Thyroid hormone augments GLUT4 expression and insulin-sensitive glucose transport system in differentiating rat brown adipocytes in culture

The effects of triiodothyronine (T3) on differentiation-dependent expression of GLUT and responses of glucose transport to insulin and norepinephrine (NE) were investigated. Precursor cells of brown adipocytes isolated from the interscapular brown adipose tissue of newborn rats were cultured in the absence or presence of various concentrations of T3. Western bolt analysis revealed that treatment with T3 resulted in an increased expression of GLUT4, in a dose-dependent manner, whereas GLUT1 contents were unchanged. In parallel with the increase in GLUT4 expression, T3 improved insulin sensitivity for glucose transport, being accompanied by an increase in maximal transport rate and a reduction of ED(50). In contrast, T3-treatment of the brown adipocytes during the differentiation process had little effect on NE-regulatable glucose transport system. These results suggest that T3 plays a predominant role in the development of insulin-sensitive glucose transport during differentiation of brown adipocytes.     

Carbon dioxide exchange of a larch forest after a typhoon disturbance

A typhoon event catastrophically destroyed a 45-year-old Japanese larch plantation in southern Hokkaido, northern Japan in September 2004, and about 90% of trees were blown down. Vegetation was measured to investigate its regeneration process and CO₂ flux, or net ecosystem production (NEP), was measured in 2006–2008 using an automated chamber system to investigate the effects of typhoon disturbance on the ecosystem carbon balance. Annual maximum aboveground biomass (AGB) increased from 2.7 Mg ha⁻¹ in 2006 to 4.0 Mg ha⁻¹ in 2007, whereas no change occurred in annual maximum leaf area index (LAI), which was 3.7 m² m⁻² in 2006 and 3.9 m² m⁻² in 2007. Red raspberry (Rubus idaeus) had become dominant within 2 years after the typhoon disturbance, and came to account for about 60% and 50% of AGB and LAI, respectively. In comparison with CO₂ fluxes measured by the eddy covariance technique in 2001–2003, for 4.5 months during the growing season, the sum of gross primary production (GPP) decreased on average by 739 gC m⁻² (64%) after the disturbance, whereas ecosystem respiration (RE) decreased by 501 gC m⁻² (51%). As a result, NEP decreased from 159 ± 57 gC m⁻² to −80 ± 30 gC m⁻², which shows that the ecosystem shifted from a carbon sink to a source. Seasonal variation in RE was strongly correlated to soil temperature. The interannual variation in the seasonal trend of RE was small. Light-saturated GPP (P_max) decreased from 30–45 μmol m⁻² s⁻¹ to 8–12 μmol m⁻² s⁻¹ during the summer season through the disturbance because of large reduction in LAI.     

Avian predation on wild and cultured sea urchin <Emphasis Type="Italic">Strongylocentrotus intermedius</Emphasis> in a rocky shore habitat

ABSTRACT:   The present study reports the annual variation in consumption of the sea urchin Strongylocentrotus intermedius by avian predators on a rocky shore where the culture of sea urchins has been conducted. Carrion crow and a few gull species were the most abundant avian predators and consumed a large number of sea urchins. Crows consumed mostly natural sea urchins, approximately 36 kg ww/ha per year on the intertidal rocky bench, but the gull species consumed mostly cultured sea urchins, approximately 100 kg ww/ha per year in the culture area. The seasonal variation in the amount of sea urchins consumed by crows was higher than that by the gull species, presumably because of the difference in foraging behavior in association with the seasonal tidal cycle. The natural sea urchins consumed are an allochthonous input from the subtidal to the intertidal habitat, and thus, crow predation may not affect the natural and the cultured populations of the sea urchin. The gull species consumed much of the cultured sea urchin, and thus, may be regarded as an effective predator causing damage to sea urchin culture. The results suggest that further studies are needed to determine why the gull species selectively feed on cultured sea urchins.     

Loss of virulence of the phytopathogen Ralstonia solanacearum through infection by φRSM filamentous phages

φRSM1 and φRSM3 (φRSM phages) are filamentous phages (inoviruses) that infect Ralstonia solanacearum, the causative agent of bacterial wilt. Infection by φRSM phages causes several cultural and physiological changes to host cells, especially loss of virulence. In this study, we characterized changes related to the virulence in φRSM3-infected cells, including (i) reduced twitching motility and reduced amounts of type IV pili (Tfp), (ii) lower levels of β-1,4-endoglucanase (Egl) activity and extracellular polysaccharides (EPS) production, and (iii) reduced expression of certain genes (egl, pehC, phcA, phcB, pilT, and hrpB). The significantly lower levels of phcA and phcB expression in φRSM3-infected cells suggested that functional PhcA was insufficient to activate many virulence genes. Tomato plants injected with φRSM3-infected cells of different R. solanacearum strains did not show wilting symptoms. The virulence and virulence factors were restored when φRSM3-encoded orf15, the gene for a putative repressor-like protein, was disrupted. Expression levels of phcA as well as other virulence-related genes in φRSM3-ΔORF15-infected cells were comparable with those in wild-type cells, suggesting that orf15 of φRSM3 may repress phcA and, consequently, result in loss of virulence.     

Identification of bovine dendritic cell phenotype from bovine peripheral blood

Dendritic cells (DCs) are professional antigen presenting cells, which initiate primary immune responses and also play an important role in the generation of peripheral tolerance. There is no reliable method established for the isolation of bovine peripheral blood DCs, and furthermore, the phenotypes and the functions of bovine DCs are still not fully clear. In the present study, we have attempted to identify bovine peripheral blood DCs by negative-selection. In bovine peripheral blood mononuclear cells (PBMC), we have newly characterized the phenotype of DCs, which is CD11c+/CD172a+. These cells display features of myeloid type DCs. In the thymic medulla, CD11c+/CD172a+ cells were also present and CD1+/CD172a+ cells were additionally detected as a population of DCs. The data suggest that one of the bovine DCs phenotypes from PBMC is derived from myeloid lineages lacking a CD1 molecule, which then drift to several tissues, and that they then may express a CD1 molecule upon their functional differentiation.     

Effects of cooling rate on post-thaw motility and fertility of Japanese pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis> spermatozoa

The post-thaw motility and fertility of Japanese pearl oyster sperm show large variances, even among sperm samples obtained from the same individuals. This study aimed to clarify the factors that cause such differences. Spermatozoa were diluted 50 times with diluent comprising 10 % methanol, 18 % fetal bovine serum, and 72 % seawater, and dispensed into 0.25 ml straws. A total of 59 straws were cooled, one by one, at 11 different heights from the surface of liquid nitrogen (LN) to −50 °C, and then immediately immersed in LN. After thawing the straws, the relationships between the cooling rate and the post-thaw motility and post-thaw fertility of the spermatozoa were examined. Both the post-thaw motility and the post-thaw fertility showed a sharp peak when the straws were cooled at around −20 °C/min. There was a strong correlation between post-thaw motility and fertility (P < 0.001). There was a large difference in the cooling rates and the post-thaw motilities and fertilities of the spermatozoa, even between straws cooled at the same height. These results indicate that the optimum range for the cooling rate of oyster spermatozoa is quite narrow, and the method of cooling straws at a fixed distance from the LN surface is unsuitable for the cryopreservation of Japanese pearl oyster spermatozoa.     

Cardiovascular effects of a phosphodiesterase III inhibitor in the presence of carvedilol in dogs

The aim of this study was to determine whether dobutamine, dopamine, or milrinone (a phosphodiesterase [PDE] III inhibitor) would support cardiac function that had been attenuated by administration of the beta-blocker, carvedilol (0.2, 0.4, or 0.8 mg/kg). Hemodynamic and cardiac parameters including the heart rate (HR), left-ventricular fractional shortening (FS), and arterial pressure were measured in six healthy dogs without cardiac disease. Carvedilol did not affect FS or arterial pressure, but decreased the HR significantly. The positive inotropic and chronotropic responses to dobutamine and dopamine were attenuated by carvedilol, whereas arterial pressure was unaffected. Milrinone did not affect the HR and decreased arterial pressure, whereas FS was significantly greater both in the control and carvedilol-treated groups. Although milrinone affect the negative chronotropic effects of carvedilol, milrinone increased FS and prevented the decrease in arterial pressure. These results suggest that inhibition of PDE III preserves cardiac contractility and hemodynamic function in the presence of carvedilol.     

Universal PCR primers for ribosomal protein gene introns of fish

Human ribosomal protein (RP) gene sequences with respect to intron/exon structures and corresponding cDNA or genomic data of fish species were obtained from the GenBank database. Based on conserved exon sequences, 128 primer pairs for 41 genes were designed for exon-primed intron-crossing (EPIC) polymerase chain reaction (PCR). In reference to the draft genome sequences of the Pacific bluefin tuna (Thunnus orientalis), 12 primer pairs expected to amplify introns of the bluefin tuna with lengths of 500–1000 bp were selected and applied to six distantly related fish species belonging to the Orders Clupeiformes, Tetraodontiformes, Pleuronectiformes, Perciformes, Scorpaeniformes, and Anguilliformes. PCR amplification was observed for at least four species in each primer pair, and all fragments were larger than those expected for intronless amplification. Single fragment amplification was observed for at least seven primer pairs per species. Fragment sizes of the bluefin tuna for nine primer pairs corresponded to those expected from the genomic data. Thus, our primer pairs are potentially applicable to a wide variety of fish species and serve as an initial step for isolating single-copy nuclear DNA sequences.     

Ex vivo tissue discrimination by visible and near-infrared spectra with chemometrics

The risk of infections from zoonotic pathogens of tissues and/or tissue-derived products has been increasing. One preventive approach in reducing infection risk is tissue decontamination, where selection and screening of highly infectious tissues are strictly followed. Therefore, the development of reliable analytical methods for rapid tissue discrimination is essentially important. In the present study, a procedure has been developed for intact tissue discrimination on the basis of multivariate analysis of visible and near-infrared (Vis-NIR) spectra of certain tissues such as brain, liver, kidney and testis of mice without any pretreatment. Transmittance spectra in the 600- to 1000-nm regions were subjected to a principal component analysis (PCA), and leave-out cross-validation was employed to develop multivariate models for tissue discrimination. The plot of PCA scores against Vis-NIR spectra of brains, kidneys, livers and testes from 11 mice portrayed reliable tissue discrimination. This result suggests that Vis-NIR spectroscopy combined with chemometrics analysis may provide a potentially useful approach for rapid non-destructive discrimination of tissues.     

A spontaneously occurring malignant ovarian Sertoli cell tumor in a young Sprague Dawley rat

Primary ovarian tumors are generally uncommon in rats used in toxicologic studies. A malignant Sertoli cell tumor was present in the ovary of a 19-week-old female Sprague Dawley rat. Macroscopically, the mass was white and firm, 10 × 13 × 17 mm in size, and located in the right ovary. Histopathologically, the mass was composed of nests of pleomorphic cells, which formed seminiferous-like tubules separated by a thin fibrovascular stroma. The tubules were lined by tumor cells, which had basally located nuclei and abundant eosinophilic and vacuolated cytoplasm. In some areas, the tumor cells were arranged in a retiform growth pattern, mimicking a rete testis/ovarii. Disseminated metastases to the surfaces of the mesentery, spleen and liver were also present. Immunohistochemically, many tumor cells were strongly positive for vimentin, estrogen receptor α and Ki 67. Some tumor cells were positive for pancytokeratin and inhibin α. These findings closely resemble those of an ovarian-derived human malignant Sertoli cell tumor. From our review of the literature, we believe this is the first report of a spontaneous malignant Sertoli cell tumor in the ovary of a young laboratory rat. This case might provide useful historical control information for rat toxicity studies.