Online-Informationssystem für die Phytotherapie bei Tieren

Die Phytotherapie gewinnt auch in der Veterinärmedizin zunehmend an Beliebtheit. Um das therapeutische Potenzial der Arzneipflanzen auszuschöpfen und deren sicheren Einsatz zu gewährleisten, bedarf es aber besonderer Kenntnisse der wirksamen Pflanzenteile beziehungsweise Zubereitungen. Unsachgemässe Anwendungen und Überdosierungen von Arzneipflanzen können toxisch sein. Deshalb haben wir mit www.phytoarznei.ch eine Entscheidungshilfe geschaffen, die online einen raschen Zugriff auf das aktuelle Wissen zu Arzneipflanzen und deren Gebrauch bei Tieren erlaubt. Dieses ­Informationssystem basiert auf der verfügbaren Fach­literatur, die nach kritischer Auswertung in eine strukturierte Datenbank eingebracht wurde. Für jede Arz­neipflanze beziehungsweise pflanzliche Droge sind therapeutische Indikationen, Anwendungsarten, organoleptische Eigenschaften, Inhaltsstoffe, pharmakologische Wirkungen, Dosierungen, Behandlungsdauer, Toxizität, gesetzliche Vorschriften bei Nutztieren sowie Dopingrelevanz aufgeführt. Zwei Suchprogramme gewährleisten einen benutzerfreundlichen Zugang, entweder durch die Eingabe des Pflanzennamens, des Drogennamens bzw. der Inhaltsstoffe der Pflanze in einem Suchfeld oder durch die Auswahl der gewünschten Tierspezies sowie therapeutischen Anwendung aus entsprechenden Dropdown-Listen. Diese Datenbank zu Arzneipflanzen ist mit der Giftpflanzendatenbank der Universität Zürich und, falls entsprechende Fertigpräparate vorliegen, mit dem Schweizerischen Tierarzneimittelkompendium sowie einem Verzeichnis von Veterinärprodukten verknüpft

Identification of regenerating island-derived protein 3E in dogs

Regenerating islet-derived protein (REG) 1A (aka pancreatic stone protein) and REG3A (aka pancreatitis-associated protein) are upregulated in humans with sepsis, pancreatitis, and gastrointestinal diseases, but little is known about this protein family in dogs. Our aim was to identify REG1 and REG3 family members in dogs. REG-family genes were computationally annotated in the canine genome and proteome, with verification of gene expression using publicly available RNA-seq data. The presence of the protein in canine pancreatic tissue and plasma was investigated with Western blot and immunohistochemistry, using anti-human REG1A and REG3A antibodies. Protein identity was confirmed with mass spectrometry. Two members of the REG3 subfamily were found in the canine genome, REG3E1 and REG3E2, both encoding for the same 176 AA protein, subsequently named REG3E. Anti-human REG3A antibodies demonstrated cross-reactivity with the canine REG3E protein in pancreas homogenates. In canine plasma, a protein band of approximately 17 kDa was apparent. Mass spectrometry confirmed this protein to be the product of the two annotated REG3E genes. Strong immunoreactivity to anti-human REG3A antibodies was found in sections of canine pancreas affected with acute pancreatitis, but it was weak in healthy pancreatic tissue. Recombinant canine REG3E protein underwent a selective trypsin digestion as described in other species. No evidence for the presence of a homolog of REG1A in dogs was found in any of the investigations. In conclusion, dogs express REG3E in the pancreas, whose role as biomarker merits further investigations. Homologs to human REG1A are not likely to exist in dogs

Development and validation of an ELISA to measure regenerating island-derived protein 3E in canine blood.

BACKGROUND Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic tissue and plasma of dogs, with high expression in pancreatitis and sepsis. OBJECTIVES We aimed to develop and validate an ELISA to measure REG3E concentrations in canine blood. METHODS An indirect sandwich ELISA was developed using recombinant canine REG3E protein and polyclonal anti-canine REG3E antibodies raised in guinea pigs and rabbits. Antibody specificity was assessed using western blot and mass spectrometric analysis of protein purified from canine plasma. Assay validation included evaluation of dilutional linearity, parallelism, spiking recovery, repeatability and reproducibility, stability, interferences, and comparison of serum and heparinized plasma. RESULTS Antibodies bound specifically to REG3E with no evidence of cross-reactivity with other proteins. The limit of detection of the ELISA was 15 ng/mL, and the lower limit of quantification was 30 ng/mL. The assay demonstrated good to excellent linearity, dilutional and mixing parallelism, and recovery, with mean observed-to-expected ratios of 104%, 107%, 102%, and 92%, respectively, and no evidence of a hook effect. Coefficients of variation were ≤8.5% for repeatability and ≤14.3% for reproducibility at three different levels. Measurements of REG3E in plasma were not significantly influenced by different storage conditions, freeze-thawing cycles, hemolysis, lipemia, or icterus. There was no significant difference between REG3E concentrations in heparinized plasma and serum samples. CONCLUSIONS The canine REG3E ELISA has acceptable precision, accuracy, linearity, and reproducibility for the measurement of REG3E in canine plasma and serum

OTU deubiquitinases reveal mechanisms of linkage specificity and enable ubiquitin chain restriction analysis

Sixteen ovarian tumor (OTU) family deubiquitinases (DUBs) exist in humans, and most members regulate cell-signaling cascades. Several OTU DUBs were reported to be ubiquitin (Ub) chain linkage specific, but comprehensive analyses are missing, and the underlying mechanisms of linkage specificity are unclear. Using Ub chains of all eight linkage types, we reveal that most human OTU enzymes are linkage specific, preferring one, two, or a defined subset of linkage types, including unstudied atypical Ub chains. Biochemical analysis and five crystal structures of OTU DUBs with or without Ub substrates reveal four mechanisms of linkage specificity. Additional Ub-binding domains, the ubiquitinated sequence in the substrate, and defined S1’ and S2 Ub-binding sites on the OTU domain enable OTU DUBs to distinguish linkage types. We introduce Ub chain restriction analysis, in which OTU DUBs are used as restriction enzymes to reveal linkage type and the relative abundance of Ub chains on substrates

Molecular characterization of Neospora caninum MAG1, a dense granule protein secreted into the parasitophorous vacuole, and associated with the cyst wall and the cyst matrix

SUMMARY: In Neospora caninum and Toxoplasma gondii, the parasitophorous vacuole (PV) is synthesized at the time of infection. During tachyzoite-to-bradyzoite stage conversion, the PV is later transformed into a tissue cyst that allows parasites to survive in their host for extended periods of time. We report on the characterization of NcMAG1, the N. caninum orthologue of T. gondii MAG1 (matrix antigen 1; TgMAG1). The 456 amino acid predicted NcMAG1 protein is 54% identical to TgMAG1. By immunoblotting, a rabbit antiserum raised against recombinant NcMAG1 detected a major product of approximately 67 kDa in extracts of N. caninum tachyzoite-infected Vero cells, which was stained more prominently in extracts of infected Vero cells treated to induce in vitro bradyzoite conversion. Immunofluorescence and TEM localized the protein mainly within the cyst wall and the cyst matrix. In both tachyzoites and bradyzoites, NcMAG1 was associated with the parasite dense granules. Comparison between NcMAG1 and TgMAG1 amino acid sequences revealed that the C-terminal conserved regions exhibit 66% identity, while the N-terminal variable regions exhibit only 32% identity. Antibodies against NcMAG1-conserved region cross-reacted with the orthologuous protein in T. gondii but those against the variable region did not. This indicates that the variable region possesses unique antigenic characteristics

Effects of radiofrequency electromagnetic fields (RF EMF) on cancer in laboratory animal studies

Background: The carcinogenicity of radiofrequency electromagnetic fields (RF EMF) has been evaluated by the International Agency for Research on Cancer (IARC) in 2011. Based on limited evidence of carcinogenicity in humans and in animals, RF EMF were classified as possibly carcinogenic to humans (Group 2B). In 2018, based on a survey amongst RF experts, WHO prioritized six major topics of potential RF EMF related human health effects for systematic reviews. In the current manuscript, we present the protocol for the systematic review of experimental laboratory animal studies (cancer bioassays) on exposure to RF fields on the outcome of cancer in laboratory animals. Objective: In the framework of WHO's Radiation Program, the aim of this work is to systematically evaluate effects of RF EMF exposure on cancer in laboratory animals. Study eligibility and criteria: WHO's Handbook (2014) for guideline development will be followed with appropriate adaptation. The selection of eligible studies will be based on Population, Exposures, Comparators, and Outcomes (PECO) criteria. We will include peer-reviewed articles and publicly available reports from government agencies reporting original data about animal cancer bioassays on exposure to RF EMF. The studies are identified by searching the following databases: MEDLINE (PubMed), Science Citation Index Expanded and Emerging Sources Citation Indes (Web of Science), Scopus, and the EMF Portal. No language or year-of-publication restrictions are applied. The methods and results of eligible studies will be presented in accordance with the PRISMA 2020 guidelines. Study appraisal method: Study evaluation of individual studies will be assessed using a risk of bias (RoB) tool developed by the Office of Health Assessment and Translation (OHAT) with appropriate considerations including sensitivity for evaluating RF EMF exposure in animal cancer bioassays. The final evaluation on the certainty of the evidence on a carcinogenic risk of RF EMF exposure in experimental animals will be performed using the OHAT Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach with appropriate considerations. The protocol has been registered in an open-source repository (PROSPERO)

Анализ эффективности применения технологии гидравлического разрыва пласта на нефтяных месторождениях Западной Сибири

Анализ технологических и геологических особенностей применения гидравлического разрыва пласта. Эффективность применения ГРП путем сравнения показателей разработки до и после введения данного метода на нефтяных месторождениях Западной Сибири.Analysis of technological and geological features of the use of hydraulic fracturing. The effectiveness of hydraulic fracturing by comparing the development indicators before and after the introduction of this method in oil fields of Western Siberia