[Show abstract][Hide abstract]ABSTRACT: Valproic acid (VPA) is a widely used antiepileptic drug, which has recently been reported to modulate the neuronal differentiation of adipose tissue-derived stem cells (ASCs) in humans and dogs. However, controversy exists as to whether VPA really acts as an inducer of neuronal differentiation of ASCs. The present study aimed to elucidate the effect of VPA in neuronal differentiation of rat ASCs. One or three days of pretreatment with VPA (2 mM) followed by neuronal induction enhanced the ratio of immature neuron marker βIII-tubulin-positive cells in a time-dependent manner, where the majority of cells also had a positive signal for neurofilament medium polypeptide (NEFM), a mature neuron marker. RT-PCR analysis revealed increases in the mRNA expression of microtubule-associated protein 2 (MAP2) and NEFM mature neuron markers, even without neuronal induction. Three-days pretreatment of VPA increased acetylation of histone H3 of ASCs as revealed by immunofluorescence staining. Chromatin immunoprecipitation assay also showed that the status of histone acetylation at H3K9 correlated with the gene expression of TUBB3 in ASCs by VPA. These results indicate that VPA significantly promotes the differentiation of rat ASCs into neuron-like cells through acetylation of histone H3, which suggests that VPA may serve as a useful tool for producing transplantable cells for future applications in clinical treatments.
[Show abstract][Hide abstract]ABSTRACT: A male crossbred calf developed a limp and pain upon deep pressure on the right hind limb and the right forelimb. The radiographic findings of affected limbs and pathological findings of bone biopsy were similar to those observed in canine panosteitis. This is the first case of suspected panosteitis reported in cattle.
Full-text Article · May 2015 · The Canadian veterinary journal. La revue veterinaire canadienne
[Show abstract][Hide abstract]ABSTRACT: Hepatoprotective effects of Rhizopus oryzae/ U-1 aqueous extract (RU) were demonstrated in carbon tetrachloride (CCl4)-induced liver-injured rats. In order to investigate the RU effects, the rats were administered RU at a dose of 10 or 100 mg/kg of body weight for 10 days before induction of the liver injury by oral administration of CCl4 (125 mg/kg body weight). (i) Pretreatment with RU caused a significant decrease in serum lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities that were increased by the administration of CCl4. (ii) RU pretreatment (100 mg/kg) increased 5-bromo-2′-deoxyuridine incorporation at 48 h after CCl4 treatment in hepatocytes. (iii) Histological hematoxylin and eosin staining of the liver showed that RU pretreatment reduced the damage induced by CCl4 administration. (iv) Reverse transcriptase PCR analysis showed RU retreatment caused a transient but significant increase in hepatocyte growth factor (HGF) and a sustained and significant increase in insulin-like growth factor-I (IGF-I) gene expression in hepatocytes injured by CCl4 treatment. From these results, we conclude that oral pre-administration of RU was effective to suppress liver injury induced by the subsequent oral CCl4 administration, and RU-induced increase in IGF-I and HGF gene expression may be, even in part, involved in biological actions of RU in rats.
[Show abstract][Hide abstract]ABSTRACT: Oral administration of Lactobacillus reuteri CP3012 or Lactobacillus acidophilus L-92 for 60 days in rats that were previously administered 3,3',4,4',5-pentachlorobiphenyl (PCB126) orally at a dose of 100 mu g/kg of body weight resulted in a significant decrease in hepatic bioaccumulation of PCB126 ( p < 0.05), with levels of 30.7 +/- 3.7 ng/g and 92.6 +/- 25.0 ng/g of liver tissue, respectively, compared with 133.1 +/- 12.7 ng/g of liver tissue in the controls. The electron paramagnetic resonance signal level of the liver PCB126-specific g = 2.49 species in rats administered L. reuteri CP3012 decreased significantly (p < 0.05). Both the bile acid concentration in the feces and total stool output increased significantly following administration of lactobacilli ( p < 0.05); however, adsorption of PCB126 onto the bacterial cells was not observed. These results suggest that these bacteria inhibit reabsorption of PCB126 with bile acid by blocking enterohepatic circulation through absorbing and/or deconjugating the bile acids in the intestinal tract and by promoting excretion of bile acids from the body, thus reducing PCB126 accumulation in the liver.
Article · Jul 2014 · Food Science and Technology Research
[Show abstract][Hide abstract]ABSTRACT: Adlercreutzia equolifaciens DSM 19450T was isolated from human feces and is able to metabolize daidzeins (soybean isoflavonoids) to equol. Here, we report the finished
and annotated genome sequence of this organism.
Full-text Article · Aug 2013 · Genome Announcements
[Show abstract][Hide abstract]ABSTRACT: Adipose tissue-derived stem cells (ADSCs) isolated from adult tissue have pluripotent differentiation and self-renewal capability. The tissue source of ADSCs can be obtained in large quantities and with low risks, thus highlighting the advantages of ADSCs in clinical applications. Valproic acid (VPA) is a widely used antiepileptic drug, which has recently been reported to affect ADSC differentiation in mice and rats; however, few studies have been performed on dogs. We aimed to examine the in vitro effect of VPA on canine ADSCs. Three days of pretreatment with VPA decreased the proliferation of ADSCs in a dose-dependent manner; VPA concentrations of 2 mM and above inhibited the proliferation of ADSCs. In parallel, VPA increased p16 and p21 mRNA expression, suggesting that VPA attenuated the proliferative activity of ADSCs by activating p16 and p21. Furthermore, the effects of VPA on adipogenic, osteogenic or neurogenic differentiation were investigated morphologically. VPA pretreatment markedly promoted neurogenic differentiation, but suppressed the accumulation of lipid droplets and calcium depositions. These modifications of ADSCs by VPA were associated with a particular gene expression profile, viz., an increase in neuronal markers, that is, NSE, TUBB3 and MAP2, a decrease in the adipogenic marker, LPL, but no changes in osteogenic markers, as estimated by reverse transcription-PCR analysis. These results suggested that VPA is a specific inducer of neurogenic differentiation of canine ADSCs and is a useful tool for studying the interaction between chromatin structure and cell fate determination.
Full-text Article · Aug 2013 · Journal of Veterinary Medical Science
[Show abstract][Hide abstract]ABSTRACT: Aqueous extracts of Rhizopus oryzae (Aq-ROU) have a broad range of physiological activity. Here we identified a new physiological effect of Aq-ROU in rat hepatocyte cell line RLN-10. Aq-ROU induced the accumulation of nitrite, a stable metabolite nitric oxide (NO), in cell culture medium and induced potent diaminofluorescein-FM diacetate staining in the cells. Real-time reverse transcriptase (RT)-PCR analysis showed marked inducible NO synthase gene expression. Additionally, markedly enhanced expression of p22(phox) and temporally increased expression of NADPH oxidase1 indicated that superoxide was produced. Nuclear translocation of nuclear factor-kappa (NF-κ) B p65 increased remarkably following Aq-ROU and following lipopolysaccharide treatment, a potent activator of NF-κB. Ammonium pyrrolidine-1-carbodithioate, an inhibitor of NF-κB, inhibited NO production following Aq-ROU treatment. Our data indicate that Aq-ROU induces NO production and potentially the production of superoxide, which may contribute to the broad range of physiological effects observed for Aq-ROU ingested by animals.
Article · Jul 2013 · Bioscience Biotechnology and Biochemistry
[Show abstract][Hide abstract]ABSTRACT: We investigated the role of nitric oxide (NO) in vascular endothelial growth factor (VEGF) expression in the rat placenta. A nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine-methyl ester (L-NAME), was constantly infused into pregnant rats 6-24 h before sacrifice on gestational day (GD) 15.5. NO production declined to about 15% of the control level as monitored by NO trapping and electron paramagnetic resonance spectroscopy. VEGF mRNA expression was temporally decreased by L-NAME, but recovered to normal levels after 24 h of treatment, whereas hypoxia inducible factor (HIF)-1α and induced NOS (iNOS) expression increased. VEGF expression decreased significantly in placental explants after 6 h of co-treatment with L-NAME and lipopolysaccharide, an iNOS inducer. Our data indicate that NO induce VEGF expression in vivo and in vitro in the rat placenta, suggesting that peaked NO production was maintained by a reciprocal relationship between NO and VEGF via HIF-1α.
Full-text Article · May 2013 · Bioscience Biotechnology and Biochemistry
[Show abstract][Hide abstract]ABSTRACT: Comparative analysis by functional categories of the gene repertoires of L. garvieae and L. lactis. The number of genes on each genome within each functional category as defined by the COG database is shown.
[Show abstract][Hide abstract]ABSTRACT: mRNA expression analysis of the genes in the capsule gene cluster of L. garvieae Lg2 and Lg2-S. Total RNA from L. garvieae Lg2 and Lg2-S was extracted and was subjected to semi-quantitative RT-PCR. The PCR products were electrophoresed, stained, and photographed.
[Show abstract][Hide abstract]ABSTRACT: Lactococcus garvieae causes fatal haemorrhagic septicaemia in fish such as yellowtail. The comparative analysis of genomes of a virulent strain Lg2 and a non-virulent strain ATCC 49156 of L. garvieae revealed that the two strains shared a high degree of sequence identity, but Lg2 had a 16.5-kb capsule gene cluster that is absent in ATCC 49156. The capsule gene cluster was composed of 15 genes, of which eight genes are highly conserved with those in exopolysaccharide biosynthesis gene cluster often found in Lactococcus lactis strains. Sequence analysis of the capsule gene cluster in the less virulent strain L. garvieae Lg2-S, Lg2-derived strain, showed that two conserved genes were disrupted by a single base pair deletion, respectively. These results strongly suggest that the capsule is crucial for virulence of Lg2. The capsule gene cluster of Lg2 may be a genomic island from several features such as the presence of insertion sequences flanked on both ends, different GC content from the chromosomal average, integration into the locus syntenic to other lactococcal genome sequences, and distribution in human gut microbiomes. The analysis also predicted other potential virulence factors such as haemolysin. The present study provides new insights into understanding of the virulence mechanisms of L. garvieae in fish.
[Show abstract][Hide abstract]ABSTRACT: Comparisons of the genomic location of the capsule gene cluster in L. garvieae Lg2 and the corresponding location of L. garvieae ATCC 49156 and L. lactis strains. Genes and their orientations are depicted with arrows. Red indicates the capsule gene cluster including IS982 elements. Gray bars indicate orthologous regions.
[Show abstract][Hide abstract]ABSTRACT: Previous studies have shown that the dilating effect of nitric oxide (NO) on the fetal ductus arteriosus (DA) is age dependent and more marked in the premature stages in rats, but the factors that mediate this effect are poorly understood. The purpose of this study is to determine the changes in the expression of NO synthase (NOS) mRNA in the fetal DA and to examine the effect of an 11-beta-hydroxylase inhibitor of corticosterone synthesis, namely, metyrapone, on NOS expression. NOS 3 mRNA expression was observed in 17.5-day-old rat fetuses; thereafter, its level significantly increased and reached its peak on day 19.5 and then decreased until the end of the gestation period (day 21.5). To inhibit corticosterone synthesis, a constant infusion of metyrapone was administered to rats; this significantly decreased the fetal plasma corticosterone concentration as well as NOS 3 mRNA expression in the DA in a time-dependent manner. These results indicate that NO is generated by NOS 3 in the DA and that the age-dependant expression of NOS 3 in the premature DA is attributable to corticosterone-associated activity.
Article · May 2010 · Journal of Veterinary Medical Science
[Show abstract][Hide abstract]ABSTRACT: We previously reported that nitric oxide (NO) is first detected in the uterus of a pregnant rat on gestational day 13.5 (GD13.5) and that NO levels peak on GD17.5. In addition, NO production in the uterus is mainly derived from the decidua and not the myometrium. The aim of the present study was to reveal the role of NO that peaked on GD17.5 of gestation in the decidua. To inhibit NO production, pregnant rats were continuously administered by an nitric oxide synthase inhibitor, N(G)-nitro-L-arginine-methyl ester (L-NAME) for 48 h. In the control group, saline was infused instead of L-NAME. After treatment, the decidua were obtained from GD13.5, GD17.5 and GD21.5 rats. Apoptosis and activated caspase-3-positive cells were observed by transferase-mediated dUTP nick-end labeling (TUNEL) assay and immunohistochemistry, respectively. The caspase-3 enzyme activity was also measured in the cell lysate from the decidua. The numbers of TUNEL-positive cells and activated caspase-3-positive cells each increased and the amount of caspase-3 activity also increased significantly in rats on GD17.5 than in rats in the control group, but no changes were observed in rats on GD13.5 and GD21.5. Furthermore, enzyme activity regarding the initiator caspases, caspase-8 and -9, upstream factors for caspase-3 in the caspase cascade, was measured simultaneously on GD17.5 under the same treatment. Caspase-8 and -9 enzyme activities increased significantly in the control group; an increment of caspase-8 activity was especially prominent. The present results indicate that an inhibitor of NO production caused apoptosis through typical apoptotic signals in the decidua on GD17.5, suggesting that an NO peak in the decidua is essential to cell survival and the maintenance of uterine formation.
Article · Apr 2010 · Experimental Biology and Medicine
[Show abstract][Hide abstract]ABSTRACT: Nitric oxide (NO) has been reported as a key mediator in enhancing hepatocyte proliferation during liver regeneration. Juvenile hepatocytes have a strong ability to proliferate while still in their undifferentiated state but the mechanism of NO production and its contribution to hepatocyte proliferation are not yet fully understood. The present study was designed to investigate NO production in the normal liver and its contribution to hepatocyte proliferation in juvenile rats. Endogenous NO production was evaluated quantitatively using a spin trap followed by electron paramagnetic resonance spectroscopy with the Fe-N, N-diethyldithiocarbamate complex as an NO-trapping reagent in the rat liver. NO production in the liver significantly peaked at 3 weeks after birth, but NO synthase (NOS) 3 expression did not change between 2 to 5 weeks after birth, while NOS 1 and NOS 2 mRNA were not detected. Hepatocyte proliferation, measured by the incorporation of 5-bromo-2'-deoxyuridine into the DNA, was found to decline significantly when endogenous NO production was inhibited by the administration of the NOS inhibitor N(G)-nitro- (L)-arginine methyl ester. These findings indicate that endogenous NO production peaked at 3 weeks after birth and hepatocyte proliferation declined significantly when NO production was inhibited. Thus, this study provides a novel insight into the contribution of NO to hepatic growth and liver maturation in juveniles.
Article · Feb 2010 · Journal of Veterinary Medical Science