Thermal stress responses of a sterile mutant of Ulva pertusa (Chlorophyta)

SUMMARY: The thermal stress responses of a sterile mutant of the marine alga Ulva pertusa were investigated at 20°C and 30°C. The amounts of the photosynthetic pigments, β-carotene, chlorophylls a and b , lutein, neoxanthin, and violaxanthin, were 1.4–2.4 times higher in the 30°C-cultivated alga than in the 20°C-cultivated alga. The free amino acids, asparagine, aspartic acid, glutamine, glutamic acid, glycine, and serine, were abundant in the 20°C-cultivated alga, and increased 1.9–10.5-fold in response to thermal stress (30°C). Total carbon and nitrogen contents also increased in the 30°C-cultivated alga. Sodium dodecylsulfate-polyacrylamide gel electrophoretic patterns of total proteins extracted from both temperature-treated algae showed the increases of 20, 25, and 90 kDa proteins in the 30°C-cultivated alga. Isozyme assays for 20 enzymes showed a different banding pattern only in the case of glutamate dehydrogenase (GDH). Although it was observed that both temperature-treated algae possessed NAD⁺- and NADP⁺-specific GDH, the 30°C-cultivated alga had an additional NADP⁺-specific GDH (NADP-GDH). These results suggest that NADP-GDH compensates for the thermally induced decreases in nitrogen assimilation efficiency and thereby regulates nitrogen metabolism under conditions of temperature stress.     

Nucleotide sequence polymorphisms of beta1‐, beta2‐, and beta3‐adrenergic receptor genes on Jinhua,Meishan, Duroc and Landrace pigs

The full amino acid coding sequences of adrenergic receptor genes beta1, beta2, and beta3 (ADRB1, ADRB2, and ADRB3)were determined for Jinhua, Meishan, Duroc and Landrace pigs. Non‐synonymous substitution of Arg458Pro was found in the porcine ADRB1 gene, resulting in a 469 amino acid sequence. Continuous substitutions of Asn29Asp and Glu30Gln were found in the porcine ADRB2 gene, resulting in a 418 amino acid sequence. Additionally, a Lys30 polymorphism of the ADRB2 gene was found in the Jinhua pigs. There were three non‐synonymous substitutions of Asn24Thr, Arg264Gln and Asn398Asp on the porcine ADRB3 gene. A thymine insertion in the ADRB3 gene, resulting in a protein with two fewer amino acids, was found in the Jinhua and Meishan pigs. To assess the effect of ADRB polymorphisms on porcine subcutaneous fat layer thickness, we calculated the genetic frequency of the variants in fatty and lean groups, each consisting of 24 pigs that were crossbreds of Duroc and Jinhua pigs. The effect of the ADRB3 gene polymorphism was not evaluated, because there was insufficient variation on the ADRB3 gene in the examined groups. Although Fisher's exact test showed no significant difference in the frequency of ADRB1 and ADBR2 variants between the two groups, the Arg458 variant of ADRB1 was higher (P = 0.11) in the lean group, and pigs in that group had a thinner fat layer than did those with the Pro458 variant. These results imply a possibility of ADRB1 polymorphism as a minor factor in porcine fat layer thickness. The Asp29 variant of ADRB2 was higher in the lean group (P = 0.11), and the Glu30 variant was higher in the fatty group (P = 0.15), but the Asp29 variant was found only in the Chinese pigs. Thus, the effect of ADRB2 polymorphisms was not clear in this study.     

Significance of CCN2 expression in bovine preimplantation development

In mammalian preimplantation development, the first cell lineage segregation occurs during the blastocyst stage, when the inner cell mass and trophectoderm (TE) differentiate. Species‐specific analyses are essential to elucidate the molecular mechanisms that underlie this process, since they differ between various species. We previously showed that the reciprocal regulation of CCN2 and TEAD4 is required for proper TE differentiation in bovine blastocysts; however, the function of CCN2 during early embryogenesis has remained otherwise elusive. The present study assessed the spatiotemporal expression dynamics of CCN2 in bovine embryos, and evaluated how changes to CCN2 expression (using a CCN2 knockdown (KD) blastocyst model) regulate the expression of pluripotency‐related genes such as OCT4 and NANOG. The conducted quantitative PCR analysis revealed that CCN2 mRNA was expressed in bovine oocytes (at the metaphase stage of their second meiosis) and embryos. Similarly, immunostaining detected both cytoplasmic and nuclear CCN2 at all analyzed oocyte and embryonic stages. Finally, both OCT4 and NANOG expression levels were shown to be significantly reduced in CCN2 KD blastocysts. Together, these results demonstrate that bovine CCN2 exhibits unique expression patterns during preimplantation development, and is required for the proper expression of key regulatory genes in bovine blastocysts.     

A case of nontraumatic gas gangrene in a common marmoset (Callithrix jacchus)

The common marmoset is widely used in neuroscience and regenerative medicine research. However, information concerning common marmoset disorders, particularly infectious diseases, is scarce. Here, we report a case of a female common marmoset that died suddenly due to gas gangrene. The animal presented with gaseous abdominal distention at postmortem, and Clostridium perfringens type A was isolated from several tissues. Vacuoles, a Gram-positive bacteremia and intravascular hemolysis were observed microscopically in the muscles, liver and lungs. On the basis of these findings, we diagnosed nontraumatic gas gangrene caused by Clostridium perfringens type A in this common marmoset.     

RNA silencing against viruses: molecular arms race between Cucumber mosaic virus and its host

Plants have developed RNA silencing as an antiviral defense mechanism. To escape from the plant host’s defenses, viruses have countered their host’s antiviral silencing by producing RNA silencing suppressor proteins (RSSs). Although the mode of action of the majority of viral RSSs has been found to be through double-stranded RNA-binding, viruses have different strategies to counteract the host’s antiviral silencing pathways. The 2b protein of Cucumber mosaic virus, which is one of the most extensively studied viral RSSs, is reviewed here to provide insights on the molecular arms race between viruses and their host plants.     

Prevalence of Bordetella hinzii in mice in experimental facilities in Japan

To reveal the current status of the prevalence of Bordetella hinzii in mice in experimental facilities in Japan, a survey of this agent was performed by culture of tracheal swabs from a total of 12,923 mice from 1699 facilities (12,192 mice from 1572 facilities in universities and research institutes and 731 mice from 127 facilities in pharmaceutical companies) in total. In the results, 195 out of 12,192 mice (1.6%) from 44 out of 1572 facilities (2.8%) in universities and research institutes were positive for B. hinzii. No B. hinzii-positive mice were found in 127 pharmaceutical companies surveyed. Gross lesions in the lungs with isolation of B. hinzii were observed in seven mice from four universities, and the lesions were identified as bronchopneumonia histopathologically. To our knowledge, this is the first report to reveal the prevalence of B. hinzii in laboratory mice.