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M K Panner Selvam, A Agarwal, R Sharma, L Samanta
Semen hyperviscosity delays the liquefaction of semen sample and is subjected to limited proteolysis by addition of α-chymotrypsin to reduce the viscosity. α-Chymotrypsin is a proteolytic enzyme involved in degradation of the proteins and polypeptides. Even though α-chymotrypsin improves the handling of hyperviscous samples, its effect on the sperm proteins is not clear. This study was aimed to evaluate the alteration in the expression of sperm functional proteins in samples treated with α-chymotrypsin...
February 6, 2018: Andrology
P Intasqui, A Agarwal, R Sharma, L Samanta, R P Bertolla
Male infertility evaluation is mainly based on semen analysis. Thus, identification of additional diagnostic methods is valuable. The aim of this study was to analyse the sperm proteome of infertile men to identify the underlying mechanisms and reliable diagnostic biomarkers. This cross-sectional study consisted of 16 infertile men and seven proven fertile men. An LC-MS/MS approach was performed in five pooled samples of each group (proven fertile men, primary infertility and secondary infertility). Differentially expressed proteins were used for functional enrichment analyses, and the most central proteins involved in altered functions in both infertile groups and the testis-specific proteins were validated using Western blotting and immunocytochemistry...
December 4, 2017: Andrologia
Parvin Sabeti, Fardin Amidi, Seyed Mahdi Kalantar, Mohammad Ali Sedighi Gilani, Soheila Pourmasumi, Atefeh Najafi, Ali Reza Talebi
BACKGROUND: Teratoasthenozoospermia (TA) is a severe form of male infertility with no clear etiology. OBJECTIVE: To compare the level of intracellular anion superoxide (O2-), heat shock protein A2 (HSPA2) and protamine deficiencies in ejaculated spermatozoa between teratoasthenozoospermic and normozoospermic men. MATERIALS AND METHODS: In this case- control study, semen samples of 20 infertile men, with TA (with normal morphology lower than 4%_ and total motility lower than 40% ) as the case group and 20 normozoospermic fertile men as the control group were evaluated for intracellular O2(-) and HSPA2 by flow cytometry and protamine deficiency by Chromomycin A3 (CMA3) test...
May 2017: International Journal of Reproductive Biomedicine (Yazd, Iran)
Lu-Lu Zhai, Qi Xie, Chun-Hua Zhou, Da-Wei Huang, Zhi-Gang Tang, Tong-Fa Ju
Heat shock-related 70-kDa protein 2 (HSPA2) is known to correlate with tumor development and progression. This work aimed to determine the expression and prognostic roles of HSPA2 in pancreatic carcinoma. Tumor and their corresponding non-tumor tissues were obtained from 80 patients with pancreatic carcinoma. HSPA2 expression in tumor and non-tumor tissues was evaluated by immunohistochemistry. Expression of vascular endothelial growth factor (VEGF) and CD31 in tumor tissues were also evaluated by immunostaining...
May 2017: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
Brett Nixon, Elizabeth G Bromfield, Jinwei Cui, Geoffry N De Iuliis
Among the numerous families of heat shock protein (HSP) that have been implicated in the regulation of reproductive system development and function, those belonging to the 70 kDa HSP family have emerged as being indispensable for male fertility. In particular, the testis-enriched heat shock 70 kDa protein 2 (HSPA2) has been shown to be critical for the progression of germ cell differentiation during spermatogenesis in the mouse model. Beyond this developmentally important window, mounting evidence has also implicated HSPA2 in the functional transformation of the human sperm cell during their ascent of the female reproductive tract...
2017: Advances in Anatomy, Embryology, and Cell Biology
Elizabeth G Bromfield, Bettina P Mihalas, Matthew D Dun, R John Aitken, Eileen A McLaughlin, Jessica L H Walters, Brett Nixon
Lipid peroxidation products, such as 4-hydroxynonenal (4HNE), are causative agents responsible for extensive protein damage within the male and female germlines. Recently, we have demonstrated that 4HNE production can initiate the proteolytic degradation of the molecular chaperone Heat Shock Protein A2 (HSPA2) in male germ cells. These events may be partially responsible for HSPA2 deficiency in the spermatozoa of patients that repeatedly fail in vitro fertilization. Given this, mechanisms that limit the production of 4HNE will be highly advantageous for the preservation of male fertility...
March 1, 2017: Biology of Reproduction
Tatiana Kisliouk, Tomer Cramer, Noam Meiri
Depending on its stringency, exposure to heat in early life leads to either resilience or vulnerability to heat stress later in life. We hypothesized that epigenetic alterations in genes belonging to the cell proteostasis pathways are attributed to long-term responses to heat stress. Epigenetic regulation of the mRNA expression of the molecular chaperone heat-shock protein (HSP) 70 (HSPA2) was evaluated in the chick hypothalamus during the critical period of thermal-control establishment on day 3 post-hatch and during heat challenge on day 10...
May 2017: Journal of Neurochemistry
Mengmeng Xu, Long Che, Zhenguo Yang, Pan Zhang, Jiankai Shi, Jian Li, Yan Lin, Zhengfeng Fang, Lianqiang Che, Bin Feng, De Wu, Shengyu Xu
Primordial follicle formation represents a critical phase of the initiation of embryonic reproductive organ development, while the primordial follicle transition into primary follicle determines whether oestrus or ovulation will occur in female animals. To identify molecular mechanism of new proteins which are involved in ovarian development, we employed 2D-DIGE to compare the protein expression profiles of primordial follicles and primary follicles of fetal ovaries in pigs. Fetal ovaries were collected at distinct time-points of the gestation cycle (g55 and g90)...
2017: BioMed Research International
Elizabeth G Bromfield, R John Aitken, Eileen A McLaughlin, Brett Nixon
STUDY QUESTION: Does oxidative stress compromise the protein expression of heat shock protein A2 (HSPA2) in the developing germ cells of the mouse testis? SUMMARY ANSWER: Oxidative stress leads to the modification of HSPA2 by the lipid aldehyde 4-hydroxynonenal (4HNE) and initiates its degradation via the ubiquitin-proteasome system. WHAT IS KNOWN ALREADY: Previous work has revealed a deficiency in HSPA2 protein expression within the spermatozoa of infertile men that have failed fertilization in a clinical setting...
February 10, 2017: Molecular Human Reproduction
Anna Slawinska, John C Hsieh, Carl J Schmidt, Susan J Lamont
Acute heat stress requires immediate adjustment of the stressed individual to sudden changes of ambient temperatures. Chickens are particularly sensitive to heat stress due to development of insufficient physiological mechanisms to mitigate its effects. One of the symptoms of heat stress is endotoxemia that results from release of the lipopolysaccharide (LPS) from the guts. Heat-related cytotoxicity is mitigated by the innate immune system, which is comprised mostly of phagocytic cells such as monocytes and macrophages...
2016: PloS One
Anders Patrik Gunnarsson, Rikke Christensen, Jian Li, Uffe Birk Jensen
The murine hair follicle contains several different keratinocyte progenitor populations within its compartments. By using antibodies against CD34, Itgα6, Sca-1 and Plet-1, we have isolated eight populations and compared their Krt10 and Krt14 expressions using fluorescence microscopy. This improved panel was used in our associated article doi:10.1016/j.scr.2016.06.002 (A.P. Gunnarsson, R. Christensen, J. Li, U.B. Jensen, 2016) [1] and the present dataset describes the basic controls for the FACS. We also used imaging flow cytometry to visualize the identified populations as control...
December 2016: Data in Brief
Anders Patrik Gunnarsson, Rikke Christensen, Jian Li, Uffe Birk Jensen
Several populations with stem cell characteristics have been identified in the mouse hair follicle, and some are characterized by specific subsets of surface markers. Here we investigate to what extend a multicolor panel of the four surface markers CD34, Sca-1, Integrin-α6 and Plet-1 is sufficient for separating previously described populations. We used global transcriptome profiling to characterize these, and we also performed transcriptome analysis of selected populations after two weeks of culturing. This has not been done before in a context of multiple populations...
July 2016: Stem Cell Research
Angelica Van Goor, Anna Slawinska, Carl J Schmidt, Susan J Lamont
Differences in responses of chicken bone marrow derived dendritic cells (BMDC) to in vitro treatment with lipopolysaccharide (LPS), heat, and LPS + heat were identified. The Fayoumi is more disease resistant and heat tolerant than the Leghorn line. Nitric Oxide (NO) production, phagocytic ability, MHC II surface expression and mRNA expression were measured. NO was induced in BMDC from both lines in response to LPS and LPS + heat stimulation; Fayoumi produced more NO with LPS treatment. Fayoumi had higher phagocytic ability and MHC II surface expression...
October 2016: Developmental and Comparative Immunology
Meizhu Bai, Dan Liang, Yinghua Wang, Qing Li, Yuxuan Wu, Jinsong Li
Tissue-specific knockout technology enables the analysis of the gene function in specific tissues in adult mammals. However, conventional strategy for producing tissue-specific knockout mice is a time- and labor-consuming process, restricting rapid study of the gene function in vivo. CRISPR-Cas9 system from bacteria is a simple and efficient gene-editing technique, which has enabled rapid generation of gene knockout lines in mouse by direct injection of CRISPR-Cas9 into zygotes. Here, we demonstrate CRISPR-Cas9-mediated spermatogenic cell-specific disruption of Scp3 gene in testes in one step...
May 20, 2016: Journal of Genetics and Genomics, Yi Chuan Xue Bao
Yonghong Tian, Lejun Li, Fengbin Zhang, Jian Xu
PURPOSE: Cell-free mRNAs (cfmRNAs) were quantitatively measured in human seminal plasma and its relationship with semen quality was investigated. METHODS: Herein, a prospectively, controlled investigation was performed to study seminal plasma HSPA2 and uPA cfmRNA alterations between 21 asthenozoospermic patients and 16 normozoospermic individuals. Standard semen analysis was performed and seminal plasma cfmRNAs content was measured by real-time quantitative PCR...
August 2016: Journal of Assisted Reproduction and Genetics
P Zambonelli, E Gaffo, M Zappaterra, S Bortoluzzi, R Davoli
Fat deposition is a widely studied trait in pigs because of its implications with animal growth efficiency, technological and nutritional characteristics of meat products, but the global framework of the biological and molecular processes regulating fat deposition in pigs is still incomplete. This study describes the backfat tissue transcription profile in Italian Large White pigs and reports genes differentially expressed between fat and lean animals according to RNA-seq data. The backfat transcription profile was characterised by the expression of 23 483 genes, of which 54...
June 2016: Animal Genetics
Wen-Hsin Chang, Bing-Ching Ho, Yi-Jing Hsiao, Jin-Shing Chen, Chien-Hung Yeh, Hsuan-Yu Chen, Gee-Chen Chang, Kang-Yi Su, Sung-Liang Yu
JAG1 is a Notch ligand that plays a critical role in multiple signaling pathways. However, the functionality of JAG1 in non-small cell lung cancer (NSCLC) has not been investigated thoroughly. By comparison of gene transcripted RNA profiles in the cell line pair with differential invasion ability, we identified JAG1 as a potential metastasis enhancer in lung cancer. Ectopic expression of JAG1 on lung cancer cells enhanced cell migration and invasion as well as metastasis in vitro and in vivo. Conversely, knockdown of JAG1 with siRNA in highly invasive cancer cells led to the reduction of migration and invasion...
2016: PloS One
Ming Liu, Min Li, Jiangfeng Liu, Hongkai Wang, Dandan Zhong, Hong Zhou, Baoxue Yang
BACKGROUND: Previous work found that urea accumulation in urothelial cells caused by urea transporter B knockout led to DNA damage and apoptosis that contributed to the carcinogenesis. The purpose of this study is to explore the potential connection between high urinary urea concentration and the bladder disorders. METHODS: A high protein diet rat model was conducted by feeding with 40 % protein diet. In-silico modeling and algorithm, based on the results of microarray and proteomics from the bladder urothelium, were used for the reconstruction of accurate cellular networks and the identification of novel master regulators in the high-protein diet rat model...
February 16, 2016: Journal of Translational Medicine
Ashok Agarwal, Rakesh Sharma, Luna Samanta, Damayanthi Durairajanayagam, Edmund Sabanegh
To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each varicocele outcome were combined to identify the proteins involved in varicocele-associated infertility in men irrespective of stage and laterality of their clinical varicocele. Expression levels of 5 key proteins (PKAR1A, AK7, CCT6B, HSPA2, and ODF2) involved in stress response and sperm function including molecular chaperones were validated by Western blotting...
March 2016: Asian Journal of Andrology
Elizabeth G Bromfield, Eileen A McLaughlin, Robert John Aitken, Brett Nixon
STUDY HYPOTHESIS: Given the importance of the chaperone Heat Shock Protein A2 (HSPA2) in the regulation of male fertility, this study aimed to identify and characterize additional proteins that may rely on the activity of this chaperone in human spermatozoa. STUDY FINDING: In view of the findings in this study we propose that angiotensin converting enzyme (ACE) and protein disulfide isomerase A6 (PDIA6) are novel interacting proteins of HSPA2 and that this multimeric complex may participate in key elements of the fertilization cascade...
February 2016: Molecular Human Reproduction
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