• Title/Summary/Keyword: protein tissues

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Alterations of mRNA and lncRNA profiles associated with the extracellular matrix and spermatogenesis in goats

  • Chen, Haolin;Miao Xiaomeng;Xu, Jinge;Pu, Ling;Li, Liang;Han, Yong;Mao, Fengxian;Ma, Youji
    • Animal Bioscience
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    • v.35 no.4
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    • pp.544-555
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    • 2022
  • Objective: Spermatozoa are produced within the seminiferous tubules after sexual maturity. The expression levels of mRNAs and lncRNAs in testicular tissues are different at each stage of testicular development and are closely related to formation of the extracellular matrix (ECM) and spermatogenesis. Therefore, we set out to study the expression of lncRNAs and mRNAs during the different developmental stages of the goat testis. Methods: We constructed 12 RNA libraries using testicular tissues from goats aged 3, 6, and 12 months, and studied the functions of mRNAs and lncRNAs using the gene ontogeny (GO) and Kyoto encyclopedia of genes and genomes (KEGG) databases. Relationships between differentially expressed genes (DEGs) were analyzed by lncRNA-mRNA co-expression network and protein-protein interaction network (PPI). Finally, the protein expression levels of matrix metalloproteinase 2 (MMP2), insulin-like growth factor 2 (IGF2), and insulin-like growth factor-binding protein 6 (IGFBP6) were detected by western blotting. Results: We found 23, 8, and 135 differentially expressed lncRNAs and 161, 12, and 665 differentially expressed mRNAs that were identified between 3 vs 6, 6 vs 12, and 3 vs 12 months, respectively. GO, KEGG, and PPI analyses showed that the differential genes were mainly related to the ECM. Moreover, MMP2 was a hub gene and co-expressed with the lncRNA TCONS-0002139 and TCONS-00093342. The results of quantitative reverse-transcription polymerase chain reaction verification were consistent with those of RNA-seq sequencing. The expression trends of MMP2, IGF2, and IGFBP6 protein were the same as that of mRNA, which all decreased with age. IGF2 and MMP2 were significantly different in the 3 vs 6-month-old group (p<0.05). Conclusion: These results improve our understanding of the molecular mechanisms involved in sexual maturation of the goat testis.

Translocalization of enhanced PKM2 protein into the nucleus induced by cancer upregulated gene 2 confers cancer stem cell-like phenotypes

  • Yawut, Natpaphan;Kaowinn, Sirichat;Cho, Il-Rae;Budluang, Phatcharaporn;Kim, Seonghye;Kim, Suhkmann;Youn, So Eun;Koh, Sang Seok;Chung, Young-Hwa
    • BMB Reports
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    • v.55 no.2
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    • pp.98-103
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    • 2022
  • Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

Hydroxyapatite/Chitosan Composites and its Application (Hydroxyapatite/키토산 복합재료 및 그 응용)

  • Chung, Yong-Sik
    • Proceedings of the Korean Fiber Society Conference
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    • 2002.04a
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    • pp.21-24
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    • 2002
  • Hydroxyapatite(HAp, Ca$\sub$10/(PO$_4$)$\sub$6/(OH)$_2$, is a compound with structural and chemical resemblance bone mineral and of particular importance in the field of biomaterials. In addition to the non-toxicity and high compatibility with hard and soft tissue, HAp exhibits strong affinity to host hard tissues and protein molecules. However, HAP is difficult to shape in the specific forms due to its hardness and brittleness. (omitted)

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Applications of Gelatin in Food and Biotechnology

  • Yang, Jae-Sung
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.263-268
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    • 1997
  • Gelatin is a high molecular weight polypeptide derived fro mcollgen, theprimary protein component of animal connective tissues, which include bone, skin and tendon. Gelatin is usually produced from two different sources of raw materials (skins or bones) which are processed in two ways(line or acid). According to this pretreatment, gelatin can be divided into elatin type A(acid) and B(lime). The market is essentially driven by thre demand sectors: food, harmaceuticals(capsules) and photography. Although there is some potential threat in the photography sector, the oher tow sectors are well placed for futher growth.

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Endo-sulfatase Sulf-1 Protein Expression is Down-regulated in Gastric Cancer

  • Gopal, Gopisetty;Shirley, Sundersingh;Raja, Uthandaraman Mahalinga;Rajkumar, Thangarajan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.641-646
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    • 2012
  • In our recent report on gene expression in gastric cancer we identified the endo-sulfatase Sulf-1 gene to be up-regulated in gastric tumors relative to apparently normal (AN), and paired normal (PN) gastric tissue samples. In the present report we investigate the protein expression levels of Sulf-1 gene in gastric tumors, AN and PN samples using tissue microarray (TMA) and immunohistochemistry. Expression data was collected from two sets of TMA's containing replicate sections of tissue samples. Scoring data from TMA set-1 revealed a significant difference in Sulf-1 immunoreactivity between tumors and "normals" (PN and AN) (p-value = 0.001928). Also, Sulf-1 expression in tumors was also significantly different from either PN (p-value = 0.019) or AN (p-value = 0.006) samples. Similar results were obtained from analysis of scoring data from the second set of arrays. Comparison of mRNA expression and protein expression in gastric tumor tissues revealed that in 6/20 (30%) tumor samples showed up-regulated protein expression concordant with over-expression of mRNA. However, a discord with mRNA being over-expressed relative to down regulated protein expression was observed in majority 14/20 (70%) of tumor samples. Our study indicates down regulation of Sulf-1 protein expression in gastric tumors relative to PN and AN samples which is discordant with mRNA over-expression seen in tumors.

PEP-1-GLRX1 protein exhibits anti-inflammatory effects by inhibiting the activation of MAPK and NF-κB pathways in Raw 264.7 cells

  • Shin, Min Jea;Kim, Dae Won;Choi, Yeon Joo;Cha, Hyun Ju;Lee, Sung Ho;Lee, Sunghou;Park, Jinseu;Han, Kyu Hyung;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.53 no.2
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    • pp.106-111
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    • 2020
  • Glutaredoxin 1 (GLRX1) has been recognized as an important regulator of redox signaling. Although GLRX1 plays an essential role in cell survival as an antioxidant protein, the function of GLRX1 protein in inflammatory response is still under investigation. Therefore, we wanted to know whether transduced PEP-1-GLRX1 protein inhibits lipopolysaccharide (LPS)- and 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced inflammation. In LPS-exposed Raw 264.7 cells, PEP-1-GLRX1 inhibited cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), activation of mitogen activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-κB) expression levels. In a TPA-induced mouse-ear edema model, topically applied PEP-1-GLRX1 transduced into ear tissues and significantly ameliorated ear edema. Our data reveal that PEP-1-GLRX1 attenuates inflammation in vitro and in vivo, suggesting that PEP-1-GLRX1 may be a potential therapeutic protein for inflammatory diseases.

Effect of Gyehyuldeung Treatments in Peripheral Nerve Regeneration of Rat (계혈등(鷄血藤)이 Rat의 말초신경 재생에 미치는 효과)

  • Lim, Seung-Min;Ahn, Jung-Jo;Jo, Hyun-Kyung;Yoo, Ho-Ryong;Kim, Yoon-Sik;Seol, In-Chan
    • The Journal of Internal Korean Medicine
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    • v.30 no.2
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    • pp.375-387
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    • 2009
  • Objective : Gyehyuldeung (GHD) has been widely used in oriental medicine for the treatments of cardiovascular and neurological disorders. Thus, its potential facilitatory activity on axonal regeneration was investigated in the rats. Methods: Sprague-Dawley rats were given crush injury at the sciatic nerve and the changes of axon growth after nerve injury on each nerve injury model were investigated with anti-NF-200 antibody, DiI, GAP-43 protein and Cdc2 protein Results : GHD-mediated enhancement of axonal regeneration after crush injury was measured in both qualitative and quantitative ways by immunofluorescence staining with anti-NF-200 antibody and retrograde tracing of fluorescence dye DiI. GAP-43 protein levels were elevated by GHD treatments in the distal injured sciatic nerve and DRG sensory neurons. The neurite outgrowth of DRG sensory neurons was facilitated by GHD treatment when co-cultured with Schwann cells and astrocytes prepared from injured sciatic nerves and injured spinal cord tissues, respectively. It was observed that Cdc2 protein was up-regulated in co-cultured Schwann cells or astrocytes and Cdc2 protein signals were co-localized to a certain extent with those of phospho-vimentin protein. Conclusions : These results suggest that GHD may play a facilitatory role in axonal regeneration by acting on the injured axons and adjacent non-neuronal cells. The current findings may be useful for the development of therapeutic targets through more specific explorations on molecular interactions between herbal components and endogenous factors.

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The Effects of Oxidative Stress Induced by Aluminum on Cellular Macromolecules in the Hippocampus and Cerebral Cortex of Rats (알루미늄을 투여한 흰쥐의 해마와 대뇌피질에서 Reactive Oxygen Species 생성으로 인한 생체거대분자의 산화적 손상)

  • Moon Chul-Jin;Koh Hyun-Chul;Shin In-Chul;Lee Eun-Hee;Moon Hae-Ran
    • Toxicological Research
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    • v.20 no.3
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    • pp.213-223
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    • 2004
  • This work aimed to study the effectiveness of cellular oxidative parameter (malondial-dehyde, protein carbonyl, and 8-hydroxy-2'deoxyguanosine). The experimental groups were aluminum treated rats and control rats. Aluminum treatd rats were given intraperitoneally aluminum nitrate nonahydrate ($Al^{3+}$, 0.2 mmol/kg) daily for 30 days except Sunday. Control rats were injected 1 ml of saline. After the dose, rats were decapitated and hippocampus and cerebral cortex were removed. The measured parameters were tissue malondialdehyde (MDA, index of lipid peroxidation), protein carbonyl (index of protein oxidation), 8-hydroxy-2'-deoxy-guanosine (8-OHdG, index of DNA oxidation), reduced glutathione (GSH) levels as well as glutathione reductase (GR) and catalase. AI concentrations in the tissues were also measured. All results were corrected by tissue protein levels. The results were as followed; 1. The concentrations of AI in the cortex and hippocampus were significantly higher in the AI-treated rats than in the control rats. 2. Antioxidative enzyme's activity, catalase and GR, were significantly higher in the AI-treated rats than the control rats. GSH levels were also higher in the AI-treated rats. 3. MDA, protein carbonyl, and 8-OHdG concentration of AI-treated rats were significantly higher than those of control rats. 4. The concentrations of antioxidants, and oxidative stress parameter were correlated with the concentrations of AI in hippocampus and cerebral cortex. Catalase and GR activity were also correlated with the concentration of AI. Based on these results, it can be suggested that intraperitoneally injected AI was accumulated in the brain and induced the increase of antioxidant levels and antioxidative enzyme activity. Also, the oxidative products of cellular macromolecules are significantly related to tissue AI concentration. Therefore MDA, protein carbonyl, and 8-OHdG are useful markers for oxidative stress on cellular macromolecules.

Isolation of GTP Binding Protein from Bovine Brain (소의 뇌로부터 GTP 결합단백질의 분리)

  • Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.360-368
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    • 1993
  • GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of $Go{\alpha}$ was revealed 39,000 dalton and $G{\beta}$ 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of $Go{\alpha}$ and $G{\beta}$.

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Changes of Protein Composition and Muscle Tissues in Top Shell Meat during Frozen Storage (바다방석고둥육의 동결저장중 단백질조성과 근육조직의 변화)

  • 송대진;김창용;박환준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.763-770
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    • 1993
  • To investigate the quality changes during frozen storage, top shell, Omphalius pfeifferi capenteri, was stored at -18$^{\circ}C$, -$25^{\circ}C$ and -3$0^{\circ}C$ immediately after shelling and water holding capacity, protein composition and histological features were examined with the lapsed period of the storage. During the storage period, amount of free drip was increased with higher frozen temperature and longer frozen period, but with the longer storage period, the lower water holding capacity was observed. The extractability and composition of muscle protein, sarcoplasmic protein and stroma protein were rather stable regardless of frozen temperature and frozen storage period. However, the extractability of myofibrillar protein was decreased with higher frozen temperature and longer frozen storage period. On the changes of muscle tissue structure, following points were observed. 1) In the muscle tissue structure of fresh sample, fine muscle fiber was closely distributed all over the tissue regardless of cross and longitudinal section. 2) In tissue structure under frozen state, it was observed that ice crystals apparently grew with the higher storage temperature. Empty spaces between muscle bundles which wee formed by aggregations of muscle fiber were observed after 3 months storage at -18$^{\circ}C$ . 3) Tissue structure in thawed state was restored satisfactorily after 1 month storage regardless of storage temperature. After 3 months storage at -3$0^{\circ}C$, muscle tissue was well restored, but at -18$^{\circ}C$, empty spaces were apparent due to incomplete restoration.

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