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Background: FertiQoL, which measures the Quality of Life (QOL) of reproductive partners, has been translated and used in 45 languages in the world. The reliability and validity of the original English version of FertiQoL have been confirmed. However, there is still no report on the reliability and validity of the Japanese version by a large-scale survey. This study aimed to examine the reliability and validity of the Japanese version of the FertiQoL scale for measuring the QOL of patients with reproductive problems. Methods: An unsigned self-filled questionnaire survey was conducted in patients undergoing infertility treatment at seven facilities in the Kanto area in Japan using the 34 items of the Japanese version of the FertiQoL scale. The study design was quantitative cross-sectional descriptive research. The investigation period was from April 2013 to April 2018. The contents of the investigation were attributes, FertiQoL scale, and distress scale. To determine the construct validity, principal component analysis, confirmatory factor analysis, and correlation analysis for each subscale were performed using SPSS Statistics Ver. 23.0 and AMOS Ver. 23.0. The study was performed after obtaining approval from the Research Ethics Review Committee of the responsible institutions to which the researchers belong as an ethical consideration. Results: The participants included 1201 patients undergoing infertility treatment and who provided valid responses. The Cronbach’s α was 0.92, and confirmatory factor analysis identified six domains with 34 items that showed the following values: goodness of fit index = 0.877, adjusted goodness of fit index = 0.855, comparative fit index = 0.893, and root mean square error of approximation = 0.059. The correlation coefficient was 0.669 (p < 0.001) with the distress scale. Conclusions: The Japanese version of FertiQoL showed adequate reliability and validity for assessing the QOL of patients with reproductive problems in Japan.

The purpose of this study was to investigate the relationship between placental oxidative stress and maternal endothelial function in pregnant women with normotensive fetal growth restriction (FGR).We examined serum concentrations of oxygen free radicals (d-ROMs), maternal angiogenic factor (PlGF), and sFlt-1, placental oxidative DNA damage, and maternal endothelial function in 17 women with early-onset preeclampsia (PE), 18 with late-onset PE, 14 with normotensive FGR, and 21 controls. Flow-mediated vasodilation (FMD) was assessed as a marker of maternal endothelial function. Immunohistochemical analysis was performed to measure the proportion of placental trophoblast cell nuclei staining positive for 8-hydroxy-2'-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage.Maternal serum d-ROM, sFlt-1 concentrations, and FMD did not significantly differ between the control and normotensive FGR groups. The proportion of nuclei staining positive for 8-OHdG was significantly higher in the normotensive FGR group relative to the control group.Our findings demonstrate that, despite the presence of placental oxidative DNA damage as observed in PE patients, pregnant women with normotensive FGR show no increase in the concentrations of sFlt-1 and d-ROMs, or a decrease in FMD.

Does the inclusion of three antioxidants (A3), acetyl-l-carnitine (ALC), N-acetyl-l-cysteine (NAC) and alpha-lipoic acid (ALA) improve human embryo development and pregnancy potential?Prospective randomized multicentre comparison of sibling oocytes. A total of 1563 metaphase II oocytes from 133 patients in two IVF centres. Day 3 embryo and day 5/6 blastocyst quality were assessed. Good embryo quality on day 3 was defined as 8 to 10 cells with even cells and low fragmentation; good quality blastocysts as 3BB or greater. Clinical outcome was assessed on transfers of fresh or vitrified-warmed blastocyst on day 5.Of the two-pronuclei, 40.7% (G-Series) and 50.2% (G-Series with A3 group) resulted in good quality embryos on day 3 (P0.05). The implantation rate by fetal sac was 39.2% and 50.6%, and by fetal heartbeat was 37.8% and 47.1% for the G-Series and G-Series with A3 group, respectively. When stratified by female patient age, patients 35-40 years had an implantation rate by fetal sac and heart of 23.5% in the G-Series compared with 57.5% (P0.05) and 50.0% (P0.05) in the A3 group. The ongoing pregnancies in patients 35-40 years were significantly higher in the A3 group (50%) compared with the control (25.8%) (P0.05).The presence of antioxidants during IVF and embryo culture for patients 35-40 years resulted in a significant increase in implantation and pregnancy rate. Supplementation of antioxidants to IVF and culture media may therefore improve the viability of human embryos in assisted reproductive technologies, plausibly through the reduction of oxidative stress.

Background: Testicular microlithiasis is said to be characterized by calcific concretions within the seminiferous tubules. There have been reports suggesting a link between testicular microlithiasis and testicular dysfunction and tumors.

Background and aims: Azoospermia factor c (AZFc) microdeletions are associated with male infertility and are caused by intrachromosal recombination between homologous repetitive sequence segments. Partial AZFc deletion (gr/gr) has been reported in male factor infertility. In the present study, we established detecting the copy number using quantitative real‐time polymerase chain reaction (qRT‐PCR) with the genome DNA, and assessed the association of the recombination pair set of gr/gr deletion and deleted in azoospermia copies. Furthermore, we determined the clinical significance of differential recombination patterns of gr/gr deletion, and compared them with azoospermia and proven fertile volunteers, with both groups having gr/gr deleted Japanese subjects.

The epigenetic status of the genome changes dynamically from fertilization to implantation. In addition, the physiological environment during the process of gametogenesis, including parental age, may affect the epigenome of the embryo after fertilization. It is important to clarify the influence of parental age on gene expression in the embryo in terms of transgenerational epigenetics to improve the techniques currently used in assisted reproductive medicine. Here, we performed single-embryo RNA-seq analysis on human blastocysts fertilized by intracytoplasmic sperm injection, including from relatively elderly mothers, to elucidate the effects of parental age on the embryonic gene expression profile. We identified a number of genes in which the expression levels were decreased with increasing maternal age. Among these genes, several are considered to be important for meiotic chromosomal segregation, such as PTTG1, AURKC, SMC1B and MEIKIN. Furthermore, the expression levels of certain genes critical for autophagy and embryonic growth, specifically GABARAPL1 and GABARAPL3, were negatively correlated with advanced paternal age. In addition, levels of transcripts derived from major satellite repeats also decreased as the maternal age increased. These results suggest that epigenetic modifications of the oocyte genome may change with parental age and be transmitted to the next generation.

Since the establishment of intracytoplasmic sperm injection (ICSI) in 1992, sperms have been observed, evaluated, and selected under an inverted microscope at 400 × magnification for ICSI. Intracytoplasmic morphologically selected sperm injection (IMSI) is a technique in which sperms are selected for micro insemination by more precisely observing their morphology, at increased magnification under an inverted microscope used for micro-insemination and enhancing the image resolution. Approximately 20 years have passed since IMSI was developed, and its efficacy has only been confirmed in cases with a high rate of sperm aneuploidy, a high rate of sperm DNA fragmentation, and repeated ICSI failures. It is well-known that the quality of sperms has a profound effect on the quality of embryos which may eventually become children. Therefore, when micro-insemination is performed, sperms should be selected by not only paying attention to their motility, but also by closely observing their morphology. Those ...

STX2 encodes a sulfoglycolipid transporter. Although Stx2 nullizygosity is known to cause spermatogenic failure in mice, STX2 mutations have not been identified in humans. Here, we performed STX2 mutation analysis for 131 Japanese men clinically diagnosed with nonobstructive azoospermia. As a result, we identified a homozygous frameshift mutation [c.8_12delACCGG, p.(Asp3Alafs*8)] in one patient. The mutation-positive patient exhibited loss-of-heterozygosity for 58.4 Mb genomic regions involving STX2, suggesting possible parental consanguinity. The patient showed azoospermia, relatively small testes, and a mildly elevated follicle stimulating hormone level, but no additional clinical features. Testicular histology of the patient showed universal maturation arrest and multinucleated spermatocytes, which have also been observed in mice lacking Stx2. PCR-based cDNA screening revealed wildtype STX2 expression in various tissues including the testis. Our results indicate that STX2 nullizygosity results in nonsyndromic maturation arrest with multinucleated spermatocytes, and accounts for a small fraction of cases with nonobstructive azoospermia.

Objectives Deletions in the azoospermia factor regions are the most common known molecular genetic cause of human male infertility involving spermatogenetic failure. Testing for these deletions in Japanese DNA samples using conventional sequence-tagged site probes occasionally lead to considerable non-specific or faint products in the Japanese population. The aim of the present study was to evaluate the sensitivity and specificity of a newly developed kit for the detection of azoospermia factor microdeletions in the Japanese population. Methods Sequence-tagged site probes were reselected and the Luminex suspension array assay was carried out. Validation was retrospectively carried out with 2014 DNA sequences with known microdeletions, which were divided into four categories. Results Category 1 deletions that corresponded to the conventional classification of azoospermia factor deletion were present in 83 men (4.2%), which can result in intrachromosomal homologous recombination. Kit data confirmed the presence of deletions of this type in DNA sequences known to harbor the azoospermia factor deletions. Category 2 deletions involved cytogenetic abnormalities in 28 men (1.4%), whereas category 3 deletions in 759 men (37.7%) were atypical classifications including the gr/gr deletion. As these deletions are thought to be a result of palindromic units and non-homologous recombination, these microdeletions might impact in the interpretation of some clinical findings. The rest of the 1145 cases (56.8%) were assigned to category 4 as normal variants (polymorphism/no deletion). Conclusions The present findings show that this new kit offers good sensitivity and specificity with the advantage of saving in terms of cost and time.

Purpose: We identified the endogenous retroviruses associated with TTYs (testis specific transcripts linked to the Y) in the AZFb region. We evaluated the relationship between endogenous retroviruses, and TTY expression patterns and function in spermatogenesis. Materials and Methods: We identified family members of TTYs in the AZFb region using computational screening. After investigating the relationship between the endogenous retrovirus genome and TTY expression patterns we screened genomic polymerase chain reaction products from TTY13 amplified from 790 Japanese men, including 275 with azoospermia, 285 with oligozoospermia and 230 who were fertile. Results: Computational screening revealed that 3 members of the TTY family, TTY9, 10 and 13, were regulated by endogenous retroviruses in the AZFb region. Homologous recombination between long terminal repeat of the TTY13 associated human endogenous retrovirus-K14C resulted in TTY13 deletion events. These deletions were more common in patients with azoospermia and oligozoospermia than in fertile males. Specifically 15.63% of the azoospermia group, 10.88% of the oligozoospermia group and 0% of fertile controls had only the deletion variant, indicating an association between the homologous recombination rate and the severity of spermatogenesis failure that was statistically significant (p

Efforts by corporations to fulfill their social responsibilities will lead to consumers' social satisfaction and a steady rise in their stock prices. For medical institutions, the establishment of a system for the fulfillment of social responsibility would lead not only to patients' trust and satisfaction, but also to improved safety of medical care. Infertility treatment plays an important role in reversing the decline in Japan's birthrate. However, since infertility treatment, unlike other medical practices, involves birth of a new life, the results of medical services provided by infertility clinics/hospitals will have a great impact on the future society. For example, in the procedures of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), sperm and an egg are once taken out of the body, and an embryo formed from the egg is returned to the uterus. Therefore, the greatest possible care must be taken to prevent mix-up of eggs and sperm. In addition, since the process of I...

金沢大学医薬保健研究域医学系, The male-specific region of Y chromosome (MSY) has accumulated a higher density of human endogenous retroviruses (HERVs) and related sequences when compared with other regions of the human genome. Here, we focused on one HERV family, HERV-K14C that seemed to integrate preferentially into the Y chromosome in humans. To identify every copies of HERV-K14C in the human genome, we applied computational screening to map precisely the locus of individual HERV-K14C copies. Interestingly, 29 of all 146 copies were located in Y chromosome, and these 29 copies were mostly dispersed in the palindromic region. Three distinct HERV-K14C-related transcripts were found and were exclusively expressed in human testis tissue. Based on our phylogenetic analysis of the solitary LTRs derived from HERV-K14C on the Y chromosome we suggested that these sequences were generated as pairs of identical sequences. Specifically, analysis of HERV-K14C-related sequences in the palindromic region demonstrated that the Y chromosomal amplicons existed in our common ancestors and the duplicated pairs arose after divergence of great apes approximately 8-10 million years ago. Taken together, our observation suggested that HERV-K14C-related sequences contributed to genomic diversification of Y chromosome during speciation of great ape lineage. © 2010 The Japan Society of Human Genetics All rights reserved., 出版社許諾要件により、2012年6月より全文公開.

Developments in assisted reproductive technology have allowed us to decrease the numbers of transferred embryos and thereby reduce the risk of multifetal pregnancies. However, even when the numbers of transferred embryos are restricted, the incidence of monozygotic multiple pregnancies is not reduced, and can be significantly higher than normal. We report two cases of monochorionic triamniotic triplet pregnancies after applying assisted reproductive technology. Three male babies were born in one pregnancy and the other pregnancy aborted at 10 weeks of gestation. Several factors, such as manipulation of the zona pellucida or extended blastocyst culture, might increase the risk of such monozygotic multiple gestations in this setting, but the actual causes remain unknown. Besides detecting predictive factors for monozygotic pregnancies, any manipulation of the zona pellucida in combination with blastocyst culture must be done cautiously to minimize the likelihood of monochorionic multiple pregnancies.

Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay (SCSA) parameters, that is, DNA fragmentation index (%DFI) and high DNA stainability (%HDS), were assessed on the fresh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation (DGC) was followed by swim-up treatment in comparison with DGC alone (P0.01). Although the %DFI increased in a time-dependent manner with incubation both at room temperature (RT) and at 37 degrees C in air, the %DFI after 24 h at RT was significantly lower than that at 37 degrees C (P0.05). Incubation with 5% CO2 was effective in maintaining sperm motility (P0.01); however, it induced further elevation of %DFI (P0.001). Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.

Numerous CpG islands containing tissue-specific differentially methylated regions (TDMRs) are potential methylation sites in normal cells and tissues. The VASA (also known as DDX4) gene is believed to be under the control of TDMRs. A total of 131 male patients with idiopathic azoospermia or severe oligospermia were evaluated histologically, and the methylation status of CpG islands in the VASA gene was screened. Genome DNAs were obtained from testicular biopsy and modified with sodium bisulfite, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied. This system is capable of analyzing both the methylated and unmethylated CpG island in the genome. The methylation analysis is conducted by an epigram as graphic data. On histological assessment, 17 of 131 patients revealed maturation arrest (MA).In all, 6 of the 17 patients showed particularly high VASA TDMR methylation rates, whereas the remaining 11 patients and controls had low methylation rates. This study may imply that the VASA TDMR methylation is significantly higher among patients with MA, in whom the VASA gene expression was silenced. This finding represents an important contribution to the molecular basis of meiotic arrest as one possible cause of idiopathic infertility. © 2009 The Japan Society of Human Genetics All rights reserved.

In conventional IVF, a sperm penetrates an ovum through natural selection, and embryologists sort sperm for ICSI. The quality of the original ovum and sperm has the greatest effect on the quality of the fertilized egg (embryo). Therefore, the selection of superior sperm is critical for ICSI. Although sperm cells are 20-fold smaller than embryos or ova, they are observed and evaluated for ICSI using an inverted microscope at ×400 magnification usually used for observation of embryos or ova. Recently, a technique called intracytoplasmic morphologically selected sperm injection (IMSI) has attracted attention. This technique uses an inverted microscope with higher magnification and resolution than that used for ICSI, which enables embryologists to precisely observe sperm morphology, particularly the presence or absence of vacuoles in the sperm head, and to sort sperm for microfertilization. Although IMSI has some technological problems, including a narrow microscopic field due to higher magnification...

Objective To evaluate the relationship between the status of homologous recombination and population-specific alleles in infertile Japanese males with azoospermia factor (AZF)-a deletions and to characterize the clinical features of these patients. Design Retrospective deletion study in infertile Japanese men. Setting University hospital and reproductive clinic. Patient(s) A total of 931 consecutive patients visiting a male-infertility clinic were genetically evaluated. Intervention(s) Patients were analyzed for Y-chromosomal microdeletions and the breakpoints of intrachromosomal homologous recombination of human endogenous retrovirus (HERV) 15qy; in addition, Y-haplogroup typing on the basis of polymerase chain reaction also was performed. Endogenous retroviruses contribute to the evolution of the host genome and can be associated with disease. Main Outcome Measure(s) Presence or absence of appropriately sized polymerase chain reaction products. Result(s) Four cases of AZFa deletions were found. All patients with AZFa deletions had an azoospermia and breakpoints in the ID2 region of HERV15qy. Three of the four cases were derived from Y-haplogroup D2b. Testicular sperm extraction procedures were performed in three of these four patients, and elongated spermatids were recovered in two. However, no pregnancies were successfully achieved. Conclusion(s) Y-haplotype D2b, specific for some Japanese clade, may be associated with HERV breakpoints that lead to intrachromosomal homologous recombination. From the clinical point of view, the testicular sperm extraction procedure is not applicable to males with complete AZFa deletions.

Background and aims Azoospermia factor c (AZFc) microdeletions are associated with male infertility and are caused by intrachromosal recombination between homologous repetitive sequence segments. Partial AZFc deletion (gr/gr) has been reported in male factor infertility. In the present study, we established detecting the copy number using quantitative real-time polymerase chain reaction (qRT-PCR) with the genome DNA, and assessed the association of the recombination pair set of gr/gr deletion and deleted in azoospermia copies. Furthermore, we determined the clinical significance of differential recombination patterns of gr/gr deletion, and compared them with azoospermia and proven fertile volunteers, with both groups having gr/gr deleted Japanese subjects.

Objective: The basic protein on Y chromosome, 2 gene (BPY2) is implicated in the spermatogenic process. Three copies (paralogs) of the BPY2 sequence lie in the AZFc region, within huge palindromic repeats consisting of Alu sequences located in 5′ flanking regions. Our objective was to screen the single nucleotide variation of BPY2 gene paralogously. Methods: Mutation screening of the exons and 5′ flanking region of the BPY2 genes was carried out by polymerase chain reaction (PCR) and sequencing in 106 infertile patients with a Sertoli cell-only (SCO) phenotype and in 126 fertile men. Results: No sequence variation was detected in any of the BPY2 exons. Paralogous A/A/A and mixed A/G genotypes are referred to the promoter of the gene. The G/G/G genotype (wild type) was observed at the rates of 89.6% (95/106) in the SCO subjects and 96.0% (121/126) in the proven-fertile subjects. The G/G/G plus mixed A/G paralogous variants and the A/A/A paralogous variant were detected in 96.2% (102/106) and 3.8% (4/106) of infertile patients with SCO, respectively. The G/G/G plus mixed A/G paralogous variants were detected in 100% (126/126) of the fertile controls, with no instances of the A/A/A phenotype, and this difference was statistically significant (P = 0.038, Fisher's exact test). Conclusion: The A/A/A genotype may be associated with the SCO phenotype. The alteration of all three copies of BPY2 within the AZFc region on the Y chromosome may affect spermatogenic process.

Objective To assess perinatal risk of major congenital anomalies in children born after embryo transfer with assisted hatching (AH). Design Retrospective cohort study. Setting Not applicable. Patient(s) Cycles registered from 2010 to 2012 and conceived via single-embryo transfer were included for the analysis. Live births, still births after 22 weeks of gestation, and selectively terminated cases because of congenital anomalies were included. Intervention(s) None. Main Outcome Measure(s) Major congenital anomaly. Result(s) AH was performed in 35,488 cycles among 72,125 included cycles (49.2%). A total of 1,046 major congenital anomalies (1.4%) were identified (1.36% in AH group vs. 1.50% in non-AH group). Overall risks for major congenital anomalies were not significantly different between AH and non-AH groups adjusting for maternal age, calendar year, fetal sex, embryo stage at transfer, and status of cryopreservation. There were 1,009 cases of twins (1.5%) and 10 cases of triplets (0.015%) among all included cycles. No specific organ system demonstrated significant association between AH and non-AH groups. Subgroup analysis demonstrated no significant association between AH and non-AH groups in intracytoplasmic sperm injection cycles or in vitro fertilization in fresh cycles. Similar nonsignificant association was observed between early-cleavage or blastocyst stage at transfer in frozen-thawed cycles. Conclusion(s) Our results suggest that AH alone does not increase the risk of major congenital anomaly.

Objective To investigate microdeletions and palindrome complexes in the azoospermic factor (AZF) c regions of the Y chromosome, by polymerase chain reaction (PCR) with sequence-tagged site (STS) markers with known physical positions, after verification of the exact physical locations of candidate STSs and exclusion of those that would give ambiguous results on PCR. Design Retrospective STS deletion study in infertile Japanese men. Setting University hospital and reproductive clinic. Patient(s) A total of 410 men with nonobstructive azoospermia and severe oligospermia (sperm concentrations of 6 /mL). Intervention(s) Polymerase chain reaction was performed for all STS markers confirmed to be physically located in the partial AZFb and AZFc regions of the Y chromosome. All STSs were retrieved from the University of California at Santa Cruz database, and their location and specificity were verified. Main Outcome Measure(s) Presence or absence of appropriately sized PCR products. Result(s) Sixty-nine markers were retrieved, 32 of which were not specific to the long arm of the Y chromosome (Yq). The markers retained for test use were contiguous deletions classified as P1+P2 (AZFc) or P3 proximal/P1 (AZFb+c). The prevalence of accurately mapped microdeletions was 5.1% (21 of 410) in this patient population. Conclusion(s) Although noncontiguous deletions were observed, considerable confusion remained until the genome sequence was finally determined because many of the STSs were either repetitive sequences or polymorphic between individuals or races.

Background Testicular microlithiasis is said to be characterized by calcific concretions within the seminiferous tubules. There have been reports suggesting a link between testicular microlithiasis and testicular dysfunction and tumors.

Although copy-number variations (CNVs) in Y-chromosomal azoospermia factor (AZF) regions have been associated with the risk of spermatogenic failure (SF), the precise frequency, genomic basis and clinical consequences of these CNVs remain unclear. Here we performed multiplex ligation-dependent probe amplification (MLPA) analysis of 56 Japanese SF patients and 65 control individuals. We compared the results of MLPA with those of conventional sequence-tagged site PCR analyses. Eleven simple and complex CNVs, including three hitherto unreported variations, were identified by MLPA. Seven of the 11 CNVs were undetectable by conventional analyses. CNVs were widely distributed in AZF regions and shared by ~60% of the patients and ~40% of the controls. Most breakpoints resided within locus-specific repeats. The majority of CNVs, including the most common gr/gr deletion, were identified in the patient and control groups at similar frequencies, whereas simple duplications were observed exclusively in the patient group. The results imply that AZF-linked CNVs are more frequent and heterogeneous than previously reported. Non-allelic homologous recombination likely underlies these CNVs. Our data confirm the functional neutrality of the gr/gr deletion in the Japanese population. We also found a possible association between AZF-linked simple duplications and SF, which needs to be evaluated in future studies.

The objective of this study was to determine whether or not abnormalities in the autosomal structure of the male partner have any influence on fertilization and early embryo development after intracytoplasmic sperm injection (ICSI). Thirty-seven couples in whom the male partners were examined by the same andrologist were included in this study. Six couples (group I) in whom the male partner possessed autosomal structural abnormality underwent seven ICSI cycles. As a control group, 31 couples (group II) in which the male partner was karyotypically normal underwent 41 ICSI cycles. Although the normal fertilization rates seen in group I were significantly higher than those in group II (P < 0.05), there was no significant difference in the cleavage rates between the two groups. We did not perform the analysis of the female partner's chromosomes, but we surmise that structural autosomal abnormalities in the male partner do not adversely influence fertilization at ICSI and early development of embryos.

We describe a 28 year old male with a pseudodicentric Y chromosome who suffered from azoospermia attributed to maturation arrest of the primary spermatocyte, as diagnosed by testicular biopsy. Chromosome analysis, using G, Q and C banding techniques, revealed an abnormal karyotype of 45,X[7]/46,X,psu dic (Y)(pter-->q11.2::q11.2-->pter)[33]. Polymerase chain reaction (PCR) DNA analysis did not detect the absence of DAZ and RBM1 which are candidates for azoospermic factor (AZF) genes. Therefore, it is suggested that the maturation arrest of the primary spermatocyte in this patient was caused either by a pairing dysfunction between the X and Y chromosomes during meiosis or by deletions in the autosomal or the Y chromosomal spermatogenesis controlling genes, excluding DAZ and RBM1.

In this report, we present the overall sexual function and clinical features of patients with Klinefelter's syndrome with the chief complaint of male infertility. The study consisted of 40 patients with a control group of 55 infertile non-azoospermic males with a normal 46,XY karyotype who visited the Reproduction Center of Toho University Hospital during the 5.5-year period between January 1991 and June 1996 with the chief complaint of male infertility. Among the 40 patients with Klinefelter's syndrome, 38 cases were pure 47,XXY, one case was 47,XXY with a pericentric inversion of chromosome 9 and one case was a mosaic of 46,XY/47,XXY(2:28). Thirty-nine of these 40 patients were azoospermic and one (47,XXY) had severe oligoasthenozoospermia. The sexual function of the patients was evaluated according to their responses to a preliminary questionnaire devised by our department. There was no significant difference in the frequency of sexual function disturbances between the patients with Klinefelter's syndrome and the control group (67.5% vs. 60.0%; chi 2 analysis; p = 0.454). The mean frequency of sexual intercourse per month in the patients with Klinefelter's syndrome was significantly higher than in the control group (4.4 +/- 2.8 vs 3.3 +/- 1.6: Welch's t-test, p < 0.05). A possible explanation for this variation may lie in the fact that many of these patients were diagnosed with azoospermia poor to the administration of the questionnaire and may have wished to continue to have relations as a couple.

The prevalence of spermatogenic failure (SF) has gradually increased during the past few decades at least in several countries. Although multiple factors would be involved in this phenomenon, one important factor would be excessive estrogen effects via estrogen receptors (ERs). Thus, we performed haplotype analysis of ESR2 encoding ERβ in 125 Japanese SF patients and 119 age-matched control males, using single nucleotide polymorphisms (SNPs) 1–9 that are widely distributed on the ∼120-kb genomic sequence of ESR2. Consequently, a linkage disequilibrium (LD) block was detected in an ∼60-kb region encompassing SNPs 2–7 in both groups, and four major estimated haplotypes were identified within the LD block. Furthermore, the most prevalent ‘TGTAGA’ haplotype was found to be significantly associated with SF, with the P-value obtained by the Cochran–Armitage trend test (0.0029) being lower than that obtained by a 100 000-times permutation test (0.0038) to cope with the problem of multiple comparisons. The results, in conjunction with our previous data indicating lack of a susceptibility factor on ESR1 encoding ERα, imply that the specific ‘TGTAGA’ haplotype of ESR2 raises the susceptibility to the development of SF.

ICSI achieves consistently high fertilization and pregnancy rates ­regardless of sperm characteristics and even azoospermic patients have fathered children by ICSI where spermatozoa were surgically retrieved. In this review, we report multicenter study results after the use of IMSI in patients with compromised semen parameters or in patients with previous ART failure using sibling oocytes. These studies, based on higher magnification screening for sperm surface irregularities, did not seem to benefit the patients’ clinical outcome in independent investigations. We conclude from these collaborative studies, IMSI does not bring any advantage in the quest to find spermatozoa devoid of surface irregularities and that it significantly lengthens the search time required.

Symptomatic women with uterine fibroid sometimes need surgical interventions. Although these procedures have been performed by laparotomy in the past, recent advances in laparoscopic techniques and...

Developments in assisted reproductive technology have allowed us to decrease the numbers of transferred embryos and thereby reduce the risk of multifetal pregnancies. However, even when the numbers of transferred embryos are restricted, the incidence of monozygotic multiple pregnancies is not reduced, and can be significantly higher than normal. We report two cases of monochorionic triamniotic triplet pregnancies after applying assisted reproductive technology. Three male babies were born in one pregnancy and the other pregnancy aborted at 10 weeks of gestation. Several factors, such as manipulation of the zona pellucida or extended blastocyst culture, might increase the risk of such monozygotic multiple gestations in this setting, but the actual causes remain unknown. Besides detecting predictive factors for monozygotic pregnancies, any manipulation of the zona pellucida in combination with blastocyst culture must be done cautiously to minimize the likelihood of monochorionic multiple pregnancies.

Aims. Dysfunction of the FSH receptor (FSHR) may be involved in some form of male infertility with azoospermia or oligozoospermia. We assessed the discrete codon combination with homo/heterozygous variation of the exon 10 in the FSHR gene. Methods. The genotype of codon 307 and codon 680 were analysed in 352 patients with idiopathic male infertility and 145 men with proven fertility. Results and Conclusion. There was no significant difference in the distributions of each homozygous codon 307 or 680 between these two groups as reported in the literature. However, the population with heterozygous combinations Thr/Ala (codon 307) and Ser/ Asn (codon 680) comprised 26% (38/146) and 44.9% (157/343) in subjects with proven fertility and idiopathic infertile men, respectively. Moreover, the heterozygous genotype Thr/Ala-Ser/Asn was significantly increased in infertile patients compared with the controls. This finding showed that the combination of heterozygous FSHR can be responsible for male infertility.

Objective To report a patient with hypogonadotropic hypogonadism of hypothalamic origin successfully treated with nasal administration of a low-dose gonadotropin-releasing hormone (GnRH) analogue. Design Case report. Setting A reproductive medical center. Patient(s) A 37-year-old man with anejaculation and infertility. Intervention(s) Nasal administration of a low-dose GnRH analogue, buserelin. Main Outcome Measure(s) Semen analysis and serum levels of gonadotropins and testosterone after nasal buserelin use. Result(s) The patient's laboratory examination showed low serum levels of gonadotropins and testosterone. After being diagnosed with hypogonadotropic hypogonadism, 15 μg of buserelin acetate spray was administrated in each nostril three times a day (total: 90 μg/day). This therapy improved semen parameters and serum gonadotropin and testosterone levels. After approximately 1 year of this treatment, the patient's serum gonadotropin and testosterone levels remained in the normal range and semen analysis showed normozoospermia. The patient and his wife were treated with intracytoplasmic sperm injection, resulting in pregnancy. Conclusion(s) A low-dose buserelin nasal spray appears to be an effective and well-tolerated therapeutic option for patients with hypogonadotropic hypogonadism of hypothalamic origin.

The basic protein on Y chromosome, 2 gene (BPY2) is implicated in the spermatogenic process. Three copies (paralogs) of the BPY2 sequence lie in the AZFc region, within huge palindromic repeats consisting of Alu sequences located in 5' flanking regions. Our objective was to screen the single nucleotide variation of BPY2 gene paralogously.Mutation screening of the exons and 5' flanking region of the BPY2 genes was carried out by polymerase chain reaction (PCR) and sequencing in 106 infertile patients with a Sertoli cell-only (SCO) phenotype and in 126 fertile men.No sequence variation was detected in any of the BPY2 exons. Paralogous A/A/A and mixed A/G genotypes are referred to the promoter of the gene. The G/G/G genotype (wild type) was observed at the rates of 89.6% (95/106) in the SCO subjects and 96.0% (121/126) in the proven-fertile subjects. The G/G/G plus mixed A/G paralogous variants and the A/A/A paralogous variant were detected in 96.2% (102/106) and 3.8% (4/106) of infertile patients with SCO, respectively. The G/G/G plus mixed A/G paralogous variants were detected in 100% (126/126) of the fertile controls, with no instances of the A/A/A phenotype, and this difference was statistically significant (P = 0.038, Fisher's exact test).The A/A/A genotype may be associated with the SCO phenotype. The alteration of all three copies of BPY2 within the AZFc region on the Y chromosome may affect spermatogenic process.

A total of 1,923 follicular human ova were collected for use in intracytoplasmic sperm injection (ICSI). Timed ovulation was induced by means of a standard ovarian stimulation protocol involving set dosages of follicle stimulating hormone (FSH), human menopausal gonadotrophin (HMG) and human chorionic gonadotrophin (hCG). Prior to performing ICSI, all of the ova were subjected to thorough examination which led to the discovery of only two parthenogenetic oocytes, also referred to as parthenogens, out of the total 1,923. These oocytes were retrieved from different patients. Although genetic analysis revealed that one of the parthenogens had a haploid number of chromosomes, it was impossible to analyze the second because it had not yet reached the metaphase stage. The 0.1 percent incidence of spontaneous parthenogenesis in this study clearly indicates that this phenomenon is extremely rare.

An azoospermic male with an unbalanced translocation between the Y chromosome and chromosome 15 was examined in the present study. Testicular biopsy found only Sertoli cells only within the seminiferous tubules of the 35-year-old patient. Chromosome analysis, using the techniques of G and C banding and fluorescent in situ hybridization revealed an abnormal karyotype of 46,XY,der(15)t(Y;15)(q12;p11). Deoxyribonucleic acid (DNA) analysis confirmed the presence of the genes such as DAZ and YRRM1 which are known to control spermatogenesis. The cause of spermatogenetic dysfunction in this particular patient therefore.

Objective: To evaluate the seminiferous tubules of nonobstructive azoospermic men, a modified version of the Johnsen score known as the seminiferous tubule score was used. Design: Retrospective clinical study. Setting: University-based urology center. Patients: One hundred twenty-eight infertile men with nonobstructive azoospermia who underwent open testicular biopsy. Intervention(s): Categorization as follows of various seminiferous tubule characteristics according to the modified seminiferous tubule score parameters: [1]a total absence of cells within the seminiferous tubule, [2] Sertoli cell only, [3] a few spermatogonia, [4] many spermatogonia, [5] a few primary spermatocytes, [6] many primary spermatocytes, [7] a few secondary spermatocytes, [8] many secondary spermatocytes, [9] a few round spermatids, [10] many round spermatids, [11] a few late spermatids and/or spermatozoa, and [12] many late spermatids and/or spermatozoa. Main Outcome Measure(s): Seminiferous tubule score. Result(s): In terms of maximum seminiferous tubule score, scores of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 corresponded with total patient numbers of 9, 90, 0, 0, 3, 10, 0, 0, 3, 1, 11, and 1, respectively. Conclusion(s): It was discovered that intracytoplasmic sperm injection (ICSI) of round spermatids could be performed in 3.1% of the patients in this study and that ICSI using late spermatids or spermatozoa could be performed in 9.4% of the patients in order to achieve fertilization.

Objective: To investigate the feasibility of using late spermatids in intracytoplasmic sperm injection (ICSI) for patients in which there is a total lack of normal and testicular sperm. Design: Clinical study. Setting: Private fertility center with adjacent laboratory facilities. Patient(s): Thirty-six males diagnosed with azoospermia underwent testicular biopsy and the resulting spermatids from nine patients were used to fertilize oocytes retrieved from their respective wives by ICSI. Intervention(s): Intracytoplasmic spermatid injection of oocytes with late spermatids obtained by testicular biopsy. Main Outcome Measure(s): Fertilization, pregnancy, and delivery rates. Result(s): Three of the cases resulted in pregnancies with childbirth occurring in all three, identical twins in one, and individual singletons in the other two cases. Conclusion(s): It is possible to achieve successful levels of pregnancy and childbirth in cases of azoospermia by intracytoplasmic injection of late spermatids.