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Gerrit Kann,1,* Junaid Owasil,1,* Karina Kuczka,2 Annette Haberl,1 Timo Wolf,1 Pavel Khaykin,1 Sebastian Harder,2 Christoph Stephan,1 Nils von Hentig1 1HIVCENTER, Medical HIV Treatment and Research Unit, Johann Wolfgang Goethe University Frankfurt, Frankfurt am Main, Germany; 2Pharmazentrum Frankfurt, Institute of Clinical Pharmacology, Johann Wolfgang Goethe University Frankfurt, Frankfurt am Main, Germany*These authors contributed equally to this workCorrespondence: Nils von HentigHIVCENTER, Medical HIV Treatment and Research Unit, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, Frankfurt am Main, 60590, GermanyTel +49-69-63017680Fax +49-69-630183442Email Hentig@em.uni-frankfurt.deBackground: In the past, protease inhibitors (PIs) and the reverse transcriptase inhibitor abacavir were identified increasing the risk for thromboembolic complications and cardiovascular events (CVE) of HIV infected patients taking a combination antiretroviral therapy (cART). Results of the previous HIV-PLA I-study lead to the assumption that platelet activation could play a substantial role in increasing CVE risks.Methods: The open label, monocentric HIV-PLA II-study investigated HIV-1-infected, therapy-naïve adults (n=45) starting with cART, consisting either of boosted PI (atazanavir, n= 6, darunavir, n=11), NNRTI (efavirenz, n=14) or integrase inhibitor (raltegravir, n=14), each plus tenofovir/emtricitabine co-medication. Main exclusion criteria were tobacco smoking, the intake of NSAIDs or abacavir or past CVE. Platelet adhesive molecule p-selectin (CD62P) and FITC anti-human Integrin α-IIb/Integrin β-3 (CD41/CD61) antibody (PAC-1) binding, monocyte CD11b/monocyte-associated CD41 expression and the endogenous thrombin potential (ETP) were assessed ex vivo-in vitro at baseline, weeks 4, 12 and 24. Therapy regimens were blinded to the investigators for laboratory and statistical analyses.Results: CD11b and ETP showed no significant changes or differences between all study groups. In contrast, the mean + SD mean fluorescence units (MFI) of CD62P and PAC-1 increased significantly in patients taking PI, indicating an enhanced potential for thrombocyte activation and aggregation.Conclusion: CD62P expression, detecting the ɑ-platelet degranulation of pro-inflammatory and pro-thrombotic factors and adhesive proteins, and PAC-1 expression, representing a marker for conformation changes of the GIIb/IIIa receptor, increased significantly in patients taking HIV protease inhibitors. The findings of this study revealed a yet unknown pathway of platelet activation, possibly contributing to the increased risk for CVE under HIV protease inhibitor containing cART.Clinical Trial Registration No.: DRKS00000288.Keywords: HIV protease inhibitors, platelets, leucocytes, PAC-1, GIIb/IIIa-receptor

From April to June 2020, during a period of increased mortality of tits (Cyanistes caeruleus, Parus major, Periparus ater) in Germany, 84 tits were pathomorphologically and microbiologically investigated in governmental veterinary investigation offices in North Rhine-Westphalia using a broad diagnostic panel. Although Suttonella ornithocola was the most frequently detected bacterium (36/84), other pathogens, in particular Chlamydia psittaci (12/65), were also identified, sometimes as concurrent infection (3/36). In conclusion, S. ornithocola-infection seems to play an important role within the epidemic tit mortality in North Rhine-Westphalia, Germany in 2020. However, these cases may be combined with sporadic and/or endemic causes of mortality (e.g., other pathogens, trauma, or predation) for these species.

Yersinia pseudotuberculosis is an important pathogen for both humans and animals. It can infect livestock, as well as pets and wild animals. During recent years, a number of reports have described the isolation of Y. pseudotuberculosis from zoo animals, mainly birds and mammals, for which the infection was mostly lethal. Between 2005 and 2019, there were at least 17 cases of deceased mammals, belonging to five different species, which suffered from a Y. pseudotuberculosis infection at the Zoo Wuppertal, Germany. Since only scarce information exists on the properties of Y. pseudotuberculosis from zoo animals, we characterized eight isolates, covering all infected species, in detail. All isolates were members of biotype 1, but belonged to five serotypes, five sequence types (STs), and seven core-genome multilocus sequence types (cgMLSTs). Using pulsed-field gel electrophoresis (PFGE) analysis and whole-genome sequencing (WGS), the seven isolates could be discriminated from each other. They differed significantly regarding their virulence genes and mobile genetic elements. While the virulence plasmid pYV existed in all serotypes (five isolates), a complete high-pathogenicity island (HPI) was detected only in the serotypes O:1a, O:1b, and O:13 (four isolates), but not in O:2a and O:2b. Similarly, the content of other plasmids and prophages varied greatly between the isolates. The data demonstrate that the deceased mammals were infected by seven individual isolates and not by a single type predominating in the zoo animals.

Glycoprotein VI (GPVI) is the major platelet receptor for collagen-mediated platelet adhesion and activation. SAR264565 is an anti-GPVI-Fab, binds to GPVI with high affinity, and blocks GPVI function in human platelets in vitro. The effect of SAR26456 on platelet responsiveness in the blood of 21 healthy male subjects was investigated using Sakariassen’s ex vivo thrombogenesis perfusion chamber model on a collagen-coated surface under conditions mimicking arterial flow. Ex vivo effects of SAR264565 (10 and 100 μg/mL) were investigated before administration of aspirin or clopidogrel to study subjects (baseline), after aspirin (2× 300 mg) administration alone, and after combined aspirin (2× 300 mg)/clopidogrel (600 mg) administration. Additional ex vivo and in vitro platelet tests were also performed. Addition of SAR264565 to the perfusion chamber dose-dependently reduced platelet and fibrin deposition, reaching statistical significance at 100 μg/mL (415 ± 67 compared to 137 ± 36 platelets/cm2, [p

Glycoprotein VI (GPVI) is the major platelet receptor for collagen-mediated platelet adhesion and activation. SAR264565 is an anti-GPVI-Fab, binds to GPVI with high affinity, and blocks GPVI function in human platelets in vitro. The effect of SAR26456 on platelet responsiveness in the blood of 21 healthy male subjects was investigated using Sakariassen's ex vivo thrombogenesis perfusion chamber model on a collagen-coated surface under conditions mimicking arterial flow. Ex vivo effects of SAR264565 (10 and 100 mu g/mL) were investigated before administration of aspirin or clopidogrel to study subjects (baseline), after aspirin (2x 300 mg) administration alone, and after combined aspirin (2x 300 mg)/clopidogrel (600 mg) administration. Additional ex vivo and in vitro platelet tests were also performed. Addition of SAR264565 to the perfusion chamber dose-dependently reduced platelet and fibrin deposition, reaching statistical significance at 100 mu g/mL (415 +/- A 67 compared to 137 +/- A 36 platelets/cm(2), [p < 0.01] and fibrin 0.095 +/- A 0.014 compared to 0.032 +/- A 0.008 mu g/cm(2), [p < 0.001]). Aspirin administration caused an additive and dose-dependent reduction of SAR264565-induced platelet and fibrin deposition. Combined aspirin/clopidogrel administration did not lead to additional SAR264565-induced inhibition of platelet or fibrin deposition. GPVI antagonism by the anti-GPVI-Fab fragment SAR264565 dose-dependently inhibits platelet adhesion and fibrin formation on a collagen surface under arterial shear. Additive inhibition is observed after prior aspirin administration with no further amplification on top of a combination of aspirin with clopidogrel. Ex vivo antiplatelet tests confirmed a selective inhibiting effect of SAR264565 on collagen-induced platelet activation.

SummaryBackground Pruritus is a common symptom associated with cholestatic liver diseases. To date only small single centre case series have suggested efficacy of nasobiliary drainage in relieving cholestatic pruritus. Aim To perform a multicentre study to evaluate the safety and efficacy of nasobiliary drainage in cholestatic pruritus. Methods This was a retrospective study of all patients treated with nasobiliary drainage for refractory cholestatic pruritus between 2006 and 2015 at five European centres. Pruritus was quantified using a visual analogue scale (VAS) and liver enzymes, serum bilirubin and total serum bile salts (TBS) were measured before (pre-NBD) and after nasobiliary drainage (post-NBD). We analysed the duration of treatment response and associated complications. Results In total, 27 patients (59% females) underwent 29 nasobiliary drainage procedures. The median duration of NBD was 7 days. NBD decreased pruritus in 89.6% of cases (VAS from 10.0 to 0.3, P < 0.0001). The median percentage decline in pruritus VAS was 94% and 33% of patients were free of pruritus within 24 h of starting drainage. The duration of treatment response was independent of duration of drainage (P = 0.12) and bile output. Significant improvements were seen in the median levels of serum alkaline phosphatase (P = 0.001) and serum bilirubin (P = 0.03) but not in serum TBS (P = 0.07). Mild post-endoscopic retrograde cholangiopancreatography pancreatitis (31%) was the most frequent complication. Conclusions Nasobiliary drainage is effective in relieving cholestatic pruritus in most patients and has favourable effect on biomarkers of cholestasis. Nasobiliary drainage may be associated with high risk of adverse events, especially pancreatitis. Prospective studies are needed to confirm our findings.

A natural reassortant influenza A virus consisting of seven genome segments from pandemic (H1N1) 2009 virus and a neuraminidase segment from a Eurasian porcine H1N1 influenza A virus was detected in a pig herd in Germany. The obvious reassortment compatibility between the pandemic (H1N1) 2009 and H1N1 viruses of porcine origin raises concern as to whether swine may become a reservoir for further reassortants of pandemic (H1N1) 2009 viruses with unknown implications for human health and swine production.

We investigated anti-FXa- and anti-FIIa-activity, thrombin generation (ETP), tissue factor pathway inhibitor (TFPI) - and D-dimer in patients exhibiting high bleeding risk in early neurological rehabilitation over 2 months in an observational study. Blood of 64 patients under LMWH administration due to therapeutic (cohort 1 [tinzaparin 90 IE/kg BID, N = 18] and 2 [enoxaparin 100 IE/kg BID; N = 15]) or prophylactic (cohort 3 [tinzaparin 4500 IE; N = 16] and 4 [enoxaparin 4000 IE; N = 15]) indication was drawn before and 4h after injection on day 7 (V1) and 2 months (follow up [V2]). Although the dose in cohort 1 and 2 was similar (median 7000 IE BID), a-FXa-activity was significantly larger under enoxaparin than under tinzaparin (e.g. median at V2: 0.70 IU/ml vs. 0.33 IU/ml). Also, prophylactic enoxaparin exhibited larger a-FXa-activity than tinzaparin (e.g. median at V2: 0.37 IU/ml vs. 0.22 IU/ml). The a-FXa/a-FIIa-ratio in plasma samples at 4h p.a. was about 4 (tinzaparin) and 8 (enoxaparin), respectively. No differences were seen for TFPI and ETP between cohort 1 and 2 or between cohort 3 and 4. D-dimer levels decreased significantly between V1 (e.g. cohort 4 median 1940 ng/ml) and V2 (median 652 ng/ml). Minimal bleeding events occurred in 6 patients (2 under tinzaparin, 4 under enoxaparin) and were associated with significantly higher anti-FXa-activity. In conclusion, although marked differences between tinzaparin and enoxaparin based on anti-FXa-activity were seen, markers of in vivo biological activity such as TFPI and D-dimer were not different. Furthermore, BID tinzaparin is a feasible option for therapeutic anticoagulation in patients with high bleeding risk.

Low-molecular-weight heparins (LMWHs) differ considerably in their influence on clotting tests and release of tissue factor pathway inhibitor (TFPI). Biosimilarity therefore becomes an issue when generic forms of LMWHs are developed. So far, no bioequivalence study with a generic LMWH has been reported. A generic enoxaparin (test) was compared with the originator (reference) in 20 volunteers after single-dose subcutaneous administration (40 mg enoxaparin sodium, 4000 IU/mL anti-factor Xa (anti-FXa; activity). Target variables were anti-FXa and anti-FIIa activity, activated partial thromboplastin time (aPTT), prothrombinase-induced clotting time (PiCT), and TFPI over 24 hours. The statistical evaluation of the anti-FXa activity profile demonstrated bioequivalence of test and reference with confidence intervals of area under the plasma concentration-time curve (AUC0-tlast) (93%-99%) and Amax (88%-95%). Confidence intervals of AUC(0-tlast) (89%-102%) and Amax (90%-103%) of anti-FIIa activity also fulfill bioequivalence criteria. The 90% confidence interval for the maximum concentration of TFPI ranged from 90% to 113%. The claim of similarity was also supported by aPTT and PiCT profiles. Bioequivalence with the originator enoxaparin could be demonstrated by ex vivo inhibition of FXa and FIIa activity, by coagulation tests (aPTT and PiCT), and by in vivo release of TFPI. Whether such data also prove biosimilarity of the generic enoxaparin needs to be determined.

Background. Self-handicapping is an attribution-related process whereby individuals create performance impediments/excuses to protect self-worth in socially evaluative environments. Thus, the prevailing motivational climate would appear to be an important factor when attempting to understand the situational self-handicapping process within school physical education. Aims. Drawing from achievement goal theory, the study examined the effect of experimentally induced conditions (viz. task vs. ego) on situational self-handicapping. Sample. Seventy British secondary school students (36 females and 34 males; M age = 11.98; SD=0.31). Method. Participants were randomly assigned to partake in a running endurance task in either an ego-involving (20 male students and 16 female students) or a task-involving (14 male students and 20 female students) condition. Prior to completing the experimental task, participants were given the opportunity to claim situational self-handicaps. Data for goal orientations, subjective climate perceptions, perceived ability and perceived task importance were also obtained. Results. After determining the effectiveness of the experimental manipulation, results revealed participants in the ego-involving condition to report significantly more situational self-handicapping claims. Further, and after controlling for individual difference variables, the results of moderated hierarchical regression analysis revealed subjective perceptions of an ego-involving climate to be the main positive predictor of situational self-handicapping. Although a weaker contributor to the percentage of variance explained, task orientation emerged as a negative predictor of situational self-handicapping. Conclusions. The findings suggest that PE teachers would be prudent to minimize ego-involving situations should they wish to reduce situational self-handicapping.

Objectives To examine the influence of perceptions of the motivational climate, self-efficacy, and perceived importance on athletes' claimed situational self-handicaps in a competitive sport setting. Perceptions of a task involving climate and self-efficacy were expected to be negatively related and perceptions of an ego-involving climate to be positively related to situational self-handicapping. Design Given the centrality of perceived ability in the self-handicapping literature, basic tenets derived from achievement goal theory provided the rationale for the present cross-sectional field study. Methods Male 70 and 70 female elite collegiate golfers completed a questionnaire prior to participating in a prestigious national golf tournament ( M age=20.61; SD=1.52) assessing claimed situational self-handicaps in the week prior to the tournament, perceptions of the team motivational climate, perceived event importance, and self-efficacy. Results Preliminary analyses revealed no significant gender differences in regard to the perceived importance of the event or situational claimed self-handicaps. Participants who perceived the event to be of low personal importance reported significantly more claimed self-handicaps during the week prior to the tournament than high importance individuals. A hierarchical multiple regression analysis revealed perceptions of a task-involving motivational climate, self-efficacy, and perceived event importance to be negatively related to claimed self-handicaps. Conclusions The findings of the present study suggest that in addition to enhancing self-efficacy, coaches should increase the salience of task-involving cues in the athletic context to attenuate the situational claimed self-handicaps of elite collegiate athletes.

The RGD cyclic pentapetide, cilengitide, is a selective inhibitor of αvβ3 and αvβ5 integrins and was developed for antiangiogenic therapy. Since cilengitide interacts with platelet αIIbβ3 and platelets express αv integrins, the effect of cilengitide on platelet pro-coagulative response and adhesion is of interest. Flow-based adhesion assays were performed to evaluate platelet adhesion and rolling on von Willebrand factor (vWf), on fibrinogen and on human umbilical vein endothelial cells (HUVECs). Flow cytometry was used to detect platelet activation (PAC1) and secretion (CD62P) by cilengitide and light transmission aggregometry was used to detect cilengitide-dependent platelet aggregation. Cilengitide inhibited platelet adhesion to fibrinogen at concentrations above 250 µM [which is the Cmax in human studies] and adhesion to vWf and HUVECs at higher concentrations under physiologic flow conditions. Platelet aggregation was already impaired at cilengitide concentrations >10 µM. Activation of αIIbβ3 integrin was inhibited by 250 µM cilengitide, whereas platelet secretion was unaffected by cilengitide. No evidence of cilengitide-induced platelet activation was found at all tested concentrations (0.01-1500 µM). At higher concentrations, platelet activation was inhibited, predominantly due to αIIbβ3 inhibition.

A modified DNA test, based on the polymerase chain reaction, was developed for the monogenic recessive disease bovine leucocyte adhesion deficiency ( BLAD ). The test was improved by the selection of new primers which facilitated the interpretation of the results. An easily scorable banding pattern makes the test useful in cattle breeding schemes and for clinical diagnosis. A total of 2381 samples was analysed over a period of three years. The carrier rate among young bulls at artificial insemination ( AI ) stations decreased from 11·6 per cent in 1993 to 9·9 per cent in the first five months of 1995. Continuous screening of young bulls before entering AI is still recommended unless both parents are proven to be genetically free of BLAD . The carrier rate among clinically suspect animals was not increased, and carriers are therefore not expected to be immunodeficient. Despite all efforts to eradicate the disease, calves with BLAD were still observed in 1995.

The 1993 National Electrical Code (NEC) cycle contained a very controversial proposal-to change the area classification system for Class I hazardous (classified) locations from a two-division to a three-division system. In general, the "user" representatives on Code Making Panel 14 supported this change, but the other representatives did not. Similarly, in 1987, a number of Western Canadian users and consultants began to develop a real awareness of the International Electrotechnical Commission (IEC) System for classified locations. Since that time, this group of Canadians has spent significant time researching the IEC System. They have developed and implemented a plan to make the necessary changes to the Canadian electrical installation codes and standards to allow Canadian users access to IEC technology. The change to permit use of the international classification system in North America has been controversial. The authors' approach to this paper was to assume that both the North American and the IEC systems for classified locations have been proven to be safe. The paper disregards the unsubstantiated claims and counter-claims made by those on both sides of the issue. Clearly, there are differences between the two systems, and this paper explores these differences. The paper attempts to emphasize the differences between the two systems to allow the readers to draw their own conclusions as to which system is best for their applications. A comparison is made between a typical North American installation and a typical IEC installation to illustrate the impact of change to a three-division system.

Je mehr die Kinematographie in bestimmten Gebieten der Zoologie zum ständig gebrauchten Handwerkzeug des forschenden Wissenschaftlers wird, um so mehr ist es nötig, daß dieser über die grundsätzlichen Fragen unterrichtet ist, die mit dem wissenschaftlichen Gebrauch dieser keineswegs einfachen Technik zusammenhängen. Die Arbeit mit dem Film ist ja nicht nur ein schwieriges, sondern unter Umständen auch kostspieliges Forschungsverfahren, und es bedarf der gleichzeitigen Berücksichtigung einer Anzahl von Regeln, um mit erträglichem Aufwand für die wissenschaftliche Verwendung wirklich befriedigende Ergebnisse zu erzielen. In Analogie zu den früher in diesen Heften gegebenen Leitsätzen zur völkerkundlichen und volkskundlichen Filmdokumentation) sollen im folgenden die Richtlinien zusammengefaßt werden, die sich im Laufe jahrelanger Erfahrungen des Instituts für den Wissenschaftlichen Film und seiner Vorgänger herausgebildet haben und die als Empfehlungen des Instituts für die Filmarbeit auf dem Gebiet der Zoologie angesehen werden können.

The membrane-anchored CX3C chemokine fractalkine (FKN) is expressed on activated endothelium and is associated with the development of atherosclerosis. The potential of FKN in mediating platelet adhesion beyond platelet activation remains unexplored to date. A flow-based adhesion assay was used to study the adhesion of platelets to immobilized FKN under physiologic flow conditions. Platelet adhesion to von Willebrand factor (VWF) was increased in the presence of FKN at 600 inverse seconds. Additional platelet adhesion to FKN coimmobilized with VWF was dependent on the FKN receptor CX3CR1 and activation of glycoprotein (GP) IIb/IIIa. The number of platelets rolling on VWF was likewise enhanced in the presence of FKN. The enhancement of rolling on FKN and VWF was insensitive to anti-CX3CR1 antibody but was fully inhibited by neutralizing GPIbα function. The extracellular domain of GPIbα was covalently coupled to fluorescent microspheres, and microsphere binding was significantly higher in the presence of FKN. Platelet adhesion to activated endothelium in vitro and to intact human arteries was substantially increased in an FKN-dependent manner. These data demonstrate that endothelial expressed FKN activates platelets via its cognate receptor CX3CR1, whereas platelet adhesion is predominantly mediated by GPIbα and independent of CX3CR1.