Gene expression profiles of CD4/CD8 double‐positive T cells in porcine peripheral blood

A characteristic subset of T cells, known as double positive T cells (DPTC) and expressing both cluster of differentiation 4 (CD4) and CD8, is observed in porcine peripheral blood. Previous studies suggested that DPTC might be memory cells. However, detailed phenotypes and functions of DPTC are yet to be fully elucidated and thus, the relatedness of DPTC with memory phenotypes remains unclear. In this study, DPTC gene expression profiles in peripheral blood were analyzed by DNA microarray in Experiment 1 and compared with those of CD4 single positive T cells (4SPTC) and CD8 single positive T cells (8SPTC). Expressions of IFNG, CCL5, NCK2, CCR2 and ITGB1 were higher than that of 4SPTC and 8SPTC. In contrast, expressions of CCR7 and SELL were lower than that of 4SPTC and 8SPTC. These results suggested that DPTC were either effector T cells or effector memory T cells (T_EM). Next, to determine whether DPTC were effector T cells or T_EM, differences in the response of DPTC and 8SPTC against immunized/primed antigens were compared (Experiment 2). While DPTC showed quick elevation of IL2 and CD25 gene expressions against in vitro stimulation of primed/immunized antigens, 8SPTC did not. These results suggest that at least some DPTC likely belong to T_EM.     

A proteomic analysis of Nipponia nippon (ID#162)

We investigated the proteome of a female Crested Ibis (Nipponia nippon, ID#162) that died on March 10, 2010 at the Sado Japanese Crested Ibis Conservation Center. Protein preparations from the brain, trachea, liver, heart, lung, proventriculus, muscular stomach, small intestine, duodenum, ovary and neck muscle were subjected to in‐solution shotgun mass spectrometry (MS)/MS analyses using an LTQ Orbitrap XL mass spectrometer. A search of the National Center for Biotechnology Information Gallus gallus databases revealed 4253 GI (GenInfo Identifier) numbers with the sum of the same 11 tissues examined in the Crested Ibis. To interpret the obtained proteomics data, it was verified in detail with the data obtained from the brain of the Crested Ibis. It has been reported that drebrin A is specifically expressed in adult chicken brain. In the shotgun proteomic analyses of the Crested Ibis, we identified drebrin A as a brain‐specific protein. Furthermore, Western blotting analysis of the protein preparations from 10 tissues of the Crested Ibis and 150‐day‐old hens using anti‐drebrin antibodies showed intensive expression of approximately 110 kDa polypeptides of drebrin in both brains. We believe firmly that the present data will contribute to initial and fundamental steps toward understanding the Crested Ibis proteome.     

Low expression of the antibacterial factor L‐amino acid oxidase in bovine mammary gland

In the mouse, L‐amino acid oxidase (LAO) produces hydrogen peroxide by utilizing free amino acids and is a proven antibacterial factor in mammary glands. Mastitis, a bacterial infection of the mammary gland, is the most frequent disease in dairy cattle. Here, we investigate whether LAO is expressed in the mammary gland of dairy cattle and is antibacterial. In dairy cattle, the expression level of LAO mRNA in the mammary gland was considerably lower than that in mice, and LAO activity was not observed in cattle milk that produced hydrogen peroxide. The expression of LAO mRNA was also low in Japanese Black cattle, the same as in Holstein cattle. A higher LAO mRNA expression was observed in the mastitis glands than in the lactating glands. Furthermore, spleen and lymph nodes expressed high levels of LAO mRNA in dairy cattle. We conclude that mammary glands in dairy cattle have lower ability to express the LAO gene compared to that in mice, which may result in a high incidence of mastitis.     

Effects of oral administration of heat‐killed Enterococcus faecium strain NHRD IHARA in post‐weaning piglets

Probiotic bacteria such as lactic acid bacteria (LAB) have recently received attention as candidates for alternative anti‐microbial feed additives. We previously isolated Enterococcus faecium strain NHRD IHARA (FERM BP‐11090, NHRD IHARA strain) and reported its probiotic efficacy. However, we have not determined the effect of oral administration of heat‐killed cells of this strain. Here, we performed two experiments to investigate the effect of oral administration of the heat‐killed NHRD IHARA strain on post‐weaning piglets. In Experiment 1, there was a significant improvement in growth performance (P = 0.04) and increase in serum immunoglobulin A (IgA) production (P = 0.03) in the group fed heat‐killed cells. These results were similar to previous results we obtained with live cells. We also found changes in serum and fecal IgA production that were unrelated to the patterns of microbiotal change. In Experiment 2, we detected a significant improvement in villus growth in the jejunum (P = 0.0002). In conclusion, oral administration of the heat‐killed NHRD IHARA strain in post‐weaning piglets had the same efficacy as administration of the live strain. The heat‐killed NHRD IHARA strain can be used as feed additives to improve pig growth and health on commercial farms.     

Shotgun proteomic analysis of porcine colostrum and mature milk

The epitheliochorial nature of the porcine placenta prevents the transfer of maternal immunity. Therefore, ingestion of the colostrum immediately after birth is crucial for neonatal piglets to acquire passive immunity from the sow. We performed a shotgun proteomic analysis of porcine milk to reveal in detail the protein composition of porcine milk. On the basis of the Swiss‐Prot database, 113 and 118 proteins were identified in the porcine colostrum and mature milk, respectively, and 50 of these proteins were common to both samples. Some immune‐related proteins, including interleukin‐18 (IL‐18), were unique to the colostrum. The IL‐18 concentration in the colostrum and mature milk of four sows was measured to validate the proteomic analysis, and IL‐18 was only detected in the colostrum (191.0 ± 53.9 pg/mL) and not in mature milk. In addition, some proteins involved in primary defense, such as azurocidin, which has never been detected in any other mammal's milk, were also identified in the colostrum.     

Photosynthetic traits and growth of Quercus mongolica var. crispula sprouts attacked by powdery mildew under free-air CO2 enrichment

To predict the performance of coppice forests with Japanese oak (Quercus mongolica var. crispula) in future changing environment, we studied the growth, photosynthesis, and powdery mildew (Erysiphe alphitoides) infection of sprouts of Japanese oak under free-air CO₂ enrichment. Elevated CO₂ reduced powdery mildew infection in both leaves of the shoot emerged in spring (1st flush) and the lammas and proleptic shoots (2nd flush) of sprouts. We observed significant increase in the net photosynthetic rate at growth CO₂ concentration (i.e., 370 and 500 μmol mol^?1 for ambient and elevated CO₂ treatments, respectively) in both 1st and 2nd flush leaves of sprouts grown under elevated CO₂. On the other hand, no significant increase in net photosynthetic rate under elevated CO₂ was found before cutting. The photosynthetic activity of 2nd flush leaves in the sprouts under ambient condition was greatly reduced by severe infection to powdery mildew. Growth of sprouts was enhanced in the elevated CO₂ condition. We conclude the growth enhancement in Japanese oak sprouts under elevated CO₂ in the present study was achieved not only by physiological response (i.e., photosynthetic stimulation) but also by disease interaction.     

Cellulose digestion by common Japanese freshwater clam Corbicula japonica

ABSTRACT:   Cellulose digestion by Corbicula japonica was investigated according to the hypothesis that without any symbiotic aid, this organism can utilize cellulose as a carbon source. Enzymatic studies revealed the complete cellulase activity of this species, and molecular cloning resulted in the isolation of cDNA with an ORF encoding a 596-amino-acid protein that shares significant homology with abalone and termite cellulases with an amino acid identity of 52.2% and 50.5%, respectively. The isolated cellulase had a carbohydrate-binding module at the N-terminal region that was also reportedly present in abalone cellulase, and its mRNA were specifically expressed in the digestive gland. These findings strongly support the assumption that C. japonica has an endogenous cellulose, as well as abalones and termites. It is further believed that C. japonica plays an important roll in decomposing cellulose, and consequently contributes to the carbon-cycle in the aquatic environment, as termites do in terrestrial forests.