Indwelling Pleural Catheters

Indwelling pleural catheters (IPC) are now being considered worldwide for patients with recurrent pleural effusions. It is commonly used for patients with malignant pleural effusions (MPE) and can be performed as outpatient based day care procedure. In malignant pleural effusions, indwelling catheters are particularly useful in patients with trapped lung or failed pleurodesis. Patients and care givers are advised to drain at least 3 times a week or in presence of symptoms i.e. dyspnoea. Normal drainage timing may lasts for 15–20 min which subsequently improves their symptoms and quality of life. Complications which are directly related to IPC insertion are extremely rare. IPC’s are being recently used even for benign effusions in case hepatic hydrothorax and in patients with CKD related pleural effusions. Removal of IPC is often not required in most of the patients. It can be performed safely as a day care procedure with consistently lower rates of complications, reduced inpatient stay. They are relatively easy to insert, manage and remove, and provide the ability to empower patients in both the decisions regarding their treatment and the management of their disease itself

Nitration of the Egg-Allergen Ovalbumin Enhances Protein Allergenicity but Reduces the Risk for Oral Sensitization in a Murine Model of Food Allergy

Nitration of proteins on tyrosine residues, which can occur due to polluted air under "summer smog" conditions, has been shown to increase the allergic potential of allergens. Since nitration of tyrosine residues is also observed during inflammatory responses, this modification could directly influence protein immunogenicity and might therefore contribute to food allergy induction. In the current study we have analyzed the impact of protein nitration on sensitization via the oral route.BALB/c mice were immunized intragastrically by feeding untreated ovalbumin (OVA), sham-nitrated ovalbumin (snOVA) or nitrated ovalbumin (nOVA) with or without concomitant acid-suppression. To analyze the impact of the sensitization route, the allergens were also injected intraperitoneally. Animals being fed OVA or snOVA under acid-suppressive medication developed significantly elevated levels of IgE, and increased titers of specific IgG1 and IgG2a antibodies. Interestingly, oral immunizations of nOVA under anti-acid treatment did not result in IgG and IgE formation. In contrast, intraperitoneal immunization induced high levels of OVA specific IgE, which were significantly increased in the group that received nOVA by injection. Furthermore, nOVA triggered significantly enhanced mediator release from RBL cells passively sensitized with sera from allergic mice. Gastric digestion experiments demonstrated protein nitration to interfere with protein stability as nOVA was easily degraded, whereas OVA and snOVA remained stable up to 120 min. Additionally, HPLC-chip-MS/MS analysis showed that one tyrosine residue (Y(107)) being very efficiently nitrated is part of an ovalbumin epitope recognized exclusively after oral sensitization.These data indicated that despite the enhanced triggering capacity in existing allergy, nitration of OVA may be associated with a reduced de novo sensitizing capability via the oral route due to enhanced protein digestibility and/or changes in antibody epitopes

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2–4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2,3,4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Cell communications decisive for the type of immune responses to dietary antigens

Allergische Reaktionen auf Lebensmittel, bis hin zum anaphylaktischen Schock, gehören zu den am häufigsten behandelten klinischen Notfällen. Wie in Kapitel 1 dieser Doktorarbeit beschrieben, sind etwa 3-4 % der Erwachsenen und 5% aller Kinder in entwickelten Ländern von Lebensmittelallergien betroffen. Zahlreiche Studien der letzten Jahren haben zur Erforschung der Faktoren, die für die Enstehung von Allergien mitverantwortlich sind, beigetragen. Das zweite Kapitel beschreibt unsere Studien über den Einfluss der Magensäurehemmer Sukralfat und Protonenpumpenhemmer kombiniert mit bakteriellem Endotoxin auf die allergische Sensibilisierung in einem Mausmodell basierend auf dem Milchallergen Casein. Dabei fütterten wir BALB/c Mäuse mit Casein kombiniert mit Magensäurehemmer(n) und/oder Endotoxin und führten anschließend Orale Provokations- und Hauttests durch. Im Rahmen dieser Untersuchungen konnten wir zeigen dass Magensäurehemmer mit Endotoxin interagieren, dabei Casein-spezifische IgG1 und IgE Antikörper induzieren und so zu allergischer Sensibilisierung beitragen können. Außerdem entwickelten Mäuse, die mit Magensäurehemmern behandelt wurden, die stärksten allergischen Reaktionen in Form von Diarrhoe, Hypothermie, und Typ-1 Hautreaktionen. Mastzellen und Basophile spielen bekanntermaßen wichtige Rollen in Allergen- induzierten anaphylaktischen Reaktionen. Das dritte Kapitel dieser Arbeit handelt von unseren Untersuchen zu den Beiträgen von Neurtrophilen, Thrombozyten, und Erythrozyten zum anaphylaktischen Schock in einem expermintellen Mausmodell für Allergen-induzierte Anaphylaxie. Dabei wurden Mäuse über die intra-peritoneale Route sensiblisiert. Verabreichung des Allergens verursachte eine anaphylaktische Reaktion. Anschließende Blutanalysen zeigten eine verringerte Zahl von Neutrophilen, Thrombozyten und Erythrozyten im Blut. Der Rückgang von Fibrinogen im Blut deutet darauf hin, dass der Rückgang der Thrombozytenzahl mit deren Aktivierung einher geht. Immunohistochemische Analysen von Milz, Lunge, Leber, Herz, und Niere zeigte eine Einwanderung von Gr-1+ Neutrophilen im pulmonären Insterstitium der Lunge. Die Ergebnisse dieser Studie deuten an, dass abgesehen von Mastzellen und Basophilen, auch Neutrophile und Thrombozyten eine wichtige Rolle in anaphylaktischen Reaktion spielen. Trotz diverser potentieller Nebenwirkungen ist spezifische Immuntherapie (SIT) momentan die wichtigste Therapiemöglichket für die Behandlung von Allergien. Im vierten Kapitel dieser Doktorarbeit werden unsere Experimente für die Entwicklung einer hypoallergenen Variante des Erdnussallergens Ara h 2 beschrieben. Dafür haben wir eine stabil-denaturierte Variante von Ara h 2 mittels chemischer Reduktion und anschließender Alkylierung hergestellt. Nach struktureller Analyse des modifizierten Allergens durch Zirkulardichroismus Analyse untersuchten wir außerdem dessen Potenzial zur Bindung von Serum IgE-Antikörpern mittels ELISA, zur Induktion von Mastzell Degranulation in vitro und zur Induktion von T-Zell Proliferation in vitro. Weiters verglichen wir das anaphylaktische Potential von denaturiertem Ara h 2 mit dem von unbehandeltem Ara h 2 in einem Mausmodell. Diese Experimente zeigten, dass denaturiertes Ara h 2 - verglichen mit dem unbehandelten Allergen - in vitro eine reduzierte IgE-Bindung aufwiest und geringere Mastzell Degranulation verursacht. Gleichzeitig stimulierte denaturiertes Ara h 2 in humanen T-Zellen die Produktion vergleichbarer Mengen von IL-4, IL-13 und Interferon-gamma wie unbehandeltes Ara h 2. Diese Ergebnisse lassen den Schluss zu, dass denaturiertes Ara h 2 ein vielversprechender Kandidat für die Immuntherapie von Erdnussallergie sein könnte. Alles in allem liefert die vorliegende Doktorarbeit neue Erkenntnisse zu Entstehung, Ablauf und Therapie von Nahrungsmittelallergien.Food allergy and anaphylaxis is one of the leading cause of anaphylactic episodes treated at the emergency clinics. As outlined in chapter 1, it affects 3-4 % adult population and 5% of the young children in westernized countries. From the last decade numerous studies have contributed to delineate the mechanisms leading to pathogenesis of allergic diseases. In chapter 2 of this PhD thesis, we studied how the different anti-acid medications, sucralfate and proton pump inhibitor together endotoxin influences the sensitization in BALB/c mice. We took milk protein, casein as our model allergen. Mice were fed casein with and without endotoxin together with either sucralfate or proton pump inhibitor and subjected to oral provocation tests and type I skin hypersensitivity. From these studies, we showed anti-acid/anti-ulcer medication interact with endotoxin to induce sensitization, by showing casein-specific IgG1, IgE antibodies. Furthermore, in oral provocation tests, anti-acid/anti-ulcer prompted the highest allergic response, by inducing either diarrheal or drop of body temperature or type I skin hypersensitivity. Mast cells and basophils have been repeatedly studied for their role in allergen- induced anaphylaxis. In chapter 3, however, we have investigated the yet less unaddressed cell types, neutrophils and platelets in an experimental mouse model of antigen-induced anaphylaxis. Mice were intraperitoneally sensitized and showed anaphylactic symptoms upon allergen challenge. Further, total blood counts revealed that upon specific allergen challenge, neutrophils, platelets and red blood cells (RBC) count dropped in total blood. Adding to this, we showed reduction of fibrinogen levels in a similar set up of anaphylaxis, showing that platelet activation correlate with the platelet reduction. We next investigated different organs, spleen, lung, liver, heart and kidney by immunohistochemistry. We observed influx of Gr-1+ neutrophils in the pulmonary interstitium. Therefore, from our data we proposed that, in addition to mast cells and basophils, neutrophils and platelets might play a role in excerbating the effects of anaphylaxis. Specific immunotherapy (SIT) is a gold standard therapy to treat allergic patients. Nevertheless, it has been shown that SIT has adverse effects. Therefore, in chapter 4 of this PhD thesis, we attempted to design a hypoallergenic variant, taking peanut allergen, Ara h 2 as our model allergen. Unfolded variant of Ara h 2 was prepared by reduction/alkylation and subjected to structural analysis by circular dichroism (CD). Unfolded Ara h 2 was studied for IgE binding capacity by ELISA, in vitro mast cell degranulation assay, T-cell proliferative response in vitro. Furthermore, anaphylactic capacity of the unfolded molecule was studied in a mouse model and splenocytes were studied for metabolic capacity and proliferative potential. In addition, human T cell proliferative capacity was assayed in vitro with native and unfolded Ara h 2. Unfolded Ara h 2 was shown to have reduced IgE binding capacity, reduced mast cell degranulation in vitro and a comparable response in human T cell proliferative capacity with similar levels of IL-4, IL-13 and IFN-[gamma]. Immunotherapy with unfolded Ara h 2 might be a safer strategy with less or no adverse effects. Together, the results in this thesis provide novel insights on the safety of food allergens in terms of natural exposure and during allergen immunotherapy.submitted by Durga KrishnamurthyAbweichender Titel laut Übersetzung der Verfasserin/des VerfassersZsfassung in dt. Sprachehttp://onlinelibrary.wiley.com/doi/10.1111/j.1365-2222.2012.04012.x/pdf ; http://onlinelibrary.wiley.com/doi/10.1111/cea.12031/pdfWien, Med. Univ., Diss., 2013OeBB(VLID)171502

Anaphylaxis Imaging: Non-Invasive Measurement of Surface Body Temperature and Physical Activity in Small Animals

<div><p>In highly sensitized patients, the encounter with a specific allergen from food, insect stings or medications may rapidly induce systemic anaphylaxis with potentially lethal symptoms. Countless animal models of anaphylaxis, most often in BALB/c mice, were established to understand the pathophysiology and to prove the safety of different treatments. The most common symptoms during anaphylactic shock are drop of body temperature and reduced physical activity. To refine, improve and objectify the currently applied manual monitoring methods, we developed an imaging method for the automated, non-invasive measurement of the whole-body surface temperature and, at the same time, of the horizontal and vertical movement activity of small animals. We tested the anaphylaxis imaging in three <i>in vivo</i> allergy mouse models for i) milk allergy, ii) peanut allergy and iii) egg allergy. These proof-of-principle experiments suggest that the imaging technology represents a reliable non-invasive method for the objective monitoring of small animals during anaphylaxis over time. We propose that the method will be useful for monitoring diseases associated with both, changes in body temperature and in physical behaviour.</p></div

Surface body temperature imaging in the milk allergy model.

<p>Mice M1-M3 were sham-treated (PBS), M4-M6 were specifically sensitized with aluminium-absorbed milk allergen casein (alum-casein), all mice were i.v. challenged with casein and monitored in the imaging cage over 20 minutes, with an image frequency of 4 frames/sec. (A) Temperature curves of individual mice; y-axis represents temperature in °C, x-axis: number of frames during 20 min. (B) The mean results of drop of the body surface temperatures over time of each mouse group were compared and differed significantly (** p<0.01).</p