Project description:DNA methylation analysis using the Infinium MethylationEPIC BeadChip arrays was applied to 40 semen-derived DNA samples (age range 24 – 58 years). The aim of the study was to discover differentially methylated sites (DMS) that correlate with age.
Project description:Identifying the type and origin of biological samples left at a crime scene is crucial in forensic investigations as it can provide important clues for crime scene reconstruction and linkages between victim/perpetrator/scene. MicroRNAs (miRNAs) are considered to be more stable than mRNA due to their small size and protection by protein and have been demonstrated to be a viable tool for body fluid identification in forensic casework. To screen reliable body-fluid specific miRNAs, ten arrays were performed in five body fluids (peripheral blood, menstrual blood, saliva, semen and vaginal secretion). Two arrays were carried out for each body fluid: three samples for the first and the other two for the second (for menstrual blood, the second array detected three samples).
Project description:<p>The age estimation of bloodstains measures the time-dependent changes in the levels of bloodstain biomolecules. Although several studies have identified bloodstain metabolites as markers for estimating bloodstain age, none has consider gender and age-related metabolomic differences or long-time bloodstain age. Therefore, we aimed to identify metabolite markers for estimating the age of bloodstains at weekly intervals within 28 d and validate them through the MRM technique. Interestingly, adenosine 5′-monophosphate (5′-AMP), choline and pyroglutamic acid were selected as markers. We validated a total of 7 metabolites, including 5 previously reported metabolites, ergothioneine, hypoxanthine, L-isoleucine, L-tryptophan and pyroglutamic acid. Choline and hypoxanthine might be used to differentiate between day 0 and day 14 at weekly intervals, while L-isoleucine and L-tryptophan might help distinguish between 7 d before and 14 d after. Evaluation of the changes in the levels of metabolites according to gender and age, revealed that the average level of all 7 metabolites was higher in women on day 0. Moreover, the level of ergothioneine was significantly higher in the elderly than the youth under all time conditions. In this study, we confirmed the potential effectiveness of metabolites in bloodstains as forensic markers, and provided a new perspective on metabolomic approaches linked to forensic science.</p>
Project description:Hair, a skin appendage, is a frequently procured biological specimen from crime scenes and has been used in forensic investigations for over a century. Proteomic analysis and identification of genetically variant peptides (GVPs) in hair samples for identification purposes is a recent but an efficiently advancing tool for forensic and archeological uses. However, the hair samples exposed to environmental insults have been seen to undergo degradation to various degrees depending on the physicochemical factors the samples have been exposed to. Therefore, one would expect that evidentiary hair samples stored for longer periods of time will undergo alterations in their proteomes and hence can affect their use on long-term samples stored as case work. To determine the degree to which age of individual or age of sample can affect the hair shaft proteome or inferred GVP profile, protein profiles of hair samples were compared from individuals of different ages as well as samples collected from same individual at different time points and stored in dry environment at room temperature for 5 - 65 years.
Project description:<h4><strong>BACKGROUND: </strong>Semen quality is negatively correlated with male age and is mainly quantified by a routine semen analysis, which is descriptive and inconclusive. Sperm proteins or semen metabolites are used as the intermediate or end-products, reflecting changes in semen quality, and hold much promise as a new biomarker to predict fertility in advanced-aged males.</h4><h4><strong>OBJECTIVES: </strong>In this study, we sought to assess whether the semen metabolome and proteome of aged males can affect semen quality and serve as biomarkers for predicting semen quality.</h4><p><strong>MATERIALS AND METHODS: </strong>We retrospectively analyzed 12825 males that underwent semen routine analysis to understand the age-dependent changes in sperm quality. To identify the difference between aged and young adults, metabolomics (n=60) analyses of semen and proteomics (n=12) analyses of sperm were conducted. Finally, integrated machine learning of metabolomics was conducted to screen biomarkers to identify aging semen.</p><p><strong>RESULTS:</strong> We discovered that male age was positively correlated with sperm concentration as well as DNA fragmentation index(DFI), and negatively with progressive motile sperm count, total sperm count, sperm volume and progressive sperm motility. The differential metabolites were significantly enriched in various metabolic pathways, and four of these differential metabolites (Pipamperone, 2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol, Arg-Pro and Triethyl phosphate) were utilized to establish a biomarker panel to identify aging semen. Proteomic analysis showed that differential proteins were significantly enriched in protein digestion and absorption and some energy-related pathways. An integrated analysis of the metabolome and proteome identified differential energy metabolism and oxidative stress-related proteins, which could explain the decreased motility and the increased DFI of aging sperm.</p><p><strong>DISCUSSION AND CONCLUSION:</strong> We provide compelling evidence that the changes in semen metabolome and sperm proteome are related to the decline of semen quality in aged males. Moreover, a biomarker panel based on four metabolites was established to identify aging semen.</p>
Project description:Genome-wide expression profiling of four kinds of body fluid samples (blood, saliva, semen and vaginal swab). The purpose of the present study was selection of specific mRNA markers for identification of the four body fluids. Results provide important information about gene expression level of each body fluid for forensic science. Total RNAs isolated from four kinds of body fluid samples (blood, saliva, semen and vaginal swab) obtained from Korean volunteers
Project description:We looked for the presence of prostatic cells and RNAs (cellular and acellular) of prostatic origin in the ejaculate semen from fertile and sterilized men and from men with prostate cancer, to ascertain the suitability of ejaculate semen for studying and monitoring prostate health. Methods. For prostatic cells, semen from fertile and sterile men and men with prostate cancer were subjected to density gradient centrifugation on a 60% cushion of Suprasperm™ and fractions containing somatic cells were washed and cytospun onto coated slides prior to fixation and immunocytochemistry with antibodies recognizing prostatic markers. Small RNAs were isolated from liquefied semen clarified of all cellular and other insoluble material by high-speed ultra-centrifugation. Sequencing libraries were prepared from fractions isolated by PAGE, corresponding to small non-coding (snc) RNAs in the 18-23 nt and 30-35 nt size ranges. Libraries were interrogated by next generation sequencing and analysed to generate lists of differentially expressed RNAs associated with cancer and sterility. Results. Cells positive for cytokeratin 18, PSMA, NKX3.1 and CD24 were observed in most ejaculate samples by indirect immunocytochemistry as either discrete cellular entities or within clumps of cell containing material (DAPI+; antigen positive and negative) and cytoplasmic fragments (DAP–; antigen positive and negative). RNA from populations of cells enriched by differential density gradient centrifugation supported their immunocytochemical designation as prostatic cells. Small RNAs (18 - 43 nt) from seminal plasma were highly heterogeneous although tRNAs and 5SRNAs were the dominant forms. A degree of overlap between the respective differential expression profiles of small RNAs from sterile and cancer samples, suggested a common feature, perhaps indicating an inflammatory response. Conclusion. This study supports a wider appraisal of ejaculate semen for studying and monitoring prostate health.
Project description:EVs are known to contain important cargo of biologically active and regulatory molecules including ncRNAs. EVs in seminal plasma, play an important role in sperm functions , including motility and fertilization. However, less is known about the comprehensive profile of spEV ncRNAs in clinical cohort and if these profiles differ by semen quality status. We assessed sperm-free spEV ncRNA profiles from semen samples of male partners receiving IVF treatment. Men were classified into having normal (n = 59, Status 0) or poor (n = 32, Status 1) semen quality based on WHO semen parameter guidelines.