Hemerythrin is required for Aeromonas hydraphlia to survive in the macrophages of Anguilla japonica.

Survival in host phagocytes is an effective strategy for pathogenic microbes to spread. To understand the mechanisms of Aeromonas hydrophila survival within host macrophages, a library of mini-Tn10 transposon insertion mutants was constructed. The M85 mutant, whose survival in host macrophages was only 23.1% of that of the wild-type (WT) strain, was utilized for further study. Molecular analysis showed that a 756-bp open reading frame (ORF) (GenBank accession No. CP007576) in the M85 mutant was interrupted by mini-Tn10. This ORF encodes for a 183-amino acid protein and displays the highest sequence identity (99%) with the hemerythrin (Hr) protein of A. hydrophila subspecies hydrophila ATCC 7966. The survival of the WT, M85 mutant, and complemented M85 (Hr) strains were compared in host macrophages in vitro, and the results showed that M85 exhibited defective survival, while that of M85 (Hr) was restored. To investigate the possible mechanisms of A. hydrophila survival in host macrophages, the expression of Hr under hyperoxic and hypoxic conditions was evaluated. The results revealed that the expression of this protein was higher under hyperoxic conditions than under hypoxic conditions, which indicates that Hr protein expression is sensitive to O2 concentration. Hydrogen peroxide sensitivity tests further suggested that the M85 mutant was more sensitive to oxidative stress than the WT and M85 (Hr) strains. Taken together, these results suggest that the Hr protein may act as an O2 sensor and as a detoxifier of reactive oxygen species, and is required for A. hydrophila survival within host macrophages.

Genetic diversity analysis of an indigenous Chinese buffalo breed and hybrids based on microsatellite data.

Chinese native buffaloes have faced the threat of extinction, along with an increase in crossbreeding with domesticated river buffaloes; consequently, conservation of local buffalo genetic resources has become a priority. A Chinese native breed, Jianghan, is often crossed intentionally and unintentionally with imported breeds from India and Pakistan, Murrah, and Nili-Ravi. A total of 128 buffaloes of the breeds Jianghan, Murrah, and Nili-Ravi and their presumed hybrid offspring were genotyped for 10 microsatellite markers. Heterozygosity and Wright's F-statistics were calculated to determine the genetic variation in those populations. The observed average heterozygosities ranged from 0.836 (Murrah) to 0.986 (Jianghan), higher than the expected heterozygosities and all the inbreeding values within the populations were negative. The genetic distances between the presumed hybrid buffaloes and the two imported river type dairy buffalo breeds (Murrah and Nili-Ravi) were lower than with the native Jianghan, indicating strong contributions of the imported breeds to this presumed hybrid buffalo population. This information will be useful for the development of rational breeding for the dairy buffalo industry and for conservation strategies for the Jianghan buffalo.

Integrating Multiple Evidence Sources to Predict Adverse Drug Reactions Based on a Systems Pharmacology Model.

Identifying potential adverse drug reactions (ADRs) is critically important for drug discovery and public health. Here we developed a multiple evidence fusion (MEF) method for the large-scale prediction of drug ADRs that can handle both approved drugs and novel molecules. MEF is based on the similarity reference by collaborative filtering, and integrates multiple similarity measures from various data types, taking advantage of the complementarity in the data. We used MEF to integrate drug-related and ADR-related data from multiple levels, including the network structural data formed by known drug-ADR relationships for predicting likely unknown ADRs. On cross-validation, it obtains high sensitivity and specificity, substantially outperforming existing methods that utilize single or a few data types. We validated our prediction by their overlap with drug-ADR associations that are known in databases. The proposed computational method could be used for complementary hypothesis generation and rapid analysis of potential drug-ADR interactions.

Embryo transfer and sex determination following superovulated hinds inseminated with frozen-thawed sex-sorted Y sperm or unsorted semen in Wapiti (Cervus elaphus songaricus).

The aim of this study was to evaluate embryo production in superovulated wapiti hinds inseminated with either Y-sorted or unsorted semen. Eighteen hinds were allocated to three treatment groups: AI following multiple ovulation (CIDR/FSH) with 10×10(6) Y-sorted frozen-thawed semen (Y group, n=6), or 10×10(6) and 100×10(6) unsorted frozen-thawed semen for the unsorted (n=6) and the control group (n=6). The embryos from the sixth day following insemination were collected and classified. Fifteen embryos from the unsorted or the control group, and four embryos from the Y group were sex determinated based on DNA analysis of the amelogenin gene. Twenty-one embryos from the Y group and 42 embryos from the unsorted or the control group were transferred into 21 and 42 synchronized recipients via standard procedures on 6th day post estrus, respectively. There were no significant differences in the number of recovered eggs, transferable embryos, degenerated embryos or unfertilized oocytes per hind among the three groups of the control (9.2±3.6, 4.7±1.9, 3.0±2.0, 1.5±1.4), the unsorted (8.2±1.9, 4.8±0.7, 1.7±1.0, 1.7±1.0) and the Y group (8.8±4.2, 4.2±1.8, 2.2±1.2, 2.5±2.1), respectively (P>0.05). The sex ratio of embryos from the Y group (4M/0F) was significantly (P<0.05) distinct from that of the unsorted and control group (8M/7F). The sex ratio of the offspring from sexed embryos (8M/0F) was deviated significantly (P<0.05) from that of the non-sexed embryos (11M/9F). In conclusion, the results suggested that the male embryos of predicted sex can be achieved with AI of sex-sorted cryopreserved sperm. PCR amplification using the amelogenin gene primers can be applied to DNA analysis of micro samples from wapiti embryo biopsies for sex identification. The male offspring can be produced after transferred with the male embryos of predicted sex.