Enzymatic Degradation of PrPSc by a Protease Secreted from Aeropyrum pernix K1

BACKGROUND: An R30 fraction from the growth medium of Aeropyrum pernix was analyzed for the protease that can digest the pathological prion protein isoform (PrP(Sc)) from different species (human, bovine, deer and mouse). METHODOLOGY/PRINCIPAL FINDINGS: Degradation of the PrP(Sc) isoform by the R30 fraction and the purified protease was evaluated using the 6H4 anti-PrP monoclonal antibody. Fragments from the N-terminal and C-terminal of PrP(Sc) were also monitored by Western blotting using the EB8 anti-PrP monoclonal antibody, and by dot blotting using the C7/5 anti-PrP monoclonal antibody, respectively. For detection of smaller peptides from incomplete digestion of PrP(Sc), the EB8 monoclonal antibody was used after precipitation with sodium phosphotungstate. Characterization of the purified active protease from the R30 fraction was achieved, through purification by fast protein liquid chromatography, and identification by tandem mass spectrometry the serine metalloprotease pernisine. SDS-PAGE and zymography show the purified pernisine plus its proregion with a molecular weight of ca. 45 kDa, and the mature purified pernisine as ca. 23 kDa. The purified pernisine was active between 58 °C and 99 °C, and between pH 3.5 and 8.0. The temperature and pH optima of the enzymatic activity of the purified pernisine in the presence of 1 mM CaCl(2) were 105 °C ± 0.5 °C and pH 6.5 ± 0.2, respectively. CONCLUSIONS/SIGNIFICANCE: Our study has identified and characterized pernisine as a thermostable serine metalloprotease that is secreted from A. pernix and that can digest the pathological prion protein PrP(Sc)

Plant Growth-Promoting Microbes from Herbal Vermicompost

Overreliance on chemical pesticides and fertilizers has resulted in problems including safety risks, outbreaks of secondary pests normally held in check by natural enemies, insecticide resistance, environmental contamination, and decrease in biodiversity. The increasing costs and negative effects of pesticides and fertilizers necessitate the idea of biological options of crop protection and production. This includes the use of animal manure, crop residues, microbial inoculum, and composts. They provide natural nutrition, reduce the use of inorganic fertilizers, develop biodiversity, increase soil biological activity, maintain soil physical properties, and improve environmental health

Встановлення поліцією причин та особливостей домашнього насильства щодо дітей

The article reveals the most pressing issues regarding the protection of children from domestic violence. The problem of protecting such children has certain difficulties. This is due to the fact that the vast majority of affected children are children from disadvantaged families, as a rule, few people want to protect this category of people. It is suggested to pay special attention to the peculiarities of the method of interviewing a child who has experienced domestic violence, to the procedure of interviewing such children. A list of grounds that are undesirable to use when communicating with children is identified, and recommendations for interviewing children affected by violence are provided. The main types of violence against children, subjects who commit violence against children, dangerous consequences for the society where domestic violence against children is actively manifested are described. It is stated that in accordance with international standards and world practice, the interview of a child who is a victim of violent crimes, domestic violence or witnessed violence should be conducted in a child-friendly atmosphere, where they feel protected and comfortable. The "green room" was described as a specially equipped room for interviewing such children. The legal status of the child is determined, the circumstances, causes and types of domestic violence against children are established. Suggestions for improving the use of tactics in the survey of this category of minors in order to effectively protect the rights and interests of the latter. The problems considered in the work relate to both theory and practice. There are two consequences for children who have experienced domestic violence, which are manifested simultaneously: harm to the victim and harm to society.У статті розкрито найбільш актуальні проблеми щодо захисту дітей від домашнього насильства. Проблема захисту таких дітей має певні складнощі. Це пояснюється тим, що переважна більшість постраждалих дітей – це діти з неблагополучних родин, як правило, захистити таку категорію осіб мало у кого виникає бажання. Звернено увагу на процедуру опитування дітей постраждалих від домашнього насильства. Встановлено, що процес отримання показань неповнолітніх, які зазнали домашнього насилля, має певні труднощі. Охарактеризовано основні види насильства над дітьми, суб’єктів, які вчиняють насильство над дітьми, небезпечні наслідки для суспільства, де активно проявляється домашнє насильство над дітьми

Helicobacter pylori outer membrane protein HopQ identified as a novel T4SS-associated virulence factor

Helicobacter pylori is a bacterial pathogen that colonizes the gastric niche of ∼ 50% of the human population worldwide and is known to cause peptic ulceration and gastric cancer. Pathology of infection strongly depends on a cag pathogenicity island (cagPAI)-encoded type IV secretion system (T4SS). Here, we aimed to identify as yet unknown bacterial factors involved in cagPAI effector function and performed a large-scale screen of an H. pylori transposon mutant library using activation of the pro-inflammatory transcription factor NF-κB in human gastric epithelial cells as a measure of T4SS function. Analysis of ∼ 3000 H. pylori mutants revealed three non-cagPAI genes that affected NF-κB nuclear translocation. Of these, the outer membrane protein HopQ from H. pylori strain P12 was essential for CagA translocation and for CagA-mediated host cellresponses such as formation of the hummingbird phenotype and cell scattering. Besides that, deletion of hopQ reduced T4SS-dependent activation of NF-κB, induction of MAPK signalling and secretion of interleukin 8 (IL-8) in the host cells, but did not affect motility or the quantity of bacteria attached to host cells. Hence, we identified HopQ as a non-cagPAI-encoded cofactor of T4SS function

Genome sequence of the ubiquitous hydrocarbon-degrading marine bacterium Alcanivorax borkumensis

Schneiker-Bekel S, dos Santos VAPM, Bartels D, et al. Genome sequence of the ubiquitous hydrocarbon-degrading marine bacterium Alcanivorax borkumensis. Nature Biotechnology. 2006;24(8):997-1004