wonderful to see you're already taking those next steps towards really kind of bringing this to a translational/clinical study. So what was the most challenging aspect of this study?   Tousif Sultan:              The challenging aspect, ponatinib is a kinase inhibitor and that was surprising for us how it's activating immune cells. Generally kinase inhibitors, inhibits all the cells like that. So that was challenging. So we repeated it many times did in vitro experiment to confirm that. So we just added, just treated in vitro immune cells with the ponatinib and confirmed it. So that was little challenging.   Cindy St. Hilaire:        So what's next? You mentioned you're going to try some clinical trials, early stage clinical trials. What's next mechanistically, what do you want to go after?   Hind Lal:                     So what we are doing next and we are very, very eagerly trying to do that. So what it was done, we used the cardiac comorbidity models, but as you know, anybody who will take ponatinib will have cancer, right? So we strongly believe that we miss one factor. There was no cancer on these. So that is very logical next step. What that will allow us to do, what rescue experiment we'll have done in this paper. So we saw, "Okay, this rescue the cardiac phenotype, which is taken care of now," but very same time, we not able to demonstrate that this is happening without hurting the cancer efficacy. So if we have the dual comorbid mouse, which have CML a real thing and we have cardiac thing, then that will allow us to demonstrate, "Okay, we got something that can take care of the cardiac problem without hurting the efficacy on the cancer." And it will be best if you also help little bit to more potentiate the cancer efficacy.   Cindy St. Hilaire:        Yes. Excellent. Well, congratulations on a beautiful study, really exciting findings. Dr Lal and Dr Sultan, thank you so much for taking the time to talk with me today.   Tousif Sultan:              Thank you so much.   Hind Lal:                     Well thank you, Cynthia. We really appreciate your time. Thank you for having us.   Cindy St. Hilaire:        Yeah, it was great.   Cindy St. Hilaire:        That's it for our highlights from the February 3rd and February 17th issues of Circulation Research. Thank you so much for listening. Please check out the Circulation Research Facebook page and follow us on Twitter and Instagram with the handle @CircRes and #DiscoverCircRes. Thank you to our guests, Dr Hind Lal and Dr Tousif Sultan. This podcast is produced by Ishara Ratnayake, edited by Melissa Stoner and supported by the editorial team at Circulation Research. Some of the copy text for the highlighted articles was provided by Ruth Williams. I'm your host, Dr Cynthia St. Hilaire, and this is Discover CircRes, you're on-the-go source for most exciting discoveries in basic cardiovascular research. This program is copyright of the American Heart Association 2023. And the opinions expressed by the speakers in this podcast are their own and not necessarily those of the editors or of the American Heart Association. For more information, please visit ahajournals.org.  

Dr Nally gives the answers to the Egg IQ quiz and answers questions about the causes of persistent elevated blood sugars while following a ketogenic or carnivorous lifestyle. Body Weight Exercise Program: https://q0y908pr.pages.infusionsoft.net Weight Management & Concierge Medicine: https://www.DocMuscles.com/membership/ #Glucose #Cytokines #BloodSugar #LeadFolloworGetOutOrMyWay #JustKeepEsterifying #Ketogenic #Keto #KetogenicLifestyle #Carnivore #DrAdamNally #DocMuscles #DocMusclesLive #DocTalk #DocsWhoLift #LiftRunShoot #DocMusclesLife #PlantBasedDietsSuck #PicoSure #Icon #SculpSure #TempSure #EMScuptNeo #EMSella #EMFace #Skin #Skincare #BeautifulSkin #SkinCarePrimer #Face #Gorgeous #NoBSsm #Muscles #Myokines #Exercise References: Li Y et al., J Diabetes Res. May 2013. Online. ID 170532, p1-8 Sonksen P, Sonksen J. Brit J. Anaesth. July 2000. 85(1): 69-79 Nieto-Vazquez, I et al., Archieves of Physiol Biochem. 2008. 114(3): 183-194 Cook CJ, Phys Behav, Sep 2004, 82:4; 751-762 Herman, J P et al. Braz Jour Med Biol Res, Mar 2012, 45:4; 292-8 McEwen, B., Neuropsychopharmacol, 2000 22:108–124. Khan S, Shafiei MS, Longoria C, Schoggins J, Savani RC, Zaki H. SARS-CoV-2 spike protein induces inflammation via TLR2-dependent activation of the NF-κB pathway. bioRxiv [Preprint]. 2021 Mar 17. Update in: Elife. 2021 Dec 06:10.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.01.04.522511v1?rss=1 Authors: Maurer, S. V., Bolton, J. L., Bilbo, S. D., Williams, C. L. Abstract: Air pollution causes widespread inflammatory changes in the body and brain. When exposure to air pollution occurs early in development, children exhibit impaired working memory ability (Sunyer et al., 2015). In addition, prenatal exposure to diesel particulate matter (DEP) increases inflammatory cytokine expression in the whole brain of embryonic day 18 (E18) males and leads to adverse long-term negative outcomes (Bolton et al., 2012). In contrast, dietary choline supplementation is negatively correlated with inflammatory cytokine production in adult rats and cultured human cells (Zhang et al., 2018; Jiang et al., 2014). When administered as a supplement to pregnant rats, choline also improves working memory in adulthood (Meck et al., 2008; Meck & Williams, 1999; 1997). The current study sought to determine if prenatal dietary choline supplementation protects against the effects of air pollution in the developing brain and in the placenta and fetal liver. These data revealed region-specific microglial morphology alterations in fetal brain and in inflammatory gene expression in the placenta and fetal liver (specifically, Tnf, Tlr2, Tlr4, and Itgam) due to maternal choline supplementation and/or maternal air pollution exposure. We found that DEP led to changes in microglial morphology in the fetal dentate gyrus of E18 male, but not female, fetuses. In the placenta and fetal liver of males, inflammatory gene expression was affected by both DEP and maternal choline supplementation. However, maternal choline supplementation alone upregulated inflammatory gene expression in females, which may indicate an alteration in maturation rate. These data further contribute to the growing literature indicating region- and tissue-specificity in the developmental immune system in the context of maternal exposures. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

In this episode we discuss: Whether PUFA causes more oxidative stress than saturated fats How PUFA intake affects testosterone Whether fruit juice should be avoided for blood sugar regulation The effects of fruit juice on inflammatory markers and endotoxin Whether fruit juice has the same effects as soda   Check out the Energy Balance Solution group coaching program: https://jayfeldmanwellness.com/solution   Click here to check out the show notes: https://www.jayfeldmanwellness.com/ep-89-the-anti-inflammatory-effects-of-fruit-juice-and-the-hormonal-effects-of-pufa-vs-sfa/    Timestamps: 4:10 – the effects of different dietary fat compositions on phospholipid and neutral lipid compositions in the testicles   18:31 – the effects of different dietary fat composition on oxidative damage and antioxidant status in the testicles 29:44 – the effects of different dietary fat composition on hormone production and metabolism in the testicles  38:26 – introducing the next study discussing the effects of orange juice on a high-fat, high-carbohydrate meal in terms of inflammation and endotoxin   44:14 – the effects of orange juice on blood glucose and insulin sensitivity  47:55 – the effects of orange juice on ROS production, TLR2 and TLR4 expression, and plasma endotoxin  51:35 – factors that could account for the benefits of orange juice   

Des chercheurs taïwanais ont découvert la clé des caillots dans le sang induits par le COVID-19. L’avancée scientifique est signée d’une équipe de recherche de l’Academia Sinica qui a identifié deux récepteurs clés qui pourraient fournir une feuille de route pour le traitement des caillots sanguins causés par le Covid-19 et les complications à long terme associées à la maladie. L’équipe dirigée par Hsieh Shie-liang (謝世良) a cherché à découvrir le mécanisme derrière la formation de « pièges extracellulaires de neutrophiles » (NET), un facteur contribuant à la thrombose chez les patients atteints de Covid-19. Cette équipe de chercheurs avait précédemment travaillé sur les personnes touchées par la Dengue, et sou la houlette du professeur Hsieh Shie-liang, ils avaient découvert que les plaquettes provoquaient la libération de vésicules extracellulaires qui amélioraient la formation de ces pièges extracellulaires de neutrophiles via deux récepteurs : CLEC5A et TLR2. Ils sont donc partis d’une hypothèse similaire : à savoir, que ce mécanisme fonctionnait également par rapport aux caillots sanguins développés chez les patients touchés par une forme grave de Covid-19 et qui présentent des niveaux élevés vésicules extracellulaires dérivées des plaquettes. Détails de cette découverte taïwanaise dans l’émission Taïwan en ébul’action de ce vendredi.

CoQ10 supplementation associated with improved trauma patient outcomes Urmia University of Medical Sciences (Iran) July 23 2021.    Findings from a trial reported on July 12, 2021 in the Journal of Nutritional Science revealed benefits for hospitalized traumapatients who were given supplements that contained coenzyme Q10.  The trial enrolled 40 men and women with traumatic injury and low plasma levels of CoQ10. Participants received a placebo or 400 milligrams CoQ10 daily for seven days. Blood samples collected at the beginning and end of the trial were analyzed for interleukin 6 (IL-6), which may be elevated during inflammation, and the oxidative stress markers malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS). Body composition was also assessed at these time points, as well secondary outcomes that included Sequential Organ Failure Assessment (SOFA) and the Glasgow Coma Scale (GCS).  While interleukin-6 levels at the beginning of the study were similar between the CoQ10 and placebo groups at an average of 175.05 pg/mL and 177.82 pg/mL, they were reduced by 76.99 pg/mL in the CoQ10 group and 17.35 pg/mL in the placebo group. MDA values averaged 232.37 picograms per milliliter (pg/mL) and 239.96 pg/mL and were lowered by 88.84 pg/ml among participants who received CoQ10 and by 26.23 pg/mL among those who received a placebo. In comparison with the placebo group, fat free mass, skeletal muscle mass and body cell mass increased among those who received CoQ10. GCS and SOFA scores, and duration of hospital stay, ICU stay and ventilator use also improved among treated patients.  “To date, no randomized clinical trial study has been conducted to evaluate the effect of CoQ10 supplementation in traumatic mechanical ventilated patients and we hypothesized that CoQ10 administration in these patients could have beneficial effects on biochemical and clinical factors,” the authors wrote. “We have shown that CoQ10 could improve some of the clinical and anthropometric parameters in patients with a traumatic injury.”     Nigella sativa (black seed) prevents covid-induced vascular damage, scientists conclude   Oriental Institute of Science and Technology (India), July 27, 2021 New research published in the journal Vascular Pharmacology shows that Nigella sativa, also known as black seed or black cumin, binds to ACE2 in the lungs, effectively stopping the Wuhan coronavirus (Covid-19) from inducing inflammation and vascular damage. Researchers out of India investigated the effects of nigellidine, an indazole alkaloid of black seed, using molecular docking for binding to different angiotensin-binding proteins, as well as the Chinese Virus spike glycoprotein. They found that nigellidine “strongly binds” to the Chinese Virus spike protein at what is known as the hinge region or active site opening, which may in turn hamper its binding to the nCoV2-ACE2 surface. “Nigellidine effectively binds in the Angiotensin-II binding site / entry pocket,” the study explains. “Nigellidine showed strong binding to mono / multi-meric ACE1.” This process of ACE blocking could, the study goes on to suggest, restore angiotensin levels and restrict vasoturbulence in Chinese Virus patients, while the receptor blocking could help to stop resulting inflammation and vascular impairment. “Nigellidine may slow down the vaso-fluctuations due to Angiotensin deregulations in Covid patients,” the paper further explains. “Angiotensin II-ACE2 binding (ACE-value -294.81) is more favorable than nigellidine-ACE2. Conversely, nigellidine-ACE1 binding-energy / Ki is lower than nigellidine-ACE2 values indicating a balanced-state between constriction-dilatation.” Nigellidine also binds to the viral spike proteins, which when taken by Chinese Virus patients, and especially those who fall in the elderly category, could greatly reduce their risk of suffering complications or death. Nigellidine impairs SARS-CoV-2 infection, “cytokine storm” through numerous mechanisms In a related study that was published last year in the journal Europe PMC, researchers learned that nigellidine inhibits the Chinese Virus infection in several other ways. It was discovered early on in the “pandemic” that many of those who tested “positive” for the virus were suffering associated “cytokine storms,” in which their immune systems were over-responding and causing more damage, or even death. Nigellidine was then studied and discovered to possess certain properties that inhibit cytokine storms, as well as impede the SARS CoV-2 virus from causing infection. It is also hepato- and reno-protective, meaning it protects against liver damage. Beyond this, nigellidine was determined to possess unique immunomodulatory and anti-inflammatory characteristics, as well as antioxidant potential strong enough to inhibit important proteins associated with the Chinese Virus. In their quest to uncover possible “drug” candidates to protect patients against hyper-inflammation and other associated problems, the researchers learned that nigellidine – and more than likely other black seed constituents – helps tremendously with preventing negative side effects. Along with nigellicine, nigellidine is found in the seed coat of Nigella sativa. Both of these constituents in their sulfated forms are extremely bioavailable, and along with thymoquinone and dithymoquinone, two other black seed components, they show strong antioxidant, antibacterial, anti-hypertensive, anti-inflammatory and immunomodulatory effects. Black seed extracts have been shown in other experiments to decrease oxidative stress, effectively lowering the risk of inflammation-related diseases. We now know that this includes the Wuhan coronavirus (Covid-19). Black seed is also recognized as a metabolic protector, helping to improve lipid and blood sugar levels. “Most importantly, in SARS CoV-2 infection ACE-2 mediated impairment of aldosterone system may be repaired by,” the study further explains, providing relevant information to the current “pandemic.” “Vasorelaxant and anti-hypertensive function of [black seed] helps in the modulation of renin angiotensin system (RAS) or the diuretic activity, which is one of the major targets of COVID. It might have great protective role during post infective secondary disorder of the peripheral vasculature namely cardiac and renal systems. In most of the instances patients die due to this organ dysfunction/failure in COVID-19 infection.” By quelling inflammation, black seed could save lives from covid Laboratory studies have found that intake of Nigella sativa significantly improves the parameters for hyperglycemia and diabetes control, as well as glycated hemoglobin and insulin resistance. Based on this, experts believe that nigellidine specifically could play an important role in fighting the Chinese Virus by “docking” to the proteins and inflammatory molecules that can cause a cytokine storm – mainly TNF-? receptors such as TNFR1, TNFR2 and IL1R. “In the experimental rat model the source of this drug Nigella sativa; black cumin seed extracts were tested for its role on antioxidant, hepatic and renal status,” the paper states. “This work will help in the urgent therapeutic intervention against COVID-19 global pandemic.” “In the current study, we have decisively shown by molecular modeling that nigellidine can bind in the active sites of several important proteins of SARS CoV 2, several host receptors specific for SARS CoV-2 induced inflammatory markers IL1, IL6, TNF-?. Moreover, the extract from black cumin seed has been shown in experimental rat to be highly antioxidative, hepato- and reno-protective. Further studies are necessary to verify the potential effects of nigellidine in in vivo laboratory experimental animal model.”   Vitamin D supplementation improves recovery time of children with pneumonia at pediatric hospital Cairo University (Egypt), July 20, 2021 According to news reporting originating from Cairo, Egypt, by NewsRx correspondents, research stated, “Despite the well-recognized effect of vitamin D in metabolism and homeostasis, there is now growing interest in its probable association with pneumonia. This study aims to supply vitamin D3 (Cholecalciferol) (100,000 IU) to pneumonic children to minimize the duration of illness and improve their outcome.” Our news editors obtained a quote from the research from Cairo University, “A double-blinded, randomized, placebo-controlled trial was conducted in a Pediatric Cairo University affiliated hospital. An intervention arm (93 children) and a control arm (98 children), who had pneumonia with an insufficient or deficient level of vitamin D and whose parental permission was obtained, were enrolled in the trial. All children were treated with antibiotics according to WHO guidelines. Children were given a single injection of 1 mL of 100,000 IU of vitamin D3 or placebo. Clinical data were recorded every eight hours for all children. Outcomes were assessed 7 days after vitamin D injection. The primary outcome variable was the change in serum level of 25(OH)D, while the secondary outcomes were the medical state of the assigned cases (improvement or death) and duration between enrollment and hospital discharge for improved cases. In the supplementation group, the percentage of patients who suffered either deficient (38.7%) or insufficient levels (61.3%) of 25 (OH)D at day one had significantly decreased in the seventh day to (11.8%) and (52.7%), respectively. Kaplan--Meier plots highlighted that the median time to recover of the placebo group was significantly longer than that of the supplementation group (Log Rank P value < .001). VDD was detected in pediatric critical care children.” According to the news editors, the research concluded: “In pneumonic children with high VDD, it is illustrated that Vitamin D supplementation is accompanied by lowered mortality risk and pSOFA scores, reduced time to recover, and improved PaO2/FiO(2).”   Physical activity could combat fatigue, cognitive decline in cancer survivors University of Illinois, July 26, 2021 A new study indicates that cancer patients and survivors have a ready weapon against fatigue and "chemo brain": a brisk walk. Researchers at the University of Illinois, along with collaborators at Digital Artefacts in Iowa City, Iowa, and Northeastern University in Boston, looked at the association between physical activity, fatigue and performance on cognitive tasks in nearly 300 breast cancer survivors. "The data suggest that being more physically active could reduce two of the more commonly reported symptoms in breast cancer survivors: fatigue and cognitive impairment," said study leader Edward McAuley, a professor of kinesiology and community health at Illinois. "Most people think, 'If I exercise, I'll become tired.' In our study, exercise actually was associated with reduced fatigue, which in turn was associated with better cognitive function." Cognitive impairment, such as memory problems or shortened attention spans, is a common complaint among cancer patients and survivors, and is thought to be similar to decline due to aging. Past Illinois research has explored the effect of physical fitness on age-related cognitive decline, so the researchers wondered whether cancer survivors would respond similarly to exercise. "Other studies of cancer survivors have relied on small samples of cancer survivors, and used self-reporting measures of physical activity and cognitive function, which can be very biased," said postdoctoral researcher Diane Ehlers, the first author of the study, which is published in the journal Breast Cancer Research and Treatment. "What makes our study novel is that we had objective measures for both physical activity and cognitive performance, and a nationwide sample of breast cancer survivors." The researchers worked with Digital Artefacts -- developer of the commercial neuroscience app BrainBaseline - to create an iPad app tailored to this study. The app included questionnaires and activities designed to measure attention, memory and multitasking skills. The researchers also sent each participant an accelerometer to track daily physical activity. "We found that higher levels of daily moderate-to-vigorous physical activity were associated with better performance on the cognitive tasks measuring attention, memory and multitasking," Ehlers said. "What was notable was that physical activity's effect on cognitive performance was mediated by fatigue. This provides evidence that physical activity interventions targeting fatigue in cancer patients and survivors might provide promising models for improving cognitive function as well." Next, the researchers plan to conduct further studies to establish causation and further explore the pathways of how physical exercise improves cognitive performance. They are working with Digital Artefacts to conduct an iPhone-based study and focusing on diverse populations of breast cancer survivors. "The message for cancer patients and survivors is, get active!" Ehlers said. "Even if it's 10-minute bouts of brisk walking. It's not a magical cure-all, but we've seen many benefits of physical activity for cancer patients and survivors."   Cannabidiol promotes oral ulcer healing by inactivating CMPK2-mediated NLRP3 inflammasome Sichuan University (China), July 26, 2021 Xingying Qi, West China Hospital of Stomatology, Sichuan University, Chengdu, China, presented the oral session "Cannabidiol Promotes Oral Ulcer Healing by Inactivating CMPK2-Mediated NLRP3 Inflammasome" at the virtual 99th General Session & Exhibition of the International Association for Dental Research (IADR), held in conjunction with the 50th Annual Meeting of the American Association for Dental Research (AADR) and the 45th Annual Meeting of the Canadian Association for Dental Research (CADR), on July 21-24, 2021. The oral ulcer is a common oral inflammatory lesion with severe pain but little effective treatment is currently available. Cannabidiol (CBD) is recently emerging as a therapeutic agent for inflammatory diseases. However, the underlying mechanisms are not fully elucidated. Qi and colleagues sought to investigate whether and how CBD could play a therapeutic role in the oral ulcer. Oral ulcer models were performed in the tongue of C57BL/6 mice by acid etching or mechanical trauma, followed by CBD local administration. Samples were harvested for macroscopic and histological evaluation. CBD oral spray on acid- or trauma-induced oral ulcers on mice tongues inhibited inflammation, relieved pain and accelerated lesions closure in a dose-dependent manner. The results show that CBD accelerates oral ulcer healing by inhibiting CMPK2-mediated NLRP3 inflammasome activation and pyroptosis, which is mediated mostly by PPARγ in nucleus and partially by CB1 in plasma membrane. This data may shed light on the development of new therapeutic strategies for oral ulcers.   Algal solution: Could Spirulina modify the microbiome to protect against age-related damage? Louvain Drug Research Institute (Belgium), July 25 2021 Spirulina might help protect against age-related liver inflammation by modifying pathways in the microbiome, say researchers. Consumption of spirulina could help protect against hepatic inflammation in the elderly, according to the new animal research published in Nutrients. Belgian researchers carried out tests on mice, which suggest that the algae Spirulina has an impact on the gut microbiota, which in turn activates the immune system in the gut and improves inflammation in the liver that is associated with ageing. Led by senior author Professor Nathalie Delzenne from the Louvain Drug Research Institute in Belgium, the team said oral feeding of Spirulina was found to modulates several immunological functions involving, among others, the TLR4 pathway in old mice. “The fact that its oral consumption can influence both gut immunity and systemic sites, such as the liver, suggests that its immune action is not confined to the gut immune system,” wrote the team – who said the findings open the way to new therapeutic tools “in the management of immune alterations in aging, based on gut microbe-host interactions.” Furthermore, they suggested that improvement of the homeostasis in the gut ecosystem ‘could be essential' during the aging process, “and, in this perspective, dietary manipulation of the gut microbiota of the elderly with Spirulina, may represent a tool for preserving a healthy gastrointestinal microbial community in addition to its beneficial effects on immune function.” Study details Delzenne and colleagues noted that while the possible cardiovascular and immune support benefits of Spirulina have been fairly widely reported, the new study brings a fresh approach by testing whether the effects could be related to a modulation of gut micrbiota. In the trial, young mice aged three months were fed a standard diet, while older mice aged 24 months were fed a standard diet either with or without 5% Spirulina for six weeks. Upton supplementation with Spirulina, the team reported several changes to gut microbiota composition, including an increase in Roseburia and Lactobacillus populations. “Interestingly, parameters related to the innate immunity are upregulated in the small intestine of Spirulina-treated mice,” said the team. “Furthermore, the supplementation with Spirulina reduces several hepatic inflammatory and oxidative stress markers that are upregulated in old mice versus young mice.” Expression of several genetic and biochemical markers of inflammation and immunity were altered by supplementation with Spirulina, said the team. In particular, the transcription factor Foxp3 – involved in the differentiation of T cells into regulatory T cells (Tregs) – and MCP1 were increased due to Spirulina supplementation in old mice. Old mice that consumed Spirulina also showed activation of several immune parameters including Foxp3 in the ileum – suggesting an improvement of the gut immune function upon Spirulina treatment in this segment, said the Belgian researchers. Furthermore, Spirulina supplementation upregulated both TLR2 and TLR4 expression in the ileum of aged mice. “In accordance with these results, a solution of Spirulina (5%) exhibited a TLR4 agonist activity similar to the one reached in old-SP mice, suggesting a direct effect of the Spirulina, itself, on the TLR4 pathway,” they added. Microbiome mechanisms While the positive effect of Spirulina on the microbiome and liver inflammation is clear, the team noted that the mechanism by which the algae could change the composition of the intestinal microbiota remains unanswered. One possible mechanism could be the presence of antimicrobial substances produced by Spirulina, they said. “On the other hand, antimicrobial peptides (AMPs) could be mediators of the nutritional modulation of the gut microbiota.” “In the present study, RegIIIγ and Pla2g2 were increased by the supplementation with Spirulina, suggesting that the host contributes to the reduction and modification of the microbial community by modulating the production of specific AMPs,” they added.

TWiV discusses the finding that the envelope (E) protein of SARS-CoV-2 is sensed by toll-like receptor 2 on cells, leading to the production of inflammatory cytokines that cause damage to cells and tissues in COVID-19. Hosts: Vincent Racaniello, Rich Condit, Kathy Spindler, and Brianne Barker Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode ASV vaccine town halls Michelson Prizes FDA authorizes Pfizer/BioNTech mRNA vaccine for adolescents (FDA) TLR2 senses SARS-CoV-2 E protein (Nat Imm) Review on CoV E protein (Virology J) Timestamps by Jolene. Thanks! Weekly Picks Brianne – Lab.Hacks Lab Assistant App Rich – Weird viral DNA spills secrets to biologists; COVAX Kathy – Planet Earth Now Vincent – Variants at ViralZone Intro music is by Ronald Jenkees Send your virology questions and comments to twiv@microbe.tv

TWiV discusses the finding that the envelope (E) protein of SARS-CoV-2 is sensed by toll-like receptor 2 on cells, leading to the production of inflammatory cytokines that cause damage to cells and tissues in COVID-19. Hosts: Vincent Racaniello, Rich Condit, Kathy Spindler, and Brianne Barker Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode ASV vaccine town halls Michelson Prizes FDA authorizes Pfizer/BioNTech mRNA vaccine for adolescents (FDA) TLR2 senses SARS-CoV-2 E protein (Nat Imm) Review on CoV E protein (Virology J) Timestamps by Jolene. Thanks! Weekly Picks Brianne – Lab.Hacks Lab Assistant App Rich – Weird viral DNA spills secrets to biologists; COVAX Kathy – Planet Earth Now Vincent – Variants at ViralZone Intro music is by Ronald Jenkees Send your virology questions and comments to twiv@microbe.tv

In this episode, Sarah and Stacy breakdown Botox - what is it, what are the side effects, what are the adverse reactions, and what does the science tell us about the impact of this procedure. Our hosts bring this thorough scientific discussion full circle and share their personal feelings towards this procedure and how they personally handle skin health. Click here to listen in iTunes If you enjoy the show, please review it in iTunes! The Paleo View (TPV), Episode 349: Must-Know Botox Info (0:41) Intro It feels like an eternity to Sarah since she last had the chance to talk to Stacy, which it basically has been since they prerecorded the latest episodes while Stacy was traveling Stacy will share about her travels on next week's episode, but she is still processing it all The trip was wonderful, and she has shared some updates via social media, but it was a family trip with many experiences and she is determining what all she wants to share and all the feels One of the things that happened while Stacy was traveling; she got a massage and it put her back in a flair Stacy did really well up until that point, which was 10 days into the trip. And she is excited to share that the first thing she did when she got home was put her Joovv to use and has been doing red light therapy on her back since returning and feeling the benefits of her Joovv (this week's episode sponsor!) While we will cover Joovv in greater detail later in the show, if you are ready to get all the details on them now, head over to joovv.com/thepaleoview, and learn all about our favorite biohack Sarah is gearing up for Paleo Fx, she leaves tomorrow - so by the show airs she will be on a stage sticking it to the man, which is what she typically does there She often finds herself in the myth-busting role at these events, but doesn't actually like confrontation and wishes she could simply have everyone read the same scientific studies she is referring to It is way easier to be on a soapbox on Skype with Stacy than it is to be in front of a few hundred or a thousand people During Sarah's workshop in February talking about different types of scientific studies became really thematic, and how do you evaluate science and what do you look for and how do you detect pseudo-science - are topics that people would love to know more about Sarah is trying to figure out how to condense these topics for the average person, as she recognizes the need for those in the health field to have access to this information Stacy notes the importance of learning not just how to read these scientific studies, but how to use and apply the information in life without bias To be able to look at the science and let that drive justification or not as to why something is improving your health or not, which is where the topic for this week's episode came from What has been interesting for Stacy as a member of this non-toxic living community is the questions she receives around the use of Botox, among many other things, and where these injections fall in the healthy living spectrum Stacy strives to help people find healthy solutions and Botox is off her list - through even just her preliminary research she has found that it is not for her So on this week's episode, she wants to answer this question from a scientific perspective Whether or not Botox is right for someone is ultimately a personal decision, but Stacy and Sarah's aim for this episode is to talk about the science and why both Stacy and Sarah would or would not make such a decision This is not a judgment show; we want to make everyone feel comfortable when they listen to the show Today's discussion is information driven and we are letting the science speak for itself that is how Stacy and Sarah are driving their decisions If Botox was safe Stacy would go out and have it done in a heartbeat Sarah wants to note that there is a lot of pressure for social influencers to look a certain way, to appear young, healthy and vibrant is a part of the credentials Botox is a fast solution and Sarah understands the allure of it and why many are driven to use it it seems like an effective way to get the results we are looking for What Stacy has personally done, in addition to more natural solutions (infrared - Joovv), is changed who is influencing her life so that she is no longer seeing only seeing a barrage of perfection, but is instead seeing people who inspire her to be herself and be real and be natural If a listener is feeling the pressure of something, Stacy suggests looking into where that pressure is coming from and how you can reduce that pressure You may not even realize where all the pressure is coming from - but ask what can you do to control it and empower yourself to value who you are and how you look Find someone who tells you that your flaws are beautiful (20:59) And Now the Science Botulinum toxin is what Botox is Botox was a carefully considered shortening of the name of a neurotoxic protein The way that it causes botulism is by preventing the release of a neurotransmitter It causes what is called flaccid paralysis, which is paralysis by muscle relaxation It is the most lethal toxin known to man A lethal dose of botulinum toxin is as low as one or two nanograms per kilogram body weight when injected into your muscle or into your bloodstream, which is an incredibly small amount of the toxin It was studied originally in research because of this interest in the ability for it to block nerves that control muscles to cause muscle relaxation There are of course medical uses where botox does have therapeutic benefit, but that is a different evaluation all together However, it is worth noting as we talk about the undesirable side effects that the therapeutic doses tend to be even higher than the cosmetic doses and so the risk of an adverse effect is much higher in the therapeutic applications So if you are dealing with one of these situations where botox may have therapeutic benefits Sarah highly recommend having a very thorough conversation with your medical provider and really understanding what all your options are and what to look for with adverse effects There are very few scientific studies that look at the effects of Botox beyond two years and very few look at the effects of multiple injections What Stacy finds interesting is all of the rebrandings of Botox, and these products aren't different they are just marketed differently So be aware of the Botox, look at the warning labels, and do the research (27:59) Side Effects and Adverse Reactions A side effect is a minor complaint that happens on the side that basically resolves on its own Whereas an adverser effect is a major problem, potentially life-threatening that requires medical intervention Both are known to happen with Botox use These are a list of the side effects: drooping eyelids uneven eyebrows a crooked smile, which can lead to drooling asymmetry swelling bruising, discomfort and inflammation in the injection area systemic effects include: fatigue headache neck pain double vision dry eyes or excessive tearing fever and chills allergic reactions (hives, rashes, asthma, etc.) Adverse reactions include: difficulty speaking difficulty swallowing severe muscle weakness loss of bladder control vision problems Then there is a gray area of in between where some studies qualify certain reactions as adverse whereas other studies define the same reactions as side effects Like vomiting, heart function, lung function, etc.; based on how severe they are they get put on either end of the spectrum What Sarah finds kind of scary is that very few studies have looked at repeated treatments and long-term effects, especially beyond two years It is estimated that there are 5 million Botox treatments a year globally and that it is a 2 billion dollars a year business right now - and this is just looking at the cosmetic use There was one study published in 2005 that looked at participants over the course of 12 years who were using Botox for both therapeutic and cosmetic reasons The study found that during the study period there were 20 cases of adverse effects in 16 of the participants, about a 1/3 of the participants This included: difficulty swallowing, droopy eyelids, neck weakness, nausea, vomiting, blurred vision, general or marked weakness, difficulty chewing, hoarseness, swelling, difficulty speaking and heart palpitations A 2015 review of the research found that there have been very few long-term studies and the risk of adverse effects seems to really increase after the 10th or 11th injection For most people this is three to four years out of doing this regularly This hasn't been studied rigorously, despite the wide use of Botox Around 2015/2016 there was a spike in studies showing problematic effects and it started to hit the news that Botox might be as safe as we think it is There have only been a handful of studies in the past few years that build on that In the grand scheme of things though, it takes three to four years to build on these ideas and complete the research, so we are essentially waiting on these research labs to come out with their follow up papers (30:30) The Latest Findings In this timeframe, 2015/2016, there were a couple of papers that showed Botox actually travels through neurons So up until 2015, it was believed that Botox could defuse a short way through the cells Now it is known that it migrates, which explains how you could get full body weakness from a Botox injections This is the explanation for these systemic adverse effects These studies haven't hit the general body of knowledge around Botox The studies Sarah referenced: Long-term botulinum toxin efficacy, safety, and immunogenicity The 2015 findings Serious and long-term adverse events associated with the therapeutic and cosmetic use of botulinum toxin Botulinum neurotoxin type A induces TLR2-mediated inflammatory responses in macrophages A 2009 study titled, “The link between facial feedback and neural activity within central circuitries of emotion--new insights from botulinum toxin-induced denervation of frown muscles” A 2010 study titled, “Cosmetic use of botulinum toxin-A affects processing of emotional language" A 2011 study titled, “Embodied Emotion Perception” 2014 study titled, “Botulinum toxin-induced facial muscle paralysis affects amygdala responses to the perception of emotional expressions: preliminary findings from an A-B-A design" A 2014 study titled, “Altered cortical activation from the hand after facial botulinum toxin treatment” 2016 study titled, “Deeper than skin deep – The effect of botulinum toxin-A on emotion processing” In addition to the impact that Botox has on the nervous system, there is also direct immune effects, which also has some concerning implications There are only a couple of studies that look at this, but Botox is basically causing an incredibly intense inflammatory response As Sarah was researching this and feeling frustrated at the lack of long-term studies, she thought that this would have been a component that would have been needed for FDA approval Sarah hit on this entire other field of research that looks at the impact of Botox of mental and emotional health An important piece to note from the latest research is that Botox is only 80% effective, so 20% of people who get Botox (whether used for therapeutic or cosmetic use) and don't actually get the benefits of Botox There is a collection of studies showing that Botox blunts emotional responses and emotional experiences Stacy's mind has been completely blown by these findings The idea of not being able to pick up on emotional queues would devastate Stacy There are 8 to 10 studies that have looked at these emotional and mental effects, so it has been fairly thoroughly looked at and it is showing the impact that Botox has on the brain stem (47:59) Closing Thoughts on Botox On a side, Sarah highly recommends the book Brainstorm There is also the potential for side effects on memory and feeling emotions in general The idea of giving up something for the benefit of another thing, in this case, the aesthetics, and does that really make the person happy In Stacy's experience, she has never known anyone who had a cosmetic procedure that said, and now I am complete and fulfilled and everything is great Stacy often hears from people that on the other side of that change you make, that it doesn't actually solve the problem, it just highlights another for you There are so many articles out there about people who become addicted to cosmetic procedures Society has come to a point where we have lost the ability to step back and look at more than just the wrinkle, and this case, all the many other things impacted by one procedure The health consequences are very problematic for Stacy, but the inflammatory response and the mental and emotional issues that come along with it make it all the more horrifying to her Sarah thinks that its this collection of facts that make Joovv such a serendipitous sponsor for this show because when Sarah thinks about the things that would bring someone to Botox her first reaction is - well, what about all the effective, safe things you can do instead Diet and hydration, exercise all have a huge impact on skin health If you are going to invest in something to improve your appearance cosmetically, Joovv red light therapy would be a route that is highly recommended There are a ton of studies showing that the two wavelengths in Joovv help to reduce inflammation and stimulate collagen production in the skin The two wavelengths used in Joovv actually make the skin physically younger, as opposed to just making it appear younger, which is what Botox does (53:34) Infrared Sauna vs. Joovv Therapy An infrared sauna is a higher wavelength that works by increasing your core temperature and forcing you to sweat, which is a detoxification pathway Joovv actually combines two different wavelengths, an infrared wavelength and a red wavelength It can still increase your core temperature if you sit in front of it for long enough, but the addition of the red light to the infrared light is what gives it the magical formula A red light goes deeper into your skin and is the main wavelength that is increasing cellular health Sarah finds Joovv to be the best of both worlds, and has actually found that their product is the only one actually delivering therapeutic doses If you want to shop Joovv, you can go to joovv.com/thepaleoview There are different sizes and different price points, and all offer the health benefits that Stacy and Sarah discussed (59:14) Final Thoughts There are also, of course, topical treatments that people can do to treat wrinkles, but a lot of the antiaging products that are on the market actually intentionally disrupt your hormones For those of us working really hard on lifestyle factors to regulate hormones, what you don't want to be doing is slathering yourself in some sort of cream that is just going to disrupt them Be careful about the products you are using Aging is not the most fun thing in the world There is not one magical thing that fixes everything, it is the little bit of benefit that we get from each choice (diet, lifestyle, the use of biohacks like Joovv) that compounds The goal for Sarah isn't to have her skin look younger, the goal is for her skin to be younger, and that is where the diet, lifestyle, and the smart use of biohacks that have scientific validity all comes together for magic (science) awesome Stacy wants to remind everyone that they are wonderful and beautiful just as you are Accepting yourself where you are and wanting to change is so important, so if there is something you want to change, Stacy suggests finding acceptance with yourself before you go on to find that next great thing because you might find that those laugh lines aren't something you actually want to change If this is something you have done already, this is not meant to be a dig on you Stacy and Sarah's goal is to simply help you be informed and make the best decisions for your health Thank you for tuning in and having patience on this deeply scientific show Thank you Sarah for pulling together all of this research and information Don't forget that you can submit follow up questions through both Stacy and Sarah's websites or on social media We will try to compile any questions received and if Stacy and Sarah need to do a follow-up show, we will as soon as we are able Stacy wished Sarah a wonderful trip to PaleoFx When Sarah returns, Stacy looks forward to discussing both of their wonderful trips

Crescentic glomerulonephritis is characterized by glomerular necrosis. Dying cells release intracellular proteins that act as danger-associated molecular patterns to activate the innate immune system. Previously, we have demonstrated that dying tubular cells release histones, which can kill endothelial cells and activate the toll-like receptor 2/4 (TLR2/4). This drives tubulointerstitial inflammation in septic or post-ischemic acute kidney injury (AKI). Furthermore, other groups have also reported that extracellular histones cause organ damage during acute lung injury, stroke, peritonitis and retinal dysfunction, and that blocking extracellular histones represents a beneficial approach during the disease progression. In this thesis, we investigated whether extracellular histones can elicit similar pathogenic effects during necrotizing glomerulonephritis. To do so, we used an animal model based on the necrotizing type of severe glomerulonephritis. Necrotic glomerulonephritis was induced in mice by a single intravenous injection of 100µl sheep anti-GBM antiserum. The impact of histone neutralization was studied by using an antibody isolated from the BWA-3 clone, which had the capacity to neutralize released extracellular histones in-vivo and in-vitro. After 7 days, mice were sacrificed and kidneys were collected for further data analysis. Proteinuria was assessed in spot urine samples. Anti-GBM treated mice showed increased proteinuria (albumin/creatinine ratio), plasma creatinine and BUN levels. This was associated with a reduced number of podocytes, increased crescentic glomeruli and the infiltration of neutrophils and macrophages into the kidney. Interestingly, neutralization of extracellular histones significantly reduced proteinuria leading to less podocyte damage. This was linked to an improved renal function defined by lower plasma creatinine and BUN levels, and with a decrease in neutrophil and macrophage infiltration and activation in kidney. Histone blockade also significantly reduced renal mRNA expression of TNF-α and fibrinogen in the glomerular capillaries, which was associated with less glomerulosclerosis, crescents and tubular atrophy. In-vitro studies demonstrated that extracellular histones and NETs-related histones kill glomerular endothelial cells, podocytes and parietal epithelial cells in a dose-dependent manner. Histone-neutralizing agents such as anti-histone IgG, activated protein C or heparin prevented this cytotoxic effect. Stimulation of BMDCs with histones upregulated the expression of the activation marker including MHC-II, CD48, CD80 and CD86 significantly as well as increased the production of TNF-α and IL-6. It has been previously reported by others including us that in biopsies from patients with ANCA-associated vasculitis showed an over-expression of the TLR2/4 receptor compared to the healthy glomeruli. Histone toxicity on glomeruli ex-vivo was also dependent on the TLR2/4 receptor axis given that the lack of TLR2/4 attenuated histone-induced renal thrombotic microangiopathy and glomerular necrosis in mice. Anti-GBM glomerulonephritis involved NET formation and vascular necrosis, while blocking NET formation via PAD inhibitor or pre-emptive anti-histone IgG injection significantly reduced all parameters of glomerulonephritis including vascular necrosis, podocyte loss, albuminuria, cytokine induction, recruitment and activation of glomerular leukocytes, and glomerular crescent formation. Finally, to evaluate histones as a therapeutic target, mice with established glomerulonephritis were treated with three different histone-neutralizing agents such as anti-histone IgG, recombinant activated protein C and/or heparin. Interestingly, all agents were equally effective in abrogating severe glomerulonephritis, while combination therapy had no additive effect. In summary, the results of this thesis indicate that NET-related histones released during glomerulonephritis elicit cytotoxic and immunostimulatory effects and that neutralizing extracellular histones, therefore, represents a potential therapeutic approach when applied during already established glomerulonephritis.

Kathleen Maguire-Zeiss, Stefano Daniele, and Hang Yin explain why drugs that inhibit signaling by pattern recognition receptors on microglia reduce neuroinflammation associated with synucleinopathies such as Parksinson's disease.

Hi guys in todays Disney Magic News we’ll go over the release of the new star wars trailer, the announcement of the new frozen short, as well as check out some new toys based on Toy Story That Time Forgot. Please be sure and hit that Subscribe button so you never miss a video. News updates every Friday at 9 am Central! Free Ebook : https://gumroad.com/l/eSHT Let's Connect : Podcast : http://goo.gl/P6NX7E Google+ : http://goo.gl/4zxZDu Twitter : http://goo.gl/1sD8Yf Instagram : http://goo.gl/P4oJF9 Hi guys andwelcome to disney magic news where every Friday we discuss the latest regarding anything Disney. I hope you had a fantastic Thanksgiving holiday I apologize that I did not have the news up. I underestimated the power of the turkey greatly. On Thursday Thanksgiving day Disney surprised us all with the release of the new Star Wars Teaser trailer,The Force Awakens. While the teaser offers little the minute and a half trailer will get your blood flowing. I have to admit I was cautious not to get to excited but after watching it one time, I repeated it pretty much all day. I am excited for this movie but I’d like to know what you think. Comment below do you think this will surpass the expectations of star wars fans or as a fan do you feel that this movie shouldn’t even be made? Let me know comment below. In other news today Disney has officially released the date for their new Frozen short film, Frozen Fever. The film is set to debut prior to the live action film Cinderella. Disney also released a brief summary regarding the new short. “It’s Anna’s birthday and Elsa and Kristoff are determined to give her the best celebration ever, but Elsa’s icy powers may put more than just the party at risk.” You can watch Frozen Fever and Cinderella when they are released on March 13 of 2015 And finally with the release of the Toy Story Holiday Special "That Time Forgot”, you’ll be able to get your hands on some special Toy Story Toys. Thinkway Toys will be releasing a Buzz and Woody action figure matching the size and color specified by Pixar. These two toys are the first to be released but not the last. Mattel will also be releasing a Buzz, Woody, Trixie, and many of the Battlesaurs from the Pixar special. For now you can find these Buzz and Woody toys at ToysRUs or Ebay. I’ll have a link to a detailed unboxing and review video down below. Alright guys that does it for this weeks Disney Magic News. Remember you can find links to all of todays stories in the description down below. As always if this is your first time visiting the channel please don’t hesitate to hit that Subscribe button so you’ll never miss a video. And be sure to check some of the other playlists on the channel such as VinylTuesdays and Let’s Play Disney Infinity. I’m your friendly mouse reporter Joseph b. Wishing you a great, night, day, weekend, or whenever you’re watching this. And as always stay magical! point to the subscriber button. and say If Links : Star Wars The Force Awakens : http://goo.gl/YKQpNE Frozen Fever : http://goo.gl/y7uplu Toy Story That Time Forgot Toys : http://goo.gl/tFXAeI

Hi guys in todays Disney Magic News we’ll discuss about the new live action filmCinderella. As well as say happy birthday to a certain mouse. lease be sure and hit that Subscribe button so you never miss a video. News updates every Friday at 9 am Central! Free Ebook : https://gumroad.com/l/eSHT Let's Connect : Podcast : http://goo.gl/P6NX7E Google+ : http://goo.gl/4zxZDu Twitter : http://goo.gl/1sD8Yf Instagram : http://goo.gl/P4oJF9 Hi guys and welcome to disney magic news where every Friday we discuss the latest regarding anything Disney. This past Tuesday Disney released their newest trailer for the live-action film of Cinderella. The film will stay true to the animated version for the most part except in some minor details. The new Cinderella film is directed by Kenneth Branagh and will star Helena Bonham Carter, Cate Blanchett, and Lily James as Cinderella. You can watch Cinderella on when it releases on March 13 of 2015. I’d like to know how you feel about all of these remakes. With Maleficent being a slight different version of sleeping beauty and now Cinderella becoming a live action film. Do you think Disney should keep this up? Or do you feel they should let animation stay animation and live action stay live action. Comment below I’d like to know what you think. In other news today Mickey Mouse had his 86 birthday this past Monday . As part of his birthday celebration disney released a new short called “Mickey Monkey”. The animated short puts Mickey back on a boat along with some of his friends that you’ll be sure to recognize. The short commemorates the original film called “Steamboat Willie” that was released on November 18,1928. Alright guys that does it for this weeks Disney Magic News. Remember you can find links to all of todays stories in the description down below. As always if you enjoyed this video please don’t hesitate to hit that Subscribe button. And be sure to check some of the other playlists on the channel such as VinylTuesdays and Let’s Play Disney Infinity. I’m your friendly mouse reporter Joseph b. Wishing you a great, night, day, weekend, or whenever you’re watching this. And as always stay magical! Links : Cinderella : http://goo.gl/smbh3h Mickey Monkey : http://goo.gl/13WctG

Die Ischämie-Reperfusion in der Niere führt zur Aktivierung des angeborenen Immunsystems mit nachfolgender steriler Entzündungsreaktion und Gewebeschädigung der Niere, das v.a. das Tubulussystem betrifft. Es werden v.a. residente dendritische Zellen aktiviert, die die größte Immunzellpopulation in der Niere darstellen.In der weiteren Signaltransduktion sind v.a. TLR2 und TLR4 involviert, die über MyD88 proinflammatorischen Zytokinen/Chemokinen induzieren. Die proinflammatorische Wirkung wird dabei u.a. über IRF5 bewirkt, das an MyD88 andockt. Diese Funktion wird von IRF4 gehemmt, das als kompetitiver Faktor IRF5 von seiner Bindungsstelle verdrängt. Die negativ regulatorische Wirkung von IRF4 schützt die Niere vor zu starker Entzündung und dadurch vor zu starker Gewebeschädigung. Dadurch wird das Ausmaß des aktuen Nierenversagens reduziert. IRF4 wird durch Sauerstoffradikale im Rahmen der Ischämie-Reperfusion induziert. Nach der Gewebeschädigung und Induktion einer Entzündung wird IRF4 erst verzögert exprimiert, um die Entzündung wieder zu begrenzen.

Die vorliegende Arbeit untersuchte die Rolle der Perizyten bei steriler Inflammation. Bisher war in diesem Zusammenhang der Einfluss der Perizyten nicht bekannt, ebenso wenig ob und wie sie zu Entzündungsreaktionen beitragen. Weiterhin war der Einfluss der Perizyten auf die interstitielle Migration myeloider Zellen in vivo unerforscht. Hier konnte gezeigt werden, dass Perizyten durch eine Vielzahl von Rezeptoren wie TLR2, TLR4, TNFR1, FPR2 in der Lage sind inflammatorische Reize zu detektieren und daraufhin einen proinflammatorischen Phänotyp annehmen. Dieser ist durch die vermehrte Expression von NLRP3 sowie des Adhäsionsmoleküls ICAM-1 und die Sekretion von Chemokinen wie CXCL1, IL8 und CCL2 gekennzeichnet. Weiterhin wird das Chemokin-ähnliche Molekül MIF von aktivierten Perizyten sowohl sezerniert als auch an der Oberfläche präsentiert. Die ausgeschütteten Chemokine beeinflussen wiederum Monozyten und neutrophile Granulozyten durch ihre chemotaktische Wirkung. Auch konnte ein anti-apoptotischer sowie aktivierender Effekt der Perizyten auf neutrophile Granulozyten gezeigt werden, was die Überlebensdauer dieser Zellen im interstitiellen Gewebe signifikant verlängert. Anhand eines Mausmodells und der 2-Photonen Mikroskopie wurde gezeigt, dass Perizyten auch in vivo einen entscheidenden Beitrag zur Rekrutierung neutrophiler Granulozyten und Monozyten zur Inflammation leisten. Zum ersten Mal wurde die Interaktion myeloider Zellen mit Perizyten in vivo visualisiert und genauer charakterisiert. Diese Interaktion beeinflusst die interstitielle Migration neutrophiler Granulozyten und Monozyten abhängig davon, ob ein Stimulus für gerichtete oder ungerichtete Migration vorliegt. Es wurde deutlich, dass Perizyten sowohl einen chemotaktischen als auch einen haptotaktischen Reiz auf myeloide Leukozyten ausüben, was an einer Polarisierung der Zellen zu erkennen ist. Ebenso tragen sie durch die Interaktion zur Aktivierung der myeloiden Zellen in vivo bei. Diese Arbeit leistet demnach einen Beitrag zur genaueren Definition der Rolle von Perizyten bei steriler Inflammation. Hierfür wurden die zellulären und molekularen Mechanismen in vitro und die in vivo ablaufenden Prozesse bei der interstitiellen Migration myeloider Zellen genauer charakterisiert. Dabei konnten Perizyten als neuer Zelltyp identifiziert werden, der Gewebeschäden detektiert und aktiv zur akuten Entzündungsreaktion beiträgt indem er die Rekrutierung und Funktionalität myeloider Leukozyten unterstützt.

Innate immune activation via IL-1R or Toll-like receptors (TLR) contibutes to acute kidney injury but its role in tissue remodeling during chronic kidney disease is unclear. SIGIRR is an inhibitor of TLR-induced cytokine and chemokine expression in intrarenal immune cells, therefore, we hypothesized that Sigirr-deficiency would aggravate postobstructive renal fibrosis. The expression of TLRs as well as endogenous TLR agonists increased within six days after UUO in obstructed compared to unobstructed kidneys while SIGIRR itself was downregulated by day 10. However, lack of SIGIRR did not affect the intrarenal mRNA expression of proinflammatory and profibrotic mediators as well as the numbers of intrarenal macrophages and T cells or morphometric markers of tubular atrophy and interstitial fibrosis. Because SIGIRR is known to block TLR/IL-1R signaling at the level of the intracellular adaptor molecule MyD88 UUO experiments were also performed in mice deficient for either MyD88, TLR2 or TLR9. After UUO there was no significant change of tubular interstitial damage and interstitial fibrosis in neither of these mice compared to wildtype counterparts. Additional in-vitro studies with CD90+ renal fibroblasts revealed that TLR agonists induce the expression of IL-6 and MCP-1/CCL2 but not of TGF-β, collagen-1α or smooth muscle actin. Together, postobstructive renal interstitial fibrosis and tubular atrophy develop independent of SIGIRR, TLR2, TLR9, and MyD88. These data argue against a significant role of these molecules in renal fibrosis.

Chitin, after cellulose the second most abundant polysaccharide in nature, is an essential component of exoskeletons of crabs, shrimps and insects and protects these organisms from harsh conditions in their environment. Unexpectedly, chitin has been found to activate innate immune cells and to elicit murine airway inflammation. The skin represents the outer barrier of the human host defense and is in frequent contact with chitin-bearing organisms, such as house-dust mites or flies. The effects of chitin on keratinocytes, however, are poorly understood. We hypothesized that chitin stimulates keratinocytes and thereby modulates the innate immune response of the skin. Here we show that chitin is bioactive on primary and immortalized keratinocytes by triggering production of pro-inflammatory cytokines and chemokines. Chitin stimulation further induced the expression of the Toll-like receptor (TLR) TLR4 on keratinocytes at mRNA and protein level. Chitin-induced effects were mainly abrogated when TLR2 was blocked, suggesting that TLR2 senses chitin on keratinocytes. We speculate that chitin-bearing organisms modulate the innate immune response towards pathogens by upregulating secretion of cytokines and chemokines and expression of MyD88-associated TLRs, two major components of innate immunity. The clinical relevance of this mechanism remains to be defined.

Thu, 20 May 2010 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/11548/ https://edoc.ub.uni-muenchen.de/11548/1/Beeck_Stefan.pdf Beeck, Stefan

Thu, 7 May 2009 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/10138/ https://edoc.ub.uni-muenchen.de/10138/1/Schickinger_Veronika.pdf Schickinger, Veronika

Thu, 4 Dec 2008 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/9435/ https://edoc.ub.uni-muenchen.de/9435/1/Herrler_Michael.pdf Herrler, Michael ddc:610, ddc:

Der Kontakt mit Mikroorganismen im frühen Kindesalter oder bereits in utero kann die Entwicklung des Immunsystems und folglich die Entstehung von atopischen Erkrankungen beeinflussen. Toll-like Rezeptoren (TLR) - wie das TLR2 und TLR4 - und das Cluster of Differentiation 14 (CD14) sind maßgeblich an der Erkennung von Mikroorganismen beteiligt. Wir stellten die Hypothese auf, dass mütterliche Allergien mit erniedrigten mRNA-Expressionsniveaus für TLR2, TLR4 und CD14 im Blut der Mütter sowie im Nabelschnurblut ihrer Kinder einhergehen. Für die vorliegende Arbeit konnten im Rahmen einer europäischen Multizentrum-Studie 185 gesunde schwangere Probandinnen aus Deutschland (n = 48), Ungarn (n = 50) und Spanien (n = 87) untersucht werden. Bei Geburt wurde peripheres Blut der Probandinnen sowie Nabelschnurblut derer Kinder gewonnen. Nach RNA-Isolation und cDNA-Synthese wurde mittels Real-Time RT-PCR die mRNA-Expression von TLR2, TLR4 und CD14 quantifiziert. Bei 42 Nabelschnurblutproben in der deutschen Subpopulation bestimmten wir außerdem den Anteil der TLR2+-, TLR4+-und CD14+-Monozyten in der Durchflusszytometrie. Zur Auswertung wurden bivariate und multivariate Regressionsanalysen durchgeführt. Mütterliche Allergien waren assoziiert mit signifikant erniedrigten mRNA-Expressionsniveaus für TLR2, TLR4 und CD14 in mütterlichem sowie im Nabelschnurblut. Ferner korrelierten die mRNA-Expressionsniveaus in mütterlichem Blut signifikant mit denen in fetalem Blut. Der durchflusszytometrisch untersuchte Prozentsatz der TLR2+-, TLR4+-und CD14+-Monozyten korrelierte mit den dazugehörigen mRNA-Expressionsniveaus für TLR2 (r = 0,5 ; p < 0,01) und TLR4 (r = 0,61 ; p < 0,01), jedoch nicht mit CD14 (r = 0,1 ; p = 0,34).

Hyper IgE syndromes (HIES) are primary immunodeficiency disorders of unknown pathogenesis. Patients are typically affected with `cold' abscesses of the skin, recurrent cyst-forming pneumonia, chronic mucocutaneous candidiasis and other less frequent features such as progressive skeletal abnormalities. Defective signaling in the Toll-like receptor (TLR) pathways has been suggested as a responsible pathologic mechanism, however, in previous reports, 10 patients revealed no defect in inflammatory cytokine responses to different TLR ligands. Here, we report the increase in pro-inflammatory cytokines TNF-alpha and IL-8, following TLR2 and TLR4 stimulation in a larger cohort of 25 additional patients with HIES, and provide a meta-analysis of the TLR data in HIES. Copyright (C) 2008 S. Karger AG, Basel.

Aspergillus fumigatus ist ein opportunistischer Krankheitserreger, der ubiquitär in der Umwelt vorhanden ist. Die schwerwiegende Krankheit, die er verursacht, ist die invasive Aspergillose, welche nur bei immungeschwächten Patienten auftritt und bis heute nur sehr schwierig zu diagnostizieren und zu heilen ist. Die Sporen von A. fumigatus können aufgrund ihrer geringen Größe bis in die Alveolen der Lunge gelangen. Dort bilden Makrophagen die erste Verteidigungslinie, indem sie die Sporen phagozytieren. Die Phagozytose ist Bestandteil der angeborenen Immunantwort und ein initialer Schritt bei der Bekämpfung von A. fumigatus-Sporen durch Makrophagen. Das Verstehen dieses Prozesses gewinnt durch die stetige Zunahme der Patienten mit invasiver Aspergillose immer größere Bedeutung und ist Gegenstand intensivster Forschung. Im Rahmen dieser Dissertation wurden die Interaktionen von murinen und humanen Makrophagen mit A. fumigatus-Sporen untersucht. Die Fragestellung wurde aus zwei unterschiedlichen Perspektiven betrachtet. Zum einen wurde die Oberfläche der A. fumigatus-Sporen analysiert; zum anderen wurden die Interaktionen von A. fumigatus mit phagozytierenden Zellen erforscht. Um die Phagozytose von A. fumigatus-Sporen in murinen und humanen Zellen genauer charakterisieren zu können, wurde in dieser Arbeit der so genannte „Biotin-Calcofluor Staining Assay“ (BCS-Assay) entwickelt. Mit Hilfe dieser Methode war es möglich, zwischen extra- und intrazellulären Sporen zu unterscheiden, ohne auf die Anwesenheit von Antikörpern angewiesen zu sein. Mit Hilfe von diversen Inhibitoren konnte der Mechanismus der Phagozytose genauer untersucht werden. So konnte gezeigt werden, dass die Aufnahme von A. fumigatus-Sporen ein Aktin-abhängiger Prozess ist und dass Makrophagen für die Phagozytose die Aktivierung der Phosphoinositid 3 Phosphat-Kinasen und von Tyrosin-Kinasen benötigen, insbesondere diejenigen der scr Familie. Butanedion Monoxim, ein Inhibitor der Myosinmotor-Aktivität, blockierte ebenfalls effizient die Sporenaufnahme. Die weiteren Untersuchungen der Phagozytoseprozesse von A. fumigatus-Sporen erfolgten u.a. mit Hilfe von Fluoreszenz- und elektronenmikroskopischer Aufnahmen. In der Immunfluoreszenz ließen sich Tyrosin-phosphorylierte Proteine in den Aufnahmestrukturen detektieren, und elektronenmikroskopische Aufnahmen infizierter Makrophagen zeigten so gennante „Ruffle“-Strukturen. Diese Tatsache deutet darauf hin, dass A. fumigatus-Sporen durch einen „Trigger“-ähnlichen Mechanismus aufgenommen werden. Im weiteren Verlauf der Arbeit wurden die Rezeptoren der Phagozytose von A. fumigatus-Sporen charakterisiert. Die Ergebnisse von Meier und ihren Kollegen zeigten bereits, dass die Erkennung von A. fumigatus durch Makrophagen mittels Toll-like Rezeptor 2 und TLR4 erfolgt. In der vorliegenden Arbeit wurde nun auch der Frage nachgegangen, welche Rolle TLR2 und TLR4 bei der Phagozytose von A. fumigatus-Sporen spielen. Hierzu wurden aus den Mausstämmen C3H/HeN (WT), C3H/HeJ (TLR4-), C3H/HeN TLR2-/- (TLR2-) und C3H/HeJ TLR2-/- (TLR2-/4-) murine Peritonealmakrophagen mittels Peritoneallavage entnommen, mit A. fumigatus-Sporen infiziert und mit Hilfe des BCS-Assays ausgewertet. Es konnte gezeigt werden, dass Toll-like Rezeptor 2 und nicht Toll-like Rezeptor 4 für eine effiziente Phagozytose benötigt wird. Dieses Ergebnis ließ sich wiederum mit Hilfe eines anti TLR2-Antikörpers bestätigen, da dieser auch die Phagozytose von A. fumigatus-Sporen, aber nicht von Kontrollbeads blockieren konnte. Des Weiteren wurde untersucht, ob der von Brown und seinen Mitarbeitern entdeckte Dectin-1 Rezeptor ein potentieller Phagozytoserezeptor von A. fumigatus-Sporen ist (Brown et al., 2001). Es konnte gezeigt werden, dass Laminarin, ein lösliches ß 1-3 Glucan, die Phagozytose von A. fumigatus-Sporen durch Makrophagen blockierte. Außerdem ließ sich mit einem anti-Dectin-1 Antikörper die Phagozytose von A. fumigatus-Sporen in Makrophagen hemmen. Zudem ließ sich Dectin-1 mit diesem Antikörper in infizierten Makrophagen in der Immunfluoreszenz detektieren. Mit einem weiteren Antikörper konnte beta-1-3 Glucan, ein wichtiger Bestandteil der pilzlichen Zellwand, auf ruhenden Sporen detektiert werden. Es zeigte sich, dass die Menge an  1-3 Glucan eine wichtige Rolle bei der Eliminierung von A. fumigatus-Sporen spielt. Vergleiche zwischen ruhenden und angeschwollenen Sporen zeigten, dass angeschwollene Sporen, welche größere Mengen an ß 1-3 Glucan auf ihrer Oberfläche besitzen, effizienter phagozytiert werden können. Auch die A. fumigatus pksP-Mutante, welche mehr  1-3 Glucan auf ihrer Oberfläche besaß, wurde effizienter phagozytiert. Betrachtet man die intrazelluläre Signaltransduktionskaskade, so deuten die Daten darauf hin, dass die Dectin-1-gesteuerte Phagozytose von A. fumigatus-Sporen abhängig von der Syk-Kinase verläuft. Die Ergebnisse der vorliegenden Arbeit zeigen, dass Dectin-1 und TLR2 für eine effiziente Phagozytose von A. fumigatus-Sporen benötigt werden. Die Ergebnisse legen allerdings nahe, dass außer Dectin-1 und TLR 2 noch weitere Rezeptoren bei der Phagozytose von A. fumigatus-Sporen beteiligt sind. Ein genaues Verständnis der bei der Phagozytose ablaufenden Erkennungsprozesse und der nachgeschalteten Signaltransduktionskaskaden könnte in Zukunft ausgenutzt werden, um die Effiziens der Phagozytose auch in immungeschwächten Patienten zu erhöhen und sie so zu schützen.

Background: Maternal atopic background and stimulation of the adaptive immune system with allergen interact in the development of allergic disease. Stimulation of the innate immune system through microbial exposure, such as activation of the innate Toll-like- receptor 2 (TLR2), may reduce the development of allergy in childhood. However, little is known about the immunological effects of microbial stimulation on early immune responses and in association with maternal atopy. Methods: We analyzed immune responses of cord blood mononuclear cells ( CBMC) from 50 healthy neonates ( 31 non-atopic and 19 atopic mothers). Cells were stimulated with the TLR2 agonist peptidoglycan (Ppg) or the allergen house dust mite Dermatophagoides farinae (Derf1), and results compared to unstimulated cells. We analyzed lymphocyte proliferation and cytokine secretion of CBMC. In addition, we assessed gene expression associated with T regulatory cells including the transcription factor Foxp3, the glucocorticoid-induced TNF receptor ( GITR), and the cytotoxic lymphocyte antigen 4 (CTLA4). Lymphocyte proliferation was measured by H-3-Thymidine uptake, cytokine concentrations determined by ELISA, mRNA expression of T cell markers by real-time RT-PCR. Results: Ppg stimulation induced primarily IL-10 cytokine production, in addition to IFN-gamma, IL-13 and TNF-alpha secretion. GITR was increased following Ppg stimulation ( p = 0.07). Ppg- induced IL-10 production and induction of Foxp3 were higher in CBMC without, than with maternal atopy ( p = 0.04, p = 0.049). IL-10 production was highly correlated with increased expression of Foxp3 ( r = 0.53, p = 0.001), GITR ( r = 0.47, p = 0.004) and CTLA4 ( r = 0.49, p = 0.003), independent of maternal atopy. Conclusion: TLR2 stimulation with Ppg induces IL-10 and genes associated with T regulatory cells, influenced by maternal atopy. Increased IL-10 and Foxp3 induction in CBMC of non-atopic compared to atopic mothers, may indicate an increased capacity to respond to microbial stimuli.

Der opportunistisch humanpathogene Hefepilz Candida albicans gehört bei vielen gesunden Menschen zur mikrobiellen Schleimhautflora, kann jedoch bei abwehrgeschwächten Patienten oberflächliche Infektionskrankheiten sowie lebensbedrohliche Organmykosen verursachen. Obwohl der Immunstatus des Wirtes für eine Infektion mit diesem Erreger von entscheidender Bedeutung ist, trägt auch eine Reihe von Virulenzfaktoren, insbesondere die sekretorischen Aspartatproteasen (Saps), zur Pathogenität von C. albicans bei. Die angeborene Immunität ist in der Lage, derartige Pathogene schon beim Erstkontakt zu erkennen und zu bekämpfen. Haupteffektoren dieser schnellen, angeborenen Immunantwort sind Makrophagen und neutrophile Granulozyten. Mitglieder der Toll-Proteinfamilie, sogenannte Toll-like Rezeptoren (TLRs), wurden kürzlich als Rezeptoren auf diesen Immunzellen in Säugern identifiziert. Sie erkennen unterschiedliche Erreger anhand von in der Evolution hoch konservierten Strukturen, den Pathogen-assoziierten molekularen Mustern (PAMPs), was zur Aktivierung des Transkriptionsfaktors NF-кB und zur Freisetzung von Zytokinen führt. Sowohl TLR2 als auch TLR 4 wurden kürzlich für die Erkennung von C. albicans diskutiert. Zielsetzung dieser Arbeit war es, die Interaktion von Makrophagen mit C. albicans im Hinblick auf die Aktivierung von Toll-like Rezeptoren und die nukleäre Translokation von NF-кB zu untersuchen. Neben dem lebenden C. albicans-Isolat wurden zudem drei weitere Präparationen untersucht: Mit den Antimykotika (AM) Amphotericin B, Nystatin und Itraconazol vorbehandelte Keime, durch Hitze inaktivierte Keime sowie Sap-inaktivierte Keime. Die Zellstimulationsexperimente wurden mit murinen Wildtyp-Makrophagen, TLR2- bzw. TLR4- defizienten Einzelknockoutmutanten und mit TLR2/4-Doppelknockoutmutanten durchgeführt. Die TLR-vermittelte Aktivierung von NF-кB wurde mit Gelshifts (EMSA) nachgewiesen. Mit Western Blots wurden die intrazellulären Signaltransduktionswege untersucht. Der Hitze-inaktivierte Stamm bewirkte keine Translokation von NF-кB in Wildtyp-Makrophagen. Eine Inhibition der Saps bewirkte keine Abschwächung der NF-кB Induktion, so dass im Umkehrschluss dieser bedeutende Virulenzfaktor die TLR-vermittelte NF-кB Aktivierung nicht beeinflusst. Der lebende Stamm benutzte sowohl TLR2 als auch TLR4 für die Induktion von NF-кB. Nach Vorstimulation der Makrophagen mit Interferon-γ ließ sich jedoch eine klare TLR2-Abhängigkeit – unabhängig von TLR4 – in der Aktivierung von NF-кB und in der Induktion von TNF-α zeigen. In beiden Fällen wurden die Makrophagen erst ab einer Candida-Dichte von 106 Zellen pro 100 µl PBS stimuliert. Für den AM-vorbehandelten Stamm ergab sich eine deutliche TLR2-Abhängigkeit in der Regulation von NF-кB, welche durch die Präinkubation der Makrophagen mit IFN-γ nicht beeinflusst wurde. AM-vorbehandelte Keime konnten NF-кB in den Makrophagen erst ab einer Dichte von 107 Zellen pro 100 µl PBS aktivieren. Zusammenfassend zeigen diese Ergebnisse, dass lebende und AM-vorbehandelte Keime im Gegensatz zur Hitze-inaktivierten Präparation und zu den Saps relevante PAMP-Strukturen für eine TLR-vermittelte NF-кB Hochregulation besitzen. Die Beteiligung beider Rezeptoren, TLR2 und TLR4, belegt beim lebenden Stamm das Konzept, dass immunkompetente Zellen sich mehrerer TLRs bedienen, um die Immunantwort möglichst spezifisch und fein zu regulieren. Beim AM-vorbehandelten Stamm scheint den Antimykotika Amphotericin B, Nystatin und Itraconazol eine besondere Rolle zuzukommen, da diese die Integrität der Pilzmembran stören und somit TLR2-aktivierende PAMPs aus Zellwand und/oder Zytosol freisetzen. Neben dem direkten Effekt auf die Pilzmembran kommt es somit zusätzlich zu einer indirekten, TLR2 vermittelten Stimulation der Makrophagen. Untersuchungen der Signaltransduktion (Stimulation von Wildtyp-Makrophagen mit dem AM-vorbehandelten C. albicans-Isolat) ergaben eine vorübergehende, zeitlich eng begrenzte Induktion von NF-кB, die durch den Inhibitor IкB-α reguliert wird. Gleichzeitig wurden im zeitlichen Verlauf der Stimulation auch MAP Kinasen (ERK, p38, JNK) und c-Jun, eine Subeinheit des Transkriptionsfaktors AP-1, phosphoryliert. Diese simultane Aktivierung beider Transkriptionsfaktoren weist auf eine feinregulierte Immunantwort der Makrophagen gegenüber C. albicans hin und legt zudem einen Cross-Talk zwischen NF-кB und AP-1 nahe.