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Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation:an appeal for performance verification

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Asian Journal of Andrology 2019年第5期21卷 438-444页

作者： Rakesh Sharma sajal gupta Ralf Henkel Ashok Agarwal American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Medical Bioscience University of the Western CapeBellville 7535South Africa

Lack of standardized,reproducible protocols and referencO values is among the challenges faced when using new or upgraded versions of instruments in reproductive laboratories and flow *** deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay combined with flow cytometry routinely used for diagnostic measurement of sperm DNA fragmentation(SDF)is a unique *** change in the setting of the standard instrument,including upgrades of hardware or software,can lead to different results and may affect clinicians'decision for ***,we compared TUNEL results of SDF obtained from a standard(C6)flow cytometer with a newer version of the same instrument(C6 Plus)and examined the cutoff,sensitivity,and specificity without calibration(adjustment)and after *** sperm preparation and matched acquisition settings were used to examine the performance of two flow *** strength of agreement of the results between the two observers was also *** adjustment of the settings,overall concordance became high and the two cytometers showed 100%positive and negative predictive value with 100%area under the *** overall correlation coefficient observed between C6 and C6 Plus was highly significant(P99%.The in terobserver agreeme nt on C6 flow cytometer for the two observers was 0.801±0.062 and 0.746±0.044 for C6 *** demonstrated a strong agreement between the samples tested on the two flow cytometers after calibration and established the robustness of both instruments.

关键词： DNA fragmentation flow cytometer oxidative stress sperm TUNEL assay

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Spermatozoa protein alterations in infertile men with bilateral varicocele

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Asian Journal of Andrology 2016年第1期18卷 43-53页

作者： Ashok Agarwal Rakesh Sharma Damayanthi Durairajanayagam Zhihong Cui Ahmet Ayaz sajal gupta Belinda Willard Banu Gopalan Edmund Sabanegh Center for Reproductive Medicine Glickman Urological and Kidney Institute Cleveland Clinic Cleveland OH USA Faculty of Medicine MARA University of Technology Selanger Malaysia Lerner Research institute Cleveland Clinic Cleveland OH USA

Among infertile men, a diagnosis of unilateral varicocele is made in 90% of varicocele cases and bilateral in the remaining varicocele cases. However, there are reports of under-diagnosis of bilateral varicocele among infertile men and that its prevalence is greater than 10%. In this prospective study, we aimed to examine the differentially expressed proteins （DEP） extracted from spermatozoa cells of patients with bilateral varicocele and fertile donors. Subjects consisted of 17 men diagnosed with bilateral varicocele and 10 proven fertile men as healthy controls. Using the LTQ-orbitrap elite hybrid mass spectrometry system, proteomic analysis was done on pooled samples from 3 patients with bilateral varicocele and 5 fertile men. From these samples, 73 DEP were identified of which 58 proteins were differentially expressed, with 7 proteins unique to the bilateral varicocele group and 8 proteins to the fertile control group. Majority of the DEPs were observed to be associated with metabolic processes, stress responses, oxidoreductase activity, enzyme regulation, and immune system processes. Seven DEP were involved in sperm function such as capacitation, motility, and sperm-zona binding. Proteins TEKT3 and TCP11 were validated by Western blot analysis and may serve as potential biomarkers for bilateral varicocele. In this study, we have demonstrated for the first time the presence of DEP and identified proteins with distinct reproductive functions which are altered in infertile men with bilateral varicocele. Functional proteomic profiling provides insight into the mechanistic implications of bilateral varicocele-associated male infertility.

关键词： bilateral varicocele bioinformatics male infertility proteomics spermatozoa proteins varicocele

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Hematological, antioxidant and protective performance of Usnea longissima on chemical induced hepatotoxicity in experimental animals

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Journal of Coastal Life Medicine 2017年第5期5卷 224-232页

作者： Pritt Verma Shravan Kumar Paswan Abhisek Raj Virendra Nath Ramesh Kumar gupta Shikhar Verma sajal Srivastava ChandanaVenketshwara Rao Pharmacognosy and Ethnopharmacology Division CSIR-National Botanical Research InstituteLucknowIndia Amity Institute of Pharmacy Amity UniversityLucknowIndia Department of Pharmaceutical Sciences Central University of RajasthanRajasthanIndia Sherwood College of Pharmacy Barabanki-225001India

Objective: To investigated the hematological, antioxidant and protective performance of Usnea longissima (U. longissima) on CCl4 induced hepatotoxicity in experimental animals. Methods: Hepatotoxicity was induced by CCl4 (1 mL/kg body weigt 1:1 CCl4 i.p.), ethanolic U. longissima extracts at a doses (200 and 400 mg/kg body weigt) were administered to and compared with Silymarin (25 mg/kg body weigt) and hematological, antioxidant and enzymatic, non-enzymatic parameters were assessed through the liver functions test. All the observation was also supplemented with histopathological examination of liver sections. Results: Phytochemical investigation showed that ethanolic extract contains poly phenolic compounds tannins, flavonoids, alkaloids and saponins and acute toxicity study shows that ethanolic extract was safe up to 2000 mg/kg body weight. The toxicant induced a rise in the plasma enzyme levels of ALT, AST, ALP and total bilirubin level. This increased level was significantly decreased by the extract at 400 mg/kg body weight than 200 mg/kg body weight. The animals were prevented (partly or fully) which was showed in the histopathological changes using ethonolic U. longissima extract. Conclusions: The outcome of this study reveals that, there is a powerful antioxidant and hepatoprotective activity of U. longissima. It is believed that the present constituents are responsible for courting the hepatic disease and alternative components have the power to act as free radical scavenging properties.

关键词： Usnea longissima Carbon tetra chloride (CCl4) Hematological Antioxidant Histopathological

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