• 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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    • v.17 no.4
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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.

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

  • Im, Kab Soo;Pak, Yong Nam
    • Journal of the Korean Chemical Society
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    • v.43 no.6
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    • pp.644-650
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    • 1999
  • Trace amount of cobalt in water samples was preconcentrated continuously with an organic precipitant and determined by flame atomic absorption spectrometry. The flow injection technique was used to preconcentrate cobalt by on-line direct precipitation with 1-nitroso-2-naphthol. The precipitation was dissolved with methyl isobutyl ketone (MlBK) and was sent to the flame. The optimum conditions for cobalt determination were determined and used to analyze Co samples. For 1.0 mL of sample, the enrichment factor was 13 and the sample throughput was about lO per hour for 0.5 ppm Co solution. The enrichment factor was increased to 68 fold for 10.0 mL. A semi-reference biologicaI sample was prepared and analyzed. The result was in good agreement with the expected value with RSD of 4%.

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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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    • v.13 no.8
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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.

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

  • Kwon, Se Ryun;Nam, Yoon Kwon
    • Korean Journal of Ichthyology
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    • v.19 no.2
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    • pp.83-87
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    • 2007
  • Edwardsiella tarda, a gram (-) pathogen causing edwardsiellosis in farmed fish, was transformed via electroporation with a plasmid expression vector driving the PhiX174 E-lysis gene under the transcriptional control by lambda PR regulatory sequence. The persistent maintenance of the plasmid vector in recombinant E. tarda was found in numerous subculture procedures over up to 6 months without any adverse effect on the original copy number of plasmids. Comparative examination based on semi-quantitative RT-PCR analysis on transcriptional efficiency of E-lysis gene between recombinant E. coli and E. tarda indicated that promoter strength and induction capacity of bacterial ghosts would be retarded in E. tarda as compared to the E. coli. However, the completeness of induction for bacterial ghosts in E. tarda was the same with E. coli, in which at least 99.99% of induction rate was possible and further the viability of recombinant bacteria was completely eliminated by a post-induction procedure including washing and freeze drying lyophilization.

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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    • v.25 no.8
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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.

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

  • Lee, Joo-Hee;Kim, Chan
    • Korean Journal of Community Nutrition
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    • v.7 no.6
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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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    • v.17 no.12
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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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    • v.15 no.2
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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.

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

  • Kim Dae-Choul;Kim Jung-Moo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.2
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    • pp.145-154
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    • 1990
  • Amounts of total suspended matter(TSM) around the Kori waters have been monitored for two years to understand the dispersal pattern of fine-grained sediment. Average concentration of the TSM is $3\~4\;mg/\iota$ in the surface and $3.5\~5\;mg/\iota$ in the near-bottom water. Semi-quantitative analysis for the four major clay minerals in TSM is also performed. The average relative abundance of each clay mineral such as illite, kaolinite, chlorite, and smectite is $65\%,\;17\%,\;17\%$, and $1\%$, respectively. It seems that the majority of recent fine-grained sediment deposited around the Kori area is derived from the Nakdong River. The texture of surface sediment is also dominated by fine-grained fraction leading up to $90\%$ of silt and clay content. The grain size decreases seaward progressively.

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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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    • v.32 no.6
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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.