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Objective@#The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. @*Methods@#Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. @*Results@#Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). @*Conclusion@#The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.
ABSTRACT
Objective@#The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. @*Methods@#Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. @*Results@#Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). @*Conclusion@#The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.
ABSTRACT
Background: Non-obstructive azoospermia [NOA] occurs in approximately 10% of infertile men. Retrieval of the spermatozoa from the testicle of NOA patients is an invasive approach. Seminal plasma is an excellent source for exploring to find the biomarkers for presence of spermatozoa in testicular tissue. The present discovery phase study aimed to use metabolic fingerprinting to detect spermatogenesis from seminal plasma in NOA patients as a non-invasive method
Methods: In this study, 20 men with NOA were identified based on histological analysis who had their first testicular biopsy in 2015 at Avicenna Fertility Center, Tehran, Iran. They were divided into two groups, a positive testicular sperm extrac-tion [TESE[+]] and a negative testicular sperm extraction [TESE[-]]. Seminal plasma of NOA patients was collected before they underwent testicular sperm extraction [TESE] operation. The metabolomic fingerprinting was evaluated by Raman spec-trometer. Principal component analysis [PCA] and an unsupervised statistical meth-od, was used to detect outliers and find the structure of the data. The PCA was ana-lyzed by MATLAB software
Results: Metabolic fingerprinting of seminal plasma from NOA showed that TESE [+] versus TESE[-] patients were classified by PCA. Furthermore, a possible subdi-vision of TESE[-] group was observed. Additionally, TESE[-] patients were in ex-treme oxidative imbalance compared to TESE[+] patients
Conclusion: Metabolic fingerprinting of seminal plasma can be considered as a breakthrough, an easy and cheap method for prediction presence of spermatogenesis in NOA
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Currently, there are 20,197 human protein-coding genes in the most expertly curated database [UniProtKB/Swiss-Pro]. Big efforts have been made by the international consortium, the Chromosome-Centric Human Proteome Project [C-HPP] and independent researchers, to map human proteome. In brief, anno 2017 the human proteome was outlined. The male factor contributes to 50% of infertility in couples. However, there are limited human spermatozoa proteomic studies. Firstly, the development of the mapping of the human spermatozoa was analyzed. The human spermatozoa have been used as a model for missing proteins. It has been shown that human spermatozoa are excellent sources for finding missing proteins. Y chromosome proteome mapping is led by Iran. However, it seems that it is extremely challenging to map the human spermatozoa Y chromosome proteins based on current mass spectrometry-based proteomics technology. Post-translation modifications [PTMs] of human spermatozoa proteome are the most unexplored area and currently the exact role of PTMs in male infertility is unknown. Additionally, the clinical human spermatozoa proteomic analysis, anno 2017 was done in this study
ABSTRACT
The human seminal fluid is a complex body fluid. It is not known how many proteins are expressed in the seminal plasma; however in analog with the blood it is possible up to 10,000 proteins are expressed in the seminal plasma. The human seminal fluid is a rich source of potential biomarkers for male infertility and reproduction disorder. In this review, the ongoing list of proteins identified from the human seminal fluid was collected. To date, 4188 redundant proteins of the seminal fluid are identified using different proteomics technology, including 2-DE, SDS-PAGE-LC-MS/MS, MudPIT. However, this was reduced to a database of 2168 non-redundant protein using UniProtKB/Swiss-Prot reviewed database. The core concept of proteome were analyzed including pT, MW, Amino Acids, Chromosome and PTM distribution in the human seminal plasma proteome. Additionally, the biological process, molecular function and KEGG pathway were investigated using DAVID software. Finally, the biomarker identified in different male reproductive system disorder was investigated using proteomics platforms so far. In this study, an attempt was made to update the human seminal plasma proteome database. Our finding showed that human seminal plasma studies used to date seem to have converged on a set of proteins that are repeatedly identified in many studies and that represent only a small fraction of the entire human seminal plasma proteome
ABSTRACT
The seminal plasma is an excellent source for noninvasive detection of spermatogenesis. The seminal plasma of normospermic and azoospermic men has been analyzed for detection of spermatogenesis. Optical spectroscopy [Attenuated Total Reflectance-Infrared spectroscopy [ATR-IR] and Fourier Transform infrared spectroscopy [FT-IR] has been used to analyze the seminal plasma and the metabolome of seminal plasma for detection of spermatogenesis. The seminal plasma of normospermic and azoospermic men has been analyzed by ATR-IR. The results show that there is a pattern variation in the azoospermic men compared to normospermic men. However, the seminal plasma is too complex to show significant pattern variation. Therefore, the metabolome which is a subcomponent of the seminal plasma was analyzed. The seminal plasma metabolome of normospermic and azoospermic men has been analyzed by FT-IR. A significant pattern change was observed. The data combined with chemometrics analysis showed that significant changes are observed at metabolome level. We suggest that FT-IR has the potential as a diagnostic tool instead of testicular biopsy
Subject(s)
Humans , Male , Spectroscopy, Fourier Transform Infrared , Semen , Azoospermia , MetabolomeABSTRACT
Chronic hypoxia exists in many diseases, including cancer. The subject of our study is analysis of molecular pathways affected in the chronically hypoxic mouse brain. Using the emPAl protocol, we performed a quantitative proteomic approach to characterize the global proteome in the mouse brain exposed to 7% O[2] for 48 hours. Utilizing the emPAl protocol to estimate protein abundance and assign molar concentrations to all proteins, we were able to identify 33 proteins with significant changes in their expression. Deregulated proteins were mainly involved in cell metabolism, apoptosis, Ca[2+] signaling, pentose phosphate pathway, 14-3-3 protein mediated signaling cascades and protein degradation. The obtained data will provide some clues for understanding mechanisms with which cells respond and adapt to chronic hypoxia
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Endometriosis is a painful reproductive disease afflicting about up to 20% of women. It is one of the most frequent benign gynaecological diseases, however, little is known about the pathological of endometriosis. Over the past decade, high-throughput proteomics technologies have evolved considerably and have become increasingly more commonly applied to the investigation of female reproductive disease, including endometriosis. In this mini-review the authors look at the application of proteomics technologies in order to find biomarker associated with endometriosis
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New advances in mass spectrometry-based proteomics technology are having a major impact on our understanding of how human spermatozoa acquire their capacity for fertilization. A complete analysis of the proteins found in the human spermatozoa is essential for understanding the events leading up to, and including, fertilization and early embryo development. In this short review, we have collected the human sperm proteome from the literature and analyzed it by the Database for Annotation, Visualization and Integrated Discovery [DAVID] software. Bioinformatics analysis demonstrated that the collected 1,300 proteins were involved in various metabolic pathways including catabolic processes. Additionally, the majority of the collected human sperm proteome belonged to cytoplasm. Application of the multi-dimensional protein identification technology [MudPIT] for obtaining a better coverage of the hydrophobic and basic proteins of the human sperm proteome is recommended
Subject(s)
Humans , Male , Proteome , Computational BiologyABSTRACT
Aldolase C as a glycolytic enzyme is associated with cellular structure at developmental stages of all cells, and this is particularly evident during the early stages of morphogenesis. It seems that expression of aldolase C can be regulated by the rate of differentiation that depends on the level of transcription or mRNA stability. There are several techniques to detect gene expression here proteomics was used for determining expression of aldolase C [as a differentiation factor] in several cell types and basal cell carcinoma [BCC] tissue. The human astrocytes were differentiated from mesenchymal stem cells, fibroblast cells were cultured as primary cell culture and BCC tissue was taken from the patient. The fibroblast cells divided into two groups including sham and exposed groups. The exposed cells are them that were exposed to continue Extremely Low-Frequency Electromagnetic Fields [ELF-EMF]. The analysis of 2DE gels, showed different expression of aldolase C in mentioned cells. The findings indicate that the amount of aldolase C expression decreases as differentiation process develops
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Proteomics concerns itself with the characterization and function of all cellular proteins, the ultimate determinants of cellular function. Mass spectrometry has emerged as the preferred method for in-depth characterization of the protein components of biological systems. Using mass spectrometry, key insights into the composition, regulation and function of molecular complexes and pathways have been gained. Now days, mass spectrometry-based proteomics has become an indispensable tool in the cellular and molecular life sciences. This review discusses current mass spectrometry-based proteomics technologies
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Tissue hypoxia occurs where there is an imbalance between oxygen supply and consumption. Growing evidence from experimental and clinical studies points to the fundamental and pathophysiologic role of hypoxia in cancer, ischemic tolerance, and stroke. Hypoxia-induced changes in ion homeostasis, erythropoiesis, angiogenesis, proliferation and differentiation. This review outlines hypoxia effect at molecular level and describes briefly hypoxia role in the physiological and pathological conditions