Atsumi Yoshida

Cytogenetic Survey of 1,007 Infertile Males

The aim of this study was to investigate the influence of a chromosome abnormality on male infertility. The subjects consisted of 1,007 males with the chief complaint of infertility. Karyotyping was conducted mainly by G banding. Major chromosome abnormalities were observed in 62 patients (6.2%) in total and consisted of sex chromosome abnormalities were observed in 62 patients (6.2%) in total and consisted of sex chromosome abnormalities in 38 patients (3.8%) and autosomal chromosome abnormalities in 24 (2.4%). Among the patients with sex chromosome abnormalities, 28 cases were 47, XXY, 3 were 47,XYY, and 7 cases had a Y chromosome abnormality. Autosomal chromosome abnormalities comprised 10 cases of reciprocal translocation, 8 cases of Robertsonian translocation, 5 cases of inversion, and 1 case of ring chromosome. In patients with a sperm density < 5 x 10(6)/ml, a total motile sperm count < 1 x 10(6), a follicle-stimulating hormone value > or = 30.1 mIU/ml, a luteinizing hormone value > or = 8.9 mIU/ml, a testosterone value < or = 2.69 ng/ml, or an average testis volume < or = 8 ml, the incidence of major chromosome abnormalities was significantly higher. These findings suggest that patients who need microinsemination should undergo chromosome analysis. We should counsel patients about obtaining adequate information on each chromosome abnormality.

Intracytoplasmic injection with late spermatids: A successful procedure in achieving childbirth for couples in which the male partner suffers from azoospermia due to deficient spermatogenesis

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.

Features of constitutive gr/gr deletion in a Japanese population

BACKGROUND The relationship between male infertility and gr/gr deletions that remove multiple genes of the Y chromosome varies among countries and populations. The aim of this study was to investigate the association between gr/gr deletions and spermatogenic phenotype in fertile and infertile Japanese men. METHODS The subjects were screened by sequence-tagged site (STS) analysis to detect gr/gr deletions, and haplogroups were assigned using eight highly informative markers. In total, 395 infertile men and 377 fertile men (controls) participated in our study. Of the 772 subjects, 260 individuals carried confirmed gr/gr deletions and were used in further analysis of deletion subtype and gene copy number, specifically loss and gain of CDY1 and DAZ copies. These 260 subjects were divided into a control group (n = 131) all with normozoospermia, and an infertile group (n = 129) with 89 infertile subjects exhibiting azoospermia (absence of sperm) and 40 exhibiting oligozoospermia (reduced sperm concentration). RESULTS There were gr/gr deletions in 33.7% (260/772) of all subjects and the deletions were widespread in haplogroup D (86.2%). There were no significant differences in the frequency of gr/gr deletions between the infertile and control groups. The gr/gr deletion subtypes were not distributed randomly among haplogroups; the CDY1a+ DAZ1/2 genes were deleted in 96.9% (217/224) of haplogroup D individuals, whereas the O lineage had a variety of gr/gr deletion types. The loss of CDY1a+ DAZ1/2 was not associated with spermatogenic impairment in haplogroup D (P = 0.33). CONCLUSIONS Taken together, gr/gr deletions in haplogroup D occur constitutively, are associated with the loss of CDY1a + DAZ1/2 and are phenotypically neutral. Further studies are needed to establish whether Y-linked compensatory factors outside the AZFc region can counteract the pathogenic effect of a gr/gr deletion in the D lineage.

Tissue-specific differentially methylated regions of the human VASA gene are potentially associated with maturation arrest phenotype in the testis

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.

Evaluation of seminiferous tubule scores obtained through testicular biopsy examinations of nonobstructive azoospermic men

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.

Semen analysis and successful paternity by intracytoplasmic sperm injection in a man with steroid 5α-reductase-2 deficiency

OBJECTIVE To report semen parameters and successful paternity by intracytoplasmic sperm injection (ICSI) in a male patient with molecularly confirmed steroid 5α-reductase-2 deficiency. DESIGN Case report. SETTING National research institute and an infertility clinic. PATIENT(S) A 29-year-old Japanese man with 5α-reductase-2 deficiency who had failed to have a child despite an ordinary conjugal life for 2 years with his wife. INTERVENTION(S) Mutation analysis, semen analysis, and execution of ICSI. MAIN OUTCOME MEASURE(S) Mutation detection, semen assessment, and production of a child. RESULT(S) Mutation analysis revealed a homozygous p.R246Q missense mutation on exon 5 of SRD5A2. Semem analysis showed oligozoospermia (semen volume 0.3 mL, sperm count 15 × 10(6)/mL, total sperm count 4.5 × 10(6), motile cells 17%, and normal morphologic sperm 8%). ICSI resulted in a production of a healthy male infant. CONCLUSION(S) The results, in conjunction with those of previously reported patients who received semen analysis and/or achieved paternity, suggest that male patients with 5α-reductase-2 deficiency, especially those with hypomorphic mutations including p.R246Q, could retain some degree of spermatogenic function and achieve paternity with and without assisted reproductive technology.

Alu sequence variants of the BPY2 gene in proven fertile and infertile men with Sertoli cell-only phenotype.

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.

A Novel Y Chromosome Microdeletion With the Loss of an Endogenous Retrovirus Related, Testis Specific Transcript in AZFb Region

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 <0.05). CONCLUSIONS Because of the finding of what are to our knowledge novel microdeletions due to endogenous retrovirus in the AZFb region, our study raises the possibility that specific variations in genomic structure may contribute to some forms of human idiopathic male infertility.

A man with hypogonadotropic hypogonadism successfully treated with nasal administration of the low-dose gonadotropin-releasing hormone analog buserelin.

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 patients laboratory examination showed low serum levels of gonadotropins and testosterone. After being diagnosed with hypogonadotropic hypogonadism, 15 mug of buserelin acetate spray was administrated in each nostril three times a day (total: 90 mug/day). This therapy improved semen parameters and serum gonadotropin and testosterone levels. After approximately 1 year of this treatment, the patients 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.

Monochorionic triamniotic triplet pregnancies with assisted reproductive technology : Two case reports

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.