- [Show abstract] [Hide abstract] ABSTRACT: To examine the accuracy and variability of identification of human germ cells and validate the previously reported diagram referable to identifying human testicular cells, which was made to improve the identification. Eighty-seven testicular cells obtained from azoospermic patients were stained with MitoTracker, and observed under phase contrast, fluorescent, and differential interference microscopy. The recorded image and movie data of phase contrast microscopy were assessed by 10 reviewers comprising embryologists and reproductive physicians 2 times, once without the diagram and 1 year later with use of it. True cell type identifications were determined as referenced by morphologic characteristics and MitoTracker staining. Variability between reviewers was assessed using multirater κ statistics, and changes of the concordance rates to the reference were examined. Multirater κ coefficients changed from 0.14 to 0.49 overall, from 0.10 to 0.34 for sperm-like cells, and from 0.044 to 0.46 in round-shaped cells before and after using the diagram, which represents a change from fair to substantial agreement overall for round-shaped cells and to moderate agreement for sperm-like cells. The concordance rates to the reference before and after the use of the diagram also significantly improved from 28.4% to 59.1% overall, from 38.9% to 54.6% for sperm-like cells, and from 19.4% to 59.1% for round-shaped cells, respectively. Identification of human germ cells by embryologists and reproductive physicians was not uniform or satisfactory. However, the diagram significantly improved identification such that it may be useful as an efficient checklist for the identification of germ cells.
Data: Figure S2[Show abstract] [Hide abstract] ABSTRACT: Analysis of PGAM4 gene expression. (A) Amplified products were analyzed by restriction enzyme digestion. A BstXI restriction site is present in PGAM4, but not in PGAM1, despite the high sequence identity between them (97.2%). The 380-bp amplification product was digested into two fragments (239 and 141 bp). (B) PCR using a human testis-specific cDNA library. M, 100-bp ladder DNA marker; C, Control. (TIF)
- [Show abstract] [Hide abstract] ABSTRACT: Background The development of novel fertilization treatments, including in vitro fertilization and intracytoplasmic injection, has made pregnancy possible regardless of the level of activity of the spermatozoa; however, the etiology of male-factor infertility is poorly understood. Multiple studies, primarily through the use of transgenic animals, have contributed to a list of candidate genes that may affect male infertility in humans. We examined single nucleotide polymorphisms (SNPs) as a cause of male infertility in an analysis of spermatogenesis-specific genes. Methods and Finding We carried out the prevalence of SNPs in the coding region of phosphoglycerate mutase 4 (PGAM4) on the X chromosome by the direct sequencing of PCR-amplified DNA from male patients. Using RT-PCR and western blot analyses, we identified that PGAM4 is a functional retrogene that is expressed predominantly in the testes and is associated with male infertility. PGAM4 is expressed in post-meiotic stages, including spermatids and spermatozoa in the testes, and the principal piece of the flagellum and acrosome in ejaculated spermatozoa. A case-control study revealed that 4.5% of infertile patients carry the G75C polymorphism, which causes an amino acid substitution in the encoded protein. Furthermore, an assay for enzymatic activity demonstrated that this polymorphism decreases the enzyme’s activity both in vitro and in vivo. Conclusion These results suggest that PGAM4, an X-linked retrogene, is a fundamental gene in human male reproduction and may escape meiotic sex chromosome inactivation. These findings provide fresh insight into elucidating the mechanisms of male infertility.
Data: Figure S1[Show abstract] [Hide abstract] ABSTRACT: Amino acid alignments for PGAM1 and PGAM4. Putative amino acids of PGAM4 has 97.2% identity with those of PGAM1. LxRHGExxxN motif for PGAM enzymatic activity was showed in grey box. (DOC)
- [Show abstract] [Hide abstract] ABSTRACT: Water avoidance stress is a potent psychological stressor and it is associated with visceral hyperalgesia, which shows degeneration of the urothelial layer mimicking interstitial cystitis. Cyclooxygenase-2 inhibitors have been recognized to ameliorate frequency both in clinical and experimental settings. We investigated the voiding pattern and cyclooxygenase-2 expression in a rat bladder model of water avoidance stress. After being subjected to water avoidance stress or a sham procedure, rats underwent metabolic cage analysis and cystometrography. Real time reverse transcription polymerase chain reaction was carried out to examine cyclooxygenase-2 messenger ribonucleic acid in bladders of rats. Protein expression of cyclooxygenase-2 was analyzed with immunohistochemistry and western blotting. Furthermore, the effects of the cyclooxygenase-2 inhibitor, etodolac, were investigated by carrying out cystometrography, immunohistochemistry and western blotting. Metabolic cage analysis and cystometrography showed significantly shorter intervals and less volume of voiding in water avoidance stress rats. Significantly higher expression of cyclooxygenase-2 messenger ribonucleic acid was verified by reverse transcription polymerase chain reaction. Immunohistochemistry and western blotting showed significantly higher cyclooxygenase-2 protein levels in water avoidance stress bladders. Furthermore, immunohistochemistry showed high cyclooxygenase-2 expression exclusively in smooth muscle cells. All water avoidance stress-induced changes were reduced by cyclooxygenase-2 inhibitor pretreatment. Chronic stress might cause frequency through cyclooxygenase-2 gene upregulation in bladder smooth muscle cells. Further study of cyclooxygenase-2 in the water avoidance stress bladder might provide novel therapeutic modalities for interstitial cystitis.
- [Show abstract] [Hide abstract] ABSTRACT: To examine the effect of solifenacin for not only overactive bladder symptoms but also sleep disturbance. Nocturia and urgency are independent factors for sleep disturbance. Fifteen male patients with overactive bladder symptoms and sleep disturbance were enrolled in this study. The overactive bladder symptoms score (OABSS) and Athens insomnia scale (AIS) were used as a subjective questionnaire for overactive bladder symptoms and insomnia. The Actiwatch-16 (Mini-Mitter-Respironics, Inc., Bend, OR) was used as an objective measurement tool for insomnia. Total sleep time, sleep efficiency, sleep latency, wake-after-sleep onset, and number of awakenings were measured by the Actiwatch. We evaluated the changes of each parameter before and 8 weeks after the administration of solifenacin. Statistical comparisons before and after the administration were made using the Wilcoxon signed-rank test. To examine the relation between OABSS and AIS, Spearman's testing was used for correlations between independent variables and P<.05 was considered statistically significant. Total OABSS and total AIS were significantly improved after administration of solifenacin. The categories of urgency and nocturia in OABSS and the categories of awakening during the night and sleep quality in AIS were also significantly improved. The Actiwatch study showed that total sleep time and sleep efficiency were significantly improved. The decrease of AIS was significantly correlated with the decrease of urgency (ρ=0.635, P=.0175) but not with nocturia. The treatment of urgency by solifenacin may improve not only overactive bladder symptoms but also sleep disturbance.
- [Show abstract] [Hide abstract] ABSTRACT: The aim of this study was to investigate the relation between lower urinary tract symptoms (LUTS), erectile dysfunction (ED) and depression in Japanese patients with late-onset hypogonadism (LOH) symptoms. The study comprised 87 Japanese patients with LOH symptoms (>27 points on the Aging Males Symptoms Scale). Thirty-four patients were diagnosed as having depression and the remaining 53 patients were diagnosed as not having depression by the Mini International Neuropsychiatric Interview. We compared the International Index of Erectile Function (IIEF) 5, International Prostate Symptom Score (IPSS), IPSS quality-of-life (QOL) index, King's Health Questionnaire (KHQ), endocrinological data, and free uroflow study between depression and non-depression patients and performed multiple logistic regression analysis. IIEF5 scores of depression patients were significantly lower than those of non-depression patients. In KHQ, only the category of general health perceptions was significantly higher in depression patients than non-depression patients. However, IPSS, QOL index, and endocrinological and uroflowmetric data showed no significant difference between the groups. Multiple logistic regression analysis revealed moderate and severe ED to be risk factors for depression. However, LUTS are not related to depression. Moderate and severe ED is correlated with depression, whereas LUTS are not related to depression in Japanese LOH patients.
- [Show abstract] [Hide abstract] ABSTRACT: There is a large individual deviation for men, which may be caused by a difference in personality characteristics, in sexual interest in response to sexual stimuli. In this study, we investigate whether attention to the sexual region in a video depends on the personality characteristics of men, assessing this with an eye-tracking system. The study included 30 healthy males with a normal psychological state, who viewed a sexual video in which the sexual region had been designated. Visual attention was measured across the designated region according to gaze duration. Ten types of personality characteristics were evaluated as a T-score by a questionnaire. By Pearson's correlation coefficient, the relations between gaze duration at the sexual region and T-scores of paranoia, psychasthenia and social introversion were found to be statistically significant. By multivariate stepwise regression analysis, only social introversion was negatively associated with the sexual region. Even normal variation of personality characteristics can affect the viewing period of the sexual region. This is the first report showing that subjects with a high degree of paranoia, psychasthenia and particularly social introversion have a tendency to view the sexual region for a shorter duration.
- [Show abstract] [Hide abstract] ABSTRACT: A 26-year-old man was admitted to our hospital with a chief complaint of left cervical lymphadenopathy. He was diagnosed as having seminoma by needle biopsy. Computed tomography revealed an aorto-caval retroperitoneal tumor. Levels of lactic deydrogenase, human beta-chorionic gonadotropin, alpha-fetoprotein were not elevated. No abnormal findings were present on palpation and ultrasonography of the testes. The patient was diagnosed as having a retroperitoneal extragonadal germ cell tumor (EGCT). Three courses of chemotherapy (bleomycin, etoposide and cisplatin) and another course of chemotherapy (etoposide, ifosfamide and cisplatin) were performed. Complete remission was achieved. Seven years later, the patient noticed an enlargement of his right scrotal contents. Neither distant metastasis nor lymph node metastasis was detected by computed tomography. Right radical orchiectomy was performed and histology revealed seminoma with yolk sac tumor components. No recurrence has been seen after 10 months.
- [Show abstract] [Hide abstract] ABSTRACT: To examine the expression profiles of the proteins translated from Acpin1 mRNA in germ cells. Northern and western blotting of various tissues and immunohistochemical analysis of germ cells were carried out in a mouse model. ACPIN1 protein was transcribed from the longer, 3' open reading frame (ORF) of Acpin1. An alternative-splicing variant, Acpin1vs, contained only the smaller, 5' ORF of the full-length Acpin1 gene. Its gene product, SAGSIN1, was expressed specifically in salivary glands. Retrotransposed regions of Acpin1 homology were also detected in various chromosomes, and intronless paralogous genes on the X chromosome were expressed in the testis and other tissues. The genomic structure of Acpin1 is highly conserved in mammals. The two ORFs on the Acpin1 mRNA are independently translated in differentiated cells. Analysis of gene Acpin1 might clarify the molecular mechanism of spermatogenesis.
- [Show abstract] [Hide abstract] ABSTRACT: To examine the correlation between testosterone and adiponectin in symptomatic late-onset hypogonadism (LOH) patients. The study included 174 patients (>40 years old) with at least one LOH symptom and an Aging Male Symptoms score >26. The correlation between serum adiponectin levels and various factors was investigated by simple and multiple regression analyses. Serum adiponectin levels before and after testosterone replacement therapy (TRT) in 43 patients with serum free testosterone <11.8 pg/mL were also compared. Serum adiponectin levels increased with increased age (P < 0.01), decreased with increased body mass index (P < 0.01), and increased with increased sex hormone-binding globulin (P < 0.01). Multiple regression analysis revealed that body mass index and sex hormone-binding globulin were factors with an influence on serum adiponectin levels. However, no association between testosterone and adiponectin was found. In the 43 patients receiving TRT, serum adiponectin levels before and after TRT did not differ significantly. Serum adiponectin is not related to serum testosterone in symptomatic LOH patients, suggesting that TRT in these subjects does not pose a higher risk of inducing a metabolic syndrome.
- [Show abstract] [Hide abstract] ABSTRACT: To investigate the possible association between variations in the PRDM9 (MEISETZ) gene and impaired spermatogenesis in humans, we screened for mutations in the human PRDM9 gene using DNA from 217 sterile male patients and 162 proven fertile male volunteers. Two single-nucleotide polymorphisms (SNPs), 17353G>T (Gly433Val) and 18109C>G (Thr685Arg), were identified, as well as an intronic SNP, 15549G>T. These SNPs were identified in the heterozygous state in separate patients who demonstrated azoospermia. Neither variant was identified in fertile subjects. Our results suggest that mutations in PRDM9 may cause idiopathic infertility in human males.
- [Show abstract] [Hide abstract] ABSTRACT: We report a case of idiopathic elephantiasis of penis. A 41-year-old man was refered to our hospital with a painful penile swelling and severe miction pain. He had no particular past or familial history, and had never been to tropical or semitropical region. Physical examination showed a prominent swelling and flection of his penis. Magnetic resonance imaging revealed a significant thickening of the penile skin and subcutaneous tissue. Due to the difficulty of voiding, an urethral catheter was placed. The penile skin biopsy showed no malignancy. Under the diagnosis of penile elephantiasis, the resection of abnormal penile skin and penile plasty with the split skin graft from his thigh was performed. Histopathological findings showed nonspecific inflammation. The skin graft was successfully adapted. He could urinate smoothly without pain after the urethral catheter was removed. There has been no recurrence 6 months after the operation. He recovered normal micturition and sexual function.
- [Show abstract] [Hide abstract] ABSTRACT: Introduction. Although men appear to be more interested in sexual stimuli than women, this difference is not completely understood. Eye-tracking technology has been used to investigate visual attention to still sexual images; however, it has not been applied to moving sexual images. Aim. To investigate whether sex difference exists in visual attention to sexual videos. Methods. Eleven male and 11 female healthy volunteers were studied by our new methodology. Main Outcome Measures. The subjects viewed two sexual videos (one depicting sexual intercourse and one not) in which several regions were designated for eye-gaze analysis in each frame. Visual attention was measured across each designated region according to gaze duration. Sex differences, the region attracting the most attention, and visually favored sex were evaluated. Results. In the nonintercourse clip, gaze time for the face and body of the actress was significantly shorter among women than among men. Gaze time for the face and body of the actor and nonhuman regions was significantly longer for women than men. The region attracting the most attention was the face of the actress for both men and women. Men viewed the opposite sex for a significantly longer period than did women, and women viewed their own sex for a significantly longer period than did men. However, gaze times for the clip showing intercourse were not significantly different between sexes. Conclusions. A sex difference existed in visual attention to a sexual video without heterosexual intercourse; men viewed the opposite sex for longer periods than did women, and women viewed the same sex for longer periods than did men. There was no statistically significant sex difference in viewing patterns in a sexual video showing heterosexual intercourse, and we speculate that men and women may have similar visual attention patterns if the sexual stimuli are sufficiently explicit.
- [Show abstract] [Hide abstract] ABSTRACT: Hormone replacement therapy (HRT) is performed for the treatment for patients with late onset hypogonadism (LOH). One of the main concerns about HRT is the influence of lower urinary tract including prostate. This study examined whether HRT for Japanese patients of LOH affected lower urinary tract symptoms. A total of 21 patients with LOH were included in this study. Changes of International prostate symptom score (IPSS), IPSS quality of life (QOL) index and King's health questionnaire (KHQ), before HRT and 3 months after HRT, were evaluated. Statistical comparisons were made using the Wilcoxon signed rank test, P < 0.05 was considered statistically significant. Total IPSS and QOL index were not significantly different from 7.43 +/- 6.56, 2.71 +/- 1.74 at baseline to 8.29 +/- 6.24, 2.91 +/- 1.13 after 3 months. There were no significant differences in any of the nine categories of KHQ. In this preliminary study, the results suggested that HRT for LOH had no short-term effects on patients with lower urinary symptoms.
- [Show abstract] [Hide abstract] ABSTRACT: We evaluated the morphological effect and alterations in gene expression caused by 1,25-dihydroxyvitamin D treatment in the mouse testis undergoing experimental cryptorchidism and subsequent orchiopexy. The mean modified Johnsen score and testicular weight were estimated after 4 weeks of treatment with a 1,25-dihydroxyvitamin D prodrug. We examined sites of vitamin D receptor and mRNA expression, and 1,25-dihydroxyvitamin D analogue accumulation in the mouse testis. Also, we compared alterations in gene expression in the cryptorchid mouse testis with or without 1,25-dihydroxyvitamin D administration by testis specific cDNA microarray. We confirmed protein synthesis of a candidate among up-regulated genes in primary cultures of Sertoli's cells by Western blotting. Mean +/- SEM Johnsen score and testicular weight were increased by 1,25-dihydroxyvitamin D treatment but not significantly (6.12 +/- 0.33 vs 5.27 +/- 0.4 and 49.3 +/- 3.8 mg vs 42.6 +/- 5.5, p = 0.13 and 0.065, respectively). Vitamin D receptor and its mRNA were positive in Sertoli's cells. The 1,25-dihydroxyvitamin D analogue accumulated mainly in Sertoli's cells. Of 2,483 testis specific genes 19 showed up-regulation by 1,25-dihydroxyvitamin D treatment. Of these genes the regulator of cellular cholesterol homeostasis Abca1 was expressed mainly in Sertoli's cells and influenced male fertility. In primary cultures of Sertoli's cells the synthesis of Abca1 protein was increased by 1,25-dihydroxyvitamin D treatment but not by follicle-stimulating hormone or testosterone treatment. We noted that 1,25-dihydroxyvitamin D contributes to spermatogenesis by up-regulating certain specific genes in Sertoli's cells. Testis specific cDNA microarray analysis and vitamin D supplementation may have implications for managing male infertility.