Effect of tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) on human sperm motility, viability and motion parameters

Male genital tract infections and non-specific inflammatory conditions may be associated with unexplained infertility. Previous studies have shown the presence of cytokines such as tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the semen of infertile men. However, the mechanism of effect of these cytokines on human sperm function is still controversial. The present study was undertaken to investigate the in-vitro effects of TNF-α and IFN-γ on human sperm motion, viability and the hypoosmotic swelling test (HOST).
Washed spermatozoa from normal volunteers ( n =9) were incubated in the presence/absence of TNF-α (1 μg/mL) plus IFN-γ (0.1 μg/mL). Sperm motility, viability, HOST, and video sequences were recorded at different time intervals (0, 30, 60 and 180 min). Sperm motion parameters were analysed using computer-assisted semen analysis. There was a time-dependent negative effect of TNF-α plus IFN-γ on sperm motility, viability, HOST, and lateral-head displacement (ALH). The maximum decrease was observed between 60 and 180 min for sperm motility (50.8 ± 5.6%), viability (52.8 ± 4.0%), HOST (38 ± 2%) and ALH (4.7 ± 0.1 μm) compared to control samples (62.2 ± 2.8, 62.4 ± 2.9, 58 ± 4, and 5.3 ± 0.4, respectively; All p  < 0.05). There was no significant effect on sperm straight-line velocity and mean linearity when compared to control.
These data suggest that the common inflammatory cytokines TNF-α plus IFN-γ have only partial detrimental effects on sperm motility, viability, membrane integrity and lateral head displacement, which may contribute to the poor fertilizing potential of human spermatozoa during inflammatory conditions.     

Detection of DNA fragmentation in human spermatozoa: correlation with semen parameters

To determine the prevalence of high levels of sperm DNA damage among infertile men with normal and abnormal semen parameters, 90 patients were subdivided into the following three groups. Group A ( n  = 30): men with normal semen parameters who acted as the controls. Group B ( n  = 30): asthenozoospermic men and group C ( n  = 30): teratozoospermic men, suffering from male infertility. DNA damage was evaluated by the rate of DNA fragmentation index (DFI) as assessed by the terminal desoxynucleotidyl transferase-mediated dUTP nick-end labelling assay. It was found that the difference was not significant between the percentage of DFI in patients with asthenozoospermia and the normospermic men (9.46% ± 8.68 and 8.19 ± 6.84 respectively, P- value not significant). The patients with teratozoospermia showed a significantly higher percentage of DNA fragmentation compared with the controls (respectively 21.37 ± 17.26% and 8.19 ± 6.84%, P  < 0.001). There was a positive correlation between abnormal sperm morphology and the DFI ( r  = 0.44, P  < 0.01) in group C. It is concluded that the impairments of sperm parameters were associated with an increase of DNA fragmentation; this association was strictly related to atypical forms.     

Anogenital distance and reproductive parameters in young men

The purpose of this study was to assess the relationship between anogenital distance (AGD) measures and semen quality and serum reproductive hormone levels in Caucasian young men from southern Spain. Two variants of AGD [from the anus to the posterior base of the scrotum (AGD_AS) and to the cephalad insertion of the penis (AGD_AP)] were assessed in 215 university students. Semen parameters (semen volume, sperm concentration, total sperm counts, motility and morphology) and serum reproductive hormones (follicle stimulating hormone, luteinising hormone, inhibin B, testosterone, calculated free testosterone, oestradiol and sex hormone‐binding globulin) were also determined. Associations between AGD measures and the semen quality and reproductive hormone levels were tested using multiple regression analyses. Overall, median sperm concentration was 44.0 × 10⁶ ml^?1 (5th–95th percentiles: 8.9–129 × 10⁶ ml^?1), median total sperm count was 121 × 10⁶ (18.0–400 × 10⁶), and mean (SD) testosterone level was 21.7 nmol/l (6.9). Mean (SD) AGD_AS and AGD_AP measures were 48.3 mm (11.6) and 128 mm (12.0) respectively. In the multivariable analysis, AGD measures were not associated with any semen parameters or any of the reproductive hormone levels, which is in contrast to results of studies of US young men or infertile men. Further research is warranted to confirm these findings.     

Low semen volume in 47 adolescents and adults with 47,XXY Klinefelter or 46,XX male syndrome

Klinefelter syndrome is characterized by progressive testicular failure causing androgen deficiency and azoospermia in most patients. The aim of this study was to evaluate semen quality in consecutive patients with an additional X chromosome as compared with healthy males. Forty-seven males with non-mosaic 47,XXY ( n  = 40) or SRY-positive 46,XX male ( n  = 7) karyotypes aged 26.1 (range: 15.0–51.7) years participated. Semen quality was compared with 2136 (control group I) men from the general population aged 18.9 (17.9–28.6) years and with 349 fertile men (control group II) aged 30.9 (22.0–43.8) years. Semen volume adjusted for duration of abstinence was significantly smaller in the patients [2.0 (0.2–5.7) mL] when compared with control group I [3.1 (0.3–12.5) mL, p  < 0.0001] and group II [3.6 (0.6–12.5) mL, p  < 0.0001]. There was no difference in semen volume between 47,XXY and 46,XX males. All patients had azoospermia except two 47,XXY males aged 29 years who had sperm concentrations of 0.5 and 1.6 million/mL, respectively. We found significantly smaller semen volume in the patients when compared with controls, and the presence of motile spermatozoa in two out of 47 patients. The small semen volume supports the notion of 47,XXY patients being androgen insufficient despite serum testosterone levels within the normal range.     

Predictors of improved seminal characteristics by varicocele repair

The aim of this study was to investigate predictors of the improvement of semen characteristics after low ligation for patients with varicoceles. The records of 97 oligozoospermic patients who underwent microsurgical left or bilateral inguinal varicocele repair were retrospectively evaluated. We assessed factors that could be predictors of the improvement of semen characteristics using logistic regression analysis. We evaluated age, testicular volume, varicocele grade, serum follicle-stimulating hormone (FSH), luteinising hormone, testosterone, sperm concentration and motility before low ligation. Among the 97 patients, sperm concentration improved from 6.4 ± 5.8 to 24.2 ± 35.1 million ml⁻¹ and sperm motility increased from 32.8 ± 24.9% to 41.0 ± 14.6% in 55 patients (57%). In logistic regression analysis, pre-operative serum FSH and testosterone concentration were predictors of sperm concentration improvement. Varicocele repair improved sperm concentration and motility. Low serum FSH and high testosterone are significant factors predicting the improvement of semen characteristics before low ligation.     

Regional differences in semen qualities in the Baltic region

Recent prospective studies of male reproductive health have shown differences between several European countries. Our objective was to evaluate the current situation in the two Baltic States Estonia and Lithuania. In 1997-99 we investigated semen parameters, levels of reproductive hormones and general health factors of 196 men from the general population in Lithuania; from Estonia, 79 men from the general population and 118 soldiers were investigated. Adjusted for interlaboratory differences and abstinence period, sperm concentration of Lithuanian and Estonian men from the general populations were shown to be 55 and 67 million/mL, respectively. The Estonian soldiers had the highest sperm concentration, 82 million/mL. The frequencies of morphologically normal spermatozoa were 6.2, 7.7 and 9.6%, respectively. In contrast to the semen qualities, highest Inhibin B levels were detected in the Lithuanians (233 pg/mL) followed by Estonian men from the general population (220 pg/mL) and Estonian soldiers (185 pg/mL). The soldiers had also the lowest level of testosterone and oestradiol. The sperm counts of the Estonian and Lithuanian men investigated here are higher than recently shown for Norwegian, Danish, Estonian and Finnish men. Comparisons should be cautiously drawn as the groups are not completely comparable. Still, even within the Baltic region, geographically close and sharing common recent social history, differences in semen quality and levels of reproductive hormones are apparent.     

Microflora of the seminal fluid of healthy men and men suffering from chronic prostatitis syndrome

Chronic prostatitis syndrome (CPS) is a common urologic condition that many clinicians find difficult to diagnose and treat effectively. The information about the composition of the flora of the seminal fluid in healthy men and patients with CPS is limited. The aim of this study was to define the microbial communities present in the seminal fluid of healthy men and patients with CPS and at in vitro detection of decomplementary activity (DCA) phenotypes of isolates along with their comparison with isolates from patients with or without CPS. The bacteriological study was carried out to 48 healthy men and 60 men with CPS. Culture specimens were spread on various selective media. Bacterial DCA was tested by measuring the decrease in complement activity (CH₅₀) under the influence of culture supernatants. The most common isolates in both groups were coryneforms, lactobacilli, coagulase-negative staphylococci, micrococci and streptococci. Enterobacteriaceae, enterococci and Staphylococcus aureus were isolated only from the CPS group. The organisms from seminal fluid of healthy men exerted DCA at 3.56 ± 2.15; 2.47 ± 1.23 and 4.36 ± 2.2 anti-CH₅₀ for staphylococci, micrococci and diphtheroids respectively. The DCA of staphylococci, coryneforms, Enterobacteriaceae, enterococci and micrococci from CPS group were 12.8 ± 2.1 ( p  <   0.05), 4.4 ± 3.6 ( p  >   0.05), 16.8 ± 2.1, 7.2 ± 1.9 and 11.6 ± 3.3 ( p  < 0.05) anti-CH₅₀ respectively. The data obtained in this study testify the microecological disorders in microbiota of seminal fluid in CPS.     

Metenkephalin in Seminal Plasma of Infertile Men

Background: Enkephalin is one of the opioids, which is expressed widely in reproductive organs. However, the function of enkephalin in male reproduction is not completely understood. The effect of metenkephalin on sperm motility remains especially controversial. In this study we examined the level of metenkephalin in seminal plasma from men with normal sperm production and patients with asthenospermia, oligospermia, and azoospermia to investigate the role of metenkephalin in seminal plasma on sperm function. We also investigated the effect of metenkephalin on sperm motility in vitro.
Methods: Sixty nine infertile patients (31 oligospermic, 21 asthenospermic, and 17 azoospermic) were included in this study. The level of metenkephalin in seminal plasma of these men was measured and the effect of the peptide on the motility of human sperm was examined in vitro. Seventeen men with normal seminograms were a control group.
Results: The level of metenkephalin in the seminal plasma of semen from asthenospermic men was significantly lower than that from the controls ( P < 0.05). No significant correlations between the level of metenkephalin and the mean pathing or progressive velocity of sperm, or serum hormone levels were observed. In the in vitro study, which used semen from the controls, treatment of sperm with metenkephalin (50–200 pg/mL) maintained sperm motility for 4 hours. On the other hand, motility of sperm incubated without metenkephalin began to decrease at 3 hours. Metenkephalin levels of 50 pg/mL in seminal plasma is considered to be necessary for maintaining sperm motility.
Conclusion: These results suggest that metenkephalin in seminal plasma is an important clue in the investigation of decreased sperm motility.     

Correlation between neutral alpha-glucosidase activity and sperm DNA fragmentation

To evaluate the association between neutral α-glucosidase (NAG) activity and sperm DNA fragmentation (DFI), ejaculates from 24 men undergoing evaluation for sperm DNA damage as a part of infertility assessment were analysed. The mean ± SD and range for the semen quality of the 24 ejaculates are as follows: volume (3.1 ± 1.3, 1.8–6.0 ml); sperm concentration (45.6 ± 41.1, 1.3–151.2 × 10⁶ ml⁻¹); sperm motility (52.8 ± 28.8, 1–95%); sperm with fragmented DNA (17.6 ± 15.4, 1.7–56.0%); sperm with immature chromatin (9.6 ± 3.8, 2.5–19.1%); NAG activity (37.9 ± 18.3, 4.4–75.3 mU ml⁻¹). The only sperm parameter significantly correlated with neutral α-glucosidase is the percentage of sperm DFI [correlation coefficient ( r ) = 0.4376, P  = 0.03].     

Endocrine and reproductive health status of men who had experienced short-term radiation exposure at Chernobyl

Hormonal and semen parameters in 416 men aged 25–45 years were examined: 328 were men who cleaned the territory around the Chernobyl nuclear reactor (called `liquidators') and 88 were healthy age-matched controls. The dose of radiation received by the liquidators was 0.16 ± 0.08 Gy. LH, FSH, prolactin, testosterone and cortisol levels were assayed using WHO -matched reagents. Semen analyses were performed according to the WHO Manual (1992 ). The mean concentration of all hormones in liquidators and controls were within the WHO-defined normal range. The mean levels of LH and cortisol in liquidators were significantly lower ( p  = 0.013 and p  < 0.001, respectively) and testosterone significantly higher ( p  = 0.023) than in controls. The variations in hormone levels in liquidators were not correlated with the acquired doses of radiation as measured by personal dosimeters (film badges). Semen parameters in a sub-group of 70 liquidators were within the normal WHO-defined range. The percentage of normal forms of spermatozoa in liquidators (35.0 ± 13.1%) was significantly lower ( p  < 0.015) than in a control group (42.8 ± 8.9%). The study has shown that exposure of men to relatively short-term radiation did not cause long-lasting disruption of their endocrine status and spermatogenesis. The study was 7–9 years retrospective and it is therefore impossible to infer what the immediate effects of the radiation exposure were on these parameters.     

Effects of indoor air purification by an air cleaning system (Koala technology) on semen parameters in male factor infertility: results of a pilot study

A number of studies indicated a clear decline in semen quality in the past 30–50 years and there is accumulating evidence that this decline might result from exposure to high levels of air pollution. To examine the impact of environment on male reproductive ability, we undertook for the first time a pilot study on semen quality of infertile men exposed to purification of indoor air. Ten subjects with a history of unexplained male infertility and poor semen quality were exposed for at least 1 year to a cleaning indoor air system (Koala technology). The key feature of this air purifier is the unique innovative multiple filtering system. The treatment of total purification of indoor air showed neither improvements in semen parameters nor variation in reproductive hormones ( P  = N.S.), but induced an evident increase ( P  < 0.03 and more) in seminal leucocytic concentrations. Within the limits due to the small sample of subjects recruited, the sole purification of indoor air does not seem enough to improve semen quality, although the increase in leucocytic concentrations could indicate an activation of the role of immunosurveillance in a purified indoor air environment.     

Relationship between semen quality and seminal plasma components: alpha-glucosidase, fructose and citrate in infertile men compared with a normospermic population of Tunisian men

The aim of this study was to evaluate the correlation between the secretory function of the male accessory glands and sperm parameters in normospermic controls and infertile patients. One hundred and fifty-nine men were investigated: they were composed of two groups: normospermic ( n  = 37) and infertile ( n  = 122) men with altered sperm characteristics. These infertile men were divided into the following groups: asthenozoospermia ( n  = 38), teratozoospermia ( n  = 40) and asthenoteratozoospermia ( n  = 44). The patients underwent semen analysis and measurements of fructose, neutral alpha-glucosidase and citric acid. The level of fructose was significantly decreased in asthenozoospermic and increased in asthenoteratozoospermic men. It was significantly correlated with semen volume, sperm count, motility and morphology. The seminal alpha-glucosidase levels were significantly correlated with semen volume and pH and citric acid was significantly correlated with pH. Thus, alpha-glucosidase and citric acid levels were associated with semen pH. The significant correlation between semen parameters, accessory glands and epididymal functions highlights the relationship between semen and normal genital tract function.     

Have sperm counts deteriorated over the past 20 years in healthy, young Japanese men? Results from the Sapporo area

Changes in semen quality of healthy men is a controversial issue throughout the world. It is suspected that many chemical endocrine disrupters may affect the quality of semen. Although exposure to them may be extensive in Japan, no evidence of changes in semen quality has been reported. In this study, changes in semen volume and sperm counts were analyzed over 20 years in the Sapporo area of Japan. Semen volume and sperm counts were measured in 254 and 457 normal, healthy volunteers who lived in the Sapporo area in 1975-1980 and 1998, respectively. Posters and handbills were used to recruit participants in both studies. Semen samples were collected by masturbation after 3 days or more of abstinence. There was no change in semen volume between 1975-1980 and 1998. Mean sperm counts were 70.9 +/- 47.3 x 10(6)/mL in 1975-1980 and 79.6 +/- 49.3 x 10(6)/mL in 1998. Sperm counts did not decline over about 20 years. No significant correlation between age and sperm counts was recognized in either study. The rates of subjects with oligozoospermia and azoospermia were the same in both studies. In the 1975-1980 study, 34 of 254 (13.4%) participants had a child, and in the 1998 study, 51 of 457 (11.2%) participants had a child. Mean sperm count was significantly (P < .02) lower in the earlier study (66.0 +/- 44.9 x 106/mL) than in the 1998 study (98.7 +/- 60.2 x 10(6)/mL). This is the first reliable report in which changes in sperm counts in Japan were studied. We conclude that there was no evidence of deterioration in sperm counts of normal healthy men who lived in the Sapporo area of Japan over 20 years. However, selection bias in the recruitment of volunteers and the issue of variable abstinence might have affected the results of these studies. Therefore, well-designed prospective studies should be performed in several different regions to extrapolate our results on sperm counts to healthy, young Japanese men in general. Key words: Fertility, endocrine disruptors, seminalysis.     

Reproductive outcome of men with azoospermia due to cryptorchidism using assisted techniques

The aetiology of cryptorchidism is still undiscernible in the majority of cases. It has long been argued that cryptorchidism reflects a primary testicular maldevelopment, where the contralateral scrotal testis also suffers from aspermatogenesis and low spermatogonia count. The aim of the study was to determine the reproductive outcome of ex-cryptorchid men with azoospermia post-orchidopexy after testicular sperm extraction (TESE) and intracytoplasmatic sperm injection (ICSI). In a retrospective analysis, we compared the sperm retrieval, fertilization, pregnancy and live birth rates after ICSI of consecutive ex-cryptorchid azoospermic patients ( n  = 15) undergoing TESE between Jan 2000 and Dec 2007 vs. non-cryptorchid azoospermic men ( n  = 142). Sperm retrieval rate of ex-cryptorchid men by TESE (66%) was comparable with non-cryptorchid men (47%) ( p  = 0.15) despite significantly higher FSH levels (30.7 ± 25.4 vs. 17.9 ± 14.8 respectively) ( p  = 0.018) and a more prevalent histopathology diagnosis of aspermatogenesis (75% vs. 40%, p  = 0.046). Fertilization (43.3%), pregnancy (30%) and live birth (20%) rates after TESE–IVF–ICSI in the ex-cryptorchid group were not different from the non-cryptorchid group (48.7, 43 and 29%, p  = 0.26, p  = 0.21, p  = 0.29 respectively). We conclude that the reproductive outcome of ex-cryptorchid men with azoospermia post-orchidopexy employing TESE–IVF–ICSI is comparable with non-cryptorchid azoospermic men.     

Efficacy and safety of dutasteride in Japanese men with benign prostatic hyperplasia

Objectives:   To assess the efficacy and safety of dutasteride in Japanese men with benign prostatic hyperplasia (BPH).
Methods:   This was a randomized, double-blind, placebo-controlled, parallel-group study. A total of 378 subjects with clinical BPH having an International Prostate Symptom Score (IPSS) of 8 points or greater, a prostate volume of 30 mL or greater, and a maximal urinary flow rate (Qmax) of 15 mL/s or less were randomized to receive placebo or dutasteride once daily for 52 weeks. Subjects were stratified according to tamsulosin use at baseline. The numbers of subjects with and without tamsulosin use were 242 and 136, respectively. IPSS, Qmax, prostate volume and drug safety were evaluated.
Results:   Continued improvement in IPSS was noted in the dutasteride group, and dutasteride significantly decreased IPSS compared with placebo. At week 52, dutasteride significantly improved Qmax and prostate volume compared with placebo. Drug-related sexual function events in the dutasteride group were infrequent and generally were not treatment limiting.
Conclusions:   Dutasteride improves urinary symptoms and flow rate and reduces prostate volume. In Japanese men with BPH, it is effective and generally well tolerated during the one-year treatment period.     

腹腔镜肝切除术中肝下下腔静脉部分阻断对中心静脉压和出血量影响研究

目的 评估肝下下腔静脉部分阻断在腹腔镜肝切除术中的有效性和安全性。方法 回顾性分析2015年9月至2020年8月中国科学院大学重庆医院肝胆外科收治的行腹腔镜肝切除术的132例病人的临床资料。根据术中是否行肝下下腔静脉阻断分为阻断组（68例）和非阻断组（64例）,分析术中出血量、中心静脉压、断肝时间、断肝面积等指标。结果 所有病人均能耐受肝下下腔静脉部分阻断;与非阻断组比,肝下下腔静脉阻断后无血流动力学不稳定且中心静脉压明显下降,差异有统计学意义［（4.4±1.6）cmH2O vs. （1.9±1.2）cmH2O,P＜0.05］。在出血量方面,阻断组总出血量和断肝出血量与非阻断组差异无统计学意义［（289.3±113.5）mL vs. （302.4±124.6）mL,（241.2±107.9）mL vs. （277.3±114.5）mL,P均＞0.05］;但在合并中重度肝硬化病人的亚组中,相较于非阻断组,阻断组中总失血量和断肝失血量明显减少,差异有统计学意义［（342.7±70.2）mL vs. （468.2±87.8）mL,（328.7±68.9）mL vs. （427.9±93.2）mL,P均＜0.05］。两组手术时间、断肝手术时间、肝断面面积、术中液体输注量以及术后肝肾功能比较差异均无统计学意义（P均＞0.05）。结论 腹腔镜肝切除术中行肝下下腔静脉阻断可减少中重度肝硬化病人术中出血量,且对肝肾功能无明显影响,安全有效。     

Programmed cell death in varicocele-bearing testes

Accelerated apoptosis is a significant factor in the pathophysiology of male infertility disorders associated with abnormal spermatogenesis. This study aimed to investigate apoptosis in varicocele-bearing testes. Sixty four men with varicocele (18 fertile and 46 infertile) were studied compared with eight men with obstructive azoospermic as controls. Apoptosis was assessed in testicular biopsy specimens using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) method as well as electron microscopy. The results demonstrated that the occurrence of apoptotic changes comprised all types of germ cells but not affecting Sertoli cells. Mean tubular apoptotic indices of fertile or infertile men with varicocele were significantly higher than controls (mean ± SD 4.55 ± 1.03%, 6.29 ± 1.82% versus 2.71 ± 0.45%, P  < 0.05). Mean Leydig cells apoptotic indices of infertile men with varicocele were significantly higher than those of fertile men without varicocele as well as controls (1.18 ± 0.38%, 0.68 ± 0.15%, 0.31 ± 0.21%, P  < 0.05). Apoptotic indices were nonsignificantly correlated with Johnsen score, testicular volume or varicocele grade. It is concluded that testicular apoptosis is increased in varicocele-associated men either fertile or infertile who may be implicated in associated spermatogenic dysfunction.     

Mast cells in the seminal plasma of infertile men as detected by flow cytometry

Increased numbers of mast cells (MCs) in the testis have been associated with testicular dysfunction, where accumulation of MCs occurs. Furthermore, it has been reported that MCs might affect sperm function as it has been demonstrated that MC-derived tryptase in the seminal fluid might reduce sperm motility. Although MCs have been detected in rat epididymis, only little is known about the presence of MCs in human seminal plasma. Thus, we analysed MC numbers in the ejaculate of men during routine semen analysis of male patients suspected for infertility ( n  = 100). MCs were detected by c-kit (CD117) expression using flow cytometry. Thereby, we detected significant numbers of MCs in the ejaculate of most patients (559 ± 525 MCs ml⁻¹, mean ± SD). However, we could neither detect a correlation with respect to MCs and sperm count, motility or morphology nor to the seminal inflammatory markers like polymorphonuclear elastase. Nevertheless, a significant correlation of MCs to spermatozoa-bound IgA ( r  = 0.5; P  = 0.03; n  = 21) was observed. It is concluded that significant numbers of MCs can be detected in the human ejaculate without necessarily influencing sperm function. A potential role of MCs in seminal plasma as well as the association between MCs and IgA on spermatozoa remains to be elucidated.     

Early, abrupt conversion of de novo renal transplant patients from cyclosporine to everolimus: results of a pilot study

Abstract:  In a single-center study, 20 kidney transplant patients without prior rejection were abruptly converted from cyclosporine to everolimus at seven wk post-transplant. All patients received basiliximab induction with maintenance enteric-coated mycophenolate sodium and corticosteroids. Biopsy-proven acute rejection had occurred in three of 20 patients (15.0%) by the end of week seven post-conversion. All episodes were mild and reversible, with subsequent recovery of renal function. Calculated glomerular filtration rate (GFR) improved significantly (51 ± 11 mL/min at time of conversion, 58 ± 12 mL/min at week seven post-conversion, 57 ± 17 mL/min at month six post-conversion; p = 0.001). No patient developed proteinuria in the nephrotic range. Twenty-two adverse events were reported in 10 patients, three of which had a suspected relationship to everolimus. Mean leukocyte and platelet count decreased significantly, and triglyceride level increased. This study suggests that kidney transplant patients without prior rejection can be converted abruptly from cyclosporine to everolimus at seven wk post-transplant, resulting in significantly improved renal function with no apparent increase, in risk of rejection and good tolerability.     

Protein tyrosine phosphorylation under capacitating conditions in porcine fresh spermatozoa and sperm cryopreserved with and without alpha tocopherol

The aim of this study was to evaluate the capacitation behaviour of fresh and α-tocopherol frozen spermatozoa. Spermatozoa frozen with or without α-tocopherol and fresh semen were incubated under capacitating conditions. Aliquots were collected at 0, 15, 30, 45, 60, 90, 120 and 180 min of incubation time. Parameters of semen quality were evaluated by optical microscopy and capacitation was determined by the epifluorescence chlortetracycline technique. Protein tyrosine phosphorylation was examined by Western immunoblotting. Motility, viability and intact spermatozoa were higher ( P  < 0.05) in fresh semen compared with frozen samples. These parameters significantly decreased, in every treatment, throughout the incubation time. Fresh semen showed a progressive increase in capacitated spermatozoa, reaching 25 ± 3% at 180 min. Cryopreserved semen had a fast increase at the beginning of incubation time (28 ± 5% at 45 min and 28 ± 3% at 30 min for samples with or without α-tocopherol, respectively). The amount of an MW 32 kDa tyrosine-phosphorilated protein, associated with capacitation, increased throughout incubation for fresh semen and spermatozoa cryopreserved with α-tocopherol. The supplementation with α-tocopherol preserved sperm plasma membrane, reflected not only in the acrosome integrity but also in a greater efficiency of energy production.