Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Sperm Retrieval”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 649 records · Page 36Linked to original sources
PubMed1999

Fertility of mouse spermatozoa retrieved from cadavers and maintained at 4 degrees C.

After male animals die, the spermatozoa within the testis and epididymis eventually disintegrate. In this study, the motility, viability and fertility of mouse spermatozoa were examined after retrieval from the epididymis at various days after death. Cadavers were maintained in a refrigerator at 4 degrees C. About 30% of the spermatozoa collected 10 days after death were viable, but they had limited ability to fertilize oocytes in vitro. However, when the spermatozoa were injected into oocytes, the fertilization rate was over 80%. Normal live fetuses were even obtained using immotile spermatozoa retrieved 20 days after death. Therefore, when valuable male animals die unexpectedly and sperm cryopreservation is not possible immediately, temporal storage of cadavers (or epididymis and vas deferens) at 4 degrees C in a regular refrigerator followed by intracytoplasmic sperm injection may help to preserve the genome of individuals. This procedure could be particularly important in endangered species.

Animals↗
PubMed1997

Relationship of human follicular diameter with oocyte fertilization and development after in-vitro fertilization or intracytoplasmic sperm injection.

The aim of this work was to evaluate the relationship between follicular size at the time of oocyte retrieval, and the subsequent oocyte competence to be fertilized and to develop in vitro. All the obtained oocytes were classified according to the corresponding volume of aspirated follicular fluid. Aspirated volume of follicular fluid <2 ml corresponded to a follicular diameter <16 mm and constituted the small size group. Volume of follicular fluid from 2 to 6 ml corresponded to a diameter from 16 to 23 mm and constituted the medium size group. The large size group contained follicles with diameter >23 mm and corresponded to an aspirated volume of follicular fluid of >6 ml. A progressive and significant increase in the rates of oocytes with a first polar body was observed from the small size group to the other groups and from the medium to the large size group: 75.3, 85.9 and 95.3% respectively. After classical in-vitro fertilization (IVF), significantly better rates of fertilization and development were obtained in the medium size group compared to the two other groups. Moreover, a positive relationship was observed between follicular diameter and rates of embryos scored as 'good' when oocytes were fertilized by intracytoplasmic sperm injection (ICSI). These results demonstrated that follicular size is positively related to the oocyte ability to be fertilized and to develop. Although oocytes from small follicles gave lower percentages of development probably due to partial oocyte incompetence, they allowed an increase in the total number of embryos scored as 'good'.

Cell Size↗
PubMed1994

Correlation of zona-binding with oocyte maturation and sperm motility in rhesus monkeys by hemizona assay.

The hemizona assay (HZA) in Rhesus monkeys was employed to study the correlation of zona-binding ability with sperm motility or with naturally developing oocytes at various maturational stages. Oocytes from unstimulated ovaries were retrieved within 2 hr from monkeys sacrificed for vaccine production (in reproductive season, but with their menstrual cycles not determined). Oocytes were divided into four groups based on their morphological maturation: 1) Oocytes surrounded by more than one cumulus layer (MC); 2) Oocytes retaining intact germinal vesicle nuclei (GV); 3) Oocytes with germinal vesicle breakdown showing distinct perivitelline space (PVS); and 4) Oocytes extruding the first polar body (PB1). The mean numbers of sperm bound to hemizona for PB1, PVS, GV, and MC groups were 132.9 +/- 12.0, 71.5 +/- 10.1, 36.1 +/- 4.0, and 20.1 +/- 2.9 (Mean +/- SE), respectively. The four groups showed significant differences from each other in sperm/egg binding ability (P < 0.01). The number of bound sperm significantly increased with oocyte maturation. The present study also showed that zona-binding ability was also affected by sperm motility. For sperm with 67.7% motility and sperm with 31.2% motility, the average numbers of bound sperm were 43.5 +/- 2.2 and 25.3 +/- 2.9 (Mean +/- SE), respectively. There was significantly higher binding ability for sperm with higher motility (P < 0.01). The results suggest that: 1) The rhesus monkey model can serve as a very sensitive model for studying sperm/egg interaction by HZA; 2) Sperm motility positively correlated with sperm/egg binding; and 3) Sperm/egg binding ability increases with oocyte maturation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Analysis of factors contributing to success in a program of micromanipulation-assisted fertilization.

OBJECTIVE: To determine factors important to clinical success in micromanipulation-assisted in vitro fertilization (IVF). DESIGN: Procedures invoked in two separate series of micromanipulation-assisted IVF cycles, one unsuccessful (series I) and the other successful (series II), were compared in an effort to identify changes that led to clinical success. SETTING: University-based IVF clinic. PATIENTS: In both IVF series involving micromanipulation, patients consisted of infertile couples who fit any of five categories of male-factor related infertility. The female patients underwent controlled hyperstimulation for oocyte retrieval and the oocytes were inseminated normally or were subjected either to partial zona dissection or subzonal sperm insertion to assist fertilization. Results in all groups were compared between the two patient series. RESULTS: In the successful series II, a noticeable improvement in fertilization rate and embryo quality was observed compared with series I. A significant increase in the percentage of patients reaching embryo transfer, the pregnancy rate per transfer, and the pregnancy rate per retrieval were noted in series II; a 25% ongoing pregnancy rate per retrieval was observed overall in this successful group, with "ongoing" defined as manifestation of at least a fetal sac on ultrasound with no detectable problems. Patients with a mixed transfer of embryos derived from manipulated and normally inseminated oocytes had a 75% rate of pregnancy per transfer in series II. Differences between the two series could not be attributed to patient selection or biases in selection of oocytes relegated to micromanipulation. However, oocyte handling, micromanipulation, and culture protocols differed significantly between the two series in that temperature and pH of oocytes was better controlled, and micromanipulation time was minimized in series II. CONCLUSION: Success in micromanipulation depends on maintenance of the oocyte in a stable and supportive environment throughout the micromanipulation procedure. It is also important to minimize trauma to the eggs by performing micromanipulation rapidly and with minimal distortion of the egg. Patients with a poor fertilization rate in standard IVF may experience a substantial increase in the likelihood of pregnancy when micromanipulation-assisted fertilization is performed on some eggs.

Cleavage Stage, Ovum↗
PubMed1999

Diagnostic epididymal and testicular sperm recovery and genetic aspects in azoospermic men.

Various procedures for sperm recovery in azoospermic men have been described, from open testicular biopsy to simple needle aspiration from the epididymis and the testis. Fifty-one obstructive and 86 non-obstructive azoospermic men were treated to compare the recovery of spermatozoa obtained by percutaneous aspiration from the epididymis (PESA) and aspiration/extraction from the testis (TESA, TESE) with histopathology. If TESA failed, the work up proceeded with TESE. All patients were karyotyped. Spermatozoa were recovered by PESA or TESA in all obstructive men (51/51 patients). In 22 out of 86 patients with non-obstructive azoospermia, testicular spermatozoa could be successfully recovered by TESA. In five additional patients TESE was successful in recovering spermatozoa where TESA had failed. In 43 patients, neither TESA nor TESE was successful. Sixteen patients chose not to proceed with TESE. Seven out of 86 patients had an abnormal karyotype in the non-obstructive group (8%), none in the obstructive group. In the non-obstructive patient group testicular histopathology showed hypospermatogenesis, incomplete maturation arrest and germ cell aplasia with focal spermatogenesis in cases where spermatozoa were recovered and complete germ cell aplasia, complete maturation arrest and fibrosis in cases where no spermatozoa were found. Spermatozoa were recovered by PESA or TESA from all patients with obstructive azoospermia and from approximately 40% of patients with non-obstructive azoospermia by TESA or TESE. Retrieval of viable spermatozoa in the infertility work-up was highly predictable for sperm recovery in subsequent ICSI cycles. TESA performed under local anaesthesia seems almost as effective as more invasive procedures in recovering testicular spermatozoa, both in obstructive and non-obstructive azoospermic men.

Epididymis↗
PubMed2001

A comparison of ICSI outcomes with fresh and cryopreserved epididymal spermatozoa from the same couples.

The published experience with frozen-thawed epididymal spermatozoa and intracytoplasmic sperm injection (ICSI) suggests that fertilization and pregnancy success rates are comparable to those achieved with freshly retrieved spermatozoa. However, no study has exactly compared clinical outcomes between the two IVF/ICSI cycles in the same couples. To formally address this issue, we assessed ICSI outcomes in couples each of whom had had two IVF/ICSI cycles: one using fresh and the second using frozen-thawed epididymal spermatozoa obtained from a single aspiration procedure. From a pool of 101 consecutive patients undergoing IVF/ICSI with epididymal spermatozoa, 19 couples initially used fresh epididymal spermatozoa and subsequently underwent a second IVF/ICSI procedure with frozen-thawed spermatozoa from the same aspiration. Normal (2PN) oocyte fertilization rates, embryo quality and pregnancy rates were compared between the two IVF/ICSI cycles for each couple. In the fresh epididymal sperm group, 58.4% of the injected oocytes fertilized normally compared with 62.0% of the injected oocytes in the frozen-thawed epididymal sperm group, revealing no statistically significant difference. Graded embryo quality also did not differ significantly between the paired IVF/ICSI cycles. The clinical pregnancy rates were 31.6% (6/19) and 36.8% (7/19) in the first and second cycles respectively. All but one pregnancy were singletons. In summary, this study provides strong evidence to support the notion that motile, cryopreserved and thawed epididymal spermatozoa are equal to freshly retrieved spermatozoa for ICSI in couples with obstructive azoospermia.

Cryopreservation↗
PubMed1997

Fertilization and pregnancy using intentionally cryopreserved testicular tissue as the sperm source for intracytoplasmic sperm injection in 10 men with non-obstructive azoospermia.

Testicular tissue extraction (TESE) to obtain spermatozoa for use with intracytoplasmic sperm injection (ICSI) has recently been employed in patients with non-obstructive azoospermia. Standard protocol is to retrieve a new sample of testis tissue on the day of oocyte recovery. Unfortunately, approximately 30% of men will possess no spermatozoa in their tissue, making ICSI an impossibility. We investigated whether testicular tissue that was intentionally obtained well before any planned ICSI cycle and cryopreserved could then serve as an efficacious sperm source in a subsequent ICSI cycle. This study reports on 10 men with non-obstructive azoospermia who did have spermatozoa found within their testis tissue at the time of TESE and who chose to use their frozen samples as the source of spermatozoa for a later cycle of ICSI. In 19 cycles the overall fertilization rate was 48%. Embryo transfer occurred in 89% of cycles. Two couples have achieved pregnancy (one ongoing, one delivered). All patients except one had multiple vials of frozen tissue remaining following their first cycle. This approach is offered as an alternative to repeated testicular tissue sampling, as the availability of spermatozoa is assured prior to the initiation of ovulation induction. This tissue can be harvested at the same time as diagnostic biopsy, thereby minimizing the number of surgical procedures.

Adult↗
PubMed2000

Testicular sperm extraction combined with cryopreservation of testicular tissue in the treatment of azoospermia.

BACKGROUND: The aim of the present study was to verify the feasibility of cryopreserving testicular tissue during the first diagnostic biopsy and then using thawed sperm to inseminate the partner's oocytes. The expected advantages are: (i) minimal risk of not having spermatozoa available at the time of intracytoplasmic sperm injection; (ii) no repeated surgical interventions, and (iii) programming the treatment cycle at the couple's convenience. MATERIALS AND METHODS: Between May 1996 and May 1998, 64 azoospermic patients underwent investigative testicular biopsy combined with cryopreservation of spermatozoa which were retrieved in a simultaneously examined fresh sample. Testicular tissue cryopreservation was carried out in 43 cases (67%) for later intracytoplasmic sperm injection attempts. RESULTS: In all, 23 couples underwent 26 assisted conception cycles; the fertilization rate was 64% with spermatozoa (139/218, 24 cycles), 40% with round spermatids (2/5, 1 cycle), and 69% with elongated spermatids (9/13, 1 cycle). The embryo cleavage rate was 84%. The mean number of embryos replaced in 24 patients was 2.7 +/- 0.7. In 2 cases, embryo quality was very poor, and they were not transferred to the patients. Eight clinical pregnancies resulted (35%/patient and 33%/transferred cycle) with an implantation rate of 14.1%; 2 patients have already delivered and 6 pregnancies are ongoing normally. CONCLUSIONS: Testicular tissue cryopreservation during the first diagnostic biopsy is an alternative to repeated surgical interventions. Patients can initiate an ovarian stimulation cycle, confident of having spermatozoa available. Moreover, since only one straw is routinely used for each intracytoplasmic sperm injection cycle, the frozen tissue remains as a sperm source for multiple attempts.

Adult↗
PubMed1994

The effect of antisperm autoantibodies in male or female partners undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To evaluate the effects of sperm bound autoantibodies on the outcome of IVF-ET. DESIGN: Couples with positive antisperm autoantibodies as determined by the immunobead test were retrospectively classified into two groups: group A, consisting of 15 couples with positive antisperm antibodies in the female sera; and group B, consisting of 16 couples with sperm antibodies bound to motile spermatozoa from the male partner. Both groups were subclassified according to pregnancy outcome, i.e., pregnant and nonpregnant cycles. PATIENTS: Thirty-one couples with positive antisperm autoantibodies were compared with 312 couples with tubal infertility undergoing IVF-ET. RESULTS: No significant correlation could be shown between the mean percent binding of any specific immunoglobulin (Ig) class (G, A, and M) nor localization of sperm binding with regard to fertilization and embryonic development among pregnant and nonpregnant cycles within groups A and B. The mean fertilization rate was 59% in the control group, compared with 62% in group A and 52% in group B. Overall, the pregnancy rate (PR) in IVF-ET cycles with positive sperm autoantibodies did not demonstrate a decreasing trend compared with controls. The PR per cycle, per oocyte retrieval, and per ET was higher in group A (47%, 50%, and 53%, respectively) compared with group B (32%, 33%, and 37%) and to controls (27%, 31%, and 34%). The implantation rate was lowest in the control group (10%) compared with the study groups (group A, 20% and group B, 14%). CONCLUSION: In vitro fertilization-embryo transfer is not significantly affected by the presence of sperm autoantibodies in female sera used to supplement the culture media or antibodies bound to inseminated sperm.

Adult↗
PubMed2004

Expression of sHLA-G in supernatants of individually cultured 46-h embryos: a potentially valuable indicator of 'embryo competency' and IVF outcome.

A retrospective cohort study was conducted on 201 women aged 28-44 years, each of whom underwent one cycle of IVF-embryo transfer with fresh, intracytoplasmic sperm injection (ICSI)-derived 7- to 10-cell embryos, transferred 72 h after oocyte retrieval. Samples of media surrounding separately cultured embryos were collected 46 h post-ICSI and stored for subsequent specific enzyme-linked immunosorbent assay. A total of 594 embryos (from own or donor oocytes) were transferred to 201 women. Group A comprised 159 recipients under 39 years and group B compromised 42 recipients aged 39-44 years. Groups A-1 and B-1 recipients had at least one embryo that tested above the geometric mean for soluble human leukocyte antigen-G (sHLA-G) ('positive expression') transferred. In groups A-2 and B-2, all embryos transferred expressed sHLA-G below the geometric mean ('negative expression'). In group A-1, 72/101 women (71%) achieved ultrasound confirmed (clinical), viable (cardiac activity observed) pregnancies. The implantation rate per embryo (IR) was 38%. In group A-2, 13/58 (22%) achieved viable clinical pregnancies. The IR was 9%. In group B-1, the viable clinical pregnancy rate was 52% (15/29) and the IR was 25% compared with a viable clinical pregnancy rate of 15% (2/13) and an IR of 5% in group B-2. The results of this study suggest that by selecting specific embryos for transfer based on their individual sHLA-G expression, pregnancy and implantation rates can be maximized while the number of embryos transferred can be reduced, thereby minimizing the incidence of high-order multiple pregnancies.

Adult↗
PubMed2001

A comparative study of three techniques for the analysis of sperm recovery: touch-print cytology, wet preparation, and testicular histopathology.

PURPOSE: The aim of this study is to evaluate the efficacy of simultaneous testicular touch-print cytology, testicular histopathology, and wet preparation in nonobstructive azoospermic (NOA) males. METHODS: Three hundred and sixty-three males with NOA underwent a multiple testicular sampling prior to ICSI for histopathologic evaluation, diagnostic testicular sperm extraction, and simultaneous touch-print cytology to evaluate sperm presence or absence. A total of 979 testicular samples were taken. RESULTS: Sperm recovery was achieved in 106 cases (29.2%). Patients with hypospermatogenesis and focal spermatogenesis needed 2.8 and 5.9 biopsies, respectively, to retrieve spermatozoa, while in patients with germ cell aplasia and maturation arrest, even after eight to nine samples no spermatozoa were recovered. Neither the FSH levels nor the testicular volume was found to be significant in the prediction of sperm recovery. If only a single testis was to be biopsied, 25% of the cases with sperm recovery would have been missed. The combination of touch-print cytology with histopathology and wet preparation increased the accuracy of spermatozoa identification. CONCLUSION: Touch-print cytology was found to be more predictive than wet preparation in the diagnosis of spermatogenesis; moreover, it was found to be a quick and easy technique providing an accurate diagnosis in prediction of sperm recovery.

Adult↗
PubMed1998

Correlation of testicular sperm extraction with morphological, biophysical and endocrine profiles in men with azoospermia due to primary gonadal failure.

To identify the predictive factors for testicular sperm extraction (TESE) and to understand the pathology associated with TESE, we carried out a prospective study in 40 consecutive men with azoospermia due to primary gonadal failure. The main outcome measure was the retrieval of at least one testicular spermatozoon. Endocrine and biophysical profiles, testicular histology, Johnsen score and testicular spermatids were used as predictors of sperm extraction. Spermatogenesis was quantified with the Johnsen score. A variable pattern of spermatogenesis was common, being present in 20 (50%) patients. Visualisation of testicular spermatids on testicular histology showed a strong association with TESE (P < 0.0001). Statistically significant differences were detected in plasma follicle stimulating hormone (FSH) and testicular volume between patients who had hypospermatogenesis and Sertoli cell-only or maturation arrest. There were no significant differences in Johnsen score, biophysical and endocrine profiles between the groups with successful and failed TESE. However, a statistically significant trend occurred with changes in histological pattern [chi2 for trend, P = 0.001; Pearson's coefficient (r) = 0.6], Johnsen score (P = 0.022; r = 0.5), testicular volume (P = 0.01; r = 0.5) and plasma FSH concentrations (P = 0.044; r = 0.4), albeit to a limited degree. Difference in the interpretation of histological patterns with different assessors was observed. The type of occupation or risk factors for azoospermia showed no association with testicular pathology or TESE. Variable histological patterns in different tubules in the same individual may explain the poor correlation of TESE with endocrine and biophysical profiles, Johnsen score and histological pattern. Differences in the amount of tissue used for TESE and histopathology, and misinterpretation of testicular histology rather than failure to quantify spermatogenesis may explain the poor correlation between histological patterns and TESE. Testicular spermatids predicted TESE. However, considerable overlap in values means that no single variable can provide a perfect discrimination between the groups with successful and failed TESE.

Adult↗
PubMed2003

A functional cytochrome P450 lanosterol 14 alpha-demethylase CYP51 enzyme in the acrosome: transport through the Golgi and synthesis of meiosis-activating sterols.

Mammalian lanosterol 14 alpha-demethylase (CYP51) is a microsomal cytochrome P450 that demethylates lanosterol to FF-MAS, an oocyte meiosis-activating sterol and late intermediate of cholesterol biosynthesis. Herein we report CYP51 unequivocally localized to acrosomal membranes of male germ cells in mouse, bull, and ram, in which it synthesizes FF-MAS in the presence of the acrosomal form of nicotinamide adenine dinucleotide phosphate reduced-P450 reductase. In the mouse, CYP51 (53 kDa) resides in endoplasmic reticulum (ER) and Golgi during all phases of acrosome development, indicating an intracellular transport from ERs through the Golgi to the acrosome. CYP51 (50 kDa) also resides on acrosomal membranes of bull- and ram-ejaculated sperm. In mouse liver, a 53-kDa CYP51 is no longer detected in trans Golgi, suggesting retrieval back to the ER and no further transport to other organelles. Glycosylated high-molecular-mass CYP51-immunoreactive proteins in acrosomal membranes of bull and ram and Golgi-enriched fractions of mouse liver indicate that mammalian CYP51s are subjected to posttranslational modifications in the Golgi. In conclusion, CYP51 is the first cytochrome P450 enzyme to be detected on acrosomal membranes. It exhibits a unique, cell-type-specific intracellular transport that is in agreement with its cell-type-specific physiological role: production of cholesterol in the liver and sterols with signaling properties in sperm. Demethylation of lanosterol to FF-MAS by the acrosomal lanosterol 14 alpha-demethylase enzyme complex demonstrates for the first time the ability of ejaculate sperm to synthesize meiosis-activating sterols.

Acrosome↗
PubMed1990

An evaluation of the hypo-osmotic sperm swelling test as a predictor of fertilizing capacity in vitro.

The ability of sperm to swell in hypo-osmotic conditions was examined in 211 semen samples from the partners of patients about to undergo oocyte retrieval for in-vitro fertilization (IVF). The test was performed using aliquots of semen, the remainder of which was then prepared for IVF. No significant difference was found, in either the percentage of swollen sperm or the type of swelling response, between samples that achieved fertilization in vitro and those that did not, or between any of the diagnostic categories of infertility (tubal damage, unexplained infertility, oligospermia). In samples which achieved fertilization in vitro there were correlations between sperm swelling and sperm motility (r = -0.51) and abnormal morphology (r = 0.33), but no such correlations were demonstrated in samples that failed to achieve fertilization. Moreover, there was no significant difference between the percentage of swollen sperm in semen (mean motility 64%), in samples immediately after preparation for IVF (mean motility 96%) or in capacitated sperm 24 h after preparation (mean motility 91%). These results demonstrate that the hypo-osmotic sperm swelling test does not assist in the prediction of the fertilizing capacity of human sperm in vitro.

Cell Membrane↗
PubMed1993

Episodic scrotal mutilation with concurrent bilateral sperm granuloma in a dog.

Bilateral sperm granuloma with diffuse spermatocele was found to be the cause of infertility in a young Golden Retriever. Anamnesis indicated that the dog had intermittent episodes of self-inflicted ulcerative scrotal dermatitis, beginning when it was 4 to 5 months old and regardless of season or environment. A complete breeding soundness examination produced consistently azoospermic ejaculates in the presence of bilaterally firm, distinct swellings of the epididymides. Because of the poor reproductive prognosis, the owner requested castration and scrotal ablation. Although scrotal trauma was initially thought to be associated with the onset of sperm granuloma formation, evaluation of the history and physical examination and laboratory findings indicated that a bilateral congenital anomaly was the most likely cause of infertility.

Animals↗
PubMed1998

Semen quality in spinal cord injured men: does it progressively decline postinjury?

OBJECTIVE: To determine if semen quality of men with spinal cord injury (SCI) undergoes a progressive decline as a function of years postinjury. DESIGN: A retrospective analysis of cross-sectional data. SETTING: University-based research center. SUBJECTS: Semen quality was examined in 638 specimens from 125 men with SCI. INTERVENTION: Penile vibratory stimulation, electroejaculation, and masturbation were used as semen retrieval methods. Routine semen analysis was performed to evaluate semen quality. MAIN OUTCOME MEASURES: Sperm concentration, total sperm count, and percent sperm motility were examined at 2-year intervals from men whose injuries had occurred 6 weeks to 26 years earlier. RESULTS: No difference in any semen parameter was found as a function of time postinjury. CONCLUSIONS: Semen quality in men with SCI does not progressively decline after the SCI. Men with SCI who are considering biologic fatherhood should be advised that the number of years after injury need not be a determinant in deciding when to start a family.

Adolescent↗
PubMed2006

Salvage microdissection testicular sperm extraction after failed conventional testicular sperm extraction in patients with nonobstructive azoospermia.

PURPOSE: TESE is considered the best procedure for identifying a tubule for spermatozoa retrieval. This technique improves the SRR to around 50%. However, it has been unclear whether it is useful in patients in whom conventional TESE has failed. We compared the outcome of microdissection TESE in patients in whom conventional TESE failed to that in patients who did not undergo conventional TESE. We also evaluated relations between the outcome of salvage microdissection TESE and the characteristics of previous conventional TESE. MATERIALS AND METHODS: A total of 46 patients with nonobstructive azoospermia in whom salvage microdissection TESE was performed after failed conventional TESE were included. Patient characteristics and the SRR were compared between these patients and 134 in whom conventional TESE had not been performed previously. The previous TESE procedure, testicular histology and interval between TESEs were also evaluated. RESULTS: Patient characteristics did not differ significantly between the groups. The microdissection TESE SRR also did not differ significantly between the groups (45.7% vs 44.0%). The possibility of successful spermatozoa retrieval by salvage microdissection TESE remained regardless of the previous failure of any other TESE procedure and regardless of testicular histology. The salvage microdissection TESE SRR was not related to the interval between TESEs. CONCLUSIONS: Because salvage microdissection TESE is effective in patients in whom conventional TESE has failed, this option should be made available to them with the understanding that extended followup after salvage microdissection TESE is necessary due to the risk of hypogonadism.

Adult↗
PubMed2002

A modified RT-PCR technique to screen for viral RNA in the semen of hepatitis C virus-positive men.

BACKGROUND: Our objective was to use an adapted RT-PCR technique to assess the presence of hepatitis C virus (HCV) in semen and also in different density gradient semen fractions collected from men with chronic viral hepatitis participating in an assisted reproduction programme. METHODS: This study included 50 semen samples from 35 HCV(+) men, with active viral replication assessed by RT-PCR, collected the day of oocyte retrieval and used for assisted reproduction. These samples were subjected to standard assisted reproduction sperm preparation conditions, using density-gradient centrifugation with 45 and 90% layers. Aliquots of semen, 45 and 90% fractions, and embryo culture media were frozen at -80 degrees C for subsequent virological analyses. All aliquots were tested with a commercially available HCV RNA assay, adapted for use with semen after a number of technical changes. This assay yielded a sensitivity of 50-100 HCV RNA copies/ml and strongly diminished the effect of seminal amplification inhibitors. RESULTS: HCV RNA was detected in 7/50 (14%) semen samples tested, 5/35 (14.3%) men. HCV RNA was found in only 1/50 45% fractions but never in the 90% fraction or embryo culture media. Sera from 3/5 men contained 3.19-7.40 x 10(5) IU/ml, while the two others had 4.5 and 11.7 x 10(6) IU/ml. However, HCV RNA was quantified at <600 IU/ml in the HCV(+) semen of these five patients. The ongoing pregnancy rate was of 20% (10/50) with one delivery at the time of the present report. No anti-HCV antibody was found in any of the women or the newborn. CONCLUSIONS: Although HCV is present at low concentrations in the semen of a few HCV(+) patients, no purified sperm fraction (i.e. 90% fraction) used in assisted reproduction was HCV(+) and no seroconversion was observed in the women and the newborn, thereby suggesting a very low risk of virus transmission. Nevertheless, because the presence of HCV in semen implies a possible risk of nosocomial contamination, safety regulations must be strictly applied in assisted reproduction laboratories.

Abortion, Spontaneous↗