Updating semen analysis: a subpopulation approach (original) (raw)
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Andrologia, 2019
The identification of idiopathic infertility cases, actually, is impossible. Among new functional tests, developed to improve the male fertility diagnosis, the evaluation of spermatic myo-inositol (MI) level, known as Andrositol ® test (AT), is one of the most interesting, considering its weak economic burden and ease of use. The aim of this study was to evaluate the predictive power of AT and its potential use for a preliminary evaluation of semen samples. To evaluate the predictive power of AT, 87 sperm samples were analysed in comparison with spermiogram and sperm chromatin dispersion (SCD) Test, the gold standard analyses for male fertility evaluation. The application of AT resulted very useful for a preliminary sample evaluation, predicting the absence of DNA fragmentation in case of Low Responder samples precisely, and the presence of DNA fragmentation in case of medium or High Responder samples with abnormal morphology, predicting SCD results with a probability of 80% for Medium Responder sample and of 96.7% for High Responder sample. Considering the predictive power of this method, we could imagine, as preliminary qualitative analysis, its application before SCD test, deepening sperm analysis, improving the daily activities of laboratory operators and maintaining a good reliability of sperm evaluation.
Human Reproduction, 1998
The impact of demographic, lifestyle, and seminal factors on the sperm chromatin structure assay (SCSA) parameters was evaluated in a population of 277 healthy Danish men. This cohort was established within the framework of a European Concerted Action on occupational hazards to male reproductive capability in order to examine the possible reproductive effects of exposure to styrene or pesticides. The SCSA measures the susceptibility of sperm DNA to in-situ acid-induced denaturation, by multiparameter flow cytometric analysis after staining with the DNA-specific fluorescent dye acridine orange. The green versus red bivariate cytogram patterns were quite variable among donors, showing a wide heterogeneity of sperm DNA denaturability. Nevertheless, in those cases where we had the possibility to measure two semen samples from the same donor, the cytogram pattern remained stable over time (0.64 < r < 0.78). Analysis of variance demonstrated that the SCSA results can be influenced by the age of the donor (P < 0.0001), smoking habits (P < 0.05), the presence of leukocytes and immature germ forms in the ejaculate (P < 0.0001), and the duration of sexual abstinence (P < 0.0001). Furthermore, the relationship between the SCSA data and sperm concentration, morphology, and vitality was weak (-0.22 < r <-0.46). Therefore, the SCSA provides independent and complementary measurements of semen quality and is thus a useful tool for epidemiological studies, but the effects of some confounders should be accounted for in the survey design and analysis.
Repeatability and variance analysis on multiple computer-assisted (IVOS*) sperm morphology readings
Andrologia, 1999
The repeatability of the Hamilton Thorne Research IVOS (version 10) semen analyser (dimension specific software, version 3) in the evaluation of sperm morphology according to strict criteria was investigated in this study. The repeat measures investigated were cell-cell (300 cells, 3 x each), intraslide (20 slides, 3 x each) and interslide (30 samples, 3 slides each), and their normal sperm morphology outcomes were recorded. Semen samples with varying normal sperm morphology percentages were obtained and sperm morphology slides prepared. The slides were stained with Diff-Quik stain. Agreements between evaluations were determined using the K statistic and average coefficients of variation. The predictive probability for an abnormal cell 'given a prior abnormal cell outcome was 9 1 %, and 89% for a similar prediction of a normal cell. The predictive probabilities for an abnormal or a normal cell given two prior abnormal or two prior normal cell outcomes were 95% and 94%, respectively. No significant bias was obtained between the repeat probabilities for normal and abnormal sperm cells. The average coefficients of variation for the intraslide trial were 9.73% and 8.30% when 100 and 200 sperm ceIls were evaluated, respectively. The average coefficient of variation for the interslide trial was 15.39%. The technical importance of good sample and slide preparation technique has once again been highlighted by this study. A uniform (spatial homogeneity), high concentration (5-10 cells per computer screen) smear must be made and the cells stained with optimal intensity (maximum contrast). In a trial in which 2000 cells were evaluated, 19 objects (0.95%) were identified as spermatozoa, but were debris. The
Current status and potential of morphometric sperm analysis
Asian Journal of Andrology, 2016
Considering the forward perspective, most studies have tried to establish functional relationships between sperm traits during the fertilization process and their performance in assisted reproductive techniques (ARTs), with regard to sperm resilience or fertility. Typically, traits such as motility, viability (either plasma-lemma integrity or the hyper-osmotic shock test [HOST] responsiveness), acrosomal integrity, or absence of abnormalities have been used as endpoints for predicting sperm fertility or, rather, discarding potentially low-fertility semen doses. 10-13 Availability of advanced techniques and hardware such as computer-assisted sperm analysis (CASA), 14-16 fluorescence probes and ultimately flow cytometry, 17,18 and new endpoints, e.g. capacitation and chromatin assessment, 19,20 have allowed more objective and faster analysis, but the predictive power of laboratory sperm assessment still needs improvement. 21 The routine evaluation of semen has traditionally included the assessment of normal sperm morphology, but the important subjective component has limited its practical use. 22 The development of automatic image-processing systems has displaced classical methods and is a major advance in sperm analysis. Computer-assisted sperm morphometric analysis (CASA-Morph) systems have been successfully used to determine the relationships between sperm shape and fertility of males 23,24 or sperm freezability. 25,26 There is a need to develop new analytical tools to capture sperm diversity better, and to improve data analysis methods. New equipment
Correlation between sperm motility and sperm chromatin structure assay parameters
Fertility and Sterility, 2003
To evaluate the association between chromatin structure and sperm motility. Design: Cross-sectional prospective study. Setting: Patient(s): One hundred seventy-one males from Danish first pregnancy planner couples (group 1) and 278 Swedish military conscripts (group 2). Main Outcome Measure(s): Sperm chromatin structure assay (SCSA) parameters, DNA fragmentation index (DFI), high DNA stainable (HDS), and sperm motility, which was evaluated manually and by use of computer-aided sperm analysis (CASA).
Asian Journal of Andrology, 2016
fragmentation in human spermatozoa. 21 Development of simple kits for the diagnosis of DNA fragmentation has increased the number of studies on the significance of DNA fragmentation in several species, 22 but there is some controversy over the diagnostic significance of the differential tests, making it difficult to decide which is the best to use. 23,24 Two commercial kids have been developed around the SCD technique: Halosperm® (Halotech, Madrid, Spain) and SDFA (ACECR, Tehran, Iran). The purpose of the present study was to compare the results from these commercial kits, by performing a morphometric analysis with the ISAS® v1 DNA fragmentation module (Proiser, Valencia, Spain). These morphometric data were used, for the first time to our knowledge, to define mathematical clusters that provide a classification matrix of different subpopulations of sperm head DNA-reacted cells. MATERIALS AND METHODS Study population Seven volunteers signed informed consent form to participate and have their semen used in the study. Semen samples were collected by masturbation after sexual abstinence for 3-5 days. Each sample was collected in a clean 60-ml wide-mouthed universal container and stored at 37°C in an incubator for 30 min to allow liquefaction.