• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1933-1942
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• 2016

A novel ${\alpha}-glucoside$ hydrolase (named PspAG97A) from glycoside hydrolase family 97 (GH97) was cloned from the deep-sea bacterium Pseudoalteromonas sp. K8, which was screened from the sediment of Kongsfjorden. Sequence analysis showed that PspAG97A belonged to GH97, and shared 41% sequence identity with the characterized ${\alpha}-glucoside$ BtGH97a. PspAG97A possessed three key catalytically related glutamate residues. Mutation of the glutamate residues indicated that PspAG97A belonged to the inverting subfamily of GH97. PspAG97A showed significant reversibility against changes in salt concentration. It exhibited halophilic ability and improved thermostability in NaCl solution, with maximal activity at 1.0 M NaCl/KCl, and retained more than 80% activity at NaCl concentrations ranging from 0.8 to 2.0 M for over 50 h. Furthermore, PspAG97A hydrolyzed not only ${\alpha}-1,4-glucosidic$ linkage, but also ${\alpha}-1,6-$ and ${\alpha}-1,2-glucosidic$ linkages. Interestingly, PspAG97A possessed high catalytic efficiency for long-chain substrates with ${\alpha}-1,6-linkage$. These characteristics are clearly different from other known ${\alpha}-glucoside$ hydrolases in GH97, implying that PspAG97A is a unique ${\alpha}-glucoside$ hydrolase of GH97.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.1
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• pp.147-154
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• 2014

This review focuses on modulation of growth hormone (GH) and its downstream actions on periparturient dairy cows undergoing physiological and metabolic adaptations. During the periparturient period, cows experience a negative energy balance implicating that the feed intake does not meet the total energy demand for the onset of lactation. To regulate this metabolic condition, key hormones of somatotropic axis such as GH, IGF-I and insulin must coordinate adaptations required for the preservation of metabolic homeostasis. The hepatic GHR1A transcript and GHR protein are reduced at parturition, but recovers on postpartum. However, plasma IGF-I concentration remains low even though hepatic abundance of the GHR and IGF-I mRNA return to pre-calving value. This might be caused by alternation in IGFBPs and ALS genes, which consequently affect the plasma IGF-I stability. Plasma insulin level declines in a parallel manner with the decrease in plasma IGF-I after parturition. Increased GH stimulates the lipolytic effects and hepatic glucose synthesis to meet the energy requirement for mammary lactose synthesis, suggesting that GH antagonizes insulin-dependent glucose uptake and attenuates insulin action to decrease gluconeogenesis.

• Journal of Nutrition and Health
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• v.30 no.6
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• pp.614-621
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• 1997

The experiments were performed to investigate the effects of protein and protein hydrolysates on lipid metabolism in the hyperlipidemic/hypercholesterolemic rat model induced by feeding fat-enriched diet. In Except 1 male rats were fed four semi-purified high fat and cholesterol diets that contained different nitrogen source, casein(C), casein hydrolysate(CH), corn gluten(G) and corn gluten hydrolysate(GH), for 6 weeks. In Expt. 2 rats were fed high fat and cholesterol diet for 4 weeks to induce hyperlipidemia and hypercholesterolemia. Then the rats were divided into 4 groups and were fed the four kinds of above experimental diets for 4 weeks consecutively. The contents of total lipid , cholesterol and triglyceride in blood, liver and feces were determined. Serum lipid concentrations of CH, G and GH were significantly lower than that of C. Serum cholesterol concentrations of hydrolysate groups(CH and GH) were significantly lower than those of intact protein groups(C and G). Serum HDL -cholesterol concentration tended to increase by hydrolysate intake. The total lipid, cholesterol contents in liver showed similarity results as above. Fecal lipid excretions of CH, G, and GH groups were significantly higher than that of C group. These results indicate that hypolipidemic and /or hypocholesterolemic effect of corn gluten or protein hydrolysates were detected in the process of inducing hyperlipidemia by high-fat and cholesterol diet or after inducing hyperlipidemia.

• Fisheries and Aquatic Sciences
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• v.1 no.1
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• pp.19-23
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• 1998

Effects of pituitary and thyroid hormones on estradiol-induced vitellogenin (VTG) induction were electrophoretically examined in primary hepatocyte cultures of rainbow trout. Hepatocytes were precultured for 2 days and then estradiol-17 $\beta$ $(E_2,\;2 \times 10^{-6}M)$>, triiodothyronine $(T_3,\;10^{-8}-10^{-6}M)$, bovine growth hormone (bGH, 10-100 ng/ml), ovine prolactin (oPRL, 100-500 ng/ml), and pituitary extract (PE) of rainbow trout (0.75PE/dish) were added to the incubation medium. The hepatocytes were cultured for 7 more days. The addition of oPRL to the incubation medium was not effective in increasing VTG production at any concentrations. The addition of PE to the incubation medium with $E_2$ was not effective in increasing VTG production. The addition of bGH to the incubation medium with $E_2$ was not effective in increasing the rate of VTG production at concentrations of 10-50 ng/ml. However, a higher concentration of bGH, 100 ng/ml, increased VTG production. The various concentrations of $T_3$ were ineffective in stimulating VTG production. These results suggest that GH could be one of stimulus factors for VTG production in rainbow trout.

• BMB Reports
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• v.40 no.5
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• pp.723-730
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• 2007

Kinesin is an ATP-driven microtubule motor protein that plays important roles in control of microtubule dynamics, intracellular transport, cell division and signal transduction. The kinesin superfamily is composed of numerous members that are classified into 14 subfamilies. Animal kinesins have been well characterized. In contrast, plant kinesins have not yet to be characterized adequately. Here, a novel plant-specific kinesin gene, GhKCH2, has been cloned from cotton (Gossypium hirsutum) fibers and biochemically identified by prokaryotic expression, affinity purification, ATPase activity assay and microtubule-binding analysis. The putative motor domain of GhKCH2, $M_{396-734}$ corresponding to amino acids Q396-N734 was fused with 6$\times$His-tag, soluble-expressed in E. coli and affinity-purified in a large amount. The biochemical analysis demonstrated that the basal ATPase activity of $M_{396-734}$ is not activated by $Ca^{2+}$, but stimulated 30-fold max by microtubules. The enzymatic activation is microtubule-concentration-dependent, and the concentration of microtubules that corresponds to half-maximum activation was about 11 ${\mu}M$, much higher than that of other kinesins reported. The cosedimentation assay indicated that $M_{396-734}$ could bind to microtubules in vitro whenever the nucleotide AMP-PNP is present or absent. As a plant-specific microtubule-dependent kinesin with a lower microtubule-affinity and a nucleotide-independent microtubule-binding ability, cotton GhKCH2 might be involved in the function of microtubules during the deposition of cellulose microfibrils in fibers or the formation of cell wall.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.12
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• pp.1696-1701
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• 2017

Objective: In the present study, we examined whether the post-prandial reduction in plasma growth hormone (GH) levels is related to the increase in plasma insulin levels in ruminants. Methods: We performed two experiments: intravenous bolus injection of insulin (0.2 IU/kg body weight) or glucose (1.0 mmol/kg body weight) was administered to increase the plasma insulin levels in male Shiba goats. Results: In the insulin injection experiment, significant (p<0.05) increase in GH concentrations was observed, 15 to 20 min after the injection; it was accompanied with a significant (p<0.01) increase in cortisol concentrations at 45 to 90 min, when compared to the concentrations in the saline-injected controls. The glucose injection significantly (p<0.05) increased the plasma GH concentration at 20 to 45 min; this was not accompanied by significantly higher cortisol concentrations than were observed for the saline-injected control. Hypoglycemia induced by the insulin injection, which causes the excitation of the adrenal cortex, might be involved in the increase in insulin levels. Conclusion: Based on these results, we conclude that post-prandial increases in plasma insulin or glucose levels do not induce a decrease in GH concentration after feeding in the ruminants.

• KSBB Journal
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• v.13 no.2
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• pp.133-140
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• 1998

Using rlFN-$\alpha$ and rhGH as the model proteins, the refolding performances of the published processes were evaluated and compared. Key engineering parameters such as the type of denaturant and this concentration, protein concentration in the refolding buffer, and pH and ionic strength of the buffer were experimentally investigated. Furthermore, the role of a co-solvent of surfactant type in aggregation reduction was also studied. Of the denaturants tested (8M urea, 6M guanidine HCI, 0.5% SDS), SDS at alkaline pH (9.5) and ambient temperature gave the highest recovery yield. The SDS process was effective in the refolding of observed where dissolution proceeded better under lower strength (10 mM) but aggregation was suppressed under higher strength (>50 mM.) When PEG-4000 and/or Tween were added as co-solvent or refolding-enhancing additive, 1.6-2 times higher yield was realized. The‘masking’of the hyrophobic patches located on the surface of the protein with the surfactant molecules was believed to be responsible for the considerable reduction in aggregation during refolding.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.2
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• pp.81-90
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• 2000

The types of $K^+$ channel which determine the pattern of spontaneous action potential (SAP) were investigated using whole-cell variation of patch clamp techniques under current- and voltage-clamp recording conditions in rat clonal pituitary $GH_3$ cells. Heterogeneous pattern of SAP activities was changed into more regular mode with elongation of activity duration and afterhyperpolarization by treatment of TEA (10 mM). Under this condition, exposure of the class III antiarrhythmic agent E-4031 $(5\;{\mu}M)$ to $GH_3$ cells hardly affected SAP activities. On the other hand, the main $GH_3$ stimulator thyrotropin-releasing hormone (TRH) still produced its dual effects (transient hyperpolarization and later increase in SAP frequency) in the presence of TEA. However, addition of $BaCl_2$ (2 mM) in the presence of TEA completely blocked SAP repolarization process and produced membrane depolarization in all tested cells. This effect was observed even in TEA-untreated cells and was not mimicked by higher concentration of TEA (30 mM). Also this barium-induced membrane depolarization effect was still observed after L-type $Ca^{2+}$ channel was blocked by nicardipine $(10\;{\mu}M).$ These results suggest that barium-sensitive current is important in SAP repolarization process and barium itself may have some depolarizing effect in $GH_3$ cells.

• Korean Journal of Medicinal Crop Science
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• v.11 no.4
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• pp.289-297
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• 2003

Using Agrobacterium-me야ated transformation method the auxin-regulated cotton GST (Gh-5) constructs were used to transform Rehmannia glutinosa L. The PCR analysis was conducted to verify transgenicity. Based on the PCR analysis, there was verified that the 988 bp DNA band had showed in transgenic plant genomes in PCR anaJysis using Gh5-1 and Gh5-2 primers. The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of Chinese foxglove (Rehmannia glutinosa L.) were investigated. Factors such as cocultivation period, use of acetosyringone, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. In vitro regeneration, the number of leaves, shoot lengths and numbers on MS medium were superior to on B5 and WPM medium, and the shoot formation rate was highest level of 95% in cultured base part containing leaf stalk. Addition of acetosyringone at concentration of $200{\mu}M$ to cocultivation medium and 3-day of cocultivation improved transformation frequencies. Exposure of explants to darkness for 4 weeks on selection medium resulted in further increased the regeneration frequency of transgenic shoots. In PCR analysis, the amplified fragments of Gh5 gene were detected (988 bp), and GST-expressing transgenic R. glutinosa L. plants had approximately three-fold higher activity in leaf extracts compared with control plant.

• Toxicological Research
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• v.22 no.4
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• pp.307-313
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• 2006

The development of in vitro assays has been recommended to screening and testing the potential endocrine disruptors (EDs). These assay systems focus only on identifying the estrogenic or antiestrogenic activity of EDs, whereas a few studies have been carried out to screen the thyroid hormone (TH) disruptors. The aim of this study was to evaluate a test system to detect TH disruptors using rat pituitary tumor $GH_3$ cells. The test system is based on the TH-dependent increase in growth rate. As expected, L-3,5,3-triiodothyronine ($(T_3)$ markedly induced a morphological change in $GH_3$ cells from flattened fibroblastic types to rounded or spindle-shaped types. $T_3$ stimulated $GH_3$ cell growth in a dose-dependent manner with the maximum growth-stimulating effect being observed at a concentration $1{\times}10^9M$. In addition, $T_3$ increased the release of growth hormone and prolactin into the medium of the $GH_3$ cells culture. Using this assay system, the TH-disrupting activities of bisphenol A (BPA) and its related compounds were examined. BPA, dimethy/bisphenol A (DMBPA), and TCI-EP significantly enhanced the growth of $GH_3$ cells in the range of $1{\times}10^{-5}M\;to\;1{\times}10^{-6}M$ concentrations. In conclusion, this in vitro assay system might be useful for identifying potential TH disruptors. However, this method will require further evaluation and standardization before it can be used as a broad-based screening tool.