• Korean Journal of Microbiology
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• v.43 no.3
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• pp.222-226
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• 2007

Proteases are degradative enzymes which hydrolyze a peptide bond between amino acids and they are abundantly applied to commercial field. In order to screen new source of pretense, bacteria secreting extracellular pretense were isolated by enrichment culture from deep sea water samples of East Sea, Korea. A bacterium, named as HJ19, showed the best growth and the largest clear zone in plates supplemented skim milk at $30^{\circ}C$. The partial DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology identified that this strain was In genus Micrococcus. The strain HJ19 could not grow at $10^{\circ}C$ but it started growth and showed pretense activity at $20^{\circ}C$. The optimal growth was at $37^{\circ}C$ and the maximal protease activity at $30^{\circ}C$ was about 480unit/ml.

• Journal of Aquaculture
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• v.19 no.2
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• pp.77-83
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• 2006

This study was conducted to investigate changes of hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD) and Heat Shock Protein 70 (HSP70) mRNA in hemolymph, hepatopancreas and gill of abalone (Haliotis sieboldii) exposed to various water temperatures. Abalones were exposed to 10, 15, 20, 25 or $30^{\circ}C$ for 0, 6, 12, 24 or 48 hours. Survival rate of abalone was 100% at 10, 15, 20 and $25^{\circ}C$, but 0% at $30^{\circ}C$. Hemolymph counts increased at lower water temperatures (10 and $15^{\circ}C$) and decreased at $30^{\circ}C$. SOD activity decreased immediately after exposure to lower or higher water temperatures compared to the control ($20^{\circ}C$) with an exception at $30^{\circ}C$ where the activity increased. At lower temperatures, SOD activity rose high after 24 hours, but decreased again at 48 hours. At $25^{\circ}C$, it decreased compared to the control. CAT activity decreased immediately after exposure to 10 or $25^{\circ}C$ compared to the control, and then was recovered to the initial level after increment. At $15^{\circ}C$, CAT activity was high after 6 hours, and then was recovered to the initial level after increment. At $30^{\circ}C$, the activity decreased throughout the experiment. The HSP70 mRNA expression in gill increased at lower temperatures compared to the control ($20^{\circ}C$) and $25^{\circ}C$. In this study, rapid change of wale, temperature caused stress response in abalone which had been raised at $20^{\circ}C$. At molecular level, HSP70 was expressed rapidly, but antioxidant enzymes like SOD and CAT were expressed later than HSP70. At 15 and $25^{\circ}C$ of water temperatures, the HSP70, SOD and CAT expression were stable with time. However, at $30^{\circ}C$, all abalone died possibly because they could not develop resistance to high temperature.

• Parasites, Hosts and Diseases
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• v.50 no.3
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• pp.243-247
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• 2012

Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at $20^{\circ}C$, $50^{\circ}C$, and $70^{\circ}C$ in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at $20^{\circ}C$ until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at $50^{\circ}C$ and day 1 at $70^{\circ}C$. At $20^{\circ}C$, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at $50^{\circ}C$ and $70^{\circ}C$, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at $20^{\circ}C$, for 3-5 days at $50^{\circ}C$, and for 2 days at $70^{\circ}C$. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.207-216
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• 2010

Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.5
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• pp.695-701
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• 1999

The hypothalamus is the classic site of synthesis of arginine vasotocin as neurohypophyseal hormone in the chicken. However, high concentrations of arginine vasotocin were also measured in ovarian tissues by radioimmunoassay. At first, we observed specific positive signal of mRNA encoding AVT in the hypothalamus by Northern hybridization. However, we could not find any specific bands in ovarian and uterine tissues. For evidence of transcription of the arginine vasotocin gene ingonadal tissues of the chicken, this study has applied the polymerase chain reaction as a highly sensitive assay. The hypothalamus, the four largest preovulatory ovarian follicles and the shell gland (uterus) were collected at 4 h and 20 h before oviposition. The ovarian follicular tissues were separated into granulose theca interns and theca externa layers. The uterine tissues were separated into myometrium and endometrium The extracted mRNA was converted to cDNA by reverse-transcriptase using oligo-$d(T)_{15}$ primer. Then, the cDNA was amplified by Vent polymerase and arginine vasotocin specific primers. The amplification reaction was incubated by 30 cycles successively, $95^{\circ}C$, $55^{\circ}C$ and $72^{\circ}C$ earth for 1 min. Te comparisons of the mRNA levels encoding arginine vasotocin between the tissues were determined by semi-quantification methods. After amplification of the cDNA, the PCR products were detected in hypothalamus, ovarian tissues and uterine tissues. The results of semi-quantification showed that the levels of arginine vasotocin mRNA in ovarian iud uterine tissues were about from 1/50 to 1/1000 when compared to that in the hypothalamus. The very low levels of mRNA encoding arginine vasotocin in ovarian and uterine tissues probably led us to conclude that arginine vasotocin may play a role of local mediate acting autocrine and/or paracrine.

• Animal cells and systems
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• v.14 no.1
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• pp.17-23
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• 2010

We isolated warm temperature acclimation-related protein 65-kDa (Wap65) cDNA from the liver of olive flounder and investigated the mRNA expression of Wap65 and HSP70 in olive flounder exposed to osmotic (17.5, 8.75, and 4 psu) and thermal stress (25 and $30^{\circ}C$). The mRNA expression of Wap65 and HSP70 was increased by thermal stress. The mRNA expression of HSP70 was also increased by osmotic stress, whereas no significant change in Wap65 expression was detected. These results indicate that Wap65 mRNA expression occurs specifically in response to increases in water temperature, but not in response to osmotic stress. Plasma cortisol levels were also increased by osmotic and thermal stress. We also utilized the stress hormone cortisol to examine whether Wap65 expression is thermal-stress-specific. Cortisol treatment increased HSP70 mRNA expression in vitro, but had no significant effect on Wap65 mRNA expression. Thus, thermal stress, but not osmotic stress, induces Wap65 expression.

• Journal of fish pathology
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• v.26 no.2
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• pp.77-88
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• 2013

Starry flounder, Platichthys stellatus (body length $4.4{\pm}0.51cm$) that became sick during an outbreak of disease at mariculture facilities at Ulsan, Korea in August of 2012, were examined to identify the cause of the disease. Diseased fish didn't show a unique sign, but the oxidase-positive and gram negative rod was isolated from moribund fish. The bacterium was revealed as Photobacterium damselae subsp. piscicida by biochemical analysis and sequence analysis of the 16S rRNA and capsular polysaccharide (CPS) genes. The isolates (AD5) was carrying susceptible to ofloxacin and gentamycin and showed high growth value at $18^{\circ}C$ and $25^{\circ}C$ compared to four other P. damsela strains.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.151-156
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• 2002

This study was performed to obtain the thermostable $\beta$-glycosidase producing bacteria from hot spring of volcanic area at Atagawa in Japan. KNOUC 202 was selected because it showed thermostable $\beta$-glycosidase activity in sodium phosphate buffer(pH 6.8) at $70^{\circ}C$ for 4h, and it was identified. The strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment-producing. Optimum growth was at $70~72^{\circ}C$, pH 7.0~7.2, and it could grow in the presence of 3% NaCl. The main fatty acids in cell were iso-15:0 and iso-l7:0. 16S rRNA sequence of KNOUC 202 showed 99.9% similarity with that of Thermus thermophilus ATCC 27634(HB8). Based on morphological, physiological, biochemical characteristics, cellular fatty acids profile and 16S rRNA sequence analysis, KNOUC 202 was identified as Thermus thermophilus.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1116-1126
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• 2008

The objective of this study was to investigate the expression and localization of heat shock protein 70 (Hsp70) and its mRNA in the heart, liver, and kidney of acutely heat-stressed broilers at various stressing times. Male AA broilers (n = 100) were randomly divided into 5 groups of 20 birds per group. After 30 d of adaptive feeding at ambient temperature, 80 experimental broilers were suddenly heat stressed by increasing the environmental temperature from $22{\pm}1^{\circ}C$ to $37{\pm}1^{\circ}C$. The 4 groups were heat stressed for 2, 3, 5, and 10 h, respectively. The localizations of Hsp70 protein and mRNA, determined by immunohistochemical staining and in situ hybridization, respectively, were demonstrated to be tissue dependent, implying that different tissues have differential sensibilities to heat stress. Intense Hsp70 staining was identified in the vascular endothelial cell of heart, liver and kidney, suggesting an association between expression of Hsp70 in vascular endothelial cell and functional recovery of blood vessels after heat shock treatment. Ante-mortem heat stress had a significant effect on the expression of Hsp70 protein and mRNA. The quantitation of Hsp70 protein and mRNA were both time and tissue dependent. During the exposure to heat stress, the heart, liver and kidney of broiler chickens exhibited increased amounts of Hsp70 protein and mRNA. The expression of hsp70 mRNA in the heart, liver and kidney of heat-stressed broilers increased significantly and attained the highest level after a 2-h exposure to elevated temperatures. However, significant elevations in Hsp70 protein occurred after 2, 5, and 3 h of heat stressing, respectively, indicating that the stress-induced responses vary among different tissues.

• Korean Journal of Ichthyology
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• v.22 no.1
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• pp.1-8
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• 2010

We determined oxidative stress caused by thermal stress in olive flounder Paralichthys olivaceus based on the altered-mRNA expression and enzymatic activity of two key antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), along with monitoring of several other biomarkers. When the fish were exposed to acute thermal change (from $20^{\circ}C$ to $25^{\circ}C$ and $30^{\circ}C$), the expression and activity of both enzymes were significantly higher at elevated temperatures ($25^{\circ}C$ and $30^{\circ}C$) than at $20^{\circ}C$. Lipid peroxidation (LPO) was also higher at $25^{\circ}C$ and $30^{\circ}C$ than at $20^{\circ}C$. In addition, the plasma $H_2O_2$ concentration was significantly increased by thermal stress. Furthermore, we investigated changes due to thermal stress by measuring levels of plasma alanine aminotransferase (AlaAT) and aspartate aminotrasferase (AspAT). Both were significantly increased by thermal stress. As an immune indicator, the lysozyme concentration was lower at $30^{\circ}C$ than at $20^{\circ}C$, indicating that thermal stress decreases immune function. Therefore, thermal stress could induce oxidative stress and suppress immune function and can cause physiological stress.