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Biomedical subjects

A Botchan

Publications and source records attributed to A Botchan.

At least 19 recordsLinked to original sources
PubMed2006

Expression profile of AZF genes in testicular biopsies of azoospermic men.

BACKGROUND: The Y-chromosome AZF regions include genes whose functions and specific roles in spermatogenesis have not been fully clarified. This study investigated the expression of several AZF (USP9Y, DDX3Y/DDX3Yt1, EIF1AY and PRY) and USP9X transcripts in testicular biopsies of 89 azoospermic men who had been classified by histology and cytology assessments. METHODS: Expression was analysed by RT-PCR, and some biopsies were evaluated by multiplex RT-PCR. Quantitative PCR was performed in some biopsies to determine the ratio of the testis-specific transcript DDX3Yt1 to the total DDX3Y transcription. RESULTS: The expression of USP9Y, USP9X and DDX3Y was found in all the specimens tested, whereas DDX3Yt1 expression was diminished or undetectable in several biopsies with impaired spermatogenesis. EIF1AY was detected in all except two of the specimens. Noteworthy, PRY expression was detected mainly in biopsies with germ cells, and this association was significant (P < 0.001). An identical expression profile was obtained by either single or multiplex RT-PCR. CONCLUSIONS: These findings suggest that PRY is usually expressed in germ cells, whereas the other transcripts are also expressed in testicular somatic cells. The absence of EIF1AY expression might sporadically contribute to azoospermia. The decreased ratio of DDX3Yt1/DDX3Y transcript in impaired spermatogenesis suggests that the DDX3Yt1 transcript is under-expressed in impaired spermatogenesis. The findings contribute to the search and selection of the most valuable gene markers potentially useful as additional tools for predicting complete spermatogenesis by multiplex expression analysis.

Azoospermia↗
PubMed2004

Seasonal variations in pre- and post-thaw donor sperm quality.

BACKGROUND: The present study was conducted to evaluate seasonal variability in the quality of pre- and post-thaw semen parameters among sperm bank donors. METHODS: The first two consecutive ejaculates during the months March (spring, 92 males), June (summer, 97 males), September (autumn, 81 males) and December (winter, 97 males) were analysed. A comparison was made between sperm parameters from the same sperm donor at different seasons. Only males who donated semen samples during at least two seasons were enrolled in the study group (n = 103). Sperm specimens were cryopreserved in aliquots with fixed range of 8-12 x 10(6)/ml of progressive motile sperm concentration after thawing. RESULTS: Differences between months were found in sperm concentration (P = 0.030) and normal morphology (P = 0.038); highest values were found in March and December, and the lowest in September. Mean specimen volume and percent of motile sperm cells did not vary throughout the seasons. The freezability of the donors' sperm dropped dramatically from March to September, as determined by the number of straws (fixed aliquots of 0.5 ml) and total thawed progressive motile sperm that were cryopreserved for each male (P = 0.017 and P = 0.002, respectively). CONCLUSIONS: Cryopreservation of donor sperm is more effective during winter and spring than during the rest of the year.

Adult↗
PubMed2001

Expression of CDY1 may identify complete spermatogenesis.

OBJECTIVE: To investigate the expression of deleted in azoospermia (DAZ), RNA-binding motif (RBM), and chromodomain y1 (CDY1) genes in the testes of men with azoospermia with variable histopathologies. DESIGN: Prospective study. SETTING: Andrology laboratory of a university-affiliated maternity hospital. PATIENT(S): Sixty-six men with azoospermia. INTERVENTION(S): Testicular sperm extraction. MAIN OUTCOME MEASURE(S): The results of gene expression in testicular tissue tested by RT-PCR were correlated with those of histopathologically and microscopically examined minced testicular tissue. Y chromosome microdeletion testing and karyotyping were performed, as was direct sequencing of CDY1-PCR products. RESULT(S): CDY1-minor expression was detected in all biopsies in which mature spermatids/spermatozoa were observed by histological analysis and/or in the minced tissue. CDY1-minor expression was also detected in two biopsies with arrest at the spermatocyte stage during which no mature spermatids/spermatozoa were observed. A previously unreported CDY1-minor alternative splicing transcript was identified. DAZ and RBM gene expressions were detected in all biopsies in which at least a few germinal cells in early stages were found and in one biopsy histologically determined as Sertoli cell only. CONCLUSION(S): Our preliminary results suggest that CDY1-minor expression might increase the prospect for complete spermatogenesis, while RBM and DAZ expression can only be indicative of the presence of germinal cells.

Adult↗
PubMed2001

Vascularity index distribution within the testis: a technique for guiding testicular sperm extraction.

Azoospermia is defined as the absence of spermatozoa in the ejaculate, although some foci of spermatogenesis may exist in the testes of these men. Currently, there are no clinical, seminal or hormonal parameters for identifying spermatogenesis within the testis sufficient for achieving genetic offspring. As a result, multiple biopsies are performed at several arbitrary sites of both testes in search of spermatozoa. We developed a power Doppler (PD) ultrasound (US) image-based technique that predicts sites with the greatest potential for spermatogenesis. PDUS images of the testes of azoospermic men were acquired at seven cross-sections to reconstruct a 3-D matrix for constructing a spatial map of preferential regions where spermatozoa are most likely to exist. This technique may obviate the need for arbitrary multiple biopsies that inflict some degree of damage upon testicular tissue, and may increase the success rate of identifying viable spermatozoa in testicular biopsies.

Biopsy↗
PubMed2001

Intrauterine insemination in male factor subfertility: significance of sperm motility and morphology assessed by strict criteria.

The study was conducted to evaluate the results of IUI treatment in a homogenous group with male factor infertility, and to assess the correlation of sperm variables, including sperm morphology by strict criteria, with pregnancy achievement after IUI. A total of 108 couples with no apparent female aetiology for infertility underwent 264 intrauterine insemination treatment cycles. A comparison was made between the sperm variables in two groups in which the achievement of pregnancy differed. The percentage of motile spermatozoa, degree of motility and normal morphology (by strict criteria) were significantly higher in the pregnant group compared with that of the nonpregnant group. A significant difference in pregnancy rates per couple after intrauterine insemination was demonstrated among three groups according to the percentage of sperm morphology, i.e. poor (< 4%), fair (4-14%) or good (> 14%) (11.1%; 36.1% and 50.0%, respectively). Intrauterine insemination is a valid mode of treatment in cases with male infertility, provided that normal morphology by strict criteria is higher than 4%.

Adult↗
PubMed2001

Effect of freezing-thawing on the expression of mannose-ligand receptors on human spermatozoa: the impact on sperm capacitation and acrosome reaction.

The aim of this study was to evaluate the change in the expression of mannose-ligand receptors following a freezing-thawing procedure, in order to assess its impact on sperm capacitation and acrosome reaction. Twenty semen samples were obtained from fertile donors. Sperm samples were divided into two equal volumes. One aliquot was cryopreserved and the other aliquot was incubated at 32 degrees C. After 2 h the frozen sample was thawed and both samples were further incubated at 32 degrees C to allow capacitation. Mannose receptors were examined following 4 and 22 h of incubation using a mannosylated-BSA-FITC probe. The expression of mannose-ligand receptors on the sperm plasma membrane was determined according to the fluorescence pattern: pattern I represents pre-capacitation, pattern II represents capacitated spermatozoa and pattern III represents acrosome-reacted spermatozoa. After 4 h incubation in capacitating medium, the percentages of patterns I, II and III were 90, 7 and 3% for fresh spermatozoa and 89, 8 and 3% for frozen-thawed spermatozoa, respectively (P > 0.05). Following 22 h of incubation, the percentages of patterns I, II and III were 84, 11 and 5 for fresh spermatozoa and 83, 11 and 6% for frozen-thawed spermatozoa, respectively (not significant at P > 0.05). The percentages of patterns II and III in fresh and frozen-thawed spermatozoa were increased by the same magnitude with longer incubation in the capacitating conditions. It was concluded that the freezing-thawing procedure for human spermatozoa does not affect the expression of mannose-ligand receptors and the dynamics of sperm pre-fertilization processes.

Acrosome Reaction↗
PubMed2001

The contribution of RNA-binding motif (RBM) antibody to the histopathologic evaluation of testicular biopsies from infertile men.

Testicular biopsies of infertile men are often characterized by mixed histologic patterns, with different types of spermatogenic impairments being found in adjacent seminiferous tubules. RNA-binding motif (RBM) is a nuclear protein expressed exclusively in the male germ cell line. We reasoned that RBM might be a useful marker to identify germ cells in testicular sections, particularly in biopsies with mixed histologic phenotype and small focal concentrations of spermatogenesis. Testicular biopsies from azoospermic men were immunohistochemically evaluated for RBM expression. RBM expression was detectable in spermatogonia, spermatocytes, and round spermatids in biopsies of men with obstructive azoospermia and normal spermatogenesis. No specific cell staining was shown in cases of Sertoli-cell-only (SCO) syndrome. In biopsies of patients with spermatogenic disorders, all the germ cells were stained up to and including the stage level of the arrest in spermatogenesis. This approach enabled identification of small focal concentrations of spermatogenesis in a biopsy previously classified as being SCO by hematoxylin and eosin staining. Thus, RBM can be a useful immunohistochemical marker for the specific identification of germ cells and provide greater accuracy in the histopathologic evaluation of testicular biopsies.

Antibodies↗
PubMed2001

Results of 6139 artificial insemination cycles with donor spermatozoa.

BACKGROUND: Artificial insemination by donor spermatozoa (AID) can prove a valuable treatment for a number of male factor disorders, although its success rate is variable. METHODS: Retrospective analysis of the results of 6139 cycles performed in 1001 women during an 18 year period is presented. Pregnancy rates per cycle are presented as a function of: female fertility history, treatment modalities, medication used for induction of ovulation, female age, year of treatment, consecutive cycle effect and the use of fresh versus frozen-thawed spermatozoa. RESULTS: Overall pregnancy rate of 12.6% and cumulative pregnancy rate after 12 months of treatment of 75% were achieved. Age was found to be the most important determinant for success rate. CONCLUSIONS: Since the establishment of AID treatments, the mean age of the population of women receiving treatment has increased each year. Consequently, success rate did not improve, even with the use of more sophisticated medical modalities.

Abortion, Spontaneous↗
PubMed2000

Zona pellucida binding improvement effect of different sperm preparation techniques is not related to changes in sperm motility characterizations.

OBJECTIVE: To study the relation between changes in sperm motion characteristics and sperm binding capacity to the zona pellucida (ZP) after different sperm preparation techniques. DESIGN: Prospective study. SETTING: Andrology laboratory at the Lis Maternity Hospital. PATIENT(S): Sixty-three infertile men. INTERVENTION(S): In vitro preparation of 57 sperm samples by pentoxifylline (PTX), 38 samples by SpermPrep I (SP1), and 27 samples by mini-Percoll (mPER). Sperm preparation was evaluated in comparison with the control washing technique. MAIN OUTCOME MEASURE(S): Motility variables and binding capacity. RESULT(S): Binding capacity after PTX, SP1, and mPER preparation methods was significantly higher compared with the control medium preparation. Computer-assisted sperm analysis (CASA) of sperm motility revealed a significant increase in a number of variables after the different treatments. A statistically significant but low correlation was observed between sperm binding capacity after control swim-up preparation and VAP (r = 0.27) as well as with VSL (r = 0.30). No significant correlations were found between the changes in the CASA and the improved sperm binding to the ZP. CONCLUSION(S): The two biologic effects of binding capacity enhancement and motility features improvement are not related. Furthermore, because the binding assay is highly correlated with fertilization capacity, these alterations in motility, as evidenced by CASA, probably have a minor effect on fertilization in vitro.

Electronic Data Processing↗
PubMed2000

Evaluation of meiotic impairment of azoospermic men by fluorescence in situ hybridization.

OBJECTIVE: To identify predictive criteria for the existence of spermatogenesis in nonobstructive azoospermic men. DESIGN: Prospective study. SETTING: Andrology laboratory at a teaching hospital. PATIENT(S): Twenty-two azoospermic men were divided into three groups by qualitative testicular histopathology and the presence of spermatozoa in minced biopsies. INTERVENTION(S): Testicular biopsies evaluation. MAIN OUTCOME MEASURE(S): The presence of spermatozoa and/or mature spermatids, the percentage of sex vesicle formation (X and Y chromosomes in proximity), and the pairing of the two 18 homologous chromosomes. RESULT(S): Spermatozoa and mature spermatids were found in 17 study patients. Whenever few mature spermatids and/or spermatozoa were found, the rates of X-Y and 18 bivalents were significantly higher (mean +/- SD, 73% +/- 13. 3% and 91% +/- 7.1%) than those in cases of spermatocyte maturation arrest (23% +/- 8.0% and 60% +/- 11.8%, respectively). CONCLUSION(S): Pairing of chromosomes during meiosis is apparently related to the progression of spermatogenesis. Consequently, high rates of bivalent formation increase the prospect of focal spermatogenesis in the testis, despite the failure to identify mature spermatids in the specific testicular biopsy under examination.

Adult↗
PubMed2000

Maturation phenotype of Sertoli cells in testicular biopsies of azoospermic men.

The involvement of Sertoli cells in different spermatogenic impairments has been studied by an immunohistomorphometric technique using cytokeratin-18 (CK-18) as a marker for immature Sertoli cells. CK-18 is known to be expressed in Sertoli cells during prenatal and prepubertal differentiation and is normally lost at puberty. Forty-nine azoospermic men were included in the current study. Quantitative measurements on testicular biopsies revealed the highest CK-18 expression in the mixed atrophy biopsies (22 men), a lower expression in the Sertoli cell-only (SCO) biopsies (12 men), and minimal residual staining in the group considered as representing normal spermatogenesis (six obstructive azoospermia patients). The cytokeratin immunopositive-stained tubules were associated either with arrest in spermatogenesis or with SCO. Examination of sections from nine men with microdeletions in the AZF region of the Y chromosome revealed that these men were either negative for CK-18 expression or showed only weak residual staining. This may suggest that the spermatogenic defect in the AZF-deleted men originates in the germ cell and has no impact on Sertoli cell maturation. The cause that determined the spermatogenic defect in the other cases of male infertility with high CK-18 expression may have damaged both the Sertoli and the germ cells.

Adult↗
PubMed2000

Follow-up of sperm concentration and motility in patients with lymphoma.

Lymphomas are a group of diseases, prevalent at reproductive age. Fertility is notoriously reduced among lymphoma patients. This study evaluates pre- and post-treatment semen concentration and motility, and factors associated with semen quality deterioration. We followed-up 33 patients with non-Hodgkin's lymphoma or with Hodgkin's disease during the years 1987-1997 who were referred for semen cryopreservation. Pretreatment semen analysis, and hormonal profile were recorded at diagnosis and at least 1 year after completion of the treatment, and compared. Medical records for disease type, disease stage and treatment protocols were related to long-term sperm outcome. Hormonal concentrations were not predictive of post-treatment sperm concentration. In patients with localized disease, initial sperm concentration and motility tended to be preserved, compared with patients with widespread disease (P: = 0. 016). In Hodgkin's disease patients, treatment with the adriamycin, bleomycin, vinblastine and dacarbazine (ABVD) protocol was superior to the mechloretamine, vincristine, procarbazine and prednisone with ABV protocol regarding germinal toxicity (P: = 0.0008). The post-treatment sperm outcome was better in patients treated with local irradiation than in those who did not undergo irradiation (P: = 0.0027). No predictive tools for post-treatment fertility were found and, therefore, every patient with a lymphoma should have his semen cryopreserved at diagnosis.

Adolescent↗
PubMed2000

Varicocele and the testicular-renal venous route: hemodynamic Doppler sonographic investigation.

The aim of this study was to quantify the anatomic and hemodynamic components of the testicular venous drainage for the purpose of understanding their mechanisms of interacting in producing mutual effects, such as "nutcracker" phenomenon, reflux, and varicoceles. Seventy-five male subjects were studied at rest and during Valsalva maneuver. Aortomesenteric distance and angle, flow velocity in different segments of the renal veins, testicular vein diameter, and flow inversion were evaluated using standard ultrasound equipment with spectral and color Doppler capabilities. The velocity of flow in the proximal segment of the left renal vein (17.5 cm/s) was found to be significantly lower than that in both the distal left renal vein (121 cm/s) and the right renal vein (37 cm/s). The flow velocity in the proximal left renal vein decreased with decrease in the aortomesenteric distance and angle. Testicular vein diameters greater than 3 mm were statistically associated with decreased superior mesenteric artery angle. A significant association also was found between the left testicular vein diameter (in Valsalva maneuver) and inversion of flow. The decrease in flow velocity in the left renal vein proximal to the bifurcation of the superior mesenteric artery from the aorta supports the "nutcracker" theory. An association was found between the decrease of superior mesenteric artery angle and the increase in testicular vein diameter. Another association exists between the presence of reflux during Valsalva maneuver and increased testicular vein diameter. These finding were significant only for testicular vein diameter values greater than 3 mm.

Adolescent↗
PubMed2000

[Immunohistochemical identification of testicular germ cells--preliminary report].

The use of testicular spermatozoa for intracytoplasmic sperm injection introduced a new treatment modality for management of male infertility. Since testicular biopsies of non-obstructive azoospermic men are not homogenous in their histological patterns, identification with certainty of focal spermatogenesis might be difficult, particularly in those with small foci of spermatogenesis. We used an immunohistochemical marker of the male germ line, an antibody generated against RBM (RNA-binding-motif), to recognize with high precision the presence of germ cells in the biopsy. Biopsies of 30 men with azoospermia, most with non-obstructive azoospermia and a few with obstruction of the vas deferens, were evaluated. Immunohistochemical staining for RBM protein contributed to the detection and accuracy of the identification of germ cells. Furthermore, this immunohistochemical technique aided the histopathologist to focus on even small foci of spermatogenesis. Absence of the protein expression confirmed the diagnosis of Sertoli-cell-only syndrome. The results indicate that expression of RBM can be a diagnostic marker for identifying the germ cells of small concentrations of spermatogenesis. This method can enhance the accuracy of histopathological evaluation of testicular biopsies that had formerly relied mainly on hematoxylin-and-eosin staining.

Biopsy↗
PubMed1999

Pre-freezing sperm preparation does not impair thawed spermatozoa binding to the zona pellucida.

The present study was conducted to assess the fertilizing potential of frozen-thawed spermatozoa, which were cryopreserved after separation on a Percoll gradient, or washed out of seminal plasma. For this purpose, binding to the zona pellucida and other characteristics of the treated sperm cells were compared with those of cryopreserved spermatozoa from the same original sample which were not manipulated before freezing. Semen specimens were obtained from 80 candidates for sperm donation. Percoll-treated sperm samples compared with the sibling, unprocessed controls had significantly higher values of sperm motility characteristics and per cent of cells with normal morphology after freezing and thawing. Sperm binding ability to the zona pellucida was not statistically different (109 +/- 8.1% and 94 +/- 6.7% in unprocessed and Percoll-treated samples respectively). Sperm specimens processed by washing had significantly higher values for motility characteristics than untreated sibling samples, but no differences were found between the treated and untreated samples for morphology and binding to the zona pellucida (hemizona index of 75 +/- 7.0% and 76 +/- 6.7% in unprocessed and washed samples respectively). These findings suggest that, judged by the binding assay, the aforementioned pre-freezing separation processes have no adverse effect upon the fertilizing potential of the thawed sperm cells. These procedures make it possible to optimize the progressive motile sperm cell concentration of the frozen specimen, which facilitates the storage of samples with good quality, even when the features of the original semen are sub-optimal.

Cell Separation↗