• Asian-Australasian Journal of Animal Sciences
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• v.31 no.6
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• pp.919-925
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• 2018

Objective: The performance, health, and behaviour of cattle can be strongly affected by climate. The objective of this study was to determine the effect of heat stress on blood parameters, blood proteins (haptoglobin [Hp]; heat shock protein 70 [HSP70]), rectal temperature (RT), heart rate (HR) and rumination time in Korean native beef calves. Methods: Thirty-two Korean native beef calves were randomly assigned to 8 groups with 4 animals per group. They were kept in environmental condition with temperature-humidity index (THI) ranging from 70.01 to 87.72 in temperature-humidity controlled chamber for 7 days. Results: Their HR, RT, and serum cortisol and HSP70 levels were increased (p<0.05) in high THI compared to those at low THI. But, serum Hp level was decreased (p<0.05) in high THI compared to these at low THI. In addition, HR, RT, serum cortisol and HSP70 were positively correlated with THI ($R^2=0.8368$, p<0.01; $R^2=0.6162$, p<0.01; $R^2=0.581$, p<0.01; $R^2=0.2241$, p = 0.0062, respectively). There was also positive association between HR and cortisol ($R^2=0.4697$, p<0.01). Similarly, RT and cortisol were positively associated ($R^2=0.4581$, p<0.01). But, THI and HR were negatively correlated with Hp ($R^2=0.2157$, p = 0.02; $R^2=0.3362$, p = 0.003). Hematology and metabolites results were different among treatment groups. Standing position was higher (p<0.05) in the high THI group compared to that in the low THI group. Conclusion: Based on these results, it can be concluded that HR, RT, blood parameters (Cortisol, HSP70, Hp) and standing position are closely associated with heat stress. These parameters can be consolidated to develop THI chart for Korean native beef calves.

• Journal of Life Science
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• v.23 no.4
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• pp.586-594
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• 2013

Post-translational O-GlcNAc modification (O-GlcNAcylation) of serine or threonine is a new protein modulation mechanism. In contrast to the classical glycosylation, O-GlcNAcylation occurs in a one-step transfer of O-GlcNAc on both nuclear and cytoplasmic proteins. In contrast to the general consensus that O-GlcNAc is a final modification, a recent paper (J Proteome Res. 2011 10:2725-2733) showed the presence of O-GlcNAc-P on a synaptic assembly protein AP180. This finding raises a fundamental question about its prevalence. To address this question, we used proteomics to identify those proteins that were phospho-signal enriched by GlcNAc kinase (NAGK). Comparison of pDsRed2-$NAGK_{WT}$-transfected HEK293T cell extract with pDsRed2-$NAGK_{D107A}$-transfected control culture revealed 15 phospho-signal increased spots. Excluding those spots that had no detectable amount of protein expression yielded 7 spots, which were selected for ID determination. Among these, two duplicate spots (two $HSP90{\beta}$ and two ENO1 spots) were shown to be O-GlcNAcylated, two (dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P) were not known to be involved in O-GlcNAcylation, and one (heat shock protein gp96 precursor or grp94) was a glycoprotein. The increase in the phospho-levels of O-GlcNAc by NAGK strongly indicates that these proteins are phosphorylated on O-GlcNAc. Our present data support the idea that O-GlcNAc is not a terminal modification.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.335-345
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• 1996

A critical role of oxygen-derived free radicals has been implicated in ischemia/reperfusion (I/R)-induced brain damage. In this study, we have produced experimental I/R to the brains of Mongolian gerbil (Meriones unguiculatus) by a transient occlusion and release of the common carotid arteries. We have attempted to determine whether the oxidative stress is generated upon I/R and whether this oxidative stress is linked to the cell damage. Since hippocampus has been suggested as one of the most vulnerable regions of the brain to the oxidative stress, we analyzed samples from hippocampus in comparison with those from cortex. In addition, we have examined the expression of heat shock protein 70kD species (HSP70) in these regions in order to evaluate a possible role of this protein in I/R-induced brain damage. To determine whether the oxidative stress is produced upon I/R, we measured the glutathione oxidation, GSSG/ (GSH + 2xGSSG), as an index of oxidative stress. We found an increase of the glutathione oxidation primarily in hippocampus upon I/R. To determine whether this oxidative stress is linked to the cell damage, we measured the degree of lipid peroxidation upon I/R. We found an increase of lipid peroxidation in both regions. However, the magnitude of increases was greater in hippocampus than in cortex. In addition, we found that changes in both the magnitude and the temporal patterns of glutathione oxidation closely correlated with those of lipid peroxidation. Our study provides biochemical evidences that the oxidative stress is generated upon I/R and this oxidative stress is linked to the oxidative cell damage. Our study also provides evidences that the degree of oxidative stress as well as oxidative cell damage is greater in hippocampus than in cortex. We could not find difference in the basal level of HSP70 expression between hippocampus and cortex, indicating that the intrinsic vulnerability of hippocampus cannot be explained by the lower level of HSP70 expression. We did find, however, that the induction of HSP70 expression upon I/R was impaired in the hippocampus. This impairment appeared to be at the transcriptional level. These results suggest that the measurement of HSP70 induction may be employed as a useful predictor of differential cellular susceptibilities to the I/R-induced brain damage.

• Journal of Plant Biotechnology
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• v.43 no.3
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• pp.359-366
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• 2016

Tradescantia is a perennial plant in the family of Commelinaceae. It is known to be sensitive to radiation. In this study, Tradescantia BNL 4430 was irradiated with gamma radiation at doses of 50 to 1,000 mGy in a phytotron equipped with a $^{60}Co$ radiation source at Korea Atomic Energy Research Institute, Korea. At 13 days after irradiation, we extracted RNA from irradiated floral tissues for RNA-seq. Transcriptome assembly produced a total of 77, 326 unique transcripts. In plantlets exposed to 50, 250, 500, and 1000 mGy, the numbers of up-regulated genes with more than 2-fold of expression compared that in the control were 116, 222, 246, and 308, respectively. Most of the up-regulated genes induced by 50 mGy were heat shock proteins (HSPs) such as HSP 70, indicating that protein misfolding, aggregation, and translocation might have occurred during radiation stress. Similarly, highly up-regulated transcripts of the IQ-domain 6 were induced by 250 mGy, KAR-UP oxidoreductase 1 was induced by 500 mGy, and zinc transporter 1 precursor was induced by 1000 mGy. Reverse transcriptase (RT) PCR and quantitative real time PCR (qRT-PCR) further validated the increased mRNA expression levels of selected genes, consistent with DEG analysis results. However, 2.3 to 97- fold higher expression activities were induced by different doses of radiation based on qRT-PCR results. Results on the transcriptome of Tradescantia in response to radiation might provide unique identifiers to develop in situ monitoring kit for measuring radiation exposure around radiation facilities.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.191-196
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• 2013

This study was aimed for development of a useful genes that has a transcript expressional specificity in the early embryonic stage of the silkworm, Bombyx mori. We constructed and analyzed a full-length cDNA library from silkworm's eggs which after a lapse of 2 ~ 6 hours post oviposit. A total 960 clones were randomly selected, and the 5' ends of the inserts were sequenced to generate 652 expressed sequence tags(EST). 334 unique ESTs were generated after the assembly of 652 ESTs. The annotation of 334 unique ESTs by BLAST search revealed that 156(47%) of the sequences represented known genes, whereas 178(53%) of the sequences has no matches in the database. Of the 156 known genes, the most abundant genes were heat shock protein hsp20.8 gene(12 times) and ubiqutin-like protein gene(11 times). The functional groups of these ESTs with matches in the database were constructed according to their putative molecular functions. Among thirteen functional categories, the largest groups were protein synthesis(9.6%) and cellular organization( 8.1%). Further defined studies on molecular functions and biological roles of their promoters will give us wellfined information and its application.

• Environmental Mutagens and Carcinogens
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• v.25 no.2
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• pp.51-59
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• 2005

Overexpression of HSP25 delayed cell growth, increased the level of $p21^{waf}$, reduced the levels of cyclin D1, cylcin A and cdc2, and induced radioresistance in L929 cells. We demonstrated that extracellular regulated kinase (ERK) and MAP kinase/ERK kinase (MEK) expressions as well as their activation (phospho-forms) were inhibited by hsp25 overexpression. To confirm the relationship between ERK1/2 and hsp25-mediated radioresistance, ERK1 or ERK2 cDNA was transiently transfected into the hsp25 overexpressed cells and their radioresistance was examined. HSP25-mediated radioresistance was abolished by overexpression of ERK2, but not by overexpression of ERK1. Alteration of cell cycle distribution and cell cycle related protein expressions (cyclin D, cyclin A and cdc2) by hsp25 overexpression were also recovered by ERK2 cDNA transfection. Increase in Bc1-2 protein by hsp25 gene transfection was also reduced by subsequent ERK2 cDNA-transfection. In addition, HSP25 overexpression reduced reactive oxygen species (ROS) and increased expression of manganese superoxide dismutase (MnSOD) gene. Increased activation of NF-kB (IkB degradation) was also found in hsp25-overexpressed cells. Moreover, transfection of hsp25 antisense gene abrogated all the HSP25-mediated phenomena. To further elucidate the exact relationship between MnSOD induction and NF-kB activation, dominant negative $I-kB\alpha(I-kB\alpha-DN)$ construction was transfected to HSP25 overexpressed cells. $I-kB\alpha-DN$ inhibited HSP25 mediated MnSOD gene expression. In addition, HSP25 mediated radioresistance was blocked by $I-kB\alpha-DN$ transfection. Blockage of MnSOD with antisense oligonucleotides in HSP25 overexpressed cells, prevented apoptosis and returned the ERK1/2 activation to the control level. From the above results, we suggest for the first time that reduced oxidative damage by HSP25 was due to MnSOD-mediated down regulation of ERK1/2.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.20 no.1
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• pp.29-40
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• 2010

This study was aimed at investigating the gene expression profile in basal ganglia of cadmium exposed rat based on cDNA array analysis. For cDNA array analysis, adult Sprague-Dawley male rats (350 ${\pm}$ 25 g) were intraperitoneally injected with 2.0 mg/kg body weight/day of CdCl2 (0.3 ml) for 5 days. For doserelated gene expression analysis rats were intraperitoneally injected with 0.0, 0.1, 0.3, 1.0 mg/kg body weight/day of CdCl$_2$ for 5 days. Control rats were injected with equal volume of saline. Cadmium concentration of brain was analyzed by atomic absorption spectrophotometer. For cDNA array, RNA samples were extracted from basal ganglia and reverse-transcribed in the presence of [${\alpha}$32P]-dATP. Membrane sets of the Atlas Rat 1.2 array II and Toxicology array 1.2 (Clontech, Palo Alto, CA) were hybridized with cDNA probe sets. RT-PCR was employed to validate the relative gene expression patterns obtained from the cDNA array. Northern blot hybridization methods were employed to assess the dose-related gene expression. Among the 2352 cDNAs, 671 genes were detected in both array sets and 63 genes of 38 classes showed significant (more than two fold) changes in expression. Thirty five of these genes were up-regulated and twenty eight were down-regulated in the cadmium exposed group. According to the dose-related gene expression analysis, heat shock 27 kDa protein (HSP27), neurodegeneration-associated protein 1 (Neurodap 1) genes were significantly up-regulated and melatonin receptor 1a (Mel1a), Kinesin family member 3C (KIF3C), novel kinesinrelated protein (KIF1D) genes were significantly downregulated even in the low-dose of cadmium exposed group (0.1 mg/kg body weight/day). Conclusions Sixty three genes detected in this study can give some more useful informations about the cadmium-induced neurotoxicity in the basal ganglia. As well as, HSP27, Neurodap1, Mel1a, KIF3C and KIF1D genes may be useful for the study of the cadmium-induced neurotoxicity because these genes showed dramatic changes of mRNA levels in response to the low dose of cadmium exposure.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1599-1607
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• 2004

This study examined the effects of pre-slaughter fasting, chasing stress and chiller ageing on objective meat quality, and their relations to the proteome profile of longissimus muscle using 20 male Korean native black pigs. Treatments were composed of two levels of pre-slaughter feed withdrawal, two levels of pre-slaughter stress and four chiller ageing times. A 15 min chasing stress immediately prior to slaughter significantly (p<0.05) decreased detectable levels of $\mu$-calpain activity during rigor development and chiller ageing, but did not have any direct effect on objective meat quality. On the other hand, pigs fed until the morning of slaughter resulted in significantly (p<0.05) higher hunter L* value and cooking loss than those which received an 18 h feed withdrawal prior to slaughter. Cooking loss and hunter L* value were constant during 7 d of chiller ageing, followed by significant increases at 14 d. The fed animals showed a significantly (p<0.05) higher hunter a* value at both 3 and 7 d, while the other group maintained a stable redness for 7 d. WB-shear force was not affected by the pre-slaughter treatments, but had significant (p<0.05) linear reduction from 1 to 7 d. A gelbased proteome analysis was performed on selected animals for low and high hunter L* values at 1 d. Ten and five spots had greater than two-fold spot densities for the low and high hunter L* groups, respectively. The ten spots included chain A, deoxyribounclease I complex with actin, heat shock protein 27 kDa, a protein similar to cardiac $Ca^{2+}$ release channel, and myosin heavy chain, while the five spots included chain A aldehyde dehydrogenase, glycerol-3 phosphate dehydrogenase, and hemoglobin alpha chain. In general, feeding until the morning of slaughter resulted in more desirable meat color, but appeared to reduce palatability due to increased cooking loss. Proteome analysis demonstrated that various proteins were concomitantly involved in the determination of final meat color. The most noticeable observation in the current study was that various isoforms for a particular protein differed in degradation and/or expression rate depending on meat quality.

• Journal of Life Science
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• v.21 no.12
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• pp.1726-1731
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• 2011

Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

• Physical Therapy Korea
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• v.13 no.3
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• pp.57-66
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• 2006

Ischemic stroke results from a transient or permanent reduction in cerebral blood flow that is restricted to the territory of a major brain artery. Thus, this study was performed to examine (1) the effects of swimming exercise on the improvement of muscle atrophy, and (2) exercise and HSP 70 expression in an ischemic stroke model induced by middle cerebral artery occlusion. The results of this study were as follows: One week after ischemic stroke was induced, changes appeared in the muscle weight of the gastrocnemius muscle due to muscle atrophy in the affected side. Group II showed statistically significant difference from group III eight weeks after ischemic stroke was induced. (p<.05). One week and eight weeks after ischemic stroke was induced there was significant decrease in the relative muscle weight of the gastrocnemius muscle in each group except Group IV, while there was statistically significant increase in group II eight weeks after ischemic stroke was induced, compared to group III (p<.05). For neurologic exercise behavior tests, Group II generally had the highest score, compared to other groups. In immunohistochemical observations, Group II showed a decrease in HSP 70. The above results suggest that swimming exercise improved muscle atrophy, changed the HSP 70 expression of ischemic stroke in rats, and contributed to the improvement of exercise function.