Docosahexaenoic acid inhibits ethanol/palmitoleic acid-induced necroptosis in AR42J cells.

Fatty acid ethyl esters (FAEEs), non-oxidative metabolites of ethanol, are the main causative agents of severe acute pancreatitis resulting from alcohol abuse. Pancreatic acinar cells exposed to ethanol in combination with the fatty acid palmitoleic acid (EtOH/POA) display increased levels of palmitoleic acid ethyl ester and cell death. Oxidative stress and acinar cell necroptosis are implicated in the pathology of severe acute pancreatitis. Docosahexaenoic acid (DHA) serves as a powerful anti-oxidant that reduces pancreatic inflammation and improves the outcomes of patients with acute pancreatitis. We investigated whether treatment of EtOH/POA, as an in vitro model of alcoholic pancreatitis, increases reactive oxygen species (ROS), necroptosis-regulating proteins, and cell death by increasing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and intracellular calcium. Also, we investigated whether DHA inhibits EtOH/POA-induced alterations in pancreatic acinar AR42J cells. As a result, EtOH/POA increased intracellular and mitochondrial ROS levels, NADPH oxidase activity, necroptosis-regulating proteins, and cell death, which was inhibited by NADPH oxidase inhibitor apocynin, the Ca2+ chelator BAPTA, and DHA. However, DHA did not reduce EtOH/POA-induced increases in Ca2+ oscillation or levels in AR42J cells. Furthermore, EtOH/POA induced mitochondrial dysfunction by reducing mitochondrial membrane polarization and hence, adenosine triphosphate (ATP) production. DHA treatment attenuated EtOH/POA-induced mitochondrial dysfunction. In conclusion, DHA inhibits EtOH/POA-induced necroptosis by suppressing NADPH oxidase activity, reducing ROS levels, preventing mitochondrial dysfunction, and inhibiting activation of necroptosis-regulating proteins in AR42J cells.

Paternal age as an independent factor does not affect embryo quality and pregnancy outcomes of testicular sperm extraction-intracytoplasmic sperm injection in azoospermia.

This study was performed to evaluate the independent influence of paternal age affecting embryo development and pregnancy using testicular sperm extraction (TESE)-intracytoplasmic sperm injection (ICSI) in obstructive azoospermia (OA) and nonobstructive azoospermia (NOA). Paternal patients were divided into the following groups: ≤30 years, 31-35 years, 36-40 years, 41-45 years and ≥46 years. There were no differences in the rates of fertilisation or embryo quality according to paternal and maternal age. However, clinical pregnancy and implantation rates were significantly lower between those ≥46 years of paternal age compared with other age groups. Fertilisation rate was higher in the OA than the NOA, while embryo quality, pregnancy and delivery results were similar. Clinical pregnancy and implantation rates were significantly lower for patients ≥46 years of paternal age compared with younger age groups. In conclusion, fertilisation using TESE in azoospermia was not affected by the independent influence of paternal age; however, as maternal age increased concomitantly with paternal age, rates of pregnancy and delivery differed between those with paternal age <41 years and ≥46 years. Therefore, paternal age ≥46 years old should be considered when applying TESE-ICSI in cases of azoospermia, and patients should be advised of the associated low pregnancy rates.

Clinical significance of subclinical varicocelectomy in male infertility: systematic review and meta-analysis.

Recent meta-analysis by the Cochrane collaboration concluded that treatment of varicocele may improve an infertile couple's chance of pregnancy. However, there has been no consensus on the management of subclinical varicocele. Therefore, we determine the impact of varicocele treatment on semen parameters and pregnancy rate in men with subclinical varicocele. The randomised controlled trials that assessed the presence and/or treatment of subclinical varicocele were included for systematic review and meta-analysis. Random effect model was used to calculate the weighted mean difference of semen parameters and odds ratio of pregnancy rates. Seven trials with 548 participants, 276 in subclinical varicocelectomy and 272 in no-treatment or clomiphene citrate subjects, were included. Although there was also no statistically significant difference in pregnancy rate (OR 1.29, 95% CI 0.99-1.67), surgical treatment resulted in statistically significant improvements on forward progressive sperm motility (MD 3.94, 95% CI 1.24-6.65). However, the evidence is not enough to allow final conclusions because the quality of included studies is very low and further research is needed.

Expression of cubilin in mouse testes and Leydig cells.

Cubilin (cubn) is a receptor for vitamins and various protein ligands. Cubn lacks a transmembrane domain but anchors to apical membranes by forming complexes with Amnionless or Megalin. In an effort to better understand the uptake of nutrients in testis, we analysed cubn expression in the developing mice testes. In testes, cubn mRNA increased from birth to adulthood. In the inter-stitium and isolated seminiferous tubules, neonatal increase in cubn mRNA until 14 days post-partum (pp) was followed by a marked increase at puberty (28 days pp). Cubn was found in the gonocytes, spermatogonia, spermatocytes and spermatids in the developing testes. In adult testes, strong Cubn immunoreactivity was found in the elongating spermatids, suggesting the role of Cubn in endocytosis during early spermiogenesis. In Sertoli cells and peritubular cells, Cubn immunoreactivity was weak throughout the testis development. In the inter-stitium, Cubn immunoreactivity was found in foetal Leydig cells, was weak to negligible in the stem cells and progenitor Leydig cells and was strong in immature and adult Leydig cells, demonstrating a positive association between Cubn and steroidogenic activity of Leydig cells. Collectively, these results suggest that Cubn may participate in the endocytotic uptake of nutrients in germ cells and somatic cells, supporting the spermatogenesis and steroidogenesis in mouse testes.

Interventions do not enhance medication persistence and compliance in patients with overactive bladder: a 24 weeks, randomised, open-label, multi-center trial.

OBJECTIVES: Anticholinergics are currently the mainstay for the management of overactive bladder (OAB). However, low drug adherence has been noted with these medications. The aim of this study was to determine whether a health education intervention (HEI) could improve drug persistence with anticholinergics in OAB patients. METHODS: We enrolled 682 OAB patients who were randomly distributed into either the HEI plus fesoterodine (HEI) group or the fesoterodine alone (control) group. The HEI consists of four education sections: understanding OAB disease, dietary control, bladder training and understanding anticholinergics. The primary end-point was the difference in drug persistence between the HEI and control groups at 24 weeks. Persistence was defined as a gap ≤ 30 days between successive prescription pills. RESULTS: Among the 682 patients, 210 (30.8%) completed 24 weeks of study. Persistence of the HEI group at 6 months was not statistically higher than that of the control group (40.4% vs. 34.9%, p = 0.181). Compliance at 6 months was also similar between the two groups (38.5% vs. 32.5%, p = 0.128). Using OAB symptom score questionnaire, the efficacy of the two groups was not different at each follow-up (p > 0.05). The global response was similar between the two groups. However, the HEI group was more satisfied with treatment than the control group (p = 0.034). The most common reason for discontinuation was satisfaction with the treatment so that they did not need to follow-up, followed by inadequate efficacy in both groups. Adverse events were reported in 12.3% of patients. CONCLUSIONS: The health education intervention was not effective to increase drug persistence in OAB patients on anticholinergics.

The efficacy and tolerability of tarafenacin, a new muscarinic acetylcholine receptor M3 antagonist in patients with overactive bladder; randomised, double-blind, placebo-controlled phase 2 study.

OBJECTIVES: To evaluate the dose-response relationship of tarafenacin, an antimuscarinic agent in development phase, for efficacy and safety, at daily doses of 0.2 and 0.4 mg for the treatment of overactive bladder (OAB) PATIENTS AND METHODS: This multicentre, placebo-controlled, randomised, double-blind, phase 2b study was conducted. Patients were randomised to tarafenacin 0.2 mg, tarafenacin 0.4 mg or placebo daily for 12 weeks. Adult patients with OAB for at least 6 months, with an average of ≥ 8 micturitions per day and ≥ 3 incontinence episodes or a total of ≥ 6 urgency episodes per 3 days were enrolled. The primary objective was to compare the mean changes in the number of micturitions per 24 h of the two doses of tarafenacin compared with placebo from baseline to 12 weeks after treatment. RESULTS: A total of 334 patients were screened, of whom 235 patients were randomised. The mean decrease in the number of micturitions per 24 h from baseline to 12 weeks was statistically higher in the tarafenacin 0.4 mg group (-2.43 ± 2.21 times per day, p = 0.033) and non-statistically significant in the tarafenacin 0.2 mg group (-1.92 ± 2.45 times per day, p = 0.393) when compared with the placebo group (-1.77 ± 2.95 times per day). There were no statistically significant differences in the mean change of urgency episodes per 24 h among three groups. The most common adverse event was dry mouth. There were no significant differences in blurred vision and constipation compared with placebo. CONCLUSIONS: Tarafenacin 0.4 mg decreased the number of micturitions in patients with OAB after 12 weeks compared with placebo, and the dose-response relationship of tarafenacin 0.2 and 0.4 mg was confirmed. Both dose levels of tarafenacin were well tolerated.

Unusual maternal uniparental isodisomic x chromosome mosaicism with asymmetric y chromosomal rearrangement.

Infertile men with azoospermia commonly have associated microdeletions in the azoospermia factor (AZF) region of the Y chromosome, sex chromosome mosaicism, or sex chromosome rearrangements. In this study, we describe an unusual 46,XX and 45,X mosaicism with a rare Y chromosome rearrangement in a phenotypically normal male patient. The patient's karyotype was 46,XX[50]/45,X[25]/46,X,der(Y)(pter→q11.222::p11.2→pter)[25]. The derivative Y chromosome had a deletion at Yq11.222 and was duplicated at Yp11.2. Two copies of the SRY gene were confirmed by fluorescence in situ hybridization analysis, and complete deletion of the AZFb and AZFc regions was shown by multiplex-PCR for microdeletion analysis. Both X chromosomes of the predominant mosaic cell line (46,XX) were isodisomic and derived from the maternal gamete, as determined by examination of short tandem repeat markers. We postulate that the derivative Y chromosome might have been generated during paternal meiosis or early embryogenesis. Also, we suggest that the very rare mosaicism of isodisomic X chromosomes might be formed during maternal meiosis II or during postzygotic division derived from the 46,X,der(Y)/ 45,X lineage because of the instability of the derivative Y chromosome. To our knowledge, this is the first confirmatory study to verify the origin of a sex chromosome mosaicism with a Y chromosome rearrangement.

A randomised, prospective double-blind, propiverine-controlled trial of imidafenacin in patients with overactive bladder.

AIM: To assess the efficacy and safety of imidafenacin compared with propiverine for treatment of overactive bladder (OAB) in Korean patients. MATERIALS AND METHODS: Patients with OAB symptoms were randomised to double-blind treatment with 0.1 mg of imidafenacin twice daily (group A) or propiverine 20 mg once daily (group B) for 12-week regimen, and assessed for efficacy and safety. The primary efficacy outcome was per cent change of weekly urgency urinary incontinence (UUI) episodes at week 12. The secondary efficacy outcomes were changes in the micturitions per day, urine volume voided per micturition, urgency episodes per day, complete disappearance of incontinence episodes and severity of urgency from baseline to week 12. Quality of life and safety profiles were also compared. RESULTS: Of 162 patients randomised, 140 completed the study protocol. The per cent change of weekly UUI episodes at week 12 was -69.1% in group A and -70.4% in group B (both p < 0.0001). The lower limit of 95% one-sided confidence interval of the difference between the groups was above the non-inferiority margin (-19.42%). Other voiding parameters and quality of life significantly improved at week 12 in both the groups. The discontinuation rates caused by adverse events were low in both the groups. While dry mouth was the most common adverse event (group A: 28.4% vs. B: 30.4%, p = 0.783), the severity of dry mouth was significantly less in the group A than B (p = 0.042) There were no significant differences in other safety profiles. CONCLUSIONS: After the 12-week treatment of imidafenacin 0.1 mg twice daily, all OAB symptoms and quality of life improved. Imidafenacin was not inferior to propiverine for the reduction of UUI episodes, and was better tolerated than propiverine in the safety profile. Our results indicate that imidafenacin is a safe and effective drug in Korean patients with OAB.

A randomised, double-blind, parallel design, multi-institutional, non-inferiority phase IV trial of imidafenacin versus fesoterodine for overactive bladder.

AIMS: Our objective was to compare the efficacy and safety of imidafenacin over fesoterodine in patients with overactive bladder (OAB). METHODS: This study is a randomised, double-blind, parallel-group, fesoterodine-controlled study in patients with continuous OAB symptoms for ≥ 3 months, daily mean voiding frequency (DMVF) ≥ 8, and daily mean urgency or urgency incontinence frequency ≥ 2. A twice-daily 0.1 mg imidafenacin with placebo, or once-daily 4 mg fesoterodine with placebo were administered for 12 weeks. The primary efficacy end-point was the difference in DMVF at 12 weeks. The secondary efficacy end-points were differences in daily mean: (i) voiding frequency at 4 and 8 weeks; (ii) urgency frequency; (iii) urgency incontinence frequency; (iv) incontinence frequency; (v) nocturia frequency; and (vi) quality of life score. The variables for safety analysis were adverse events, vital signs, residual urine volume and clinical laboratory tests. An efficacy analysis was conducted in per-protocol patients and the safety analysis was conducted in all randomised patients. RESULTS: The differences in DMVF at 12 weeks were -3.38 ± 3.63 and -2.45 ± 3.73 in the imidafenacin and fesoterodine groups, respectively, and the difference was not significant between the two groups. Imidafenacin was non-inferior to fesoterodine, and the lower limit of 95% two-sided confidence intervals was -0.53. The other six secondary end-points and variables for safety analysis showed no difference between the two groups. CONCLUSIONS: Imidafenacin was non-inferior to fesoterodine in terms of efficacy, and showed no significant difference in terms of safety.

Temperature dependence of the electronic structure of two-dimensional Na gas on the Si(111)-7 × 7 surface.

The temperature dependence of the irreversible phase transition from a two-dimensional gas to an ordered zero-dimensional solid on the Si(111)-7 × 7 surface was studied using photoemission spectroscopy. With increasing Na coverage, the two-dimensional Na gas, which is a state of highly mobile Na atoms, undergoes a phase transition into ordered zero-dimensional magic nanoclusters at room temperature. The critical Na coverage of the phase transition was found to increase with reduced temperature. This was used to develop a gas-solid phase diagram of Na atoms on the Si(111)-7 × 7 surface as a function of Na coverage and sample temperature based on the electronic structure. The temperature dependence of the phase transition can be ascribed to the suppression of the thermal energy that is required to overcome the energetic barrier between the two-dimensional gas and the zero-dimensional solid at low temperature, where three different hopping mechanisms are related to the phase transition.

Molecular and clinical characteristics of 26 cases with structural Y chromosome aberrations.

Structural abnormalities include various types of translocations, inversions, deletions, duplications and isochromosomes. Structural abnormalities of the Y chromosome are estimated to affect less than 1% of the newborn male population and are particularly hazardous for male reproductive function. The objective of this study was to characterize a group of patients with structural abnormalities of the Y chromosome. All patients who visited our laboratory between 2007 and 2010 underwent cytogenetic investigations. Among these, we detected 26 patients with structural abnormalities of the Y chromosome. To confirm the structural Y chromosome alterations, we used special bandings, FISH and multiplex PCR to detect Y chromosome microdeletions. Of the 26 patients presented here, 11 had an isodicentric Y chromosome, 7 had an inversion, 3 had a translocation, 2 had a derivative, 2 had a Yqs and 1 had a deletion. Sixteen were diagnosed with azoospermia, 8 as normal fertile males and 1 as a man who was unable to donate semen due to mental retardation. One of the patients having 45,X/46,X,idic(Y) was reported to be phenotypically female with primary amenorrhea and without uterus. Deletions of the AZFbc region were correlated with the sperm concentration (p < 0.05), but no correlation with the levels of FSH, LH, testosterone, prolactin and estradiol were found. The present report shows that the precise identification of structural Y chromosome aberrations may be clinically important for genetic counseling and assisted reproductive technology treatment.

Patient-reported goal achievement after antimuscarinic treatment in patients with overactive bladder symptoms.

AIM: Standardised traditional outcome measures may fail to address factors that are important to patients and address irrelevant factors. Aim of this study was to assess patient-reported goals and goal achievement (GA) in the antimuscarinic treatment for overactive bladder (OAB) patients. METHODS: Men and women aged ≥ 18 years with OAB symptoms were eligible for the study. Treatment began with a dose of 10 mg oxybutynin, to be increased if necessary to 30 mg. Before treatment, each patient's primary treatment goal was identified. After 12-week treatment, patients reported GA using a Likert scale from 0 (no achievement) to 5 (complete achievement). Successful achievement was defined as a score of 4 or 5. Traditional outcome measures including voiding diaries, the OAB questionnaire short form, patient perception of bladder condition, and treatment benefit and satisfaction were assessed. Baseline characteristics affecting GA and the correlation between GA and traditional outcome measures were evaluated. RESULTS: A total of 303 goals were identified from 303 patients (51 men, 252 women). Of those, 72.3% addressed symptom relief and frequency as the most common target symptom. Other goals addressed were improving quality of life (13.5%) and eliminating coping behaviours (14.2%). After treatment, 42% had a successful GA with a median score of 3 (interquartile range; 2-4). Age had a negative effect on GA. Goal achievement was the outcome measure most correlated with treatment benefit and satisfaction. DISCUSSION AND CONCLUSIONS: Goal achievement can be a valuable outcome measure in OAB patients, addressing individual treatment goals and reflecting treatment benefit and patient satisfaction.

Optical spectroscopy of PPV-based block copolymers of nanostructured supramolecular organic semiconductor.

The third-order nonlinearity of a PPV-based nanostructured supramolecular organic semiconductor (DBAB), with an electron donor (D) connected to an electron acceptor (A) via nonconjugated and flexible bridge (B) units, was investigated in this work at both near-resonant (532 nm) and nonresonant (1064 nm) wavelength by using degenerate four-wave mixing. The second hyperpolarizabilities of D, A, and DBAB at 532 nm were found to be approximately 2.42 x 10(-43) m2/V2, 7.75 x 10(-44) m2/V2, and 1.80 x 10(-43) m2/V2 in copolarization geometry, and approximately 1.59 x 10(-43) m2/V2, 2.59 x 10(-44) m2/V2, and 1.18 x 10(-43) m2/V2 in orthogonal polarization geometry, respectively. The second hyperpolarizabilities of DBAB at 1064 nm were approximately 1.66 x 10(-46) m2/V2 and approximately 8.77 x 10(-47) m2/V2 for parallel and orthogonal polarization cases.

Nonlinear optical properties of mushroom-shaped CdSe/CdS coreshells.

The third-order nonlinear optical susceptibilities of mushroom-shaped CdSe/CdS coreshells as a function of concentration have been investigated using polarization- and concentration-resolved degenerate four-wave mixing in a resonant region. The effective third-order nonlinear optical susceptibilities, /chi(3)xxxx/ and /chi(3)xyyx/ of CdSe/CdS coreshells were estimated to be approximately 1.86 x 10(-21)-1.03 x 10(-20) m2/V2, and approximately 0.45 x 10(-21)-6.15 x 10(-21) m2/V2, respectively, for various concentrations of approximately 0.64 x 10(-3)-4.95 x 10(-3) mol/m3. The second hyperpolarizabilities, /xxxx/ and /xyyx/, of CdSe/CdS coreshells were extracted to be approximately 2.37 x 10(-41) m5/V2 and approximately 1.29 x 10(-41) m5/V2, respectively.

Relationship between ambient temperature and heat flux in the scrotal skin.

Excessive scrotal heating or cooling may lead to the cessation of spermatogenesis. Data regarding heat exchange rates in scrotal skin can be used to control testicular temperature within the appropriate range. Heat flux (HF) in the scrotal skin surface is generated based on the surrounding environment. This study aims to elucidate the HF of scrotal skin by varying ambient temperature. Twenty college students including seven varicoceles volunteered as the subjects (mean age: 22.95 +/- SD 1.96 years; height: 175.00 +/- 5.17 cm; weight: 68.40 +/- 8.65 kg; body mass index: 22.28 +/- 2.15), and participated in the experiments from September 11 to October 4, 2006. The environmental temperature was controlled at 20 degrees C and 25 degrees C in the first and second experiment respectively. The HF and skin temperature on both sides of the scrotal surface were measured for 60 min in the environmental chamber. The results revealed that the HF was 87.64 +/- 12.69 W/m(2) and 78.91 +/- 12.09 W/m(2) in the left and right side of the scrotum respectively. The scrotal skin temperature (SST) was 30.28 +/- 0.75 degrees C and 30.24 +/- 0.62 degrees C on the left and right side of the scrotum in the 20 degrees C environment respectively. In the 25 degrees C environment the HF was 53.54 +/- 8.86 W/m(2) and 45.25 +/- 8.32 W/m(2), and the SST was 32.29 +/- 0.61 degrees C and 32.07 +/- 0.36 degrees C on the left and right side of the scrotum respectively. The cooling source power to decrease testicular temperature is suggested at 290 W/m(2). This suggested value could be adopted a cooling device as clinical therapy for a heat stress patient to decrease testicular temperature affecting spermatogenesis.

Efficacy and safety of solifenacin succinate in Korean patients with overactive bladder: a randomised, prospective, double-blind, multicentre study.

PURPOSE: We assessed the efficacy and safety of solifenacin compared with tolterodine for treatment of overactive bladder (OAB) in Korean patients. MATERIALS AND METHODS: The study was randomised, double-blind, tolterodine-controlled trial in Korea. Patients had average frequency of >or= 8 voids per 24 h and episodes of urgency or urgency incontinence >or= 3 during 3-day voiding diary period. Patients were randomised to 12-week double-blind treatment with either tolterodine immediate release (IR) 2 mg twice daily (TOL4) or solifenacin 5 mg (SOL5) or 10 mg (SOL10) once daily. The outcome measure was mean change in daily micturition frequency, volume, daily frequency of urgency incontinence, urgency and nocturia from baseline to week 12. Quality of life was assessed using the King's Health Questionnaire. RESULTS: A total of 357 were randomised and 329 were evaluated for efficacy. All voiding parameters recorded in micturition diary improved after treatment in all three groups. Mean changes in volume voided were 19.30 ml (26.69%) in TOL4, 30.37 ml (25.89%) in SOL5 and 37.12 ml (33.36%) in SOL10 group (p = 0.03). Speed of onset of SOL10 efficacy on urgency incontinence was faster than that of SOL5 and TOL4. Quality of life improved in all three groups. Dry mouth was the most common adverse event; its incidence was the lowest in SOL5 group (7.63%, compared with 19.49% and 18.64% in SOL10 and TOL4 groups respectively). CONCLUSIONS: Solifenacin succinate 5 and 10 mg once daily improve OAB symptoms with acceptable tolerability levels compared with tolterodine IR 4 mg. Solifenacin 5 mg is a recommended starting dose in Korean patients with OAB.

The effect of overactive bladder syndrome on the sexual quality of life in Korean young and middle aged women.

Some reports showed that urinary incontinence (UI) or female lower urinary tract symptoms (LUTS) affect life quality and sexual activity. In clinical practice, it is commonly found that not only the symptoms of UI but also overactive bladder (OAB) syndrome affect daily lifestyle and sexual activity, especially in women in the most active era in their social and personal life. However, there is lack of data proving the effect of OAB syndrome on sexual activity or sexual life quality in sexually active age group. This study aimed at evaluating the effect of OAB syndrome and UI on the sexual activity and on the sexual quality of life (QoL) of Korean women age from 20s to 40s. We investigated 3372 women aged between 20 and 49 y, enrolled via a multicenter internet survey. A structured questionnaire was used to collect data about their LUTS and sexual activities. The prevalence of OAB syndrome and UI in 3372 women was 12.7 and 21.0%, respectively. Mean subject age was 26.4+/-4.8 y and 79.5% of subjects were 20-29 y old. Having OAB syndrome or UI were found to be significant predictors of sexual life problems (OAB syndrome: OR=5.08, 95% CI=3.68-7.01; UI: OR=4.16, 95% CI=3.06-5.67). Sexual activity was significantly reduced in OAB syndrome and UI versus the asymptomatic group (OAB syndrome: OR=4.8, 95% CI=3.14-6.83; UI: OR=3.9, 95% CI=2.81-5.27). This study is the first internet-based study concerning the sexual QoL in UI and OAB syndrome. In this study, OAB syndrome was found to cause a greater deterioration in the sexual QoL than UI. These results suggest that these symptoms have a significant impact upon women's personal and social lives and markedly affect the QoL.

Staurosporine mobilizes Ca(2+) from secretory granules by inhibiting protein kinase C in rat submandibular acinar cells.

Staurosporine was previously shown to mobilize Ca(2+) from the thapsigargin-insensitive Ca(2+) store in rat submandibular acinar cells. However, the nature of the store is not yet known. Therefore, in the present study, the staurosporine-releasable intracellular Ca(2+) store was characterized. Staurosporine increased the cytosolic Ca(2+) concentration ([Ca(2+)](c)) after the inositol 1,4,5-trisphosphate (IP(3))-sensitive Ca(2+) store was depleted. Ionomycin caused only small increases in [Ca(2+)](c) after the depletion of the IP(3)-sensitive Ca(2+) store, whereas ionomycin+monensin caused large increases. However, ionomycin+monensin did not increase [Ca(2+)](c) when added after [Ca(2+)](c) was increased by staurosporine, indicating that the acidic Ca(2+) store was the main source of Ca(2+). The acidic Ca(2+) store appeared to be associated with secretory granules, since ionomycin+monensin- and staurosporine-induced [Ca(2+)](c) increases were significantly reduced when the acinar cells were degranulated. The effect of staurosporine on [Ca(2+)](c) was mimicked by other protein kinase C inhibitors. Therefore, we conclude that staurosporine mobilizes Ca(2+) from secretory granules, probably through the inhibition of protein kinase C in rat submandibular acinar cells.

Predictive factors of successful testicular sperm recovery in non-obstructive azoospermia patients.

Recovery of testicular spermatozoa from non-obstructive azoospermic patients for intracytoplasmic sperm injection (ICSI) is a recent advance in the treatment of male infertility. The purpose of this study is to identify predictive factors for sperm recovery in non-obstructive azoospermic patients. A total of 178 men with non-obstructive azoospermia had multiple testicular sperm extraction (TESE) procedures to recover spermatozoa for intracytoplasmic sperm injection (ICSI) from June 1996 to February 1999. Testicular volume, serum follicle stimulating hormone (FSH) level and testicular histology were examined as positive predictive factors for sperm recovery. Testis biopsies were categorized as severe hypospermatogenesis, maturation arrest and Sertoli cell-only syndrome based on the most advanced pattern of spermatogenesis seen on histology. Sperm retrieval success rates for the patients in three histopathological categories were compared. Spermatozoa were successfully recovered in 94 of 178 (52.8%) men. Sperm were retrieved in 13 of 80 (16.3%) with Sertoli cell-only syndrome, 15 of 24 (62.5%) with maturation arrest, and 66 out of 74 (89.2%) with severe hypospermatogenesis. Spermatozoa recovery has no correlation with testicular volume or serum FSH level. When compared against Sertoli cell-only syndrome, the odds of sperm retrieval success rate was 44.3 times higher in severe hypospermatogenesis and 8.4 times in maturation arrest. These results demonstrate meaningful correlation between successful testicular sperm recovery and testis histopathology. Only testicular histopathology can be used as a predictor of successful sperm recovery.

Zn2+-induced ERK activation mediated by reactive oxygen species causes cell death in differentiated PC12 cells.

Recent studies have provided evidence that Zn2+ plays a crucial role in ischemia- and seizure-induced neuronal death. However, the intracellular signaling pathways involved in Zn2+-induced cell death are largely unknown. In the present study, we investigated the roles of mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinase (JNK), p38 MAPK and extracellular signal-regulated kinase (ERK), and of reactive oxygen species (ROS) in Zn2+-induced cell death using differentiated PC12 cells. Intracellular accumulation of Zn2+ induced by the combined application of pyrithione (5 microM), a Zn2+ ionophore, and Zn2+ (10 microM) caused cell death and activated JNK and ERK, but not p38 MAPK. Preventing JNK activation by the expression of dominant negative SEK1 (SEKAL) did not attenuate Zn2+-induced cell death, whereas the inhibition of ERK with PD98059 and the expression of dominant negative Ras mutant (RasN17) significantly prevented cell death. Inhibition of protein kinase C (PKC) and phosphatidylinositol-3 kinase had little effect on Zn2+-induced ERK activation. Intracellular Zn2+ accumulation resulted in the generation of ROS, and antioxidants prevented both the ERK activation and the cell death induced by Zn2+. Therefore, we conclude that although Zn2+ activates JNK and ERK, only ERK contributes to Zn2+-induced cell death, and that ERK activation is mediated by ROS via the Ras/Raf/MEK/ERK signaling pathway.