Association between Sperm Mitochondrial DNA Copy Number and Concentrations of Urinary Cadmium and Selenium.

Elevated sperm mitochondrial DNA copy number (mtDNAcn) is associated with damage to sperm and poorer measures of semen quality. Exposure to cadmium (Cd) can increase oxidative stress and damage sperm mitochondria. The adverse effects of Cd can potentially be reduced by sufficient selenium (Se). The objective of this study was to examine the associations between sperm mtDNAcn and urinary concentrations of Cd and Se, as well as the Cd/Se molar ratio. Participants were recruited from patients who sought infertility treatment at two hospitals in Japan. Urine and semen specimens and self-administered questionnaires were collected on the day of recruitment. Sperm mtDNAcn was measured in extracted sperm DNA by multiplex real-time qPCR. Urinary Cd and Se concentrations were measured using inductively coupled plasma mass spectrometry, and their molar weights were calculated to obtain the Cd/Se molar ratio. Linear regression was used to estimate associations after adjusting for age, body mass index, smoking, drinking, exercise, varicocele, and hospital of recruitment. Sperm mtDNAcn showed statistically insignificant associations with creatinine-adjusted concentrations of urinary Cd (ß = 0.13, 95% CI -0.18, 0.44) and Se (ß = -0.09, 95% CI -0.54, 0.35), and Cd/Se molar ratio (ß = 0.12, 95% CI -0.13, 0.37). The current study found no evidence of an association between mtDNAcn and urinary concentrations of Cd or Se, or the Cd/Se molar ratio.

The ratio of 2nd to 4th digit length and reproductive function of infertile male patients.

Purpose: To investigate the relationship between the ratio of index to ring finger lengths (2D:4D ratio), reflecting androgen exposure in utero, and reproductive function among men. Methods: Male patients (N = 180) who consulted for fertility issues participated in the study. The palms of both hands were scanned, and the 2D:4D ratio was calculated. Data on semen volume, sperm concentration and total motility, total and motile sperm counts, and serum hormone concentrations were obtained. Spearman correlation coefficients between the 2D:4D ratio and hormone and semen quality parameters were calculated. Results: The total sperm count was significantly negatively correlated with the 2D:4D ratio of the left hand (r = -0.154, p  = 0.039) but not with that of the right hand (r = -0.045, p  = 0.548). Testosterone showed weak negative correlations with the 2D:4D ratio in the left (r = -0.142, p  = 0.058) and right (r = - 0.149, p  = 0.046) hands. Follicle-stimulating hormone levels were negatively correlated with the 2D:4D ratios of the left (r = -0.173, p  = 0.020) and right (r = -0.164, p  = 0.027) hands. Other semen quality parameters or luteinizing hormone levels showed no significant correlation with the 2D:4D ratios. Conclusions: No clear associations were observed between the 2D:4D ratios and reproductive function.

Analysis of the correlation between gene copy deletion in the AZFc region and male infertility in Japanese men.

Deletion of the azoospermia factor c (AZFc), located on the long arm of the Y chromosome, is a cause of male infertility. The structure of the Y chromosome is diversified by the copy number of various genes, such as deleted in azoospermia (DAZ), basic protein Y2, chromodomain Y1, testis-specific transcript Y-linked 4, and Golgi autoantigen golgin subfamily a2 like Y, located in the AZF region. In this study, we investigated the deletion of each gene copy and analyzed its relationship with Japanese male infertility. Deletions of single nucleotide variants of each gene copy in 721 proven fertile men as controls, 139 patients with non-obstructive azoospermia (NOA), and 56 patients with oligozoospermia (OS) were analyzed via polymerase chain reaction-restriction fragment length polymorphism analysis. Their association with infertility was analyzed using logistic regression analysis adjusted for the Y-chromosome haplogroup, D1a2a. Deletions of DAZ/II in the r1 region and DAZ/V in the r1 and r2 regions showed significant associations with NOA (odds ratio [OR] = 4.15, 95 % confidence interval [CI] = 1.18-14.6, P = 0.026; OR = 4.19, 95 % CI = 1.19-14.7, P = 0.025, respectively). They did not show any association with OS. Partial deletion of the AZFc region affects spermatogenesis in Japanese male.

Effects of antioxidant co-supplementation therapy on spermatogenesis dysfunction in relation to the basal oxidation-reduction potential levels in spermatozoa: A pilot study.

Purpose: In this pilot study, the authors compared the effects of antioxidant co-supplementation therapy and methylcobalamin therapy in patients with impaired semen quality. Methods: Eighty-four subjects who visited male infertility clinics and showed abnormal semen test results were randomly subjected to one of the two therapies: antioxidant co-supplementation therapy with vitamin C, vitamin E, coenzyme Q10, and flaxseed oil or methylcobalamin therapy. The oxidation-reduction potential (ORP) and 8-hydroxy-2'-deoxyguanosine levels were used as indicators of oxidative stress levels in semen. Semen analysis was also performed. Results: The authors obtained results from 67 patients who had completed 3 months of treatment. Neither antioxidant co-supplementation therapy nor methylcobalamin therapy changed the semen parameters significantly (except for the sperm concentration, which was increased by the latter therapy). When the pre-treatment ORP value in semen was higher than the cutoff value, both therapies significantly increased the sperm concentration. The 8-hydroxy-2'-deoxyguanosine level did not yield any meaningful predictive value with regard to increased sperm concentrations. Conclusions: Both antioxidant co-supplementation therapy and methylcobalamin therapy increased the sperm concentration in patients with impaired semen quality when the basal ORP levels in their semen were elevated.

Light-guided renal cyst fenestration during laparoscopic ureterocalicostomy in a patient with left ureteropelvic junction obstruction.

INTRODUCTION: Laparoscopic ureterocalicostomy is a useful alternative to laparoscopic pyeloplasty for treating ureteropelvic junction obstruction under certain conditions. One concern regarding this technique is the inevitability of amputation of the renal lower pole to expose the lower renal calyx. CASE PRESENTATION: A 43-year-old man who presented ureteropelvic junction obstruction and multiple renal cysts underwent laparoscopic pyeloplasty, which could not be performed because of the intrarenal ureteropelvic junction. We switched the surgical technique to modified laparoscopic ureterocalicostomy, wherein amputation of the lower renal pole was substituted with fenestration of a renal cyst under the guidance of ureteroscopic light. Computed tomography performed 2 months postoperatively showed good patency of the anastomosis. CONCLUSION: Light-guided laparoscopic renal cyst fenestration followed by ureterocalicostomy is feasible in patients with ureteropelvic junction obstruction and lower pole renal cysts adjacent to the lower renal calyx.

Laparoscopy-assisted vasovasostomy for post-herniorrhaphy vas deferens obstruction.

INTRODUCTION: Repair of obstructive azoospermia caused by childhood herniorrhaphy may be difficult. Therefore, intracytoplasmic sperm injection using testicular sperm is performed. However, vasovasostomy combined with laparoscopic surgery is challenging. CASE PRESENTATION: A 42-year-old man underwent inguinal hernia repair at age 3. He had normal testicular size, azoospermia, normal hormone levels (follicle-stimulating hormone, luteinizing hormone, and testosterone), absence of Y chromosome micro deletion, and karyotype:46XY, t(1:21)(p34.1:q22.3). He was diagnosed with obstructive azoospermia. Repeated intracytoplasmic sperm injections using testicular sperm resulted in miscarriages. Vasovasostomy combined with laparoscopic surgery was subsequently performed. Postoperative semen analysis result was almost normal. After intracytoplasmic sperm injection of ejaculated sperm, his wife got pregnant. CONCLUSION: Even if patients have chromosomal abnormalities, performing microsurgical re-anastomosis first is recommended. To our knowledge, this is the first case of a laparoscopy-assisted vasovasostomy for post-herniorrhaphy vas deferens obstruction in Japan.

Relationship between Semenogelins bound to human sperm and other semen parameters and pregnancy outcomes.

BACKGROUND: Semenogelins (SEMGs) are major components of human seminal vesicle secretions. Due to SEMG's sperm-motility inhibitor, a significant negative correlation between sperm motility and the proportion of SEMG-bound spermatozoa (SEMG+) was found in asthenozoospermic patients. SEMGs also show intrinsic inhibitory capability for sperm capacitation; however, studies on actual clinical specimens have not been conducted. METHODS: To reveal the relationship between SEMGs and the fertilizing capacity of sperm from male infertile patients who are not restricted to asthenozoospermia, we measured the proportion of SEMG+ in the spermatozoa of 142 male infertile patients. The pregnancy outcomes in partners of these patients were retrospectively analyzed using questionnaires. RESULTS: Among examined semen parameters, only the total SEMG-unbound sperm count showed a tendency to be different between the spontaneous pregnancy or intra-uterine-insemination-pregnancy groups and in-vitro-fertilization- or intracytoplasmic-sperm-injection-pregnancy groups. It was elevated in the former group, which includes patients who used in vivo fertilization. CONCLUSIONS: The total SEMG-unbound sperm count would be a relevant parameter for in vivo fertilization. This result suggests that SEMGs inhibit ectopic capacitation before sperm reach the fertilization site and that the number of total SEMG-unbound sperm is a parameter directly linked to the possibility of in vivo fertilization.

CONTEXTE: Les séménogélines (SEMG) sont des composantes principales des sécrétions des vésicules séminales humaines. En raison de la présence sur SEMG d'un inhibiteur de la mobilité des spermatozoïdes, une corrélation significative a été rapportée entre la mobilité des spermatozoïdes et le pourcentage de spermatozoïdes liés à SEMG chez des patients asthénozoospermiques. Les SEMG possèdent aussi une capacité intrinsèque d'inhibition de la capacitation; aucune étude n'a cependant été réalisée sur des échantillons de sperme utilisés en pratique clinique quotidienne. MATÉRIEL ET MÉTHODES: De façon à mettre en évidence une relation entre les SEMG et la capacité fécondante de spermatozoïdes d'hommes inféconds qui ne soient pas seulement des patients asthénozoospermiques, nous avons mesuré la proportion de spermatozoïdes SEMG+ (liés à SEMG) chez 142 patients inféconds. L'issue des grossesses chez les partenaires de ces patients a été rétrospectivement analysée à partir de questionnaires. RÉSULTATS: Parmi les paramètres spermatiques analysés, seul le nombre total de spermatozoïdes non liées à SEMG montre une tendance à être différent entre le groupe de grossesses obtenues spontanément ou par insémination intra-utérine et le groupe de grossesses obtenues par fécondation in vitro ou injection intra-cytoplasmique d'un spermatozoïde. Ce nombre total était élevé dans le premier groupe qui incluait des patients utilisant une fécondation in vivo. CONCLUSIONS: Le nombre total de spermatozoïdes non liés à SEMG pourrait être un paramètre pertinent pour la fécondation in vivo. Ces résultats suggèrent que les SEMG inhibent la capacitation ectopique avant que les spermatozoïdes n'atteignent le site de fécondation, et que le nombre total de spermatozoïdes non liés à SEMG est. un paramètre directement lié à la possibilité de fécondation in vivo. MOTS-CLÉS: Protéine plasmatique séminale, Infécondité masculine, Séménogélines, IIU, FIV, ICSI, Issues de grossesses.

Progressing management of non-obstructive azoospermia in the era of microdissection testicular sperm extraction.

Previously, it was absolutely impossible for azoospermic men to reproduce except in some obstructive azoospermic cases for whom reconstruction of the seminal pathway was successful. However, nowadays, intracytoplasmic sperm injection and microdissection testicular sperm extraction have brought about chances of biological paternity in some non-obstructive azoospermic men. It is almost 15 years since the first trials of testicular sperm retrieval using surgical microscopy for non-obstructive azoospermia were reported. In this manuscript, the progress and outcomes of these novel techniques since then are reviewed, the controversial points are discussed and the latest research to achieve pregnancies in tough non-obstructive azoospermic couples are introduced. Not only the bright side of the renovations, but the underlying concerns are also discussed.

Intratubular trophoblasts in the contralateral testis caused elevation of serum human chorionic gonadotropin following complete remission of stage II testicular tumor: a case report.

We report the case of a 22-year-old male who had a history of metastatic right testicular tumor successfully treated with chemotherapy and surgery. Twenty-one months after the initial treatment, the serum human chorionic gonadotropin started to increase gradually, but whole body imaging including the left testis revealed no abnormal finding except testicular microlithiasis. A biopsy of the left testis revealed intratubular germ cell neoplasia, unclassified type. After the human chorionic gonadotropin level reached 6.6 mIU/ml, he underwent left high orchiectomy. Histology demonstrated a small malignant germ cell tumor as well as intratubular germ cell neoplasia, unclassified type, both of which were negative for human chorionic gonadotropin staining. Besides these lesions, there were tiny foci of human chorionic gonadotropin-immunoreactive intratubular trophoblasts. Serum human chorionic gonadotropin normalized immediately after the orchiectomy, and he had no sign of recurrence at 6 months. The present case will provide new insight into the diagnosis of testicular tumor recurrence with isolated elevation of a serum tumor marker.

Successful management by provocative angiography and endovascular stent of Ureteroarterial fistula in a patient with a long-term indwelling ureteral stent.

We present the clinical course of a ureteroiliac arterial fistula in a patient who had been managed by ureteral stenting for 8 years for severe ureteral stricture after abdominoperineal resection with pelvic irradiation for advanced rectal cancer. A multidisciplinary team approach including provocative angiography and an endovascular stent saved the life of the patient. Ureteroarterial fistula is a rare complication of a long-term indwelling ureteral stent that is potentially fatal unless a prompt diagnosis and adequate therapy are provided. Heightened awareness and a high index of suspicion for this condition are required to make an early diagnosis.

Glucosamine induces rapid desensitization of glucose transport in isolated adipocytes by increasing GlcN-6-P levels.

We have examined the hypothesis that glucosamine (GlcN) can rapidly induce insulin resistance through an allosteric mechanism. When insulin-treated adipocytes were exposed to 2mM GlcN, glucose uptake was rapidly reduced by approximately 60% with a T(1/2) of 2 min. We also observed an increase in intracellular GlcN-6-P (at 5 min) from undetectable levels to approximately 260 nmol/g. Continued GlcN treatment resulted in additional accumulation of GlcN-6-P (>1200 nmol/g at 2h), but caused no further decrease in glucose uptake. Although the acute inhibitory action of GlcN could be completely reversed by removing extracellular GlcN, a slow and progressive decrease in insulin-stimulated glucose transport was observed with longer treatment times (T(1/2) of 45 min, 62% loss by 5h). From these data, we conclude that: (1) GlcN elevates intracellular GlcN-6-P levels within minutes, resulting in desensitization of the glucose transport system through allosteric inhibition of hexokinase; (2) prolonged treatment elevates GlcN-6-P to levels that cannot be effectively lowered by cell washing; and (3) residual levels of GlcN-6-P continue to allosterically inhibit glucose uptake, resulting in a slower rate of desensitization that is temporally similar to glucose-induced desensitization, but mechanistically different.

Glucosamine-induced activation of glycogen biosynthesis in isolated adipocytes. Evidence for a rapid allosteric control mechanism within the hexosamine biosynthesis pathway.

Enhanced flux through the hexosamine biosynthesis pathway (HBP) induces insulin resistance and facilitates lipid storage through the up-regulation of enzyme mRNA levels. Both actions occur over several hours and require gene expression. We now identify a regulatory arm of the HBP that involves rapid allosteric activation of glycogen synthase (GS) and stimulation of glycogen biosynthesis (GBS). When insulin-pretreated adipocytes were exposed to 2 mM GlcN, incorporation of [14C]glucose into glycogen doubled by 10 min (t(1/2) of <5 min), whereas UDP-glucose levels were concomitantly decreased during this time (t(1/2) of 1.4 min; >90% depletion). Stimulation of GBS and depletion of UDP-glucose both correlated with an early and rapid rise in the levels of glucosamine-6-phosphate (GlcN-6-P), a known activator of GS. The lowering of GlcN-6-P levels by removing extracellular GlcN (>80% reduction by 45 min) was accompanied by the restoration of UDP-glucose levels. Prolonged GlcN treatment (20 min to 2 h) inhibited GBS, which corresponded to a massive intracellular accumulation of GlcN-6-P (t(1/2) of approximately 32 min; >1,400 nmol/g). From these data, we conclude the following. 1) GlcN treatment elevated intracellular GlcN-6-P levels within minutes, resulting in allosteric activation of GS, stimulation of GBS, and a reduction in steady-state levels of UDP-glucose due to increased precursor utilization. 2) Prolonged treatment with high concentrations of GlcN caused massive accumulation of GlcN-6-P that adversely affected cellular metabolism and reduced GBS. 3) The biphasic actions of GlcN on GBS may explain many of the discrepant reports on the role of the HBP in glycogen metabolism.

Dynamic actions of glucose and glucosamine on hexosamine biosynthesis in isolated adipocytes: differential effects on glucosamine 6-phosphate, UDP-N-acetylglucosamine, and ATP levels.

Glucose and glucosamine (GlcN) cause insulin resistance over several hours by increasing metabolite flux through the hexosamine biosynthesis pathway (HBP). To elucidate the early events underlying glucose-induced desensitization, we treated isolated adipocytes with either glucose or GlcN and then measured intracellular levels of glucose-6-P (G-6-P), GlcN-6-P, UDP-Glc-NAc, and ATP. Glucose treatment rapidly increased G-6-P levels (t((1/2)) < 1 min), which plateaued by 15 min and remained elevated for up to 4 h (glucose ED(50) = 4mm). In glucose-treated cells, GlcN-6-P was undetectable; however, GlcN treatment (2 mm) caused a rapid and massive accumulation of GlcN-6-P. Levels increased by 5 min ( approximately 400 nmol/g) and continued to rise over 2 h (t((1/2)) approximately 20 min) before reaching a plateau at >1,400 nmol/g (ED(50) = 900 microm). Thus, at high GlcN concentrations, unrestricted flux into the HBP greatly exceeds the biosynthetic capacity of the pathway leading to a rapid buildup of GlcN-6-P. The GlcN-induced rise in GlcN-6-P levels was correlated with ATP depletion, suggesting that ATP loss is caused by phosphate sequestration (with the formation of GlcN-6-P) or the energy demands of phosphorylation. As expected, GlcN and glucose increased UDP-GlcNAc levels (t((1/2)) approximately 14-18 min), but greater levels were obtained with GlcN (4-5-fold for GlcN, 2-fold for glucose). Importantly, we found that low doses of GlcN (<250 microm, ED(50) = 80 microm) could markedly elevate UDP-GlcNAc levels without increasing GlcN-6-P levels or depleting ATP levels. These studies on the dynamic actions of glucose and GlcN on hexosamine levels should be useful in exploring the functional role of the HBP and in avoiding the potential pitfalls in the pharmacological use of GlcN.