• 제목/요약/키워드: accessory protein

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Monocrotophos induced inhibition of the activities of testis and accessory reproductive organs in male mice

  • Patill, Saraswati B;Malashetty, Vijaykumar B
    • Advances in Traditional Medicine
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    • 제7권3호
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    • pp.304-310
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    • 2007
  • Monocrotophos was administered orally to adult male albino mice at dose level of 3.0 mg/kg body weight/day/mice for 50 days. The treatment has found to affect spermatogenesis as well as the endocrine functions of the testis as indicated by gravimetric, histopathological and biochemical changes. The treatment has caused degenerative changes in the seminiferous tubules and Leydig cells of the testis and regression of the epididymis, seminal vesicle, vas deferens, prostate, Cowper's gland and levator ani. Similarly, cauda epididymal sperm count and sperm motility have shown significant reduction. There was a significant reduction in the protein, glycogen, sialic acid, acid and alkaline phosphatase and increase in cholesterol in the testis of monocrotophos treated mice compared with the control. The causative factors for these changes due to monocrotophos administration were discussed.

Cytochemical and Immunocytochemical Study on the Cellulase Activity in the Accesssory Glands of the Digestive System of the Oriental Land Snail, Nesiohelix samarangae

  • Jeong, Kye-Heon;Lee, Yong-Seok;Shim, Yun-Bo
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.538-538
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    • 2000
  • Histochemical, cytochemical, and immmunocytochemical investigations were conducted to find out the cellulase activities in the accessory glands of the digestive system in the oriental land snail Nesiohelix samarangae under the LM, SEM, and TEM. It was found that the Type 1 and Type 3 cells out of five types of the epithelial cells of the digestive gland were labeled with the immunogold(protein-A gold) particles. Otherwise, none of epithelial cells of the mucus gland and the salivary gland were not labeled with the immunogold particles.

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동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합 (Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell)

  • 김혜영;윤보현;장효일
    • 한국미생물·생명공학회지
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    • 제39권4호
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    • pp.337-344
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    • 2011
  • 이전 연구에서, bacteriophage ${\Phi}FC1$이 Enterococcus faecalis KBL703에서 UV induction을 통해 분리 동정되었으며, ${\Phi}FC1$은 phage attachment site인 attP와 bacterial attachment site인 attB 사이에서 site-specific integration을 촉매하는 integrase를 가지고 있다는 것을 밝혀냈으며 이를 MJ1이라 명명하였다. 이 연구에서는 이를 바탕으로 MJ1에 의한 site-specific integration의 효율을 Escherichia coli와 NIH3T3 cell에서 확인 하기 위해 attP, attB, MJ1을 각각의 벡터에 삽입하였다. MJ1 인테그라제에 의한 재조합을 수행하기 위해서 기질 벡터 pABLP를 $DH5{\alpha}$에 형질전환시킨 후, LB 배지에서 $37^{\circ}C$ 1시간 배양한 후 암피실린(ampicillin)과 테트라싸이클린(tetracycline) 항생제 플레이트로 pGMJ1과 pABLP 같이 가지고 있는 colony 들을 선별하여, LacZ 유전자가 불활성화 된 흰색 콜로니 개수를 세고 통계를 낸 결과 integration의 frequency가 99% 이상인 것으로 나타났다. 또한, 실제로 재조합이 일어났는 지를 확인하기 위해서 콜로니 PCR을 수행하여 재조합의 산물인 attL 150 bp을 확인하였다. PCR 산물은 염기서열분석을 통해 정확한 site-specific integration이 일어났음을 확인하였다. MJ1에 의한 integration을 보이기 위해 attP와 attB를 가지고 있는 vector를 MJ1 expression vector와 함께 NIH3T3 cell에 cotransfection 했으며 GFP를 reporter로 사용해 그 activity를 관찰하였다. NIH3T3 cell에서 GFP의 발현을 형광 현미경을 통해 알아본 결과, MJ1에 의한 sitespecific integration이 다른 accessory protein의 도움 없이 일어난다는 것을 볼 수 있었다. 마찬가지 방법으로, attR과 attL 간의 excision을 GFP로 알아본 결과, GFP는 발현하지 않았으며, 이는 MJ1에 의한 excision이 일어나지 않았음을 보여주었다. 이와 같은 결과로 볼 때, MJ1의 host만이 아니라 넓은 범위안에서도 integration을 수행할 수 있다는 것을 보여주었다. 따라서 MJ1을 이용한 site-specific integration system의 개발은 gene therapy를 위한 gene delivery system의 구축에 있어서 좋은 시작이 될 수 있다.

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

  • Khalilpour, Akbar;Santhanam, Amutha;Lee, Chun Wei;Saadatnia, Geita;Velusamy, Nagarajan;Osman, Sabariah;Mohamad, Ahmad Munir;Noordin, Rahmah
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권3호
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    • pp.1635-1642
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    • 2013
  • Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

CTLA-4 항원의 활성 T 세포내 발현의 특성: 세포질내 단백복합체 구성분자의 동정 (Characterization of CTLA-4 Antigen Expression: Identification of Molecules Composing Intracellular CTLA-4 Multiprotein Complex)

  • 임대철;정용훈
    • IMMUNE NETWORK
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    • 제2권1호
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    • pp.35-40
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    • 2002
  • Background: CTLA-4 (Cytotoxic T Lymphocyte associated Antigen 4, CD152) has been known as a homologue of CD28, an accessory molecule providing a key costimulatory signal for successful antigen-driven activations of T lymphocyte. Most of biochemical and cell biological characteristics of the CD152 protein remain unknown while those of CD28 have been characterized in detail. Methods: In this study CD152 expression in both $CD4^+$ and $CD8^+$ PBLs was studied by using flow cytometry. And intracellular CD152 multiprotein complex was purified and used for generating antibodies recognizing proteins composing of intracellular CTLA-4 multi protein complex. Results: Level of surface expression of this molecule was peaked at 2 days of PHA stimulation in flow cytometric analysis. 40~45% of PHA blast cells were $CD152^+$ in both of two subsets at this stage and the level of expression were equivalent in both two subsets. Contrary to this surface expression, intracellular expression was peaked at day 3 and it was preferentially induced in $CD8^+$ cells and about 60% of $CD8^+$ cells were $CD152^+$ at this stage. High molecular weight (>350 kD) intacellular CD152 protein complex purified by using preparative electrophoresis were immunized into rabbits and then 3 different anti-P34PC4, anti-P34PC7 and anti-P34PC8 antibodies were obtained. Using these 3 antibodies two unknown antigens associated with intracellular CD152 multiprotein complex were found and their molecular weights were 54 kD and 75 kD, respectively. Among these, the former was present as 110 kD homodimer in non-reducing condition. Conclusion: It seemed that 34 kD intracellular CD152 molecule forms high molecular weight multiprotein complex at least with 2 proteins of 75 kD monomer and 110 kD homodimer.

Combined transcriptome and proteome analyses reveal differences in the longissimus dorsi muscle between Kazakh cattle and Xinjiang brown cattle

  • Yan, XiangMin;Wang, Jia;Li, Hongbo;Gao, Liang;Geng, Juan;Ma, Zhen;Liu, Jianming;Zhang, Jinshan;Xie, Penggui;Chen, Lei
    • Animal Bioscience
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    • 제34권9호
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    • pp.1439-1450
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    • 2021
  • Objective: With the rapid development of proteomics sequencing and RNA sequencing technology, multi-omics analysis has become a current research hotspot. Our previous study indicated that Xinjiang brown cattle have better meat quality than Kazakh cattle. In this study, Xinjiang brown cattle and Kazakh cattle were used as the research objects. Methods: Proteome sequencing and RNA sequencing technology were used to analyze the proteome and transcriptome of the longissimus dorsi muscle of the two breeds of adult steers (n = 3). Results: In this project, 22,677 transcripts and 1,874 proteins were identified through quantitative analysis of the transcriptome and proteome. By comparing the identified transcriptome and proteome, we found that 1,737 genes were identified at both the transcriptome and proteome levels. The results of the study revealed 12 differentially expressed genes and proteins: troponin I1, crystallin alpha B, cysteine, and glycine rich protein 3, phosphotriesterase-related, myosin-binding protein H, glutathione s-transferase mu 3, myosin light chain 3, nidogen 2, dihydropyrimidinase like 2, glutamate-oxaloacetic transaminase 1, receptor accessory protein 5, and aspartoacylase. We performed functional enrichment of these differentially expressed genes and proteins. The Kyoto encyclopedia of genes and genomes results showed that these differentially expressed genes and proteins are enriched in the fatty acid degradation and histidine metabolism signaling pathways. We performed parallel reaction monitoring (PRM) verification of the differentially expressed proteins, and the PRM results were consistent with the sequencing results. Conclusion: Our study provided and identified the differentially expressed genes and proteins. In addition, identifying functional genes and proteins with important breeding value will provide genetic resources and technical support for the breeding and industrialization of new genetically modified beef cattle breeds.

Effects of the Antitumor Component, F-D-P, Isolated from Elfvingia applanata on the Immune Response

  • Kim, Young-So;Ryu, Ku-Hyun;Mo, Young-Keun;Lee, Chong-Kil;Han, Seong-Sun
    • 생약학회지
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    • 제25권4호통권99호
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    • pp.348-355
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    • 1994
  • An antitumor component, F-D-P, was purified from the hot water extract of the carpophores of Elfvingia applanata by precipitation with ethanol, dialysis, and passage through a column of DEAE-cellulose ion exchange. F-D-P inhibited the growth of Sarcoma 180 in mice showing the tumor inhibition ratio of 88.3% in doses of 20 mg/kg for ten days. Chemical analysis of F-D-P showed that it was composed of polysaccharide(65.3%) and protein(6.5%0, and that the monosaccharides consisting of the polysaccharide was glucose(89.1%) and mannose(10.9%). The immunomodulatory activities of F-D-P were explored by determining its effect on the proliferation of the whole and subpopulations of lymphocytes, and on the generation of natural killer(NK) cell activity in vitro. F-D-P was mitogenic to total lymphocytes and B cells, but not to purified T cells, even in the presence of accessory cells. F-D-P did not increase NK cell activity when added to cultures of resting lymphocytes. From these results, it is clear that F-D-P modulates primarily the humoral immune responeses.

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Overexpression of a Rice Diacylglycerol Kinase Gene OsBIDK1 Enhances Disease Resistance in Transgenic Tobacco

  • Zhang, Weidong;Chen, Jie;Zhang, Huijuan;Song, Fengming
    • Molecules and Cells
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    • 제26권3호
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    • pp.258-264
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    • 2008
  • A rice diacylglycerol kinase (DGK) gene, OsBIDK1, which encodes a 499-amino acid protein, was cloned and characterized. OsBIDK1 contains a conserved DGK domain, consisting of a diacylglycerol kinase catalytic subdomain and a diacylglycerol kinase accessory subdomain. Expression of OsBIDK1 in rice seedlings was induced by treatment with benzothiadiazole (BTH), a chemical activator of the plant defense response, and by infection with Magnaporthe grisea, causal agent of blast disease. In BTH-treated rice seedlings, expression of OsBIDK1 was induced earlier and at a higher level than in water-treated control seedlings after inoculation with M. grisea. Transgenic tobacco plants that constitutively express the OsBIDK1 gene were generated and disease resistance assays showed that overexpression of OsBIDK1 in transgenic tobacco plants resulted in enhanced resistance against infection by tobacco mosaic virus and Phytophthora parasitica var. nicotianae. These results suggest that OsBIDK1 may play a role in disease resistance responses.

Morphine induced inhibition of the activities of accessory reproductive ducts in male rats

  • Londonkar, Ramesh L.;Sharangouda, Sharangouda;Patil, Saraswati B.
    • Advances in Traditional Medicine
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    • 제8권1호
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    • pp.67-72
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    • 2008
  • Adult male albino rats were treated with 0.5 mg and 0.75 mg morphine/100 g body weight intraperitoneally for 30 days. All the animals were autopsied on $31^{st}$ day. Epididymis and vas deferens were dissected out, weighed and processed for histological and biochemical studies. Morphine has caused a reduction in the weight of epididymis and vas deferens in both the doses of drug treated groups. The total cholesterol content is increased while protein, DNA and RNA contents and epididymal sperm counts are decreased. The acid phosphatase content is decreased 10.12 $\pm$ 0.11 in caput, 9.26 $\pm$ 0.30 in cauda of epididymis and in vas deferens 8.14 $\pm$ 0.15 in 0.5 mg treated groups and in 0.75 mg treated rats shows 9.52 $\pm$ 0.27 in caput, 9.14 $\pm$ 0.18 in cauda of epididymis and in vas deferens 7.84 $\pm$ 0.11is decreased, whereas alkaline phosphatase is increased. The surface epithelial cell height of these ducts is reduced and secretory activity is inhibited with the disruption of epithelial cell projections. The gravimetric and histometric changes of epididymis and vas deferens may be due to non-availability of androgens in morphine treated rats.

Spermatogenic index and hormonal profile in the rats received chromatographic fractions of ethanol extract of Crotalaria juncea L. seeds

  • Malashetty, Vijaykumar B.;Patil, Saraswati B.
    • Advances in Traditional Medicine
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    • 제6권2호
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    • pp.86-95
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    • 2006
  • The ethanol extract of the Crotalaria juncea seeds, which showed promising antispermatogenic and antiandrogenic activities in albino mice, was taken up further for the isolation of the active fractions present in it. Two fractions that were obtained from thin layer chromatography were subjected for testing to know their antispermatogenic and antiandrogenic activities. After preliminary trials the fraction I showed maximum antifertility activity at the dose level of 200 mg/kg body weight when administered orally to the rats for 50 days. The fraction I was found to affect spermatogenesis as well as the endocrine functions of the testis as indicated by gravimetric, histopathological and biochemical changes. Further this fraction has caused degenerative changes in the seminiferous tubules and Leydig cells of the testis. The accessory reproductive organs like epididymis, seminal vesicles, vas deferens, prostrate, Cowper's gland and Levator Ani muscle showed significant malfunction. Cauda epididymal sperm count and sperm motility were reduced significantly. The treatment has also resulted in increase in the cholesterol level and alkaline phosphatase activity, and decrease in protein, glycogen, sialic acid contents and acid phosphatase activity in testis. It is noteworthy that RIA studies have shown significant reduction in serum FSH, LH and testosterone. Scanning electron microscopic observations revealed abnormalities in sperm structure.