• Title/Summary/Keyword: semi-quantitative analysis

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(-)-Epigallocatechin-3-gallate Modulates the Differential Expression of Survivin Splice Variants and Protects Spermatogenesis During Testicular Torsion

  • Al-Ajmi, Nada;Al-Maghrebi, May;Renno, Waleed Mohammed
    • The Korean Journal of Physiology and Pharmacology
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    • 제17권4호
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    • pp.259-265
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    • 2013
  • The anti-apoptotic effect of (-)-epigallocatechin-3-gallate (EGCG) during unilateral testicular torsion and detorsion (TT/D) was established in our previous study. In mice, the smallest inhibitor of apoptosis, survivin, is alternatively spliced into three variants, each suggested to have a unique function. Here, we assessed how EGCG exerts its protective effect through the expression of the different survivin splice variants and determined its effect on the morphology of the seminiferous tubules during TT/D. Three mouse groups were used: sham, TT/D+vehicle and TT/D treated with EGCG. The expression of the survivin variants (140 and 40) and other apoptosis genes (p53, Bax and Bcl-2) was measured with semi-quantitative RT-PCR. Histological analysis was performed to assess DNA fragmentation, damage to spermatogenesis and morphometric changes in the seminiferous tubules. In the TT/D+vehicle group, survivin 140 expression was markedly decreased, whereas survivin 40 expression was not significantly different. In parallel, there was an increase in the mRNA level of p53 and the Bax to Bcl-2 ratio in support of apoptosis induction. Histological analyses revealed increased DNA fragmentation and increased damage to spermatogenesis associated with decreased seminiferous tubular diameter and decreased germinal epithelial cell thickness in the TT/D+vehicle group. These changes were reversed to almost sham levels upon EGCG treatment. Our data indicate that EGCG protects the testis from TT/D-induced damage by protecting the morphology of the seminiferous tubules and modulating survivin 140 expression.

1-Nitroso-2-naphthol 침전제를 사용한 연속흐름 선농축법에 의한 코발트의 정량분석 (Quantitative Analysis of Cobalt with Continuous Flow Preconcentration Using 1-Nitroso-2-naphthol as an Organic Precipitant)

  • 임갑수;박용남
    • 대한화학회지
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    • 제43권6호
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    • pp.644-650
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    • 1999
  • 본연구에서 유기 침전제를 사용하여 수용액중의 코발트를 on-line으로 연속 침전시켜 선농축하는 방법을 연구하였다. 침전제로 1-nitroso-2-naphthol을 사용하여 라인내에서 직접 침전시킨 뒤 MlBK(Methyl lsobutyl Ketone)로 용해시켜 불꽃 원자흡수 분광기로 보내어 분석하였다. 최적조건을 구하고 농축배율을 연구한 결과 대개 0.5 ppm의 코발트 시료 1.0 mL를 사용하였을 때 13배의 농축율을 얻었으며 시료 처리 속도는 시간당 I0개 정도 이었다. 부피를 10.0 mL로 증가시키면 시료의 처리속도는 늦어지지만 농축비를 68배로 높일 수 있었다. 본 연구의 정확성을 알기 위해 합성된 준 표준 생체시료를 분석한 결과 4%의 상대 표준편차를 가지고 예상치에 근접하는 좋은 결과를 얻을 수 있었다.

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Transmembrane Protein 166 Expression in Esophageal Squamous Cell Carcinoma in Xinjiang, China

  • Sun, Wei;Ma, Xiu-Min;Bai, Jing-Ping;Zhang, Guo-Qing;Zhu, Yue-Jie;Ma, Hai-Mei;Guo, Hui;Chen, Ying-Yu;Ding, Jian-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.3713-3716
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    • 2012
  • Objective: Transmembrane protein 166 (TMEM166) expression in esophageal squamous cell carcinoma (ESCC) and remote normal esophageal tissues was examined to assess any role in tumour biology. Methods: TMEM166 mRNA expression in 36 cases with ESCC (36 tumour samples, 36 remote normal esophageal tissue samples) was detected by RT-PCR. TMEM166 protein expression was analysed in paraffin-embedded tissue samples from the same cases by immunohistochemistry. Results: Semi-quantitative analysis showed TMEM166 mRNA expression in ESCCs to be significantly lower than in remote normal esophageal tissues ($0.759{\pm}0.713$ vs. $2.622{\pm}1.690$, P=0.014). TMEM166 protein expression was also significantly reduced (69.4% vs. 94.4%, P<0.01). Conclusion: TMEM166 mRNA and protein expression demonstrated significant reduction in ESCCs compared with remote esophageal tissues, albeit with no correlation with tumour size, differentiation, stage, and lymph node metastasis, suggesting a role in regulating autophagic and apoptotic processes in the ESCC.

어류 병원성 세균 Edwardsiella tarda의 형질전환 및 재조합 ghost 세균에서의 E-lysis 유전자의 전사 발현 특징 (Transformation of Edwardsiella tarda and Transcriptional Characteristics of E-lysis Gene in Recombinant Bacterial Ghosts)

  • 권세련;남윤권
    • 한국어류학회지
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    • 제19권2호
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    • pp.83-87
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    • 2007
  • 어류 에드워드감염증에 대한 예방 재조합 ghost 백신을 개발하기 위한 연구의 일환으로 어류 병원성 세균인 Edwardsiella trada를 대상으로 플라스미드 형질전환을 실시하고 형질전환 안정성을 평가하였으며, 형질 도입된 재조합 ghost 세균의 E-lysis 유전자 발현을 분석하였다. E. tarda를 대상으로 한 ghost 유도는 대장균에 비해 상대적으로 장시간의 반응 시간이 요구되며 lysis의 개시가 지연된 점을 고려 시 발현된 E protein의 용해 능력 또는 E-gene의 전사발현 양이 E. tarda에서 다소 약화되는 것으로 나타났다. 그러나 대장균에 비해 ghost 유도 속도가 다소 낮음에도 불구하고 반응이 완성되었을 시점에서의 E. tarda의 ghost 효율은 대장균과 전혀 차이가 없이 99.99% 이상의 유도효율을 나타내었다.

Expression Profiling of WSSV ORF 199 and Shrimp Ubiquitin Conjugating Enzyme in WSSV Infected Penaeus monodon

  • Jeena, K.;Prasad, K. Pani;Pathan, Mujahid Khan;Babu, P. Gireesh
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권8호
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    • pp.1184-1189
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    • 2012
  • White spot syndrome virus (WSSV) is one of the major viral pathogens affecting shrimp aquaculture. Four proteins, WSSV199, WSSV 222, WSSV 249 and WSSV 403, from WSSV are predicted to encode a RING-H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimp can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, a time course expression profile analysis of WSSV Open Reading Frame (ORF) 199 and Penaeus monodon ubiquitin conjugating enzyme (PmUbc) was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge by semi-quantitative RT-PCR as well as Real Time PCR. EF1${\alpha}$ was used as reference control to normalize the expression levels. A significant increase in PmUbc expression at 24 h post infection (h.p.i) was observed followed by a decline till 72 h.p.i. Expression of WSSV199 was observed at 24 h.p.i in WSSV infected P. monodon. Since the up-regulation of PmUbc was observed at 24 h.p.i where WSSV199 expression was detected, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. However, further studies need to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this chaos in the shrimp culture industry.

통영지역 해녀의 식습관 및 영양소섭취 실태조사 (A Nutritional Survey of Women Divers in Tongyoung Area)

  • 이주희;김찬
    • 대한지역사회영양학회지
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    • 제7권6호
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    • pp.741-748
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    • 2002
  • The purpose of this study was to investigate the nutritional intake and dietary habits of women divers in Tongyoung area and to support the guideline for the improvement of their dietary life style by a validation of a Computerized Semi-quantitative Food Frequency Questionaire. Compared to the women at Jeju Island, the Tongyoung divers showed similar heights (154.6 $\pm$ 0.8 cm), weights (53.5 $\pm$ 1.0 kg) and BMIs (23.3 $\pm$0.4) , but they showed especially high levels of body fat (33.8 $\pm$ 0.8) . Their dietary habits showed they usually skipped lunch, but ate too much food at once, which may represent the dietary habits shown by obese people. Over 40% of the divers were taking some nutritious food or complementary food for health, without realizing the efficacy of these foods. The analysis of nutritional intake showed that all of average daily nutrients intakes of those below 50 years were higher than those of the Korean RDA. In the case of those above 50 years, it was shown that the average daily nutrients intakes were also higher than those of the Korean RDA except for energy (94.5%) and calcium (82.1%) . However, some individual divers showed lower nutrients intakes than 75% of RDA for calcium, iron, Vitamin A and Vitamin B$_2$. Also, excessive intakes of phosphorous and sodium could be a dietary problem. There was no correlation between BMI or percentage of body fat and energy, CHO, protein or fat. These results can provide the nutritional information for this special community, women divers, to improve their health.

Developmental Gene Expression of Antimicrobial Peptide PR-39 and Effect of Zinc Oxide on Gene Regulation of PR-39 in Piglets

  • Wang, Y.Z.;Xu, Z.R.;Lin, W.X.;Huang, H.Q.;Wang, Z.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권12호
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    • pp.1635-1640
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    • 2004
  • Two experiments were conducted to evaluate developmental gene expression of antimicrobial peptide PR-39 and effect of zinc oxide on gene regulation of PR-39 in piglets using semi-quantitative RT-PCR analysis. In experiment 1, fifteen female Tai-Hu pigs (a local breed in China) in five groups, each of three pigs at 1, 14, 28, 42 and 56 days of age were used to determine effect of age and weaning on mRNA expression of PR-39. In experiment 2, nine groups of pigs (total seventy-two female 36 days-age weanling Tai-Hu piglets) were assigned to three treatments (${ZnO}_0$, ${ZnO}_{100}$ and ${ZnO}_{3000}$). The feeding experimental period lasted 15 days. After feeding experiment, nine pigs with three animals in each treatment were chosen to determine the effect of ZnO on PR-39 mRNA expression of pigs. The results showed that PR-39 mRNA levels increased steadily in postnatal day 1-28 (preweaning), and weaning significantly decreased PR-39 mRNA expression of piglets (p<0.05). ${ZnO}_{3000}$ (3,000 mg zinc/kg diet) significantly increased PR-39 mRNA expression (p<0.05) when piglets were feed ${ZnO}_{3000}$ diet for 15 days. ${ZnO}_{100}$ (100 mg zinc/kg diet) also increased PR-39 gene expression, but the result was not statistically significant (p>0.05). The result was in accordance with the effect of ${ZnO}_{3000}$ and ${ZnO}_{100}$ on weight gain of piglets and prevention of diarrhea.

Mechanisms of Hela Cell Apoptosis Induced by Abnormal Savda Munziq Total Phenolics Combined with Chemotherapeutic Agents

  • Zhang, Yun-Xia;Abliz, Guzalnur;Ye, Wei-Jun;Mutalipu, Zuohelaguli;Li, Xiao-Wen;Wang, Hai-Qin;Buranjiang, Gulimire;Upur, Halmurat
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권2호
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    • pp.743-747
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    • 2014
  • Objective: To investigate the effects of abnormal Savda Munziq (ASMq) total phenolics combined with cisplatin and docetaxel on the Hela cell growth. Methods: In vivo cultured Hela cells were treated with cisplatin, docetaxel, total phenolics, cisplatin+total phenolics or docetaxel+total phenolics. MTT was performed to assess inhibition of cell proliferation, flow cytometry to detect apoptosis, and semi-quantitative RT-PCR to test for survivin and Bcl-2 expression. Results: The total phenolics, cisplatin and docetaxel had significant inhibitory and apoptosis-promoting effects on Hela cells (P<0.05), with the early apoptotic rates of $12.8{\pm}0.70%$, $18.9{\pm}3.79%$ and $15.8{\pm}3.8%$; the total phenolics, cisplatin and docetaxel significantly decreased the expression of Bcl-2 and survivin (all P<0.01), especially when used in combination. Conclusion: ASMq total phenolics, combined with cisplatin and docetaxel, could promote the apoptosis of Hela cells possibly through reducing the expression of Bcl-2 and survivin.

고리 부근 현탁물중 점토광물의 분포에 관한 연구 (Distribution of Suspended Clay Minerals around the Kori Waters)

  • 김대철;김정무
    • 한국수산과학회지
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    • 제23권2호
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    • pp.145-154
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    • 1990
  • 고리 해역 현탁물의 총량은 표층수에서 $3\~4\;mg/\iota$ 정도이고 저층수에서는 $3.5\~5\;mg/\iota$ 정도로 표층보다 높으며 특별한 계절변화는 없는 것으로 나타났다. 표층퇴적물은 주로 실트와 점토가 $90\%$이상인 세립질 퇴적물로 되어 있으며 미약하나마 외양으로 갈수록 세립화되는 경향이 나타난다. 현탁물중 점토광물의 상대비를 보면 illite가 평균 $65\%$ 정도로 가장 많고 kaolinite와 chlorite는 각각 $170\%$ 정도로 거의 비슷하고 smectite는 $1\%$ 내외의 미량이었다. 고리 부근 현생퇴적물의 주 공급원은 낙동강으로 생각되지만 대마해류에 의해 유입된 동지나해와 남지나해 세립질 퇴적물의 영향도 고려해야 할 것으로 사료된다.

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Overexpression of a Pathogenesis-Related Protein 10 Enhances Biotic and Abiotic Stress Tolerance in Rice

  • Wu, Jingni;Kim, Sang Gon;Kang, Kyu Young;Kim, Ju-Gon;Park, Sang-Ryeol;Gupta, Ravi;Kim, Yong Hwan;Wang, Yiming;Kim, Sun Tae
    • The Plant Pathology Journal
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    • 제32권6호
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    • pp.552-562
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    • 2016
  • Pathogenesis-related proteins play multiple roles in plant development and biotic and abiotic stress tolerance. Here, we characterize a rice defense related gene named "jasmonic acid inducible pathogenesis-related class 10" (JIOsPR10) to gain an insight into its functional properties. Semi-quantitative RT-PCR analysis showed up-regulation of JIOsPR10 under salt and drought stress conditions. Constitutive over-expression JIOsPR10 in rice promoted shoot and root development in transgenic plants, however, their productivity was unaltered. Further experiments exhibited that the transgenic plants showed reduced susceptibility to rice blast fungus, and enhanced salt and drought stress tolerance as compared to the wild type. A comparative proteomic profiling of wild type and transgenic plants showed that overexpression of JIOsPR10 led to the differential modulation of several proteins mainly related with oxidative stresses, carbohydrate metabolism, and plant defense. Taken together, our findings suggest that JIOsPR10 plays important roles in biotic and abiotic stresses tolerance probably by activation of stress related proteins.