Search

Lipid-based nanoparticles for RNA delivery (LNP-RNA) are revolutionizing the nanomedicine field, with one approved gene therapy formulation and two approved vaccines against COVID-19, as well as multiple ongoing clinical trials. As for other innovative nanopharmaceuticals (NPhs), the advancement of robust methods to assess their quality and safety profiles—in line with regulatory needs—is critical for facilitating their development and clinical translation. Asymmetric-flow field-flow fractionation coupled to multiple online optical detectors (MD-AF4) is considered a very versatile and robust approach for the physical characterisation of nanocarriers, and has been used successfully for measuring particle size, polydispersity and physical stability of lipid-based systems, including liposomes and solid lipid nanoparticles. However, the unique core structure of LNP-RNA, composed of ionizable lipids electrostatically complexed with RNA, and the relatively labile lipid-monolayer coating, is more prone to destabilization during focusing in MD-AF4 than previously characterised nanoparticles, resulting in particle aggregation and sample loss. Hence characterisation of LNP-RNA by MD-AF4 needs significant adaptation of the methods developed for liposomes. To improve the performance of MD-AF4 applied to LNP-RNA in a systematic and comprehensive manner, we have explored the use of the frit-inlet channel where, differently from the standard AF4 channel, the particles are relaxed hydrodynamically as they are injected. The absence of a focusing step minimizes contact between the particle and the membrane, reducing artefacts (e.g. sample loss, particle aggregation). Separation in a frit-inlet channel enables satisfactory reproducibility and acceptable sample recovery in the commercially available MD-AF4 instruments. In addition to slice-by-slice measurements of particle size, MD-AF4 also allows to determine particle concentration and the particle size distribution, demonstrating enhanced versatility beyond standard sizing measurements.

Hintergrund: Toxinbildende Clostridium-difficile-Stamme sind weltweit die haufigsten Erreger von Antibiotika-assoziierten Darmerkrankungen und nosokomialer Diarrho. In den letzten Jahren wurden vermehrt Clostridium-difficile-Infektionen (CDI) mit besonders schweren klinischen Verlaufen und damit einhergehender erhohter Letalitat beobachtet. Verlaufe mit multiplen Rezidiven werden insbesondere bei Vorliegen einer Immunsuppression beobachtet. Methoden: Ziel dieser retrospektiven kontrollierten Beobachtungsstudie war die Charakterisierung einer Immunsuppression als unabhangiger Risikofaktor fur CDI. Dazu wurden Symptome und klinische Merkmale von 105 Patienten mit Clostridium-difficile-Toxinnachweis im Stuhl (detektiert mittels EIA) sowie positivem Antigen-Test oder positiver Kultur, die sich zwischen 2006 und 2009 in stationarer Behandlung der Kliniken fur Innere Medizin I und IV des Universitatsklinikums Halle (Saale) befanden, erfasst und ausgewertet. 55 immunsupprimierte Patienten wurden 50 Patienten ohne Immunsuppression gegenubergestellt (Durchschnittsalter 59,3 Jahre ± 16,2 vs. 69,2 Jahre ± 15,0). Fur die multivariate statistische Analyse mittels binarer logistischer Regression wurde eine Kontrollgruppe von insgesamt 105 Patienten ohne CDI herangezogen. Diese Gruppe umfasste 62 Patienten ohne Immunsuppression und 43 immunsupprimierte Patienten (Durchschnittsalter 66,9 Jahre ± 12,4 vs. 56,0 Jahre ± 13,7). Ergebnisse: Bei immunsupprimierten Patienten mit CDI fand sich signifikant haufiger eine Kolonisation durch Clostridium difficile ohne klinisch fassbare Manifestation (22 vs. 2 %, p = 0,003), wahrend es vergleichbar viele moderate (46 vs. 52 %, p = 0,503), aber weniger schwere Krankheitsverlaufe (27 vs. 40 %, p = 0,167) gab. Rezidive wurden in der Gruppe der Immunsupprimierten haufiger beobachtet (14,5 vs. 6 %, p = 0,153). Die multivariate Analyse mittels logistischer Regression identifizierte das Vorliegen einer Immunsuppression als unabhangigen Risikofaktor fur das Erleiden einer CDI (OR = 2,75). Daruber hinaus waren vorangegangene Antibiotikatherapie (OR = 10,15) und PPI-Einnahme (OR = 2,93) unabhangige Risikofaktoren. Schlussfolgerung: Neben dem wichtigsten Risikofaktor Antibiotikatherapie ist somit auch eine Immunsuppression als unabhangiger Risikofaktor fur eine CDI zu betrachten. Eine immunsuppressive Behandlung erhoht das Kolonisations- und Infektionsrisiko fur toxinbildende Clostridium-difficile-Stamme und fuhrt zu einer erhohten Rezidivrate bei Patienten mit CDI.

Introduction: Low Carbohydrate High Protein diet represents a popular strategy to achieve weight loss. Objective: The aim of this study was to characterize effects of low carbohydrate, high protein diet (LCHP) on atherosclerotic plaque development in brachiocephalic artery (BCA) in $apoE/LDLR^{-/-}$ mice and to elucidate mechanisms of proatherogenic effects of LCHP diet. Materials and Methods: Atherosclerosis plaques in brachiocephalic artery (BCA) as well as in aortic roots, lipoprotein profile, inflammation biomarkers, expression of SREBP-1 in the liver as well as mortality were analyzed in Control diet (AIN-93G) or LCHP (Low Carbohydrate High Protein) diet fed mice. Results: Area of atherosclerotic plaques in aortic roots or BCA from LCHP diet fed mice was substantially increased as compared to mice fed control diet and was characterized by increased lipids and cholesterol contents (ORO staining, FT-IR analysis), increased macrophage infiltration (MOMA-2) and activity of MMPs (zymography). Pro-atherogenic phenotype of LCHP fed $apoE/LDLR^{-/-}$ mice was associated with increased plasma total cholesterol concentration, and in LDL and VLDL fractions, increased TG contents in VLDL, and a modest increase in plasma urea. LCHP diet increased SCD-1 index, activated SREBP-1 transcription factor in the liver and triggered acute phase response as evidence by an increased plasma concentration of haptoglobin, CRP or AGP. Finally, in long-term experiment survival of $apoE/LDLR^{-/-}$ mice fed LCHP diet was substantially reduced as compared to their counterparts fed control diet suggesting overall detrimental effects of LCHP diet on health. Conclusions: The pro-atherogenic effect of LCHP diet in $apoE/LDLR^{-/-}$ mice is associated with profound increase in LDL and VLDL cholesterol, VLDL triglicerides, liver SREBP-1 upregulation, and systemic inflammation.

The elution behavior of branched macromolecules during their separation by size exclusion chromatography (SEC) was studied. The elution behavior of branched polymers was investigated using samples of randomly branched polystyrene and star branched poly(benzyl methacrylate) of different levels of branching by means of a SEC chromatograph coupled with a multiangle light scattering detector. Abnormal SEC elution behavior was found to be typical for highly branched polymers. After a normal elution at small elution volumes the molar mass and root mean square radius of the eluting molecules increased with increasing elution volume. Several SEC experiments were carried out to find explanation for this effect and SEC separation was compared with the separation by thermal field flow fractionation. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1588–1594, 2001

Finiteness, own resurrection and the polities of differance Employing Derrida's notion of diffirance, the author approaches the conviction of life after death (-own resurrection -) as a conviction which has been typically marginalized and even disallowed by modernity. The politics of differance, which could be regarded as a postmodern strategy for rehabilitating those aspects of reality which were marginalized by modernity, is shown to have critical consequences for the linear (modern) understanding of finiteness and death. Rather than understand finiteness and own resurrection as being in linear relation, the author appeals to the postmodern persuasion of the siclical nature of reality: death is own resurrection. Death is a life giving gift: The Self dies so that Others may live - and the Self is resurrected when it lives on in the lives of Others. Thus parting company with the empirical understanding of history, death and resurrection, the author leaves us with the tadt question: is this not the way the resurrection of Christ is to be understood? He lives on in us, as we will live on in Others. Wie dood is, is dood, en wie dood is, sallewe - Georges Bataille

Toxigenic Clostridium difficile strains are known as the most common infectious cause of antibiotic-associated intestinal disease and nosocomial diarrhoea. The increased incidence of hypervirulent strains gives rise to worldwide concern. In particular, courses with multiple recurrences are observed in the presence of immunosuppression.In this retrospective controlled observational study we aimed to determine immunosuppression as an independent risk factor for symptomatic CDI and to identify characteristics and differences of immunocompromised patients with respect to disease severity, disease progression, intestinal manifestations, recurrence rates and other factors. We compared symptoms and clinical features of 55 immunosuppressed patients with confirmed CDI with those of 50 patients without immunosuppressive medication (mean age 59.3 years ± 16.2 vs. 69.2 years ± 15.0) who were treated at the Departments of Internal Medicine I and IV of the University Hospital Halle (Saale), Germany, between 2006 and 2009. Multivariate analysis using binary logistic regression was performed for a control group of 105 patients without CDI. In this group, there were 62 patients without evidence of immunosuppression and 43 immunosuppressed patients (mean age 66.9 years ± 12.4 vs. 56.0 years ± 13.7).The clinical courses of the two groups differed considerably. Immunosuppressed patients were significantly more frequently colonised with Clostridium difficile without clinically detectable manifestation or only mild clinical symptoms not requiring therapy (22 vs. 2 %, p = 0.003), while there were similar numbers of moderate (46 vs. 52 %, p = 0.503) but less severe CDI cases (27 vs. 40 %, p = 0.167). Relapses were observed more frequently in the group of immunosuppressed patients (15 vs. 6 %, p = 0.153). Multivariate analysis using logistic regression identified immunosuppression as an independent risk factor for CDI (OR = 2.75), in addition to prior antibiotic therapy (OR = 10.15) and PPI intake (OR = 2.93).We conclude that immunosuppression has to be regarded as an independent risk factor for CDI. Immunosuppressive treatment increases the risk of colonisation and infection with Clostridium difficile and leads to a higher relapse rate in patients with CDI.

The development of new “pre-MS” methods for protein isolation/separation from complicated biological samples is a fundamental requirement for successful, MS-based approaches to proteomics. Flow field-flow fractionation (F4) has been applied as pre-MS step for proteomics. Hollow-fiber F4 (HF5) is the innovative, microcolumn version of F4. HF5 employs a piece of hollow fiber as separation channel and current HF5 shows performance comparable to commercial F4. The reduced channel volume and flowrates, the low dilution of fractionated analytes, and the potentially disposable usage to exclude contaminations or carryover then make HF5 ideally suited as online or offline pre-MS separation step for the characterization of complex protein samples in native form. We have developed different applications based on HF5 in combination with MS or LC-MS for the fractionation, purification and characterization of native proteins, and for proteomic analysis of complex protein samples.

A novel series of2-benzothiophenealkylpiperazine derivatives 11(a-d) with potential affinity at 5-HT 1A serotonin receptors have been synthesized via solvent-free, microwave-promoted Michael addition of benzo[b]thiophene piperazine derivatives 6(a-c) to substituted benzo[b]thiophen-2-yl propenones 10(b,c).

In families with one of the Mendelian hereditary polyposes, one can predict the proportion of patients at risk and thus obtain a denominator against which colonoscopy, barium enema, and fecal occult blood can be validated. Colonoscopy is more sensitive than barium enema. There were 42 positive colonoscopies, 12 positive barium enemas, both being positive in 10 of these. There were 141 negative enemas, 133 negative colonscopies, and 118 negative for both. Occult blood was positive in 30% of patients with polyposis, five to seven times more frequently than in those without evident polyposis. Colonoscopy detected polyposis in 30% of the progeny of affected people. The shortfall, compared with the 50% expected under the Mendelian hypothesis, is readily explained by removal of affected cases from the study by surgery or death from cancer.

Post-infectious sequelea such as Guillain Barré syndrome (GBS), reactive arthritis (RA), and inflammatory bowel disease (IBD) may arise as a consequence of acute Campylobacter-enteritis (AE). However, reliable seroprevalence data of Campylobacter-associated sequelae has not been established. The objectives of this study were, first, to identify the most specific and sensitive test antigen in an optimized ELISA assay for diagnosing a previous Campylobacter-infection and, second, to compare the prevalence of anti- Campylobacter antibodies in cohorts of healthy blood donors (BD), AE, GBS, RA, and IBD patients with antibodies against known GBS, RA and IBD triggering pathogens. Optimized ELISAs of single and combined Campylobacter-proteins OMP18 and P39 as antigens were prepared and sera from AE, GBS, RA and IBD patients and BD were tested for Campylobcter-specific IgA and IgG antibodies. The results were compared with MIKROGEN™-recomLine Campylobacter IgA/IgG and whole cell lysate-immunoblot. Antibodies specific for Helicobacter pylori, Mycoplasma pneumoniae, Yersinia enterocolitica, and Borrelia afzelii were tested with commercial immunoblots. ROC plot analysis revealed AUC maxima in the combination of OMP18 and P39 for IgA and in the P39-antigen for IgG. As a result, 34–49 % GBS cases, 44–62 % RA cases and 23–40 % IBD cases were associated with Campylobacter-infection. These data show that Campylobcater-seropositivity in these patient groups is significantly higher than other triggering pathogens suggesting that it plays an important role in development of GBS and RA, and supports the hypothesis that recurrent acute campylobacteriosis triggers IBD. Introduction peerReviewed

Die Buchhaltung ist nicht allein fur den Kaufmann, sondern, wie schon oft hervorgehoben, fur jedes privatwirtschaftliche Unternehmen geeignet. Gibt sie doch in groser Einfachheit dem Unternehmer das Mittel an die Hand, seine Unternehmung im ganzen Umfange von allen anderen ihm fremd gegenuberstehenden Unternehmen genau abzugrenzen, so das Mein und Dein jederzeit streng voneinander geschieden werden konnen, und innerhalb der Gemarkung seines Besitzstandes eine zuverlassige Kontrolle uber die durch den Geschaftsbetrieb hervorgerufenen Formund Wertveranderungen in allen Teilen seines Vermogens herbeizufuhren. Um dieser Vorzuge willen hat sich Vor allem die systematische Buchhaltung den Eingang in alle groseren Geschaftsbetriebe erzwungen und erobert sich taglich neue Gebiete. Denn einmal wird es jeder gewissenhafte Unternehmer als seine Pflicht erachten, sich selbst uber den Stand und Erfolg seines Geschaftsbetriebes Rechenschaft zu geben. Dazu kommt aber noch ein zweites, die Verantwortlichkeit gegenuber allen denjenigen Personen, mit welchen er in Geschaftsverbindung tritt.

Diesem Teile des Werkes fallt die Aufgabe zu, die wirtschaftlichen Vorgange, welche die Buchhaltung zu bearbeiten hat, auf ihre einfachsten Grundformen zuruckzufuhren, um sie zunachst mit Hilfe mathematischer Gleichungen darzustellen, diese hierauf in die buchhalterische Kontenform uberzufuhren und auf dieser Grundlage die Gesetze und Regeln der Buchhaltung abzuleiten. Dabei bedienen wir uns der knappsten Thesenform und fugen nur die fur das Verstandnis notwendigsten Erklarungen bei.

Nachdem wir die Buchhaltung vom theoretisch-mathematischen Standpunkte aus betrachtet haben, erwachst uns die Aufgabe, die praktische Anwendung im Wirtschaftsleben zu besprechen, um zu zeigen einmal, in welchem Zusammenhang die Buchhaltung zu dem Wirtschaftsbetrieb steht, sodann wie das System allen verschiedenen Wirtschafts- und Unternehmungsformen angepast werden kann. Zu diesem Zwecke schicken wir eine Definition des Wirtschaftsbetriebes voraus.