Jianghui Wang

Characterization of the antimicrobial peptide attacin loci from Glossina morsitans

The antimicrobial peptide Attacin is an immune effector molecule that can inhibit the growth of gram‐negative bacteria. In Glossina morsitans morsitans, which serves as the sole vectors of African trypanosomes, Attacins also play a role in trypanosome resistance, and in maintaining parasite numbers at homeostatic levels in infected individuals. We characterized the attacin encoding loci from a Bacterial Artificial Chromosome (BAC) library. The attacin genes are organized into three clusters. Cluster 1 contains two attacin (attA) genes located in head‐to‐head orientation, cluster 2 contains two closely related genes (attA and attB) located in a similar transcriptional orientation, and cluster 3 contains a single attacin gene (attD). Coding and transcription regulatory sequences of attA and attB are nearly identical, but differ significantly from attD. Putative AttA and AttB have signal peptide sequences, but lack the pro domain typically present in insect Attacins. Putative AttD lacks both domains. Analysis of attacin cDNA sequences shows polymorphisms that could arise either from allelic variations or from the presence of additional attacin genomic loci. Real time‐PCR analysis reveals that attA and attB expression is induced in the fat body of flies per os challenged with Escherichia coli and parasitized with trypanosomes. In the midgut, expression of these attacins is similarly induced following microbial challenge, but reduced in response to parasite infections. Transcription of AttD is significantly less relative to the other two genes, and is preferentially induced in the fat body of parasitized flies. These results indicate that the different attacin genes may be differentially regulated.

Heat Stress in Dairy Cattle Alters Lipid Composition of Milk

Heat stress, potentially affecting both the health of animals and the yield and composition of milk, occurs frequently in tropical, sub-tropical and temperate regions. A simulated acute heat stress experiment was conducted in controlled-climate chambers and milk samples collected before, during and after the heat challenge. Milk lipid composition, surveyed using LC-MS, showed significant changes in triacylglycerol (TAG) and polar lipid profiles. Heat stress (temperature-humidity index up to 84) was associated with a reduction in TAG groups containing short- and medium-chain fatty acids and a concomitant increase in those containing long-chain fatty acids. The abundance of five polar lipid classes including phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine, lysophosphatidylcholine and glucosylceramide, was found to be significantly reduced during heat stress. Lysophosphatidylcholine, showing the greatest reduction in concentration, also displayed a differential response between heat tolerant and heat susceptible cows during heat stress. This phospholipid could be used as a heat stress biomarker for dairy cattle. Changes in TAG profile caused by heat stress are expected to modify the physical properties of milk fat, whereas the reduction of phospholipids may affect the nutritional value of milk. The results are discussed in relation to animal metabolism adaptation in the event of acute heat stress.

Seasonal Variation of Triacylglycerol Profile of Bovine Milk

Milk contains 3–6% of fat, of which the dominant component is triacylglycerol (TAG). Over 100 TAG groups can be readily detected in any non-enriched milk sample by LC-MS; most TAG groups contain several isomers (TAG molecules with different fatty acid composition), which cannot be fully resolved chromatographically by any single stationary phase. TAG profile of mature milk from 19 cows was surveyed in this study for eight consecutive months using RP-LC-Orbitrap MS. It was found that TAG profile of milk was not constant throughout the milking season and the seasonal pattern varied with TAG groups. The overall unsaturation level of TAG was stable from October 2013 to January 2014, decreased in February/March 2014 and then increased from April and peaked in May 2014. In addition to the seasonal fluctuation in TAG profile, the proportion of different isomeric species within a TAG group also changed substantially across seasons. However, the proportion of different positional isomers within a given TAG group does not seem to vary during the milking season. To our knowledge, this is the first report on the seasonal change of milk lipid at the TAG group and isomer level.

Milk-derived ribonuclease 5 preparations induce myogenic differentiation in vitro and muscle growth in vivo

Ribonuclease 5, also known as angiogenin, is a stable and abundant ribonuclease in milk whey protein, which is able to regulate several cellular functions, including capillary formation, neuron survival, and epithelial cell growth. Ribonuclease 5 is important for protein synthesis directly stimulating rRNA synthesis in the nucleolus. Here, we show that biologically active RNase5 can be purified from bovine milk. Furthermore, we show that milk-derived RNase5 directly stimulates muscle cell differentiation in vitro, inducing C2C12 cell differentiation and myogenesis. When supplemented into the diet of healthy adult mice, milk-derived RNase5 preparations promoted muscle weight gain and grip strength. Collectively, these data indicate that milk-derived RNase5 preparations exhibit a novel role in skeletal muscle cell function.