Oligozoospermic Patients Research Articles

Study on the relationship between partial deletions in the azzospermia factor C region of Y chromosome and male infertility

To investigate the relationship between the partial deletions in the azzospermia factor(AZFc) region of Y chromosome and male infertility. Multiplex PCR technology was performed to screen the partial deletions in the AZFc region in 158 azoospermia, 160 severe oligozoospermia and 135 oligozoospermia patients and 236 men with normal spermatogenesis.For samples with gr/gr, b2/b3 recombinogenic deletions, author applied RFLP method to identify which DAZ gene doublet deletion was involved. The gr/gr and b2/b3 were two types of common deletions detected. There were significant differences in the b2/b3 deletion in patients with oligozoospermia and severe oligozoospermia compared to the controls (both P< 0.05). However, there was no difference for the gr/gr deletion between the patients and controls. The results suggested that the b2/b3 deletion might be a risk factor to spermatogenic impairment and might lead to male infertility.

Male infertility in China: laboratory finding for AZF microdeletions and chromosomal abnormalities in infertile men from Northeastern China

To investigate the frequencies of AZF microdeletions and chromosomal abnormalities in infertile men from Northeastern China. Moreover, to compare the prevalence of these abnormalities with other countries and regions in the world. 305 infertile men were enrolled. A complete semen analysis and reproductive hormones were measured according to standard methods. Multiplex polymerase chain reaction (PCR) amplification using nine specific sequence-tagged sites (STS) were used to detect AZF microdeletions. Karyotype analyses were performed on peripheral blood lymphocytes with standard G-banding. Of the 305 infertile men, 28 (9.2%) had AZF microdeletions and 26 (8.5%) had chromosomal abnormalities. The most frequent microdeletions were in the AZFc+d, followed by AZFc, AZFb+c+d and AZFa. A total of 19 patients (82.6%) had Klinefelter's syndrome (47, XXY) in the azoospermic group. The freqencies of AZF microdeletions and chromosomal abnormalities in infertile men from Northeastern China were comparable with infertile men from other countries and regions. However, there was a slightly higher prevalence rate of AZF microdeletions in oligozoospermic patients than reported in previous studies.

Screening for Y chromosome azoospermia factor region microdeletions using modified multiplex PCR

Objective To establish an universal primer-multiplex PCR system for diagnosis of Y chromosome AZF region microdeletions in 262 patients with non-obstructive azoospermic and severe oligozoospermic male infertility. Methods In each panel of multiplex PCR, YUP and YCP containing a fragment of non-human DNA sequence at their 5' ends were designed. The universal primers and chimiric primers were employed for the amplification at the same multiplex PCR system to screen for the Y chromosome AZF region ( a, b and c) microdeletions in 262 non-obstructive azoospermic and severe oligozoospermic male infertility patients. Results Thirty-three out of 262 patients (12. 60% ) were detected with Y chromosome AZF microdeletions. Among them, 27 cases were AZF c microdeletions and 6 ones were AZF b + c microdeletions. These results were in agreement with the results from EMQN method. There was no false-positivity. The gel electrophoresis for detection of multiple STS from both methods showed that the sY84,sY86, sY127, sY134, sY254, sY255, SRY bands were homogeneous and clear with similar brightness. Conclusion The modified multiplex PCR is suitable for screening of Y chromosome AZF microdeletions in non-obstructive azoospermic and severe digozoospermic male infertility patients. Key words: Infertility,male; Y chromosome; Chromosome deletion; Polymerase chain reaction

Cytogenetic abnormalities detected in patients with non-obstructive azoospermia and severe oligozoospermia

To find the frequency and types of major chromosomal abnormalities with nonobstructive azoospermia and severe oligozoospermia to give appropriate genetic counseling before assisted reproduction techniques in Isparta (South of Turkey), and to investigate the general characteristics in this infertile male population. A total of 115 infertile males (92 were azoospermic, 23 severe oligospermic) were studied for the cytogenetic evaluation prior to use of assisted reproduction techniques. Also, 60 fertile males as a control group were studied. Karyotyping was performed on peripheral blood lymphocytes according to the standard methods. Levels of luteinising hormone, follicle-stimulating hormone (FSH), testosterone and prolactin were obtained and a testicular sonography examination was conducted. The total prevalence of chromosomal abnormalities was found to be 4.3% (5/115), including 4 patients with Klinefelter's Syndrome and 1 patient with gonadal dysgenesis (46XX). All of them were azoospermic males, corresponding to a frequency of 5.4% (5/92 patients). Oligozoospermic males and control males had no chromosomal abnormalities. There was a significant difference in serum FSH, LH, mean testicular volume and smoking when comparing patients (both azoospermic and oligozoospermic) and control groups (p<0.05). Also, there was a significant difference in serum FSH, LH and mean testicular volume when compared with azoospermic and oligozoospermic patients (p<0.05). The occurrence of chromosomal abnormalities among infertile males strongly suggests the need for routine genetic testing and counseling prior to the employment of assisted reproduction techniques.

The Effects of Severity of Oligozoospermia on Intracytoplasmic Sperm Injection (ICSI) Cycle Outcome

The objective of this study was to explore the relationship between the severity of oligozoospermia and the development of embryos and clinical outcome in patients undergoing ICSI. A total of 908 intracytoplasmic sperm injection cycles involving women of ≤37 years of age were included in this study. The patients were divided into four treatment groups according to the results of an analysis of their husbands' semen: (A) mild oligozoospermia that ranged from 10×106/ml to <20×106/ml (n=283), (B) mild to severe oligozoospermia that ranged from 5×106/ml to <10×106/ml (n=192), (C) severe oligospermia that ranged from 1×106/ml to <5×106/ml (n=259), and (D) very severe oligozoospermia that ranged from 0 to <1×106/ml (n=174). Two pronuclei (PN) oocytes at MII were injected and the development of high quality embryos on day 2, as well blastocyst formation rate on day 5, the implantation rate, clinical pregnancies, and fetal loss, were examined. A lower percentage of two pronuclei (2PN) oocytes in the very severe oligozoospermia group was observed, however, there was no difference in clinical outcome when the oligozoospermic patients were divided by sperm concentration. In addition, no significant difference was detected in zygote production or clinical outcome between spermatozoa with a motility of <40% and spermatozoa with a motility of ≥40%. The results of this study emphasize the importance of selecting good quality sperm for oocyte injection, especially in cases involving very severe oligozoospermia.

Follicle-Stimulating Hormone Receptor Gene Haplotypes and Male Infertility in Estonian Population and Meta-Analysis

Follicle-stimulating hormone (FSH) is crucial for male fertility and it exerts its effects via a gonad-specific receptor (FSHR). In the present study, the common G-29A, A919G, and A2039G polymorphisms in the FSHR gene were analysed in 150 (36 non-obstructive azoospermia and 114 individuals with oligozoospermia) patients and 208 normozoospermic men. The results showed that the FSHR polymorphisms were not associated with either azoo- or oligozoospermia as the distributions of alleles, genotypes, and haplotypes among patients and controls were similar. Amongst normozoospermic men, those carrying at least one minor A allele (GA and AA genotypes) of the G-29A polymorphism had a smaller mean testicular volume compared to men with GG homozygosity (25.8 ml vs. 27.4 ml, respectively; P=0.013). In a subsequent meta-analysis combining our data with previous studies, the G-29-A919-A2039 haplotype was shown to be more prevalent in normozoospermic men than in azoospermic patients (38.4% vs. 33.9%, respectively; χ2test, P=0.045), indicating that this haplotype may be a protective factor against male sterility. In conclusion, we suggest that FSHR haplotypes are not considerable risk factors for spermatogenic failure. The protective nature of G-29-A919-A2039 haplotype cannot be concluded without additional studies.

“One sperm, one embryo, one baby”?

“One sperm, one embryo, one baby”?

A decade of experience emphasizes that testing for Y microdeletions is essential in American men with azoospermia and severe oligozoospermia

To evaluate the benefit of Y microdeletion testing. Retrospective analysis. University-based male fertility clinic and genetics laboratory. A total of 1,591 men with sperm concentrations less than 5 million sperm/mL. Semen analysis, Y microdeletion testing, microdissection testicular sperm extraction (TESE). Sperm concentration, incidence and nature of Y microdeletions, microdissection TESE outcome. We identified 149 microdeletions (9.4%). 10.4% of azoospermic men and 10.1% of men with sperm concentrations >0-1 million sperm/mL harbored microdeletions. Two-thirds of microdeletions in azoospermic men were AZFa, AZFb, AZFb+c, or complete Yq deletions. Virtually all microdeletions in oligozoospermic patients were AZFc deletions. Seven hundred eighteen patients underwent microdissection TESE, including 41 with microdeletions. Microdissection TESE failed in all patients with AZFa, AZFb, AZFb+c, and complete Yq deletions. Sperm were retrieved in 15/21 AZFc deleted patients (71.4%). The presence of an AZFc deletion was associated with increased likelihood of sperm retrieval when compared with the 48.8% retrieval rate in 385 idiopathically azoospermic men who consecutively underwent microdissection TESE at our institution during the study period. Clinical pregnancy was achieved in 10/15 azoospermic AZFc deleted patients for whom sperm were successfully retrieved. Of azoospermic and severely oligozoospermic American men, 10% harbor Y microdeletions that alter prognosis for surgical sperm retrieval and are vertically transmissible. Y microdeletion testing is essential for genetic and preoperative counseling in these patients.

Alterations in sperm DNA methylation patterns at imprinted loci in two classes of infertility

To evaluate the associations between proper protamine incorporation and DNA methylation at imprinted loci. Experimental research study. Research laboratory. Three populations were tested-abnormal protamine patients, oligozoospermic patients, and fertile donors. The CpG methylation patterns were examined at seven imprinted loci sequenced: LIT1, MEST, SNRPN, PLAGL1, PEG3, H19, and IGF2. The DNA methylation patterns were analyzed using bisulfite sequencing. The percentage of methylation was compared between fertile and infertile patients displaying abnormal protamination. At six of the seven imprinted genes, the overall DNA methylation patterns at their respective differentially methylated regions were significantly altered in both infertile patient populations. When comparing the severity of methylation alterations among infertile patients, the oligozoospermic patients were significantly affected at mesoderm-specific transcript (MEST), whereas abnormal protamine patients were affected at KCNQ1, overlapping transcript 1 (LIT1), and at small nuclear ribonucleoprotein polypeptide N (SNRPN). Patients with male factor infertility had significantly increased methylation alteration at six of seven imprinted loci tested, with differences in significance observed between oligozoospermic and abnormal protamine patients. This could suggest that risk of transmission of epigenetic alterations may be different with diagnoses. However, this study does not provide a causal link for epigenetic inheritance of imprinting diseases, but does show significant association between male factor infertility and alterations in sperm DNA methylation at imprinted loci.

Semen parameters and hormonal profile in obese fertile and infertile males

To study the changes in semen parameters, gonadotropic and sex hormones, and serum leptin in obese fertile and infertile oligozoospermic men. Prospective. University hospital. Forty-two obese fertile and 80 obese infertile oligozoospermic men (body mass index [BMI] >30). The BMI calculation, semen analysis, and serum FSH, LH, T, E(2), PRL, and leptin estimation. Semen and hormonal profiles. Obese oligozoospermic patients had significant increase in mean BMI, serum FSH, LH, E(2), PRL, and leptin compared with obese fertile controls. The BMI had significant positive correlation with abnormal sperm morphology, LH, serum leptin and significant negative correlation with sperm concentration, sperm motility, serum T. Serum leptin demonstrated significant positive correlation with patients' age, abnormal sperm morphology, serum FSH, LH, PRL and significant negative correlation with sperm concentration, sperm motility, and serum T. Serum leptin mediates a link between obesity and male infertility.

Negative influence of paternal age on clinical intracytoplasmic sperm injection cycle outcomes in oligozoospermic patients

To evaluate the effect of male age on clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles, according to sperm concentration. Retrospective, observational study. Assisted reproduction center. The study included 1,024 couples undergoing ICSI cycles with fresh spermatozoa. The influence of paternal age on ICSI outcomes of oligozoospermic and normozoospermic patients was evaluated. Rates of high-quality embryos, pregnancy, implantation, and miscarriage were evaluated through linear logistic regression analyses. When the sperm concentration was abnormal, paternal age influenced implantation (regression coefficient value = -0.7009) and pregnancy rates (odds ratio = 0.95, 95% confidence interval 0.91-0.99). However, in normozoospermic patients, no influence of paternal age was observed on implantation (regression coefficient value = 0.0566) or pregnancy rates (odds ratio = 1.00, 95% confidence interval 0.97-1.03). For couples in which the men are oligozoospermic, the implantation rate could be impaired by increased paternal age. In these couples, the chance of pregnancy decreased 5% for each year of paternal age. When men are normozoospermic, this effect is not observed.

Expansion of CAG repeats in the spinocerebellar ataxia type 1 (SCA1) gene in idiopathic oligozoospermia patients

The lengths of CAG repeats in two spinocerebellar ataxia genes, SCA1 and SCA3, were analyzed to determine whether such repeats exist in higher numbers in infertile males. Blood samples were collected from healthy controls, oligozoospermia patients, and azoospermia patients. DNA fragments containing target CAG repeats were amplified by PCR with template DNA purified from the blood samples. CAG repeats in PCR fragments were determined, using ABI PRISM 310 Gene Analyzer. In SCA1, the distribution of CAG repeats in oligozoospermic males was different from that of the control group: More alleles had a repeat number that exceeded 32. Conversely, for SCA3, the examined oligozoospermia and azoospermia patients exhibited no differences in distribution of CAG repeats in comparison with the control group. SCA1 in a subset of oligozoospermia patients has an increased number of CAG repeats.

Evaluation of the acrosome in oligozoospermic patients by simplified triple stain technique.

The human acrosome, especially the acrosome reaction, is an important parameter for evaluation of sperm function. By histochemical staining, the acrosome is visualized at the light microscopic level. However, a high number of cells are lost due to the multiple centrifugation steps that are required for preparation of spermatozoa. In the present study, semen samples from oligozoospermic patients were stained by an improved method. Compared to the conventional triple-stain technique, optimal staining of the acrosome and a lower number of cells lost during sperm preparation were observed (P<0.025). Differences in the percentage of live acrosome-reacted spermatozoa were not statistically significant.

Seminal plasma magnesium, calcium and inorganic phosphate concentration in normozoospermic and subfertile men.

The relationship of seminal plasma Mg, Ca and inorganic phosphate concentration to various parametres of seminal quality was evaluated in 41 normozoospermic and 102 dysspermic men with or without accessory gland infection. Ca concentration in asthenozoospermic and oligozoospermic patients was lower than in normozoospermic men and was positively correlated with Mg concentration irrespectively of seminal quality or the presence of infection. Mg concentration in normozoospermic or in oligozoospermic patients with infection was lower than in normozoospermic men without infection but higher in azoospermic men. Mg was negatively correlated to pH and both Mg and Ca concentrations were related to acid phosphatase levels. Inorganic P was higher in asthenozoospermic but lower in azoospermic patients than in normozoospermic men. In astheno-, oligo- and azoospermic men, P was positively correlated with fructose. In patients with accessory gland infection, successful treatment resulted in a significant concentration rise of all three elements. It is concluded that, from a diagnostic point of view, estimations of these elements, particularly of Mg and P, in seminal plasma reflect satisfactorily the functional capacity of the accessory glands of the genital system and the response to treatment in dysspermia associated with infection.

Testosterone and dihydrotestosterone seminal plasma levels in varicocele patients.

Testosterone (T) and Dihydrotestosterone (DHT) were measured in seminal plasma levels of forty-four varicocele patients and in seventeen normal men. T values were not significantly different from controls, while DHT values were significantly lower in varicocele patients. When we grouped the patients according to the sperm count, we observed a decrease of DHT similar to that reported by other authors in oligozoospermic patients. However in the patients with normal sperm count, where the motility was the only parameter significantly lower, DHT in the ejaculate showed again a marked decrease. In contrast to that observed in the normal subjects, in the latter group, with normal sperm count, we observed a negative correlation between the age of the patients and DHT in the ejaculate. This suggests that the duration of varicocele per se could affect DHT seminal plasma levels.

Improvement in Semen Quality in Infertile Males after Treatment with Tamoxifen

The effect of the anti-oestrogen Tamoxifen on the seminal quality of 131 men was studied. Parameters studied before and after treatment were sperm density, total ejaculate count, percentage progressive motility, progressively motile ejaculate count, percentage total motility and total motile ejaculate count. In a group of 38 males, the effect on serum LH, FSH, testosterone, oestradiol and prolactin was also studied. Tamoxifen significantly improved (p less than 0.05) the progressive motility in all patient groups where there was reduced pretreatment motility. Sperm density was also significantly improved in oligozoospermic patients. Elevations in the basal serum levels of FSH was noted, even in those patients where the basal level was elevated before treatment. Increases were also observed in the serum levels of the four other hormones studied.

Differentiation of ejaculates showing reactive oxygen species production by spermatozoa or leukocytes.

Differences between subgroups and correlations between reactive oxygen species (ROS), sperm motility, concentration of leukocytes and viability in semen samples from 143 men were investigated. Patients with azoospermia or leukocytospermia were excluded from the study. Spermatozoa were separated by means of glass wool filtration. Reactive oxygen species were determined by means of luminol chemiluminescence before and after sperm separation; thereafter, normozoospermic and oligozoospermic patients were divided into three subgroups using the mean of all patients investigated (17462 count 10(-7) viable spermatozoa) as cut-off value as follows: G1--high reactive oxygen species production in native semen and after glass wool filtration; G2--high production of reactive oxygen species only in native semen; G3--low levels of reactive oxygen species before and after glass wool filtration. In general, reactive oxygen species were significantly higher in oligozoospermic samples than in normozoospermic samples. In men with normal sperm count, ROS production correlated significantly with the number of leukocytes in the ejaculate before glass wool filtration, but not thereafter. Glass wool filtration is useful to distinguish between sperm samples in which reactive oxygen species are generated by leukocytes and those in which reactive oxygen species are excessively generated by the spermatozoa themselves.

L-dopa and oligozoospermia.

A clinical investigation of L-Dopa on idiopathic oligozoospermia is carried out. Thirty nine male oligozoospermic patients with primary sterility were divided in two groups. The first group of 25 patients was treated with 500 mg a day during two months. The second group of 14 patients received 750 mg a day for the same time. In the first group we find 14 improvements, 9 at the first month and 5 at the second month. In the second group 8 improvements were observed, 5 in the first month and 3 in the second months. It would seem that L-Dopa has a stimulatory effect on spermatogenesis possibly through hypophysis when it is administered at these dosis and for these periods of time, and it was also observed similar results for longer terms of treatment.

Survey of carnitine content of human semen using a semiquantitative auxanographic method: decreased semen total carnitine concentration in patients with azoospermia or severe oligozoospermia.

A microbiological method, using the carnitine-requiring yeast, Torulopsis bovina ATCC 26014, was developed to identify samples of human semen which contained low levels (less than 250 micron M) of total carnitine. Of 399 semen samples from a male infertility clinic which were tested, 30 (7.5%) were low in carnitine. Of these, 14 were azoospermic and 16 were severely oligozoospermic. Some azoospermic samples (19 = 58%) and severely oligozoospermic samples (51 = 79%) did not give evidence of low carnitine concentrations. These results indicate that decreased total carnitine concentration in semen occurs in certain classes of azoospermic and severely oligozoospermic patients.

Superoxide anion production and some sperm-specific enzyme activities in infertile men.

The relationship between superoxide anion production and the activities of sperm specific NADH-diaphorase and isozyme LDH-C4 has been investigated in 25 normozoospermic and 17 oligozoospermic semen samples. The samples were separated into high and low density sperm populations on discontinuous, two step (40%:80%) Percoll gradients prior to analysis. Superoxide anion generation was significantly higher (P < 0.001) and diaphorase activity significantly increased (P < 0.01) in both low and high density sperm fractions from oligozoospermic patients compared to the values found in normozoospermic men. A significant positive correlation (P < 0.01) between diaphorase activity and superoxide anion production was established in the oligozoospermic samples of low density sperm fraction. A tendency of statistically insignificant increase in LDH-C4 isozyme activity was observed in the oligozoospermic samples. The activity of this isozyme appears to have no association with superoxide anion generation or diaphorase activity. These findings seem to suggest that excessive superoxide anion production along with increased diaphorase activity point to biochemical abnormalities which may affect the functional status of sperms. Hence, determination of these parameters may provide useful data in male infertility of unknown etiology.