• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.124-124
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• 2017

Cadmium (Cd) is of particular concern because of its widespread occurrence and high toxicity and may cause serious morpho-physiological and molecular abnormalities in in plants. The present study was performed to explore Cd-induced morpho-physiological alterations and their potentiality associated mechanisms in Sorghum bicolor leaves at the protein level. Ten-day-old sorghum seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CdCl_2$, and different morpho-physiological responses were recorded. The effects of Cd exposure on protein expression patterns in S. bicolor were investigated using two-dimensional gel electrophoresis (2-DE) in samples derived from the leaves of both control and Cd-treated seedlings. The observed morphological changes revealed that the plants treated with Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cd was markedly increased by treatment with Cd, and the amount of Cd taken up by the shoots was significantly and directly correlated with the applied level of Cd. Using the 2-DE method, a total of 33 differentially expressed protein spots were analyzed using MALDI-TOF/TOF MS. Of these, treatment with Cd resulted in significant increases in 15 proteins and decreases in 18 proteins. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. Proteomic results revealed that Cd stress had an inhibitory effect on carbon fixation, ATP production and the regulation of protein synthesis. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. Our study provides insights into the integrated molecular mechanisms involved in response to Cd and the effects of Cd on the growth and physiological characteristics of sorghum seedlings. The upregulation of these stress-related genes may be candidates for further research and use in genetic manipulation of sorghum tolerance to Cd stress.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.141-141
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• 2017

The ubiquitin-proteasome pathway is the major regulatory mechanism in a number of cellular processes for selective degradation of proteins and involves three steps: (1) ATP dependent activation of ubiquitin by E1 enzyme, (2) transfer of activated ubiquitin to E2 and (3) transfer of ubiquitin to the protein to be degraded by E3 complex. F-box proteins are subunit of SCF complex and involved in specificity for a target substrate to be degraded. F-box proteins regulate many important biological processes such as embryogenesis, floral development, plant growth and development, biotic and abiotic stress, hormonal responses and senescence. However, little is known about the F-box genes in wheat. The draft genome sequence of wheat (IWGSC Reference Sequence v1.0 assembly) used to analysis a genome-wide survey of the F-box gene family in wheat. The Hidden Markov Model (HMM) profiles of F-box (PF00646), F-box-like (PF12937), F-box-like 2 (PF13013), FBA (PF04300), FBA_1 (PF07734), FBA_2 (PF07735), FBA_3 (PF08268) and FBD (PF08387) domains were downloaded from Pfam database were searched against IWGSC Reference Sequence v1.0 assembly. RNA-seq paired-end libraries from different stages of wheat, such as stages of seedling, tillering, booting, day after flowering (DAF) 1, DAF 10, DAF 20, and DAF 30 were conducted and sequenced by Illumina HiSeq2000 for expression analysis of F-box protein genes. Basic analysis including Hisat, HTseq, DEseq, gene ontology analysis and KEGG mapping were conducted for differentially expressed gene analysis and their annotation mappings of DEGs from various stages. About 950 F-box domain proteins identified by Pfam were mapped to wheat reference genome sequence by blastX (e-value < 0.05). Among them, more than 140 putative F-box protein genes were selected by fold changes cut-offs of > 2, significance p-value < 0.01, and FDR<0.01. Expression profiling of selected F-box protein genes were shown by heatmap analysis, and average linkage and squared Euclidean distance of putative 144 F-box protein genes by expression patterns were calculated for clustering analysis. This work may provide valuable and basic information for further investigation of protein degradation mechanism by ubiquitin proteasome system using F-box proteins during wheat development stages.

• Applied Microscopy
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• v.25 no.3
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• pp.51-62
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• 1995

It has been well known that ischemia and reperfusion injury to skeletal muscle following an acute arterial occlusion causes significant morbidity and mortality. The skeletal muscle, which contains high energy phosphate compounds, has ischemic tolerance. During the ischemia, the ATP is catalyzed to hypoxanthine anaerobically and hypoxanthine dehydrogenase is converted to xanthine oxidase. During reperfusion, the hypoxanthine is catalyzed to xanthine by xanthine oxidase under $O_2$, presence and that results in production of cytotoxic oxygen free radicals. These cytotoxic free radicals, $O_2^-,\;H_{2}O_2,\;OH^-$, are toxic and make lesions in skeletal muscle during reperfusion. The authors perform the present study to investigate the effects of allopurinol, the inhibitor of xanthine oxidase, on reperfused ischemic skeletal muscles by observing the ultrastructural changes of the muscle fibers. A total of 48 healthy Sprague-Dawley rats weighing from 200 g to 250 g were used as experimental animals. Under urethane(3.0mg/kg., IP) anesthesia, lower abdominal incision was done and the left common iliac artery were ligated by using vascular clamp for 1, 2 and 6 hours. The left rectus femoris muscles were obtained at 6 hours after the removal of vascular clamp. In the allopurinol pretreated group, 50mg/kg of allopurinol was administered once a day for 2 days and before 2 hours of ischemia. The specimens were sliced into $1mm^3$ and prepared by routine methods for electron microscopic observations. All preparations were stained with uranyl acetate and lead citrate, and then observed with Hitachi -600 transmission electron microscope. The results were as follows: 1. In 1 hour ischemia/6 hours reperfused rectus femoris muscles of rats, decreased glycogen particles and electron density of mitochondrial matrix and dilated terminal cisternae are seen. In 2 hours ischemia/6 hours repersed rectus femoris muscles of rats, mitochondria with electron lucent matrix, irregularly dilated triad and spheromembranous bodies are observed. In 6 hours ischemia/6 hours reperfused rectus femoris muscles of rats, irregularly arranged myofibrils, and many spheromembranous bodies, fat droplets and lysosome are seen. 2. In 1 hour ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol, decreased glycogen particle and dilated cisternae of sarcoplasmic reticulum and triad are observed. In 2 hours ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol decreased electron density of mitochondrial matrix and spheromembranous bodies are seen. In 6 hours ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol, mitochondria with electron lucent matrix, spheromembranous bodies and dilated cisternae of sarcoplasmic reticulum and terminal cistern are observed. The results suggest that the allopurinol attenuates the damages of the skeletal muscles of rats during ischemia and reperfusion.

• Korean Journal of Poultry Science
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• v.14 no.2
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• pp.137-143
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• 1987

This study was carried out to compare the changes in the extractability, biological activity, and solubility of myofibrillar proteins and actomyosins during storage period at 4$^{\circ}C$ and -20$^{\circ}C$in pectoral. and leg muscle of broiler meat. 1. The results obtained are as fellows ; The extractabilities of myofibrillar proteins in pectoral and leg muscle were increased gradually to 7-days during storage at 4$^{\circ}C$ and decreased slightly during frozen storage at -20$^{\circ}C$. The extractabilities of actomyosins in pectoral and legmuscle were not greatly changed during cold storage and decreased gradually during frozen storage. 2. The Ca$\^$2+/-ATP ase activities of myofibrillar proteins in the both muscles were not greatly changed to 7-days during cold storage, and in the case of frozen storage, those were highest on the 2nd week, thereafter decreased with storage period. The Ca$\^$2+/-ATPase activities of actomyosins in pectoral and leg muscle were decreased sightly only frist day during cold storage and decreased gently during frozen storage. 3. Myofibrillar proteins in the both muscles were solubilized completely at 0.20M KCl in fresh meat, at 0.25M (pectoral) and 0.30M KCl (leg) in the cold storage, and at 0.30M KCl in the frozen storage. Actomyosins of both muscles were solubilized completely at 0.40M KCl in fresh meat, cold and frozen storage.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.13-22
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• 2000

목적 : 본(本) 연구(硏究)는 배기음(排氣飮)이 인간(人間)의 장관내(腸管內)에서 산화물(酸化物)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡) 및 DNA의 손상(損傷)을 방지할수 있는지를 검증(檢證)하기 위한 실험(實驗)이다. 방법 : 배양(培養)된 인체장관(人體腸管) 세포계열(細胞系列)인 Caco-2 세포(細胞)에서 세포(細胞)의 사망(死亡)은 trypan bile의 소실정도에 의해서 평가했으며, DNA의 손상(損傷)은 double stranded DNA의 파괴정도를 측정하여 평가하였다. $H_2O_2$는 표본(標本) 산화제(酸化劑)로 사용되었다. 결과 : $H_2O_2$에 노출된 세포들의 세포사망(細胞死亡) 정도는 노출시간과 용량에 비례하여 증가하는 양상을 보였다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)되는 세포방지를 방지하였고, 0.05-1%의 농도범위에 걸쳐서는 그 효과가 용량에 비례하여 증가하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 세포손상(細胞損傷)은 catalase(hydrogen peroxide scavenger enzyme)와 deferoxamine(iron chelator)에 의해 억제되었다. 그러나 강력한 항산화제(抗酸化劑)인 DPPD는 $H_2O_2$에 의해 유발(誘發)되는 세포손상(細胞損傷)에는 영향을 주지 못했다. $H_2O_2$에 의해 유발(誘發)된 지질(脂質)의 과산화(過酸化)는 배기음(排氣飮)과 DPPD에 의해 억제되었다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상(損傷)은 배기음(排氣飮)에 의해 방지되었으며 용량에 의존하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상은 catalase와 deferoxamine에 의해 억제되었지만 DPPD는 억제시키지 못했다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)된 ATP의 소실을 회복시켰다. 이러한 실험결과 $H_2O_2$에 의해 유발(誘發)된 세포(細胞)의 손상(損傷)은 지질(脂質)의 과산화(過酸化)와는 다른 독립적인 기전에 의해 일어남을 나타낸다. 결론 : 이러한 결과들로 볼 때 Caco-2 세포(細胞)에서 배기음(排氣飮)이 항산화작용(亢酸化作用)보다는 다른 기전을 통하여 Caco-2 세포안에서 산화제(酸化劑)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡)와 DNA의 손상(損傷)을 방지할 수 있다는 것을 가리킨다. 따라서 본 연구(硏究)는 배기음(排氣飮)이 반응성산소기(反應性酸素基)에 의해 매개된 인체(人體) 위장관질환(胃腸管疾患)의 치료(治療)에 사용할 수 있을 가능성(可能性)이 있음을 제시하고 있다.

• The Journal of Korean Society of Virology
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• v.28 no.2
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• pp.175-182
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• 1998

We investigated the rate of hepatitis G virus infection among 50 patients who were not infected with the hepatitis C virus but showed symptoms of hepatitis. Viral RNA was extracted from the patients' sera and cDNA was synthesized and amplified by RT-PCR (reverse transcription-polymerase chain reaction) using random hexamer and 5 primers (470-20-1-77F, 470-20-1-211R, 470-20-1-211R-biotin, GV57-4512MF, GV57-4657MR). The amplified PCR products were confirmed by electrochemiluminescence (ECL), liquid hybridization (LH) and Southern blotting (SB). Among the 50 PCR products, by means of ECL, we found 4 samples to be positive and 5 samples to be indeterminate. The GV45-89M probe (5'-CYCGCTGRTITGGGGTGTACfGGAAGGC-3') was end-labelled with gamma-$^{32}P$ ATP and used for liquid hybridization with the PCR products. By using liquid hybridization, we detected specific bands from 4 positive sera and also from one indeterminate serum as determined by ECL. An 1.5% agarose gel electrophoresis of the 9 PCR products which were HGV positive or indeterminate as determined by ECL showed a 160bp band from 4 positive and one indeterminate serum. The 5 PCR products proved to be positive when SB was applied with the GV45-89M probe as well as when LH was applied. LH and SB were shown to have higher sensitivity and specificity than ECL. Two cases among 5 positive cases had relatively high SGOT, SGPT, ALP values when compared with other 48 cases. In summary, we confirmed hepatitis G virus infection in 5 cases among 50 Korean patients showing symptoms of viral hepatitis.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1562-1569
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• 2004

The objective of this experiment was to determine the effects of n-3 polyunsaturated fatty acids (n-3 PUFAs)-enriched diet supplemented with different levels of $\alpha$-tocopherol on the activities of hepatic lipogenic-related enzymes and the contents of liver and plasma lipid fractions in laying Tsaiya ducks. A total of 180 30-wk-old laying Tsaiya ducks, at the beginning of peak production, were allotted into 6 treatments with 3 replicates each. Ducks were fed one of the 6 experimental diets, containing 4% tallow (control), and 4% fish oil supplemented with graded levels of $\alpha$-tocopheryl acetate ($\alpha$-tocopherol) at 0, 100, 200, 300 and 400 mg/kg, respectively, for 6 wks. Feed and water were supplied ad libitum throughout the experimental period. The results indicated that the n-3 PUFAsenriched diet supplemented with different levels of $\alpha$-tocopherol did not affect (p>0.05) egg weight, feed intake, body weight change or liver and abdominal fat weights. Egg production, egg mass and feed efficiency significantly (p<0.05) improved as dietary $\alpha$-tocopherol levels increased. The activities of hepatic lipogenic-related enzymes including acetyl-CoA carboxylase (EC 6. 2. 1. 3; ACC), glucose-6-phosphate dehydrogenase (EC 1. 1. 1. 49; G-6-PDH), ATP-citrate cleavage enzyme (EC 4. 1. 3. 8; CCE), NADP-malate dehydrogenase (EC 1.1.1.40; NADP-MDH) and fatty acid synthetase (FAS) were higher (p<0.05) in birds fed with the tallow diet than in those fed with fish oil diets and increased with increasing dietary $\alpha$-tocopherol levels. None of the dietary treatments significantly affected the contents of triglyceride and total cholesterol in the liver, or total cholesterol, phospholipid and total lipid in the plasma. However, the contents of phospholipid and total lipid in the liver, and triglyceride in the plasma increased as dietary $\alpha$-tocopherol levels increased. Increasing dietary $\alpha$-tocopherol levels decreased the non-esterified fatty acid (NEFA) content in the plasma and trended to decrease the cholesterol contents in the egg yolk. The lipid metabolism of laying Tsaiya ducks was influenced not only by the dietary fat but also by the supplementation levels of $\alpha$-tocopherol.

• BMB Reports
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• v.36 no.6
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• pp.545-551
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• 2003

A cDNA of bovine brain glutamate dehydrogenase (GDH) was isolated from a cDNA library by recombinant PCR. The isolated cDNA has an open-reading frame of 1677 nucleotides, which codes for 559 amino acids. The expression of the recombinant bovine brain GDH enzyme was achieved in E. coli. BL21 (DE3) by using the pET-15b expression vector containing a T7 promoter. The recombinant GDH protein was also purified and characterized. The amino acid sequence was found 90% homologous to the human GDH. The molecular mass of the expressed GDH enzyme was estimated as 50 kDa by SDS-PAGE and Western blot using monoclonal antibodies against bovine brain GDH. The kinetic parameters of the expressed recombinant GDH enzymes were quite similar to those of the purified bovine brain GDH. The $K_m$ and $V_{max}$ values for $NAD^+$ were 0.1 mM and $1.08\;{\mu}mol/min/mg$, respectively. The catalytic activities of the recombinant GDH enzymes were inhibited by ATP in a concentration-dependent manner over the range of 10 - $100\;{\mu}M$, whereas, ADP increased the enzyme activity up to 2.3-fold. These results indicate that the recombinant-expressed bovine brain GDH that is produced has biochemical properties that are very similar to those of the purified GDH enzyme.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.5
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• pp.701-708
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• 1996

To evaluate the possibility of using a by-products of skipjack canning as a food or feedstuff, the proximate composition, total and free amino acids, total lipid composition, and nucleotide related compounds were analyzed. The crude protein was highest in dark muscle, while lipid was highest in head. The important total amino acids in by-products were founded to be glycine, glutamic acid, alanine and histidine. The important free amino acids from dark muscle and head were taurine, histidine and anserine. The amounts of histidine, anserine and carnosine in dark muscle was higher than those of cooking drips, head, and raw vicera. The major fatty acids in by-products were palmitic, stearic, oleic and docosahexaenoic acid (DHA). The inosine and hypoxanthine were important nucleotide related compounds in by-products. The results suggests that by-products from skipjack can be used as food sources and feedstuffs especially for marine fish culture.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.138-148
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• 2018

Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.