Podcasts about elisas

Velkommen til en spennende episode hvor vi dykker dypt ned i bønnens historie i både Gamle og Nye Testamentet. Vår taler begynner med å lese fra Salme 65 og reflekterer over hvordan Guds folk har bedt gjennom årtusener, med en trygg forvissning om at Gud hører våre bønner. Vi utforsker forskjellene og likhetene mellom bønnene i Gamle Testamentet og i Nye Testamentet. Hva er det som skiller bønnene til profeten Elisas lærlinger fra de kristnes bønner i Jerusalem og dagens Norge? Dette er en av de mange interessante spørsmålene vi dykker ned i. Videre tar vi for oss to av de mest kjente lovsangene i Lukas-evangeliet: Marias lovsang og Sakarias lovsang. Disse lovsangene, sammen med Simeons lovsang, har hatt en fast plass i kirkens gudstjeneste og daglige bønneliv. Vi får også et innblikk i livet til Sakarias og Elisabeth, og deres fromme liv i troens rettferdighet og bønn. Gjennom hele episoden vil vi reflektere over hvordan bønn har vært en sentral del av Guds folks liv, og hvordan det fortsatt er en viktig del av våre liv i dag. Bli med oss på denne åndelige reisen og la oss sammen utforske dybden og rikdommen i bønnens historie. Tema i 2024; «Ikke filosofenes, men Abrahams, Jakobs og Isaks Gud» Du kan lytte til bibeltimene som ligger her gratis, men som en takk ville vi sette pris på om du støttet menigheten DELK Bergens arbeid med en gave på kontonummer 3624 63 90015 (Sparebanken vest) eller Vipps: 103439 (DELK Bergen)

Many autoimmune neuromuscular disorders are reversible with prompt diagnosis and early treatment. Understanding the potential utility and limitations of antibody testing in each clinical setting is critical for practicing neurologists. In this episode, Teshamae Monteith, MD, FAAN speaks with Divyanshu Dubey, MD, FAAN, author of the article “Autoimmune Neuromuscular Disorders Associated With Neural Antibodies,” in the Continuum® August 2024 Autoimmune Neurology issue. Dr. Monteith is the associate editor of Continuum® Audio and an associate professor of clinical neurology at the University of Miami Miller School of Medicine in Miami, Florida. Dr. Dubey is an associate professor in the departments of neurology and laboratory medicine and pathology at the Mayo Clinic in Rochester, Minnesota. Additional Resources Read the article: Autoimmune Neuromuscular Disorders Associated With Neural Antibodies Subscribe to Continuum: shop.lww.com/Continuum Earn CME (available only to AAN members): continpub.com/AudioCME Continuum® Aloud (verbatim audio-book style recordings of articles available only to Continuum® subscribers): continpub.com/Aloud More about the American Academy of Neurology: aan.com Social Media facebook.com/continuumcme @ContinuumAAN Host: @headacheMD Guest: @Div_Dubey Transcript Full episode transcript available here Dr Jones: This is Dr Lyell Jones, Editor-in-Chief of Continuum, the premier topic-based neurology clinical review and CME journal from the American Academy of Neurology. Thank you for joining us on Continuum Audio, which features conversations with Continuum's guest editors and authors who are the leading experts in their fields. Subscribers to the Continuum journal can read the full article or listen to verbatim recordings of the article and have access to exclusive interviews not featured on the podcast. Please visit the link in the episode notes for more information on the article, subscribing to the journal, and how to get CME.   Dr Monteith: This is Dr Teshamae Monteith, Associate Editor of Continuum Audio. Today, I'm interviewing Dr Divyanshu Dubey about his article on autoimmune neuromuscular disorders associated with neural autoantibodies, which is part of the August 2024 Continuum issue on autoimmune neurology. Welcome to the podcast. How are you?   Dr Dubey: Hi, Dr Monteith. Thank you for inviting me to be a part of this podcast. I'm doing well.   Dr Monteith: Well, why don't you introduce yourself to the audience? And, call me Tesha.   Dr Dubey: I'm Divyanshu Dubey (please, call me Div). I'm one of the autoimmune neurology consultants here at Mayo Clinic Rochester. I'm an Associate Professor of neurology, as well as lab medicine and pathology. My responsibilities here are split - partly seeing patients (primarily patients with autoimmune disorders, including neuromuscular disorders), and then 50% of my time (or, actually, more than 50%), I spend in the lab, either doing research on these autoimmune disorders or reporting antibodies in a clinical setting for various antibody panels which Mayo's neuroimmunology lab offers.   Dr Monteith: That's a nice overlap of subspecialty area. How did you get into this work?   Dr Dubey: I think a lot of it was, sort of, by chance. Meeting the right people at the right time was the main, sort of, motivation for me. Initially, I trained in India for my medical school and didn't really got much exposed to autoimmune neurology in India. I think our primary concern in my training was sort of treating TB meningitis and cerebral malaria - that was my exposure to neurology, including stroke and some epilepsy cases. As a part of application for USMLEs and coming here to residency, I did some externships, and one of the externships was at Memorial Sloan Kettering Cancer Center, and that's when I worked a few weeks with Dr Posner and got introduced to the idea of paraneoplastic neurological syndrome working with him. And that sort of started - I wouldn't call it vicious cycle - but my interest in the area of autoimmune neurology and paraneoplastic neurological disorders, which subsequently was refined further through residency and fellowships.   Dr Monteith: That's interesting. I actually rotated through - I did a externship also at Sloan Kettering, and I had a clinic with Dr Posner. And I thought, at the time, he was such a rock star, and, like, I took a picture with him, and I think he thought it was insane. And I didn't go into autoimmune neurology. So, you know, interesting pathways, right?   Dr Dubey: Yes. And I think he's inspired many, many people, and sort of trained a lot of them as well.   Dr Monteith: So, why don't you tell us what you set out to do when writing this article?   Dr Dubey: So, I think, given my background and training in various subspecialties in neurology, I was, sort of, formally did fellowships in autoimmune neurology, as well as neuromuscular medicine. One of the areas in these areas that I focus on is in my clinical practice, as well as in my sort of lab work, is autoimmune muscular disorders - and that to, specifically, autoantibodies and their clinical utility for autoimmune muscular disorders. So, that's what I wanted to focus on in an article. When I was invited to write an article on autoimmune muscular conditions in general, I thought it was very difficult to pack it all in one chapter or one article, so I narrowed my focus (or tilted my focus) towards antibody-positive disorders and trying to understand how we as neurologists can firstly sort of identify these conditions (which may end up being antibody-positive) – and then, on the other hand, once we get these antibody results, how we can find the utility in them or find them useful in taking care of our patients. At the same time, I also wanted to kind of highlight that these antibodies are not perfect, they do have certain limitations – so, that's another thing I sort of highlighted in the article.   Dr Monteith: So, why don't we just start with a very broad question - what do you believe the role of autoantibodies is in the workup of neuropathies and then neuromuscular disorders? Obviously, when we think of myasthenia gravis, but there are some presentations that you may not necessarily think to first order autoantibody tests. So, what is the role, and where does it fit in the paradigm?   Dr Dubey: I think it's extremely crucial, and it's evolving as time goes on, and it's becoming more and more clinically relevant. Let's say three, four decades ago, the number of biomarkers which were available were very limited and only a handful - and there has been a significant increase in these biomarkers with growing utilization of newer techniques for discovery of antibodies, and more and more people jumping into this field trying to not only discover, but try and understand and validate these biomarkers (what they truly, clinically mean). These antibodies, like you pointed out, ones for myasthenia (such as acetylcholine receptor-binding antibodies, or MuSK antibodies), they can be extremely helpful in clinical diagnosis of these patients. We all know the importance of EMG in managing our patients with neuromuscular disorders. But, oftentimes, EMG nerve conduction studies are often not available at every center. In those scenarios, if you have antibodies with very high clinical specificity, and you're seeing a patient on examination whom you're seeing ptosis (fatigable ptosis), double vision, you're suspecting myasthenia, you send antibodies, and they come back positive. It brings you closer to the answer that may, in turn, require you to refer to a patient to a place where you can get high-quality EMGs or high-quality care. In addition to getting to the diagnosis, it also, sometimes, leads you in directions to search for what is the trigger. A good example is all these paraneoplastic neurological syndromes (which we started our conversation with), where once you find a biomarker (such as anti-Hu antibodies or CRMP5 antibodies) in a patient with paraneoplastic neuropathies, it can direct the search for cancer. These are the patients where, specifically, these two antibodies, small-cell lung cancer is an important cancer to rule out - they require CT scans, and if those are negative, consider doing PET scan – so, we can remove the inciting factor in these cases. And then, lastly, it can guide treatment. Depending upon subtypes of antibodies or particular antibodies, it can give us some idea what is going to be the most effective treatment for these patients.   Dr Monteith: I think paraneoplastic syndromes are a very good example of how autoantibodies can help guide treatment. But, what other examples can you provide for us?   Dr Dubey: Yeah, so I think one of the relatively recent antibody tests which our lab started offering is biomarkers of autoimmune neuropathies - these are neurofascin and contactin, and those are great examples which can target or guide your treatment. I personally, in the past, have had many CIDP patients before we were offering these testings, where we used to kind of start these patients on IVIG. They had the typical electrodiagnostic features, which would qualify them for CIDP. They did not show any response. In many of these cases, we tried to do sort of clinical testing or sort of research-based testing for neurofascin and contactin back in the day, but we didn't have this resource where we can sort of send the blood, hopefully, and within a week, get an answer, whether these patients have autoimmune neuropathy or not. Having this resource now, in some of these cases, even before starting them on IVIG, knowing that test result can guide treatments, such as considering plasma exchange up front as a first-line therapy, followed by rituximab or B-cell depleting therapies, which have been shown to be extremely beneficial in these conditions. And it is not just limited to neurofascin or contactin (which are predominantly IgG4-mediated condition), but the same concept applies to other IgG4-mediated diseases, such as MuSK myasthenia, where having an antibody result can guide your treatment towards B-cell depleting therapies instead of sort of trying the typical regimen that you try for other myasthenia gravis patients.   Dr Monteith: And you mentioned where I was reading that, sometimes, nerve conduction studies and EMG can be useful to then narrow the autoantibody profiles. Oftentimes, in the inpatient service, we order the autoantibodies much faster, because it's sometimes harder to access EMG nerve conduction studies - but talk about that narrowing process.   Dr Dubey: Yeah. And it goes back to the point you just made where we end up sending, sort of, sometimes (and I'm guilty of this as well), where we just send antibodies incessantly, even knowing that this particular patient is not necessarily likely to be an autoimmune neurological disorder, and that can be a challenge, even if the false-positive rate for a particular test is, let's say 1% - if you send enough panels, you will get that false-positive result for a particular patient. And that can have significant effects on the patient - not only unnecessary testing or imaging (depending on what type of antibody it is), but also exposure to various immunotherapies or immunosuppressive therapies. It's important to recognize red flags – and that's one of the things I've focused on in this article, is talking about clinical, as well as electrodiagnostic, factors, which make us think that this might be an autoimmune condition, and then, subsequently, we should consider autoantibody testing. Otherwise, we can be in a situation - that 1% situation - where we may be sort of dealing with a false-positive result, rather than a true-positive result. In terms of EMGs, I think I find them extremely useful, specifically for neuropathies, distinguishing between demyelinating versus exonal, and then catering our antibody-ordering practices toward specific groups of antibodies which are associated with demyelinating neuropathies (if that's what the electrophysiology showed) versus if it's an exonal pathology (considering a different subset of antibodies) - and that's going to be extremely important.   Dr Monteith: You're already getting to my next question, which is what are some of the limitations of autoantibody testing? You mentioned the false-positivity rate - what other limitations are there?   Dr Dubey: So, I think the limitations are both for seropositive, as well as seronegative, patients. As a neurologist, when we see patients and send panels, we can be in a challenging situation in both of those scenarios. Firstly, thinking about seropositives - despite the growing literature about neurology and antibodies, we have to be aware, at least to some extent, about what methodologies are being utilized for these antibody tests. And what I mean by that is knowing when you're sending a sample to a particular lab, the methodology that they're utilizing - is that the most sensitive, specific way to test for certain antibodies? We've learned about this through some of the literature published regarding MOG and aquaporin-4, which has demonstrated that these antibodies, which we suspect are cell surface antibodies, not only generate false-positive, but also false-negative results if they are tested by Western blots or ELISAs. Similar can be applied to some of the cell surface antibodies we are investigating on the autoimmune neuromuscular side (we have some sort of unpublished data regarding that for neurofascin-155). Secondly, it's also kind of critical when you're getting these reports to kind of have a look at what type of secondary antibodies are being utilized, an example being we talked about neurofascin-155, and I mentioned these are IgG4-predominant diseases, so testing for neurofascin IgG4 and knowing that particular patient is positive IgG4 rather than neurofascin pan-IgG. That's an important discrimination, and important information for you to know, because we have seen, at least in my clinical practice, that patients who are positive for neurofascin IgG4 follow the typical story of autoimmune neuropathies - the ones who are not (who are just neurofascin-155 IgG-positive), oftentimes can have wide-ranging phenotypes. The same applies to neurofascin-155 IgMs. And then (not for all antibodies, but for some antibodies), titers are important. A good example of that is a3 ganglionic receptor antibodies, which we utilize for when we're taking care of patients who have autoimmune dysautonomia - and in these cases, if the titers of the antibodies are below .2 nmol/L, usually, those don't have a high specificity for AAG diagnosis. So, I get referred a lot of patients with very low titers of a3 ganglionic receptor antibodies, where the clinical picture does not at all look like autoimmune autonomic ganglionopathy. So, that's another thing to potentially keep in mind. And then, on the seronegative front, it's important to recognize that we are still sort of seeing the tip of the iceberg as far as these antibodies or biomarkers are concerned, specifically for certain phenotypes, such as CIDP. If you look at the literature, depending upon what demographics we're looking at or sort of racial profiles we're looking at, the frequency of these autoimmune neuropathy biomarkers range from 5% to 20%, with much higher frequency in Asian patients - so, a good chunk of these diseases are still seronegative. In the scenario where you have a very high suspicion for an autoimmune neuromuscular disorder (specifically, we'll talk about neuropathies, because that's why we utilize tissue immunofluorescence staining on neural tissues), I recommend people to potentially touch base with that tertiary care lab or that referral lab to see if they have come across some research-based antibodies which are not clinically validated, which can give you some idea, some additional supportive idea, that what you're dealing with is an autoimmune neuromuscular disorder. So, we have to keep the limitations of some of these antibody panels and antibody tests in mind for both positive, as well as negative, results.   Dr Monteith: So, you've already given us a lot of good stuff, um, about titer seronegativity and false-positive rates. And, you know, also looking at the clinical picture when ordering these tests, utilizing EMG nerve conduction studies, give us a major key point that we can't not get when reading your article.   Dr Dubey: I think the major key point is we are neurologists first and serologists later. Most of these patients, we have to kind of evaluate them clinically and convince ourselves at least partly that this might be an autoimmune neuromuscular disorder before sending off these panels. Also, I find it useful to narrow down the phenotype, let's say, in a particular neuropathy or a muscle disease or a hyperexcitability syndrome. So, I have a core group of antigens, autoantigens, or autoantibodies, which I'm expecting and making myself aware of - things beyond that will raise my antenna - potentially, is this truly relevant? Could this be potentially false-positive? So, clinical characterization up front, phenotypic characterization upfront, and then utilizing those antibody results to support our clinical decision-making and therapeutic decision-making is what I've tried to express in this article.   Dr Monteith: And what is something that you wish you knew much earlier in your career?   Dr Dubey: It's a very challenging field, and it's a rapidly evolving field where we learn many things nearly every year, and, sometimes, we learn things that were previously said were incorrect, and we need to kind of work on them. A good example of that is initial reports of voltage-gated potassium-channel antibodies. So, back in the day when I was actually in my medical school and (subsequently) in my residency, voltage-gated potassium-channel antibodies were closely associated with autoimmune neuromyotonia, or autoimmune peripheral hyperexcitability syndromes. Now, over time, we've recognized that only the patients who are positive for LGI1 or CASPR2 are the ones who truly have autoimmune neuromuscular disorders or even CNS disorders. The voltage-gated potassium-channel antibody by itself, without LGI1 or CASPR2, truly doesn't have a very high specificity for neurological autoimmunity. So, that's one example of how even things which were published were considered critical thinking or critical knowledge in our field of autoimmune neuromuscular disorders has evolved and has sort of changed over time. And, again, the new antibodies are another area where nearly every year, something new pops up - not everything truly stands a test of time, but this keeps us on our toes.   Dr Monteith: And what's something that a patient taught you?   Dr Dubey: I think one of the things with every patient interaction I recognize is being an autoimmune neurologist, we tend to focus a lot on firstly, diagnosis, and secondly, immunotherapy - but what I've realized is symptomatic and functional care beyond immunotherapy in these patients who have autoimmune neurological disorders is as important, if not more important. That includes care of patients, involving our colleagues from physical medicine and rehab in terms of exercise regimen for these patients as we do immunotherapies, potentially getting a plan for management of associated pain, and many other factors and many other symptoms that these patients have to deal with secondary to these autoimmune neurological conditions.   Dr Monteith: I think that's really well said, because we get excited about getting the diagnosis and then getting the treatment, but that long-term trajectory and quality of life is really what patients are seeking.   Dr Dubey: Yeah, and as you pointed out, most of the time, especially when we are in inpatient service, or even when we're seeing the patients upfront outpatient, we are seeing them, sometimes, in their acute phase or at their disease not there. What we also have to realize is, what are the implications of these autoimmune neurological conditions in the long term or five years down the line? And that's one of the questions patients often ask me and how this can impact them even when the active immune phase has subsided - and that's something we are actively trying to learn about.   Dr Monteith: So, tell me something you're really excited about in your field.   Dr Dubey: I think, firstly (which is pretty much the topic of my entire article), is novel antibodies and new biomarker discoveries. That's very exciting - we are actively, ourselves, involved in the space. The second thing is better mechanistic understanding of how these antibodies cause diseases, so we can not only understand diseases, we can also try and understand how to target and treat these diseases - this is being actively done for various disorders. One of the disorders which continue to remain a challenge are T-cell mediated diseases, where these antibodies are just red flags or biomarkers are not causing the disease, but it's potentially the T-cells possibly attacking the same antigen which are causing disease process, and those are often the more refractory and harder-to-treat conditions. I'm hoping that with some of the work done in other fields (such as rheumatology or endocrinology for type one diabetes), we're able to learn and apply the same in the field of autoimmune neurology and autoimmune neuromuscular medicine. And then, the final frontier is developing therapies which are antigen specific, where you have discovered that somebody has a particular antibody, and if that antibody is pathogenic, can I just deplete that antibody, not necessarily pan-depleting the immune system. And there is some translational data, there's some animal model data in that area, which I find very exciting, will be extremely helpful for many of my patients.   Dr Monteith: So, very personalized targeted therapies?   Dr Dubey: Correct. Without having all the side effects we all have to kind of take care of in our patients when we start them on, let's say, cyclophosphamide, or some of these really, really, significantly suppressive immunosuppressive medications.   Dr Monteith: Well, thank you so much. I learned a lot from reading your article to prepare for this interview, but also just from talking to you. And it's clear that you're very passionate about what you do and very knowledgeable as well, so, thank you so much.   Dr Dubey: Thank you so much. Thank you for inviting me to do this. And thank you for inviting me to contribute the article.   Dr Monteith: Today, I've been interviewing Dr Divyanshu Dubey, whose article on autoimmune neuromuscular disorders associated with neural autoantibodies appears in the most recent issue of Continuum on autoimmune neurology. Be sure to check out Continuum Audio episodes from this and other issues. And thank you to our listeners for joining us today.   Dr Monteith: This is Dr Teshamae Monteith, Associate Editor of Continuum Audio. If you've enjoyed this episode, you'll love the journal, which is full of in-depth and clinically relevant information, important for neurology practitioners. Use this link in the episode notes to learn more and subscribe. AAN members, you can get CME for listening to this interview by completing the evaluation at Continpub.com/AudioCME. Thank you for listening to Continuum Audio.

Part Two of our taboo listeners erotic fantasy. Elisas passion has been laid bare.Unable to resist she has succumbed to Brix's charmYet in the back of her mind she knowsHis still the best friend of her ex husband.Is this all another one of Brandon's gamesor could this chemistry be real?

Minu seekordseks vestluskaaslaseks on Elisa Eesti tegevjuht Andrus Hiiepuu. Elisas töötatud aja jooksul on Andrus töötanud nii turundusdirektorina kui ka seejärel juhtinud juhatuse liikmena eri perioodidel Äri- ja Erakliendiüksuseid. Ma ei tundnud Andrust varem, kuid seda ägedam oli tundma õppida tema vaateid juhtimisfilosoofiale, kus ta rõhutab, kuidas juhtimine on võimekus mobiliseerida organisatsiooni energiat. Arutleme ka selle üle, kuidas Elisa on oma juhtimis- ja organisatsioonistruktuuri kohandanud agiilsuspõhimõtetele vastavaks, et käia ühte sammu pidevalt muutuva ärimaailmaga. Andrus jagab ka oma mõtteid organisatsioonikultuuri ja psühholoogilise turvalisuse tähtsusest, töötajate võimestamisest ning sellest, kuidas erinevad juhtimisstiilid mängivad olulist rolli erinevates olukordades. "Juhtimine on võimekus mobiliseerida ja suunata organisatsioonilist energiat. Minu nägemuses peitub eestvedamise roll suures osas organisatsioonilise energia valla päästmises. Võib-olla toon võrdluse Einsteiniga ja tema tuntud võrrandiga E=mc2, kus isegi ühes väikeses terakeses peitub tohutu energia sees. Samamoodi on lugu inimestega: see energia tuleb lihtsalt üles leida ja vabastada. Selleks on vaja leida õige tasakaal meeskonda suunava ja kaasava juhtimise vahel, et luua dünaamiline, kuid samas stabiilne organisatsioon. Edukas juhtimine on justkui tants erinevate juhtimisstiilide vahel, pidevalt kohandudes erinevate olukordade nõudmistega." - Andrus Hiiepuu Kuulake ikka ...

Wir sprechen mit Elisa Baş über die Unstimmigkeiten zwischen FFF Deutschland und FFF International, die deutschen Kontroversen rund um Greta Thunberg und ihre Solidarität mit Palästina, sowie Elisas eigene Erfahrungen mit Springer-Hetzkampagnen und der folgenden Behandlung durch FFF Deutschland. Vizualizing Palestine: https://www.visualizingpalestine.org/ Wir sind 99 ZU EINS! Ein Podcast mit Kommentaren zu aktuellen Geschehnissen, sowie Analysen und Interviews zu den wichtigsten politischen Aufgaben unserer Zeit.#leftisbest #linksbringts #machsmitlinks Wir brauchen eure Hilfe! So könnt ihr uns unterstützen: 1. Bitte abonniert unseren Kanal und liked unsere Videos. 2. Teil unseren content auf social media und folgt uns auch auf Twitter, Instagram und FB 3. Wenn ihr Zugang zu unserer Discord-Community, sowie exklusive After-Show Episoden und Einladungen in unsere Livestreams bekommen wollt, dann unterstützt uns doch bitte auf Patreon: www.patreon.com/99zueins 4. Wir empfangen auch Spenden unter: https://www.paypal.com/donate/?hostedbuttonid=NSABEZ5567QZE

Thu, 07 Sep 2023 16:30:00 +0000 https://chilisincarne.podigee.io/17-new-episode eb9611727574dffc0114018c4b08a8af Elisa von Die Spriesserie gibt eine geballte Ladung an Wissen weiter zum Thema Kaltkeimer, Wintergemüse und gärtnern im Herbst. **Elisa, was geht in diesem Jahr noch so alles im Garten oder im Balkonkasten - und geht da überhaupt noch was? **Diese Frage habe ich Elisa von DIE SPRIEßERIE gestellt und war selbst total überrascht, dass der Herbst genauso viel Pflanzpotenzial bietet wie der Frühling. Zudem ist es viel leichter im Herbst zu pflanzen bzw. zu säen (also auch was für faulere HobbygärtnerInnen ;-) ). Wusstest du zum Beispiel, dass du deinen Balkon oder Garten bereits jetzt fürs nächste Jahr vorbereiten kannst und im Frühjahr es dann einfach los geht mit dem Sprießen? Oder, dass du jetzt noch bestimmte Gemüsesorten aussäen kannst, um sie - halte dich fest - noch in diesem Jahr zu ernten? Wir sprechen neben den Pflanz- und Aussaat Tipps für Stadtgärten und Balkone über: die Ernte von Wintergemüse, wie du im Herbst aussäst (auch etwas für Faulere), 3 Tipps, wo du auch im Herbst noch Gemüse-, Obst- und Kräuterpflänzchen online einkaufen kannst, Erdbeeren: damit du im nächsten Jahr easy ernten kannst, was du jetzt für deine letzte Tomatenernte noch tun kannst, wie du kranke Pflanzenteile am besten entsorgst, Elisas kommender Workshop: "Pflanzen und Aussäen im Herbst", Winterbepflanzung von Balkonkästen, wie du Kräuter gut trocknest u.n.v.m. **Hier kannst du auch im Herbst noch Pflänzchen kaufen: ** (unbezahlte Werbung) https://dachgemuese.com/ https://biogartenversand.de/ https://www.bio-kraeuter.de/ **Elisas Workshops **findest du auf ihrer Webseite. Die Teilnehmerzahl ist begrenzt. https://die-spriesserie.de/pages/veranstaltungen-im-dachgarten-studio Den **Pflanzkalender **zum Download gibt es hier: Aussaat und Pflanzkalender Dieser Podcast lebt von positiven Bewertungen. Freue mich daher nen Kullerkeks, wenn du mir eine Bewertung dalässt

Bibeltime ved Jens Jensen. Emne: "Elisas kald" ud fra 1. Kong. 19,19-21.

Franco Michienzi has been looking after people in Vancouver, British Columbia, Canada for many years. He has earned his place in the top tier of wine professionals in the city, In fact he was recently named Vancouver Magazine's 2023 Sommelier of the year. Today you'll finds him at pouring wine at Elizas Steak House. https://www.instagram.com/francomichienzi/?hl=en Follow Over a Glass https://www.instagram.com/overtheglasspod  Host Shanteh Wale https://www.instagram.com/shantehwale/?hl=en Executive Producer Rob Locke https://www.instagram.com/foodwinedine/ Executive Producer Anthony Huckstep https://www.instagram.com/huckstergram/ LISTEN TO OUR OTHER FOOD PODCASTS https://linktr.ee/DeepintheWeedsNetwork Over a Glass is a wine & drinks podcast with Shanteh Wale exploring the personalities, stories and landscape of the wine and drinks business. An Australian Wine and Drinks Podcast from the Deep in the Weeds Network.

Franco Michienzi has been looking after people in Vancouver, British Columbia, Canada for many years. He has earned his place in the top tier of wine professionals in the city, In fact he was recently named Vancouver Magazine's 2023 Sommelier of the year. Today you'll finds him at pouring wine at Elizas Steak House. https://www.instagram.com/francomichienzi/?hl=en Follow Over a Glass https://www.instagram.com/overtheglasspod  Host Shanteh Wale https://www.instagram.com/shantehwale/?hl=en Executive Producer Rob Locke https://www.instagram.com/foodwinedine/ Executive Producer Anthony Huckstep https://www.instagram.com/huckstergram/ LISTEN TO OUR OTHER FOOD PODCASTS https://linktr.ee/DeepintheWeedsNetwork Over a Glass is a wine & drinks podcast with Shanteh Wale exploring the personalities, stories and landscape of the wine and drinks business. An Australian Wine and Drinks Podcast from the Deep in the Weeds Network.

Welcome to the Why Science initiative where we explore the reasons why life science researchers and industry professionals are driven to do the work they do and why the general public should understand and care about their discoveries and breakthroughs. I'm your host Kyle Valgardson and in every episode, we will strive to unravel the motivations behind science and try to inspire you as the listener to better understand the world we live in. Before we get into the good stuff, I wanted to introduce you to the mission of our podcast. The why science initiative aims to bridge the gap between scientific research and the public by highlighting individuals and findings that really demonstrate the impact of good science on our lives.  In each episode, we will speak with a researcher or industry professional to explore their backgrounds and allow them to explain what they are doing to help our communities. We believe that everyone should have access to scientific knowledge which historically has been shared through complicated terminology making it hard to understand the impacts that scientific breakthroughs have in our lives. We will strive to ask the right questions so you will understand the work no matter what background you have. Our guests come from diverse backgrounds which make for some fun and fast discussions around a breadth of topics.This podcast series is sponsored by my employer Quansys Biosciences where we strive to develop and provide multiplex ELISA tools to help further therapeutic discovery and development. So if you need some help with your multiplexed ELISAs don't be shy and feel free to reach out through our website or podcast email. Stay tuned as we explore the scientific research world searching for the importance and impacts of science in our daily lives. And don't forget to subscribe so you can be notified of future episodes, also leave a review so others can find our show and join us on this journey.  --Links--For more information on Quansys Biosciences visit our website https://www.quansysbio.com/Please direct any questions or inquiries for The Why Science initiative to podcast@quansysbio.com

Es kommt ja jetzt so langsam die Zeit, wenn die Schafe sich von ihrer Wolle verabschieden müssen und geschoren werden. Wusstet ihr, dass Elvis Eifel eine Schafherde zu Hause hat? Zumindest kümmert er sich darum, dass seine kleinen Pulloverschweinchen von einer Hundefriseurin frisiert werden sollen.

When will we get to a place of reliable protein biomarkers as predictors or indicators of disease? Ole Vorm is a founder and director at Evosep, producing separation products between the patient sample and a mass spectrometer.What is the state of those analyses and what are the obstacles yet to be overcome? The first obstacle in terms of the proteome itself is that in any sample - urine, blood etc.- there are a few abundant proteins that represent the bulk of the material. It's likely that the proteins of interest (if we can find them) are a small portion of the total and any changes in quantity or relevant modifications may be subtle. But of the currently FDA approved biomarkers typically analyzed by ELISA assays, about 50 can be detected by simple injection of a protein digest into a mass spec. That's good but not a lot.From a physician's perspective in the clinic, …they never start out, uh, from a blank sheet of paper. They have an idea what they're looking for and asking for quantitation of say, two or three biomarkers, I mean, proteins basically,would typically suffice to say, okay, we're in this direction, versus going in that direction. And that is very cheaply, automatically done with ELISAs running on a fully automated, robotic platform in the clinical biochemistry labs.The current ability to detect 300 proteins out of the mix is way more than a physician could use right now, but probably does not go deep enough to uniquely detect a disease. In many sports, this is called no man's land.Nevertheless between improvements in detection, robustness and AI to help analyze the data, Ole sees the field moving dramatically over the next few years.In terms of analysis, we may find that the changes we see in a patient's proteome, may not directly identify targets for therapy. Rather they might reflect secondary effects of a disease. In this case, where treatments can be found, those biomarkers may serve to monitor progress during treatment as opposed to being used for diagnosis.Beyond the need for improved robustness (a clinical analyzer needs to run hundreds of samples per day without human intervention), there aren't enough mass spectrometers to analyze the possible volume of samples. Using proteomics on top of genomic or metabolic analysis seems to be a more likely strategy.Ole closed with this idea:I do think things are progressing and, and I think, you know, what is really, really needed in our field would be sort of like the first success story. I mean that, from my perspective, that's what we really need to somebody, some group coming out and saying, so now we have this, this protein pattern is the detection of this and this. Whatever the success story basically is, I think it is just important that we begin to see some success stories and that will fuel the fire and get, more things going.I'm interested in your thoughts. Do you know of any success stories already? Where do you see proteomics taking off?Chat with Chris about custom content for your life science brand. This is a public episode. If you would like to discuss this with other subscribers or get access to bonus episodes, visit cclifescience.substack.com

In dieser Geschichte geht es um einen jungen Dinosaurier namens Elisa, der sich auf ein Winterabenteuer begibt und dabei auf eine Vielzahl von Herausforderungen und Hindernissen stößt. Komm, wir begleiten Elisa bei ihrem Abenteuer. P.S.: Noch mehr tolle Geschichten findest du auf onkelguido.de

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.12.05.518134v1?rss=1 Authors: Sullivan, M. A., Lane, S. D., Ball, S. R., Sunde, M., Neely, G. G., Moreno, C., Werry, E. L., Kassiou, M. Abstract: Background: Widescale evidence points to the involvement of glia and immune pathways in the progression of Alzheimers disease (AD). AD-associated iPSC-derived glial cells show a diverse range of AD-related phenotypic states encompassing cytokine/chemokine release, phagocytosis and morphological profiles, but to date studies are limited to cells derived from PSEN1, APOE and APP mutations or sporadic patients. The aim of the current study was to successfully differentiate iPSC-derived microglia and astrocytes from patients harbouring an AD-causative PSEN2 (N141I) mutation and characterise the inflammatory and morphological profile of these cells. Methods: iPSCs from three healthy control individuals and three familial AD patients harbouring a heterozygous PSEN2 (N141I) mutation were used to derive astrocytes and microglia-like cells and cell identity and morphology were characterised through immunofluorescent microscopy. Cellular characterisation involved the stimulation of these cells by LPS and A{beta}42 and analysis of cytokine/chemokine release was conducted through ELISAs and multi-cytokine arrays. The phagocytic capacity of these cells was then indexed by the uptake of fluorescently labelled fibrillar A{beta}42. Results: AD-derived astrocytes and microglia-like cells exhibited an atrophied and less complex morphological appearance than healthy controls. AD-derived astrocytes showed increased basal expression of GFAP, S100{beta} ; and increased secretion and phagocytosis of A{beta}42 while AD-derived microglia-like cells showed decreased IL-8 secretion compared to healthy controls. Upon immunological challenge AD-derived astrocytes and microglia-like cells show exaggerated secretion of the pro-inflammatory IL-6, CXCL1, ICAM-1 and IL-8 from astrocytes and IL-18 and MIF from microglia. Conclusion: Our study showed, for the first time, the differentiation and characterisation of iPSC-derived astrocytes and microglia-like cells harbouring a PSEN2 (N141I) mutation. PSEN2 (N141I)-mutant astrocytes and microglia-like cells presented with a primed phenotype characterised by reduced morphological complexity, exaggerated pro-inflammatory cytokine secretion and altered A{beta}42 production and phagocytosis. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Welcome to the 1st episode in English. This podcast is about suicide. People who are affected by suicide tell their story, talk about their experiences and everything that goes with their respective situation. My name is Elisa Roth and I am someone who is affected myself. My mother took her own life in 2002 when I was 27. Since the topic of suicide and talking about it is still a taboo, I came up with the idea for this podcast. Because those affected want to talk. They want to tell their story. And they should - loud and clear and audible for everyone. I would like to request you listeners, if you are also directly affected by suicide, send me an email so that you too can tell me your story. Send your email to mail@selbstwort.com In the first 3 episodes I will tell you my story. I started writing it down almost 5 years ago. I will read this text in the first few episodes so that you can find out who I am and what my story is. 2 years ago I created exactly this podcast in German. And what I triggered with it is still overwhelming and can hardly be put into words. Those of you who are interested in the German version can find it under the title “Selbstwort” wherever there are podcasts. I'm not a native speaker. A long long time ago I lived in England for a few years and studied there. I was in a relationship with an English guy for 5 years..Unfortunately, my English is a bit rusty now and not as fluent and practiced as it was then. But I hope you'll forgive me for that. Before you hear my story, consider the subject of this podcast. Since I keep mentioning my mother's chosen method of suicide and also recounting how I saw her again before the funeral, people who are sensitive to it might want to refrain from listening to this episode. Also in all future episodes everything is often told down to the smallest detail- as it really was and still is today. Because in my opinion this is the only way to contribute to clarification of the subject „suicide“ and the removal of taboos. This is the first of three parts of my story.

In der heutigen Folge habe ich Elisa-Marie Schütz zu Gast.Wir sprechen über ihren besonderen #nft Einstieg, NFT #fotografie und Elisas #community Gedanken. Also viel Spaß mit der heutigen Folge von #allinnftElisa-Marie Schütz:https://elisamarieschuetz.dehttps://www.linkedin.com/in/elisa-marie-sch%C3%BCtz-2b29a0213/https://www.instagram.com/elisamarieschuetz/ALL IN NFT Homepage:https://www.allinnft.de/ALL IN NFT Genesis NFT Mint Seite/ Kauf:https://smartminty.io/all-in-nft-genesisALL IN NFT T-Shirt Mint Seite/ Kauf:https://smartminty.io/all-in-nft-tshirtsOpensea Kollektion ALL IN NFT T-Shirtshttps://opensea.io/de-DE/collection/all-in-nft-tshirtsOpensea Kollektion ALL IN NFT:https://opensea.io/assets/ETHEREUM/0x71608b3551895385637d59c3713e7369ecac2e97/0Social Media, Discord und offizielle Affiliate Links von All in NFT:Linktree :https://linktr.ee/mic_sebDiscord: https://discord.gg/4js6Pg7FJkTikTok: www.tiktok.com/@allinnft77Instagram: Mic_Sebhttps://instagram.com/mic_seb?igshid=...LinkedIn:https://www.linkedin.com/in/sebastian-michelsTwitter: @Mic_Seb91https://twitter.com/MicSeb91Twitch: https://www.twitch.tv/allinnft77YouTube: All in NFThttps://youtube.com/user/VRK226Bei den oben genannten Themen handelt es sich um keine Anlageberatungen. Der Podcast dient lediglich der Unterhaltung.Erläuterung zu Affiliate Links:Die mit ** gekennzeichneten Links sind sogenannte Affiliate Links. Kommt über einen solchen Link ein Einkauf zustande, werde ich mit einer Provision beteiligt. Für Dich entstehen dabei keine Mehrkosten. Wo, wann und wie Du ein Produkt kaufst, bleibt natürlich Dir überlassen.** weitere freiwillige Unterstützung:https://www.patreon.com/user?u=86396667Newsletter Barista Insights by Mic_Sebhttps://tab728b31.emailsys1a.net/247/735/e3ee874dc3/subscribe/form.html?_g=1695576545ALL IN NFT Homepage:https://www.allinnft.de/ALL IN NFT Merchandising Shop:https://all-in-nft.myshopify.com/Opensea Kollektion ALL IN NFT:https://opensea.io/assets/ETHEREUM/0x71608b3551895385637d59c3713e7369ecac2e97/0Linktree :https://linktr.ee/mic_sebDiscord: https://discord.gg/4js6Pg7FJkTwitter: @MicSeb91https://twitter.com/MicSeb91Twitch: https://twitch.tv/mic_sebYouTube: All in NFThttps://www.youtube.com/@Mic_Seb**Blockpit Code für deine einfache Steuererklärung mit Krypto und NFT Wallets:https://blockpit.cello.so/v7peWE3ssIIKontakt:Sebastian@allinnft.deGM Coffee Handy:+491733544148Bei den oben genannten Themen handelt es sich um keine Anlageberatungen. Der Podcast dient lediglich der Unterhaltung.Die mit ** gekennzeichneten Links sind sogenannte Affiliate-/ oder Unterstützungs-Links. Kommt über einen solchen Link ein Einkauf zustande, werde ich mit einer Provision beteiligt. Für Dich entstehen dabei keine Mehrkosten. Wo, wann und wie Du ein Produkt kaufst, bleibt natürlich Dir überlassen. --- Send in a voice message: https://podcasters.spotify.com/pod/show/sebastian-michels7/message

We earlier isolated 10 single-domain antibodies (VHHs) that specifically bind intact (146S) FMDV particles but not the (12S) degradation products of serotype A strain A/TUR/14/98. Such 12S particles are much less immunogenic than 146S particles. As a result, the 146S particle specific VHHs are useful tools for quality control of FMDV antigens used in vaccines. Conventional monoclonal antibodies (mAbs) were shown to bind at least 5 independent antigenic sites by competition ELISAs and sequencing of mutants that are resistant to neutralization by mAbs. Escape mutants of serotype A binding 146S specific mAb 9 had mutations in VP3, near the 2-fold symmetry axis where separate 12S particles join into a 146S particle. This suggests that 146S specificity is caused by the epitope lying on two adjacent 12S pentamers that is separated by 146S particle dissociation. However, cryo EM structural analysis the serotype O 146S specific VHH M170 showed that it recognizes a loop on VP3 that has a different 3D structure in the 12S particle compared to the 146S particle. This change in conformation most likely causes the 146S particle specificity. To further understand the mechanism of 146S specificity, we mapped the antigenic sites of serotype A binding VHHs by competition ELISAs, VNTs and cross-linking mass spectrometry. By Haozhou Li

Heute möchte ich dich einladen, gemeinsam mit mir 2. Könige 13, 14-25 zu lesen und gleichzeitig spirituell, kulturell und historisch zu beleuchten.

Ihr wollt nichts über Masturbation wissen? Na gut, dann müsst ihr halt mit Elisas und Paul langweiligen Partystories leben. Heute gibt es einen Abriss über die Partynacht, die zum Zeitpunkt der Aufnahem nicht einmal 24 Stunden zurückliegt. Wieso sich der Abend komplett anders entwickelt hat, als der unschuldige Paul vermutet hatte, und weshalb Elisa komplett die Schnauze voll hat, erfahrt ihr in der heutigen Episode.

Heute möchte ich dich einladen, gemeinsam mit mir 2. Könige 2, 19-25 zu lesen und gleichzeitig spirituell, kulturell und historisch zu beleuchten.

PROPHETEN UND KÖNIGE mit Pastor Mag. Kurt Piesslinger 2.Serie - PROPHETEN DES NORDREICHES Sein Nachfolger Elisa wirkt besonders in den Prophetenschulen, in denen die Studenten eifrig die Heiligen Schriften vervielfachen. Seine Wundertaten erinnern stark an die Wunder Jesu. 2.9.Die Berufung Elisas Mitten in den Wirren der Reformation unter dem Propheten Elia kommt der Auftrag Gottes an den Propheten seinen Nachfolger zu salben. Elisa, Sohn eines reichen Bauern in der Jordanebene, tritt sofort willig in die Nachfolge und hinterlässt dadurch bleibende Spuren bis in unsere Tage. Gottes Segen! Für Videoaufnahme: vimeo.com/105156267

Heute möchte ich dich einladen, gemeinsam mit mir 1. Könige 19, 19-21 zu lesen und gleichzeitig spirituell, kulturell und historisch zu beleuchten.

Willkommen zu einer neuen Staffel »Bibelstund hat Gold im Mund« – deine tägliche Dosis Bibellesen mit Sascha. Heute wollen wir uns das Setting und die Hintergründe Elias und Elisas anschauen, um morgen gut gewappnet in den Text einsteigen zu können.

Jason Elias, author, acupuncturist, herbalist, is this week’s guest on the Energy Stoners™ Cafe podcast.  He discusses his latest book “The Seven Graces of Ageless Aging, How to die young-as late in life as possible”  with host Toni Quest.   Host: Toni Quest  toniquesttv@gmail.com Guest:  Jason Elias  jasonforhealth@aol.com                                  www.fiveelementhealing.net Energy Stoners™ Cafe is the property of Toni Quest and James H. Brooks, producers.

Tiden går fort när man har roligt, redan dags för deltälvling 3! Den här veckan har vi dessutom otroligt roligt i schlagerstudion. Både åt diverse låttexter som tävlar denna veckan, men också åt att Elisas nummer beskrivs i vårt pressmaterial med bland annat ord som "fnissel". Roligt har vi även förstås med våra härliga gäster som denna veckan är Emil Assergård som verkar gilla jägershots, Alvaro Estrella som repar framför oimponerad son och storfavoriten Tusse som kanske inte är helt nöjd med att vara just det. Det pratas minnen från en av våra bästa efterfester och tävlas i Schlagerschlag, denna veckan utsätts Tusse för det tvivelaktiga nöjet. Häng med!Support this show http://supporter.acast.com/schlagerfesten. See acast.com/privacy for privacy and opt-out information.

P. Juan Carlos (Ecuador)-Una de las características principales de san José es que era tierno vio a Jesús progresar día tras día en sabiduría en estatura en gracia ante Dios y ante los hombres. Jesús vio la ternura de Dios en José, como un padre siente ternura por sus hijos así el señor siente ternura por los que nos sentimos hijos suyos.

A new pan FMDV das antigen Elisa using multi-serotype reactive monoclonal antibodiesThe wide range of hosts, rapid replication, high levels of viral excretion and multiple forms of transmission make FMD difficult to control and eradicate. As FMD can spread fast, rapid and specific identification of the agent is required, as FMD is clinically indistinguishable from other vesicular diseases such as vesicular stomatitis (VS) and swine vesicular disease (SVD).In this study, we describe the preliminary performance evaluation of a Double Antibody Sandwich (DAS) ELISA for FMDV detection.Reactivity profile of different panFMD Mabs was investigated by indirect ELISAs. The Mabs that showed the wider spectrum of recognition were selected and submitted to further testing for their ability to capture and reveal the different virus strains in DAS ELISA format.Inclusivity was assessed by testing different serotypes including O, A, Asia1, SAT1 and SAT2. The analytical sensitivity was evaluated by testing serial dilutions of these viruses. Results obtained with the new panFMDV DAS ELISA were compared to commercially available techniques: a Mab –based ELISA kit produced by reference laboratories and a lateral flow device test based on the well described 1F10 Mab.The new panFMDV DAS ELISA was able to detect all strains tested. SVD virus was not detected. Interestingly, SAT1 and SAT2 strains were very well detected, whereas a very low signal was observed with the other DAS ELISA. The new panFMDV DAS ELISA showed an improvement in analytical sensitivity (up to 10-fold) compared to the other techniques, and seemed to have a wider spectrum of detection.Further testing on more strains is ongoing to better characterize the Mabs recognition pattern and the usefulness of their use in viral detection tests. This new panFMD DAS ELISA allows for rapid and specific FMDV detection. It could be a useful tool for detecting and controlling FMDV outbreaks.Comtet, Loïc1 ; Carpentier, Alix1 ; Donnet, Fabien1 ; Roche, Mickaël1 ; Pourquier, Philippe 1IDVET, 310 rue Louis Pasteur, 34790 GRABELS, FranceEuFMD Open Sessionwww.eufmdvirtual.com

A. Capozzo - Indirect Elisas bases on purified viral particles that measure different aspects by European Commission for the Control of FMD

S. Baselli - Serological ELISAs based on monoclonal antibodies as diagnostic tools for LSD by European Commission for the Control of FMD

In this episode, we review three hematology cases. One case illustrates the work-up and treatment of immune thrombocytopenia (ITP). Another case demonstrates how to diagnose and manage heparin-induced thrombocytopenia (HIT). And the final case is a patient who presented with anemia, a new mitral valve murmur, and mild splenomegaly. Host David H. Henry, MD, reviews these cases with three residents from Pennsylvania Hospital in Philadelphia – Sheila De Young, DO; Ronak Mistry, DO; and Debika Shinohara, MD, PhD. Case 1: Suspected ITP with Sheila De Young, DO Patient presentation: A 50-year-old female with no past medical history and incidental platelet count of 4,000/microL (normal 150,000-450,000/microL [150-450 x 109/L]). On physical exam, there was no lymphadenopathy, and the spleen was nonpalpable. She had obvious petechiae on her legs. A urine pregnancy test was negative. Her hemoglobin and white blood cell counts were normal via complete blood count. ITP definition: Acquired thrombocytopenia caused by autoantibodies against platelet antigens. One of the most common causes of thrombocytopenia in otherwise asymptomatic adults. To consider: Increased destruction, decreased production, and pseudothrombocytopenia To ensure the platelet count is not falsely low (in the case of pseudothrombocytopenia), looking at a peripheral smear is helpful. If red blood cells and white blood cells appear normal, we can exclude pseudothrombocytopenia. Work-up: We need to rule out secondary causes of thrombocytopenia such as HIV, hepatitis C, chronic lymphocytic leukemia, systemic lupus erythematosus, etc. Management/treatment: In the acute setting, the treatment for ITP is intravenous immunoglobulin and steroids. Long-term management of ITP includes steroids, splenectomy, thrombopoietin receptor agonists (romiplostim/eltrombopag), and rituximab. Case conclusion: This patient was found to have ITP. Shared decision-making led to the patient receiving a thrombopoietin receptor. Case 2: Possible HIT with Ronak Mistry, DO Patient presentation:  A male with ischemic leg and creatinine phosphokinase greater than 4,000 units/L. His platelet count was 101,000/microL on admission, 70,000/microL on the second day, and 60,000/microL on the third day. The patient was on prophylactic subcutaneous heparin for 48 hours, so the surgery team considered HIT to explain the drop in platelets. HIT definition: A life-threatening complication of exposure to heparin. Results from autoantibody directed against endogenous platelet factor 4 (PF4) in complex with heparin. To consider: Determine baseline platelet count, what type of heparin the patient received, and look at when the heparin was administered in relation to when the platelet count dropped. HIT is far less common in patients who receive subcutaneous heparin versus intravenous heparin. Typically, we see a 50% decrease in platelet count 5-10 days following exposure to heparin. Work-up: In the inpatient setting, it is important to consider other causes that predispose patients to thrombocytopenia (i.e., critical illness, medications). Thrombocytopenia can represent a consumptive process of platelets secondary to tissue injury in the setting of elevated creatine phosphokinase. Diagnosis: Enzyme-linked immunosorbent assays (ELISAs) can detect the presence of PF4-heparin antibody. ELISA should be followed by a confirmatory test. The serotonin release assay is preferred among diagnostic tests for HIT. Management/treatment: Stop heparin immediately. Giving more platelets is not the solution. It increases a person’s risk for thrombotic events. The patient needs to be placed on different anticoagulation, such as argatroban or fondaparinux, to carry them through this procoagulant time frame. Case conclusion: HIT was ruled out in this patient. Case 3: Anemia case with Debika Shinohara, MD, PhD Patient presentation: A female, age 45 years, with a 4-month history of intermittent fevers and unintentional weight loss. Her hemoglobin was 8 g/dL, but she had otherwise unremarkable blood work. On physical exam, she was found to have a new mitral valve murmur and mild splenomegaly. To consider: Increased destruction versus decreased production of red blood cells. Low reticulocyte count (

Monopolet: Entertainer Casper Christensen, bestyrelsesformand Søren Pind og iværksætter Andrea Elisabeth Rudolph. Vært: Mads Steffensen. 1. Kathrine er i tvivl, om hun kan kalde sin datter det samme navn som sin kærestes eks. 2. Lone ved ikke, hvornår hun skal fortælle sin datter om sit biologiske ophav. 3. Mettes søn er blevet venner med en dreng fra klassen, der stjæler. 4. Jannie har en ferie-affære med en mand i Berlin. 5. Prebens datter er lige blevet 18 år og har bedt ham tegne en tatovering. 6. Elisas søster er meget syg og erklæret terminal - Elisa og hendes søskende er i tvivl, om søsterens datter på 4 år skal se sin mor, når hun dør 7. Lars er frustreret over, at hans søster og svoger kun ønsker sig pengegaver 8. Marianne har haft en affære med en kollega og god ven siden foråret 9. Katinka ved ikke, om hun skal fortælle sin veninde, at hun lugter af sved 10. Mikkel føler sig overvåget af sine venners digitale gadgets, når han er på besøg hos dem

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.26.314567v1?rss=1 Authors: Van Belle, S. C., de Lange, A., Tomes, H., Lucas, R., Naidoo, V., Raimondo, J. V. Abstract: Human cysticercosis is a disease caused by larvae of the cestode Taenia solium. It is the most common cause of adult-acquired epilepsy world-wide where it exacts a debilitating toll on the health and well-being of affected communities. It is commonly assumed that the major symptoms associated with cysticercosis are a result of the direct presence of larvae in the brain. As a result, the possible effect of peripherally located larvae on the central nervous system are not well understood. To address this question, we utilised the Taenia crassiceps intra-peritoneal murine model of cysticercosis, where larvae are restricted to the peritoneal cavity. In this model, previous research has observed behavioural changes in rodents but not the development of seizures. Here we used ELISAs, immunoblotting and the Evans Blue test for blood-brain barrier permeability to explore the central effects of peripheral infection of mice with Taenia crassiceps. We identified high levels of parasite-targeting immunoglobulins in the serum of Taenia crassiceps infected mice. We show that the Taenia crassciceps larvae themselves also contain and release host immunoglobulins over time. Additionally, we describe, for the first time, significantly increased levels of IgG within the hippocampi of infected mice, which are accompanied by changes in blood-brain barrier permeability. However, these Taenia crassiceps induced changes were not accompanied by alterations to the levels of proinflammatory, pro-seizure cytokines in the hippocampus. These findings contribute to the understanding of systemic and neuroimmune responses in the Taenia crassiceps model of cysticercosis, with implications for the pathogenesis of human cysticercosis. Copy rights belong to original authors. Visit the link for more info

PROPHETEN UND KÖNIGE mit Pastor Mag. Kurt Piesslinger 2.Serie - PROPHETEN DES NORDREICHES Sein Nachfolger Elisa wirkt besonders in den Prophetenschulen, in denen die Studenten eifrig die Heiligen Schriften vervielfachen. Seine Wundertaten erinnern stark an die Wunder Jesu. 2.9 Die Berufung Elisas Mitten in den Wirren der Reformation unter dem Propheten Elia kommt der Auftrag Gottes an den Propheten seinen Nachfolger zu salben. Elisa, Sohn eines reichen Bauern in der Jordanebene, tritt sofort willig in die Nachfolge und hinterlässt dadurch bleibende Spuren bis in unsere Tage. Gottes Segen! Weitere Infos unter: vimeo.com/105156267

Elisa hebt den Mantel des Propheten Elia auf. Nach der Himmelfahrt Elias ist er alleine. Er ist ein Werkzeug in der Hand des himmlischen Meisters. Wie kann er in einer Zeit der "vergifteten Quellen" Gottes Ruf folgen - und wie wird er sich bewähren? Bibelstelle: 2 Kö 2,14. Predigt von Jobst Bittner, vom Sonntag den 28.06.2020 16 Uhr

Elisa hebt den Mantel des Propheten Elia auf. Nach der Himmelfahrt Elias ist er alleine. Er ist ein Werkzeug in der Hand des himmlischen Meisters. Wie kann er in einer Zeit der "vergifteten Quellen" Gottes Ruf folgen - und wie wird er sich bewähren? Bibelstelle: 2 Kö 2,14. Predigt von Jobst Bittner, vom Sonntag den 28.06.2020 16:00 Uhr.

Episode is Fully in Spanish! El Episodio es Completamente en Español. Gracias a mi hermano Elias Hernandez, creador dominicano y unos de los fundadores del grupo christiano llamados "Revolucionarios Music." En este episodio hablamos sobre la trayectoria desde un proyecto llamado "La Ofrenda" hasta lo que es hoy, un movimiento espiritual, su nueva canción "Dueño" con Loammy Bido . Con el complejo uso de la música urbana, Elias usa sus letras para impactar las vidas de personas que quizás aún no han escuchado del evangelio. Pueden seguir su página de instagram Revolucionario Music También pueden apoyar el movimiento: Album: La Ofrenda en Itunes Su página de Youtube: "Revolucionarios Music." Twitter FaceBook SoundCloud Gracias a Mi hermano Elisas por su presencia en esta plataforma, gracias a su esposa por apoyarlo en sus momentos más difíciles, y gracias a dios por darle el talento necesario para seguir sus sueños y edificar las vidas de otras personas.

On January 29th 2013, 21 year old college student Elisa Lam vanished while on a solo trip to the West Coast during her stay at a hotel in downtown Los Angeles. During the investigation into Elisas disappearance, police publicly released a surveillance video from inside a hotel elevator showing Elisa acting strangely before she vanished. Then stories of the hotels dark past began to surface - stories of suicides, violence that happened inside its walls and serial killers who once lived there. Then, several weeks after she disappeared, hotel guests began complaining of a foul taste and smell in the hotel water. A maintenance man looking inside one of the rooftop water tanks found Elisas naked and decomposing body. After an autopsy the official cause of her death was ruled a drowning which did not answer the biggest questions of the case. How did she get to the hotel rooftop -which was inaccessible to guests? how did she end up naked inside of a water tank that was 8 feet tall, without a ladder, and then open the lid that was supposedly closed and weighed close to 80lbs? And what was with her bizarre behavior caught on the surveillance video? Was she playing what is known as the elevator game? This remains one of the most discussed cases on the World Wide Web with unlimited theories as to what events led to her demise. Links to research used in this episode: Civil case article: https://laist.com/2015/10/02/ elisa_lam_wrongful_death_suit.php. Join us and discuss this case on Twitter : https://mobile.twitter.com/DisappearedThey --- Support this podcast: https://anchor.fm/theydisappeared/support

Uuenenud saate “Tööelu” kuuenda saate külaliseks oli Elisa personalijuht Kaija Teemägi, kes tõstis pisut loori sellelt, kuidas Eesti ühe suurima mobiilioperaatori köögipool välja paistab. Lisaks sellele räägiti saatejuht Martin Hansoniga sellest, kuidas läks käima Elisas juurutatud paindliku tööaja ja töökoha projekt. “Meil nüüd juba ehk viis aastat töös olev paindliku tööaja ja paindliku töökoha projekt on tõstnud nii töötajate efektiivsust, ettevõtte kasumlikkust ning parandanud meie töökeskkonda. Tuleb tunnistada, et kui inimene töötab kodust on töötaja palju distsiplineeritum, kiirem ja töötab rohkem,” nendib Teemägi. Personalijuht lisab, et ka nende inimeste tööle võtmine käib natukene teistel alustel, kui ehk tavaliselt firmades. Nimelt ei otsita Elisa paljudesse ametitesse väga konkreetse oskusega inimesi, vaid inimesi kellel silm särab, kellel on soov õppida ja areneda. “Ameti saab õppida, hoiakuid ja suhtumist mitte. Olgugi, et esimese pilgu uue ametikoha täitmiseks viskame me oma majas sees olevatele inimestele, siis väljastpoolt otsime enda juurde neid, kes soovivad, midagi ära teha,” lisab Teemägi. Kõik kommentaarid, küsimused ja ettepanekud saates räägitu osas palume saata aadressile tooelu@delfi.ee!

En ettermiddagsprat med Elisa Røtterud og hennes  reiseeventyr får TinTin til å fremstå som en kjedelig nesepiller. I denne podcasten får du høre Elisas spinnville reise fra fashion til fjell, hva som skjedde da guruen stappet en finger i rumpa, og hvorfor ble Elisa helt besatt av fjelljuvelen Ama Dablam? Hva skjedde egentlig når Elisa bæsjet på et muldyr?  Visste du at sherpaene ikke skjønner ironi? Og at Elisa vet svaret på hvorfor Everestkøen er blitt så massiv? Og at Elisa serverer dohistorier på et fat :)  Ja, også en ting til - Elisa har gifta seg!!! Og det er bare begynnelsen..Og hvorfor har Elisa alarmen på  kl på 00.00 i disse tider? Og hvorfor mener Elisa at du bør søke om å få lov til å kjøpe El-sykkel? Og hvorfor er Elisa så sliten når hun kommer på jobb? Herregud for et vandrende eventyr på to bein!   

Terminando nossos especiais do Diversão Offline 2019, temos agora o Balbi encarnando um repórter de campo, enrolado em fio de microfone, caçando e sendo caçado por companheiros, autores, criadores e fãs de RPG para trocar uma idéia e fazer aquela social que cria tantos laços de amizade e companheirismo. A gravação foi bem caótica e o episódio refletirá isso, não só na qualidade do áudio mas também no volume às vezes errático, mas são ossos do ofício: a idéia é mostrar de forma crua o tipo de encontro randômico do mais desejável que ocorre. Espero que te faça voltar nos próximos ou considerar conhecer caso não tenha ido ainda. É como encontrar amigos de infância, gente que divide a mesma paixão. Lista dos entrevistados, em ordem caótica.- Carlos Silva, do Regra da Casa - Gustavo Tertoleone além do Regra da Casa, participa do Câmara Obscura e do zine Vomitations of the Grotesque Princess - Fabio Balbinot, criador do Conectron - Café com Games (???) - Cesar Capacle, do Not a Giraffe Studio, co-autor com Fabio Silva de Gatos Espaciais - Bruno Cobbi da Roleplayers, companhia de mestres profissionais - Desaventureiros, web série da Maré Geek - Aya e Pam do Mestre dos Magos RPG, um imenso servidor de Discord de RPG - Diogo Nogueira do Pontos de Experiência, autor de Sharp Swords & Sinister Spells e Solard Blades & Cosmic Spells - DM Arthas (você sabe todos os links pro DM Arthas) e sua ajudante (Ramon Mineiro e Carol "Escaroles" Batista) - Matheus do Forja do Mestre - Julio Matos com Goddess Save the Queen e Áureos - Luciano Jorge do Jornal Empoderado - Luís da Orgutal - BsB - Rafael "Ramon" Amon, do Perdidos no Play com a ativação do D&D 5e pela Galápagos Jogos - Xezz, Capusso e Kazz do Tear dos Mundos - Vinzão e Elisas do Game Chinchila - Ximu do Casa Velha RPG com a ativação do D&D 5e pela Galápagos Jogos

Du kan lytte til bibeltimene som ligger her gratis, men som en takk ville vi sette pris på om du støttet menigheten DELK Bergens arbeid med en gave på kontonummer 3624 63 90015 (Sparebanken vest) eller Vipps: 103439 (DELK Bergen)

Idag, kommer era öron trilla av, för Eliza gnäller på sitt jobb. Hon är också glad över att någon verkar ha smugit i henne lite uppåttjack. Karin är i vanlig ordning glad över att ha lämnat det sjunkande skeppet. Är vårdpersonal gnälligare än andra eller har vi bara mer att gnälla över? Elisas syrra Claudia gästar en liten snutt och berättar vad som verkligen är värt att gnälla över. Trevlig lyssning! See acast.com/privacy for privacy and opt-out information.

Join us in this webinar featuring Dr. Omonse Talton who will guide you through developing a "fool proof" enzyme-linked immunosorbent assay (ELISA). In this webinar, you will learn: - When and why you should use the different types of ELISAs– direct, indirect, sandwich, competitive/inhibition ELISA - Major considerations for developing your ELISA - Tried and tested tips for you to perform a successful ELISA The ELISA is one of (if not the most) common techniques in biology and biochemistry laboratories. You can use an ELISA to detect minute amounts of protein for medical diagnostics, for testing food for common allergens, and in toxicology screens screen for certain drugs. While the overall premise of an ELISA is simple, as with any assay, its success hinges on the experimental set up. In this webinar, Omonse will give you a crash course on that experimental set up—from start to finish. She will discuss the different types of ELISAs, the major considerations in development of the assay, and troubleshooting—including tips and tricks. With this webinar, you will be able to ensure that your ELISA provides reproducible and publishable results!

Einleitung: Zur in-vitro Diagnostik der Tuberkulose (TB) und latenten TB Infektion (LTBI) im Kindesalter werden derzeit zunehmend „Interferon-gamma (IFN-γ) release assays“ (IGRA) verwendet. Die Sensitivität der IGRA insbesondere im Kindesalter ist umstritten. Auch eine diagnostische Unterscheidung zwischen TB und LTBI ist mittels IGRA und anderen bisherigen basierten Testverfahren nicht möglich. In vorangegangen Studien zeigte sich der Biomarker „IFN-γ-inducible-protein-10“ (IP-10) als viel versprechend zur Diagnose der TB und LTBI. Ziele: In der vorliegenden Dissertationsschrift wurden die IP-10-Plasmakonzentrationen von Kindern mit TB, LTBI, Atemwegsinfektion (AWI) oder nicht-tuberkulöse Mykobakterien (NTM)-Erkrankung ohne Stimulation der Proben, nach unspezifischer Mitogen-Stimulation und nach spezifischer Mycobacterium tuberculosis (M. tuberculosis)-Antigen-Stimulation bestimmt. Die Antigen-stimulierten IP-10-Plasmakonzentrationen der TB- und LTBI-Gruppe wurden mit denen der NTM- und AWI-Gruppe verglichen. Darüberhinaus wurde beurteilt, ob eine Unterscheidung zwischen TB und LTBI anhand der IP-10-Plasmakonzentration möglich ist. Außerdem wurde die Konkordanz und Korrelation zwischen dem IP-10-ELISA und QuantiFERON® -TB Gold In-Tube (QFT-IT) Test beurteilt und untersucht, ob der Biomarker IP-10 altersabhängig sezerniert wird. Material & Methoden: 48 Kinder wurden in die Studie eingeschlossen. Das mittlere Alter der Studienteilnehmer war 54 Monate. Alle Studienteilnehmer wurden zuvor in Deutschland entweder mit einer TB (n=11), LTBI (n=14), NTM (n=8) oder AWI (n=15) diagnostiziert. Unabhängig von der vorliegenden Studie wurden bei allen teilnehmenden Kindern IFN-γ-Werte mittels des QFT-IT-Testes bestimmt. Im Rahmen des QFT-IT wurden die für den Test notwendigen Blutproben entweder nicht stimuliert, mit einer unspezifischen Mitogenen-Substanz oder mit spezifischen M.tuberculosis-Antigenen stimuliert. Die jeweiligen Plasma-Überstände wurden asserviert und zur Bestimmung von IP-10 verwendet. Die IP-10-Konzentrationen wurden, in Zusammenarbeit mit dem Klinischen Forschungszentrums der Universität von Kopenhagen, mittels einem zu Forschungszwecken entwickelten ELISAs gemessen. Ergebnisse: Die IP-10-Plasmakonzentrationen ohne Stimulation, mit unspezifischer Mitogen- und spezifischer Antigen-Stimulation betrug für die TB-Gruppe 704 pg/ml, 12.966 pg/ml und 12.702 pg/ml; für die LTBI-Gruppe 366,5 pg/ml, 10.232 pg/ml und 9.109 pg/ml; für die NTM-Gruppe 309 pg/ml, 11.197 pg/ml und 97 pg/ml; und für die AWI-Gruppe 694 pg/ml, 5.401 pg/ml und 84 pg/ml. Es konnte kein signifikanter Unterschied zwischen der IP-10-Konzentration der TB- und LTBI-Gruppe festgestellt werden (p-Wert= 0,24). Die IP-10- und IFN-γ- Plasmakonzentrationen der Kinder mit TB und LTBI korrelierten stark miteinander (rsp=0,65; p-Wert = 0,03 und rsp=0,79; p-Wert < 0,001). Der IP-10-ELISA und QFT-IT Test zeigten ebenso eine hohe Konkordanz (κ =0,96). Die IP-10-Sekretion war 18fach höher im Vergleich zur IFN-γ-Sekretion. Es konnte keine Korrelation zwischen dem Alter und der Mitogen-stimulierten IP-10-Konzentration nachgewiesen werden. Schlussfolgerungen: Die IP-10-Plasmakonzentration von Kindern mit TB und LTBI im Vergleich zu Kindern mit NTM und AWI ist signifikant nach spezifischer M.tuberculosis-Antigen-Stimulation erhöht (p-Wert > 0,001). Die qualitativen und quantitativen Testergebnisse des IP-10-ELISAs korrelieren stark mit denen des QFT-IT-Testes. Im Vergleich zu IFN-γ scheint IP-10 in höheren Konzentrationen und möglicherweise unabhängig vom Alter sezerniert zu werden. Das legt die Vermutung nahe, dass IP-10 zur Diagnose der TB und LTBI im Kindesalter in Zukunft Verwendung finden könnte.

Desoxyribonuklerinsäure (DNA), die Erbinformation, definiert die Struktur und Funktion jeder Zelle. In Eukaryoten ist sie um Oktamere aus den Histonen H2A, H2B, H3 und H4 gewunden. Sie bilden mit der DNA höhere Strukturen, das sog. Chromatin. Die Struktur des Chromatins beeinflusst direkt die Aktivität der gebundenen DNA. Eukaryoten besitzen daher viele molekulare Mechanismen zu ihrer Veränderung, z.B. posttranslationale Modifizierungen (PTMs) von Histonproteinen oder den Austausch von kanonischen Histonen mit Histonvarianten (z.B. H3.1, H3.2, H3.3, u.a.). Für einige Varianten sind ihnen eigene, charakteristische PTMs beschrieben, z.B. die Phosphorylierung des Serins 31 der Variante H3.3 (H3.3S31ph). Die Histone Code Hypothesis postuliert, dass PTMs von Histonen in festen Mustern vorliegen können. Die Switch Hypothesis beschreibt die Regulation von Bindemolekülen an Histone durch benachbarte PTMs. Auf ihrer Grundlage wurde die These der Doppelmodifizierung einer bekannten Trimethylierung der Aminosäure (AS) Lysin 79 mit einer mutmaßlichen Phosphorylierung der AS Threonin 80 auf Histon H3 aufgestellt (H3K79me3T80ph). Neben der Etablierung eines einfachen Systems zur Identifizierung bisher unbeschriebener Phosphorylierungen bestand ein zweites Ziel dieser Arbeit im Nachweis der Phosphorylierung von H3 Threonin 80 in vivo. Eine weitere Zielsetzung lag in der genaueren Charakterisierung der bereits beschriebenen Varianten-spezifischen PTM H3.3S31ph, deren Expression zwar eng umschrieben ist, über deren spezifische Funktionen, Kinase und mögliche Effektorproteine aber wenig bekannt ist. Um einen Überblick über Phosphorylierungen verschiedener Histonroteine zu gewinnen, wurden unterschiedliche Polyacrylamidgel-Elektrophoresen Verfahren (PAGE) etabliert. Zum Einsatz kamen A/U-, T/A/U- und 2D-T/A/U-PAGE Verfahren. Sie ermöglichten in Übereinstimmung mit der Literatur die Auftrennung bekannter PTM und stehen nun für weiterführende Studien zur Verfügung. Der massenspektrometrische Nachweis der putativen PTM H3K79me3T80ph in vivo gelang nicht. Trotz optimierter Versuchsbedingungen konnte die Phosphorylierung weder in der MALDI-ToF, noch in der Orbitrap MS/MS nachgewiesen werden. Initiale Antikörperdaten wurde aufgrund einer aufgedeckten Kreuzreaktivität in Frage gestellt. Obgleich nicht mit letzter Sicherheit gesagt werden kann, dass H3K79me3T80ph in vivo nicht existiert, wurde die These letztlich verworfen. Die molekularbiologische Untersuchung der Varianten-spezifischen PTM H3.3S31ph ergab bei verifizierter Überexpression und Chromatin-Integration punktmutierter Histone übereinstimmend Hinweise auf einen Effekt der PTM auf die Zellteilung. Es konnte gezeigt werden, dass Serin 31 bzw. ihre PTM H3.3S31ph sowohl Zellzyklus, als auch Wachstums- und Proliferationsgeschwindigkeiten von HeLa Zellen beeinflusst. Dies argumentiert für eine aktivierende Funktion von H3.3S31ph in der Mitose. Weiter konnten mithilfe eines ELISAs fünf potentielle Proteinkinasen für H3.3S31ph identifiziert werden. Vorrangig kommt dabei die nukleäre Kinase PIM1 in Betracht. Zur weiteren Untersuchung potentieller Effektorproteine wurde in vitro ein molekularbiologisches Modellsystem etabliert. Es steht nun für weiterführende Studien zur Verfügung. Erste Vorarbeiten konnten bereits zeigen, dass hier evtl. Interaktionen mit dem Linkerhiston H1 eine wichtige Rolle spielen.

Quorum Sensing (QS) is a process that enables bacteria to communicate via chemical signalling molecules, which are called autoinducers (AI). When a threshold concentration of QS molecules is reached, the bacteria start their QS regulated gene expression e.g. bioluminescence (Vibrio fischeri), virulence factor secretion (Pseudomonas aeruginosa), biofilm formation (Burkholderia cepacia), sporulation, and mating. It was found that many Gram-negative bacteria use acylated homoserine lactones (AHLs or HSLs) as autoinducers. Due to the broad biological functions of HSLs, the interest in detection and analysis of HSLs is increasing for medical, biotechnological and agricultural applications. In the past years, numerous analytical methods have been developed for HSLs. Conventional analysis, which usually combines chromatography, mass spectrometry (MS), and nuclear magnetic resonance (NMR) has been very successfully applied for identification and quantification of HSLs. But normally, conventional analysis requires many steps of sample preparation, e.g. extraction, pre-concentration and optimisation of conditions to separate individual HSL molecules. In addition, many sensitive bioreporter assays have been developed using different LuxR responsive promoters, which contain LuxR family functional proteins but lack the HSL synthase. A combination of different bioassays is strongly recommended, since no bioreporter is sensitive to all HSLs. Alternatively, in this study, an anti-HSL antibody based immunochemical detection method has been successfully developed and established. HSL molecules consist of a homoserine lactone ring and an acyl side chain (4-18 carbon atoms), and they differ only from side chain length and substitution at C3 atom. Regarding the variation of the molecule structures, four HSL haptens, named HSL1, HSL2, HSL3 and HSL4, were designed for antibody and assay development. HSL1 and HSL3 have a long chain (C11-COOH), but HSL1 has an -oxo and HSL3 has an –OH functional group at the C3 position. In comparison, HSL2 (C5-COOH) and HSL4 (C9-COOH) have shorter side chains and no substitution on the C3 atom. The haptens were synthesised and were covalently coupled to the C-terminal COOH-group of the NH2-residues (lysines) of the carrier proteins (BSA/OVA). Using these HSL hapten-conjugates, rat and mouse anti-HSL monoclonal antibodies (mAbs) were produced, screened and further characterised with enzyme-linked immunosorbent assays (ELISAs). Corresponding to hapten structures, the antibodies showed different selectivities to HSLs with different substitution on C3 position and chain length. Eight mAbs (HSL1-1A5, HSL1-8E1, HSL1/2-2C10, HSL1/2-4H5, HSL4-4C9, HSL4-5H3, HSL4-5E12 and HSL4-6D3) were selected from about 200 mAbs and characterised in detail using coating antigen and enzyme tracer formats. It was demonstrated that the new assays have HSL detection ranges from nM to low µM, which is sensitive enough for detection of HSLs in natural samples according to literature. Interestingly but not surprisingly, AHLs mAbs have at least 20 times higher sensitivity against hydrolysed HSLs (named HSs) than original HSLs, because the conjugation and immunisation conditions, e.g. pH and temperature, for mAb development resulted in HSL hydrolysis. This property of antibodies additionally offers a new sensitive method to detect quorum quenching (QQ) relevant homoserines (HSs), which are important degradation products of HSLs. Comparable results have been obtained by Biacore and Aqua-Optosensor biosensors for HS (L) characterisation. Based on these properties of the mAbs, a detection method of HSLs and HSs in biological samples has been developed and optimised. With the comparison of the real samples before and after hydrolysis treatment, the assays could simply present the relative HSL- and HS- contents in the samples. Similar to bio-reporters, the identification or quantitation of single HSL molecules is not possible only using immunoassay due to the broad recognition of HSLs and HSs. For this purpose, a combination with conventional chemical analysis is a must. However, as a novel sensitive HSL and HS detection method, the developed immunoassays have the advantages of being fast, cost effective and having low sample volume requirement. Using the direct or indirect fluorescence signals of fluorophore labelled anti-HSL mAbs, the in situ experiments with modified Burkholderia cepacia biofilm on ibidi slide (plastic flow chamber from ibidi GmbH) and Pseudomonas putida inoculated barley root have been carried out. Unfortunately, the in situ tests were not successful, mainly due to remaining unspecific background signals. Nevertheless, a few steps, e.g. fluorophore labelling, biofilm formation, and surface blocking have been optimised. The door of HSL in situ tests with the antibodies is still open, if a suitable specific visualising detection method could be found in the future. Certainly, the antibodies can also be broadly applied for many other immunochemical techniques, such as immunosensors or immunoaffinity columns for characterisation or pre-screening of HSLs/HSs, as have been demonstrated successfully.

Über einen Zeitraum von 1,5 Jahren wurden von 1013 Schaf-, 503 Ziegen- und 555 Rindern Blutproben aus 100 bayerischen Betrieben gezogen und auf das Vorhandensein von Pestivirus-Antikörpern bzw. BVDV/ BDV untersucht. Zum Antikörpernachweis wurden kommerziell erhältliche ELISAs verwandt: Ein indirekter ELISA (Svanovir BDV-Ab-ELISA, Svanova) und ein Blocking-ELISA (Priocheck BVDV-Ab-ELISA, Prionics) für Seren kleiner Wiederkäuer und ein weiterer indirekter ELISA (Svanovir BVDV-Ab-ELISA, Svanova) für Rinderseren. Weiterhin wurden 325 Pestivirus-Isolate aus bayerischen Rindern mittels Multiplex real time RT-PCR (Cador BVDV Type 1 / 2 BDV RT-PCR, Qiagen) typisiert. Die Einzeltierprävalenz betrug bei Schafen 8,2% und bei Ziegen 1,2%. Bezogen auf 91 schafhaltende Betriebe lag die Prävalenz bei 12,0%, in einem von 17 Betrieben wurden Pestivirusantikörper bei Ziegen gefunden. Mit Kreuzneutralisationstesten wurden die Antikörper typisiert: Bei den Schafseren waren 24,1% BVDV-1-, 1,2% BVDV-2-, 4,8% BVDV-1- oder 2- und 48,2% BDV-spezifisch. Die BDV-spezifischen Seren stammten alle aus einem Betrieb. Die verbleibenden 21,7% konnten wegen nur geringer Titerunterschiede zu den SNT-Stämmen nicht typisiert werden. Alle sechs Ziegenseren stammten aus einer Herde, sie wiesen jeweils den höchsten Titer gegen BVDV-2 auf, in zwei Fällen mit einem mehr als vierfachen Abstand zu BVDV-1 und BDV. Eine signifikant höhere Anzahl an seropositiven kleinen Wiederkäuern (p= 0,003) stammte aus Betrieben mit Rinderhaltung. BVDV-Infektionen von Rindern zu kleinen Wiederkäuern wurden beobachtet, wohingegen keine Hinweise für die Übertragung von Pestiviren von kleinen Wiederkäuern zu Rindern oder für die Persistenz von BVDV in kleinen Wiederkäuern vorhanden waren. Die Genotypisierung der 325 Pestivirus-Isolate aus Rindern lieferte folgendes Ergebnis: 91,4% erwiesen sich als BVDV-1 und 8,6% als BVDV-2, BDV wurde nicht gefunden. Anhand von 128 Schafseren mit neutralisierenden Antikörpern wurden Sensitivitäten von 81% für den indirekten ELISA und 94% für den Blocking-ELISA bestimmt. Die Spezifitäten lagen jeweils bei 93% and 100% (N=200 SNT-negative Seren).

Thu, 12 Mar 2009 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/10009/ https://edoc.ub.uni-muenchen

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

Early detection and elimination of cattle persistently infected (PI) with bovine viral diarrhoea virus (BVDV) is a key element for eradication programs. Testing dried skin biopsies derived from ear tagging might be useful for detection of BVDV in newborn calves. The aims of this study were the development of methods for antigen solubilization and RNA preparation, the investigation of the stability of these viral components and the comparison of the analytical sensitivity of different tests. Commercial antigen capture ELISAs for NS3, Erns and mixed antigens, a blocking ELISA for BVDV antibodies and two real time RT-PCR assays for 5’UTR were used as BVDV specific tests. Ear biopsies were collected from nine BVDV antibody free PI animals (infected with BVDV-I CR4043) after slaughter and from twelve PI calves (fetal infection with BVDV-I PT810) after euthanasia at the age of 5-13 days in the time of colostral antibodies. For solubilization of the BVDV antigens the detergents dodecyl sulfate, sodium deoxycholate and EMPIGEN were inappropriate. Out of the suitable detergents (CHAPS, Triton X100, Nonidet P-40, Tween 20, n-Octyl-ß-D-glucopyranoside and Digitonin) Triton X100 in a concentration of 1% was chosen for antigen solubilization. Best RNA yield was obtained using a mixer mill and a guanidine thiocyanate containing buffer, followed by RNA isolation with Qiagen RNeasy® kits (Fibrous Tissue Mini Kit and Lipid Tissue Mini Kit). RNA isolation kits provided by Roche were less efficient. NS3-ELISAs were of low analytical sensitivity for ear biopsies, maternal antibodies led to negative results. In addition NS3 epitopes are very heat sensitive. In contrast Erns-ELISAs showed high analytical sensitivity. Samples of PI animals without maternal antibodies gave mean titers above 30. Maternal antibodies had limited effects. In samples of nine PI animals with colostral antibodies the lowest titer was 13. Relevant temperatures by sample drying and storage led to minor titer reductions. The real time RT-PCR resulted in a sensitivity more than 104 fold over the detection limit, even in calves in the time of neutralizing antibodies. Bacterial or fecal contamination before sample drying had no relevant influence on the stability of Erns and 5’UTR RNA. Erns-ELISAs and real time RT-PCR seem to be suitable for detecting BVDV in dried ear notch samples of PI animals. Before appliance in BVDV eradication programs, the diagnostic sensitivity and specificity of tests using ear biopsies have to be evaluated by studies in the field with an adequate number of samples and an appropriate monitoring.

Ziel der vorliegenden Studie war es, abzuklären, wie verlässlich die Serotypisierung von HIV-1 die in einem ostafrikanschen Land vorkommenden, seit Jahren nebeneinander existierenden, unterschiedlichen Genotypen bestimmen kann. Hierzu wurden die genetischen Subtypen von 86 AIDS-Patienten aus Mbeya-Stadt im südwestlichen Tansania durch die Nukleinsäuresequenzierung eines env-Abschnitts bestimmt. Die Daten wurden mit den Ergebnissen der V3-Serotypisierung verglichen, welche durch 4 verschiedene Testverfahren erhoben wurden. In den zur Verfügung stehenden Patientenproben konnten 4 genetische Subtypen identifiziert werden: A (25,29%), C (47,55%), D (13,15%) und G (1,1%). Im Vergleich der serologischen Tests untereinander konnte gezeigt werden, dass die Sensitivität und Spezifität der verwendeten ELISAs beträchtlich variierte. Weiterhin konnten verschiedene Aminosäurereste im V3-loop identifiziert werden, die größte Bedeutung für eine erfolgreiche und gleichzeitig spezifische Antikörperbindung haben. Abweichungen hiervon waren in hohem Maße mit einer serologischen Fehlklassifizierung verbundn. Die Ergebnisse haben gezeigt, dass die Tests zumindest auf Populationsebene die Subtypenverteilung in den richtigen Proportionen vorharsagen. Auf individueller Ebene ist die Vorhersagekraft jedoch nicht ausreichend. Die Schuld dafür ist in den extrem ähnlichen Aminosäuresequenzen der prävalenten genetischen Subtypen zu suchen, die eine Unterscheidung von A und C bzw. D und C in vielen Fällen unmöglich machten. Um in groß angelegten Studien die genetischen HIV-1-Subtypen untersuchen zu können, sind modifizierte Algorithmen nötig, mit deren Hilfe die durch regionale Besonderheiten der Viren verursachten Schwierigkeiten der serologischen Tests erkannt und korrigiert werden können.

SUMMARY Immunological monitoring in the context of gene therapy of feline fibrosarcomas Cytokine kinetics and humoral response to vector and transgene In the context of an adenoviral gene therapy of the fibrosarcoma of the cat we introduced the human interleukin 2 (huIL 2)-gene and the feline Interferon g (feIFN g)-gene, one or the other or a combination of both. The aim of these studies is to evaluate the toxic risk and the therapeutic potential of multiple injections of two adenoviral vectors (type 5) expressing immunostimulating genes as adjuvant therapy in cats with spontaneous cancers. Several immunological parameters such as cytokine concentration, antibodies of different classes against vector and transgene and their neutralizing capacity were examined in the thesis at hand in connexion with the questions of adenoviruses in cats, pre-existing immunity and influence of inflammatory context and way of application. The kinetic of the production of IL 2 in different therapeutic approaches was evaluated. A dose-dependency was shown. The levels found in the group treated with a combination of both genes were lower than in the group treated with just AdhuIL 2. Prospects of explications are the shutdown of the viral CMV-Promoter due to the presence of interferon, the rivalry of the two transgenes to transduce a cell and the antiproliferative and antiviral effect of IFN g which directly reduces vectors and infected cells and indirectly lowers the IL 2-production by this means. More than that the application mode played an important role: The plasmic cytokine concentration was more than doubled after intratumoral injection compared to a postoperative use to the tumour’s bed. The serum peak 24 hours after vector application, which quickly redescended, is no decisive factor for the course of cytokine concentration observed locally at the tumor site. Literature shows rather a consistent local secretion for a period of nine days with intensity ten times higher than measured in blood circulation. There are indices that the serum level of IL 2 the day after the first gene transfer may be predictive for the appearance of recidivism within the first year. The tendency of infected cells in?vitro to express more IL 2 without the presence of IFN g is the same. The possibility to dose feIFN g per ELISA is presently not given. Furthermore the development of anti-huIL 2-antibodies was surveyed for a period of one year following the gene-carrying vector application. The immunoglobulins appeared early in the group of treatment with IL 2 alone (day 30), and a tardily rise of Ig G was detected in the therapeutic group with association to IFN g (day 90 to 180). By measuring the existing antibodies capable to detect or to neutralize as well the used adenovirus in ELISAs and biological assays the influence of the transgene on the antiadenoviral immune response could be proofed as well as the influence of the lieu of administration in which the vector is applied. Antiviral Ig M showed up day 14 and Ig G day 30, but their occurrence was much higher in animals treated with interferon (solitary or in association). The neutralizing capacity reached high percentages even in elevated dilutions after just one week. Moreover the immunological response to the vector depends on the application mode in terms of considerably reduced neutralizing antibodies after intratumoral injection compared to the postoperative inoculation to the tumor-surrounding tissues. The parallel-lessened defence to the transgene in this case may be assumed. Altogether the described methods are convenient to ensure an immunological monitoring of veterinary patients during an adenoviral gene therapy and to document the reactions to the applied viral vector and the expressed transgene. RESUME Surveillance immunologique du chat lors de la thérapie génique du fibrosarcome félin Cinétique des cytokines et réponse humorale contre vecteur et transgène Dans le contexte d´une thérapie génique adenovirale du fibrosarcome du chat nous avons introduit les gènes de l´interleukine 2 humaine (huIL 2) ou de l´interféron g félin (feIFN g), seuls ou en combinaison. Le but de ces études est d’évaluer le risque toxique et le potentiel thérapeutique d´injections multiples de deux vecteurs adénoviraux (type 5), contenant des gènes immunostimulateurs, comme thérapie adjuvante dans des cancers spontanés des chats. Plusieurs paramètres immunologiques comme la concentration des cytokines, des anticorps de classes différentes contre le vecteur ou le transgène et leur capacité de neutralisation ont été examinés dans la thèse présentée en rapport avec la question d´adénovirus chez le chat pour déterminer l’impact de l´immunité anti-adénovirale préexistante, l´influence d´un contexte inflammatoire et le mode d´administration. La cinétique de la production d´IL 2 dans différentes approches thérapeutiques a été évaluée. Une relation dose-réponse a été démontrée. Les niveaux trouvés dans le groupe traité avec une combinaison de deux gènes étaient plus bas que dans le groupe traité avec AdhuIL 2 seul. Ces résultats peuvent être expliqués par la coupure du promoteur viral CMV due à la présence de l´interféron dans le tissu, la compétition des deux vecteurs à transgènes différents à transduire une cellule et l´effet antiproliferatif et antiviral de l´IFN g qui limite la persistance de l’infection et engendre de ce fait une baisse de la production d´IL 2. De plus, le mode d´administration joue un rôle important: La concentration de cytokines dans le sérum a plus que doublé après une injection intratumorale, contrairement à une application postopératoire dans le lit tumoral. Le pic de concentration de cytokines dans le sérum, qui apparaît 24 heures après l´administration du vecteur et qui disparaît très vite, n´est pas un facteur décisif pour l’évolution de la concentration de cytokine observée localement au site de la tumeur. La littérature montre plutôt une sécrétion locale continue pour une période de neuf jours, avec une intensité dix fois plus élevée que celle mesurée dans la circulation sanguine. Certaines indications montrent que le niveau d´IL 2 dans le sérum, un jour après le premier transfert génétique, peut refléter les risques d’une récidive au cours de la première année. KeywordsBitte tragen Sie hLa tendance des cellules infectées in?vitro d´exprimer plus d´IL 2 sans la présence d´IFN g est la même. La possibilité de doser feIFN g par ELISA n´est actuellement pas donnée. En outre l´ apparition d´anticorps anti-huIL 2 a été surveillée pendant un an, après administration du vecteur porteur des gènes. Ces immunoglobulines se produisent tôt dans le groupe de traitement avec l’IL 2 seul (jour 30), et une hausse tardive des Ig G a été détectée dans le groupe thérapeutique avec l´association d´IFN g (jour 90 à 180). En mesurant la quantité d’anticorps capables de détecter ou de neutraliser même l´adénovirus utilisé, par des tests d´ELISA et biologiques, l´influence du transgène sur la réponse immunitaire anti-adénovirale et aussi l´influence de l´endroit de l´application dans lesquels les vecteurs sont injectés ont été démontrées. Les Ig M antiviraux se manifestaient jour 14 et les Ig G jour 30, mais étaient, en l´occurrence, beaucoup plus forte chez des animaux traités avec l´interféron (seul ou en association). La capacité de neutralisation atteint des pourcentages élevés à des dilutions importantes après une semaine de traitement. De plus, la réponse immunologique contre le vecteur dépend de la voie d´ administration: la quantité d’anticorps neutralisants est nettement réduite après injection intratumorale, contrairement à une inoculation postopératoire dans les tissus autour de la tumeur. La défense contre le transgène peut être soupçonnée d´être diminuée en parallèle. En tout, les méthodes décrites sont adéquates pour garantir une surveillance immunologique des patients vétérinaires lors d´une thérapie génique adénovirale et pour témoigner les réactions au vecteur viral appliqué et au transgène exprimé.

Plasminogen wird durch verschiedene Proteasen proteolytisch gespalten. Zu den Enzymen, von denen bekannt ist, daß sie Plasminogen an definierten Peptidbindungen prozessieren können, gehören Elastase aus polymorphnukleären Neutrophilen (PMN) und Metallo-Elastase aus Makrophagen. Ein Spaltprodukt ist Miniplasminogen, das die proteolytische Domäne und den Kringel 5 umfaßt. Die Spaltstelle bei Val441 ist charakterisiert und ein Sandwich-ELISA gegen die Neodeterminante ist etabliert. Das dabei entstehende Counterpart besteht aus den Kringeln1-4. Es wird Angiostatin genannt, weil es hemmend auf die Neubildung von Gefäßen wirkt. Im Rahmen dieser Arbeit sollte untersucht werden, ob Miniplasminogen unter pathologischen Bedingungen wie Sepsis, Peritonitis und Tumorerkrankungen auftritt, um daraus einerseits den Einfluß der Elastase abzuleiten und Einblicke in die pathophysiologischen Abläufe bei Entzündung und Tu-morgeschehen unter Betrachtung der Plasminogenspaltprodukte in Korre-lation zu anderen Parametern zu gewinnen. In Vorversuchen konnte gezeigt werden, daß unter dem Einfluß von aktivierten polymorph-nukleären Neutrophilen Miniplasminogen aus Plas-minogen generiert wird. Von den potentiell an der Proteolyse beteiligten En-zymen konnte in vitro nur bei dem Einsatz von PMN-Elastase Mini-plasminogen nachgewiesen werden, nicht jedoch von MMP-2, -8 und -9. In Untersuchungen zur Stabilität von Miniplasminogen unter verschiedenen Blutabnahmebedingungen waren Citrat-Proben den anderen Systemen (EDTAPlasma, Serum) überlegen. In Proben von gesunden Probanden konnte in keinem Fall Miniplasminogen über der Nachweisgrenze des ELISAs gemessen werden.Es wurden Proben aus klinischen Studien zu Sepsis, Peritonitis und Mammakarzinom ausgewählt, die auf Grund einer hohen PMN-Elastase-Konzentration ein Entstehen von Miniplasminogen erwarten lassen konnten. Ein Ausscheiden von Miniplasminogen über die Niere konnte durch Messungen im Urin ausgeschlossen werden. In Citratplasma-Proben von Peritonitispatienten war kein Miniplasminogen nachweisbar, in Peritonitisexsudaten desselben Patientenkollektivs waren Miniplasminogenwerte bis 90 ng/ml meßbar. Es zeigte sich allerdings keine Korrelation zu anderen Parametern (Elastase-Konzentration, Plasminogenkonzentration). Signifikante Unterschiede der Miniplasminogenkonzentrationen konnten zwischen den Mittelwerten der Proben der Patientengruppe, bei der therapeutisch Fresh-Frozen-Plasma intraabdominell appliziert wurde, und der Kontrollgruppe, sowie zwischen den Gruppen mit und ohne Tumorerkrankung nachgewiesen werden. Bei der Evaluierung von Serumproben aus einem Mammakarzinom-Kollektiv wurden Werte bis 52 ng/ml gemessen. Eine Korrelation mit anderen Parametern oder signifikante Unterschiede in den verschiedenen Subgruppen konnten auch hier nicht gezeigt werden. Ein Zusammenhang zwischen der proteolytischen Kapazität in den Exsudaten und der MPlg-Entstehung ließ sich nicht zweifelsfrei beweisen. MPlg ist daher – im Gegensatz zu dem Elastase-spezifischen Spaltprodukt des Fibrinogens (FEP) (Gippner-Steppert, 1991) - als ein spezifisches Spaltprodukt des Plg nicht für den indirekten Nachweis der proteolytischen Aktivität der PMNElastase geeignet. Erfolgversprechend könnten ggf. immunhistochemische Untersuchungen von Tumormaterial in Hinblick auf das lokale Entstehen von Miniplasminogen sein.

Samuel predikar om att växa i uppenbarelse bland annat utifrån Elisas liv.