Isotretinoin administration improves sperm production in men with infertility from oligoasthenozoospermia: a pilot study.

There is currently no effective medical therapy for men with infertility due to oligoasthenozoospermia. As men with abnormal sperm production have lower concentrations of 13-cis-retinoic acid in their testes, we hypothesized that men with infertility from oligoasthenozoospermia might have improved sperm counts when treated with isotretinoin (13-cis-retinoic acid). We conducted a single-site, single-arm, pilot study to determine the effect of therapy with isotretinoin on sperm indices in 19 infertile men with oligoasthenozoospermia. Subjects were men between 21 and 60 years of age with infertility for longer than 12 months associated with sperm concentrations below 15 million sperm/mL. All men received isotretinoin 20 mg by mouth twice daily for 20 weeks. Subjects had semen analyses, physical examinations, and laboratory tests every 4 weeks during treatment. Nineteen men enrolled in the study. Median (25th, 75th) sperm concentration increased from 2.5 (0.1, 5.9) million/mL at baseline to 3.8 (2.1, 13.0) million/mL at the end of treatment (p = 0.006). No significant changes in sperm motility were observed. There was a trend toward improved sperm morphology (p = 0.056). Six pregnancies (three spontaneous and three from intracytoplasmic sperm injection) and five births occurred during the study. Four of the births, including all three of the spontaneous pregnancies, were observed in men with improvements in sperm counts with isotretinoin therapy. Treatment was well tolerated. Isotretinoin therapy improves sperm production in some men with oligoasthenozoospermia. Additional studies of isotretinoin in men with infertility from oligoasthenozoospermia are warranted.

Intratesticular 13-cis retinoic acid is lower in men with abnormal semen analyses: a pilot study.

Intratesticular retinoic acid is necessary for spermatogenesis, but the relationship between intratesticular retinoic acid and sperm quality in man has not been studied. We hypothesized that intratesticular concentrations of retinoic acid would be lower in men with abnormal semen analyses compared to men with normal semen analyses. We recruited men requiring scrotal or penile surgery in a pilot observational study examining the relationship between sperm quality and intratesticular and serum retinoic acid. Twenty-four men provided two pre-operative blood and semen samples, and underwent a testicular biopsy during surgery. Serum and tissue all-trans and 13-cis retinoic acid and reproductive hormones were measured by LC/MS/MS and radioimmunoassays, respectively. Seven men had abnormal semen analyses by at least one WHO criteria and 17 men were normal. In men with abnormal semen, the median (25th, 75th percentile) intratesticular 13-cis retinoic acid was 0.14 (0.08, 0.25) pmol/gram tissue compared with 0.26 (0.18, 0.38) pmol/gram tissue in men with normal semen (p = 0.04). There were no significant differences in intratesticular all-trans retinoic acid or serum reproductive hormones between men with normal and abnormal semen analyses. Intratesticular 13-cis retinoic acid is significantly lower in men with abnormal semen analyses compared to men with normal semen analyses. Lower intratesticular 13-cis retinoic acid concentrations may be due to decreased biosynthesis or increased metabolism in the testes. Further investigation of the relationship between intratesticular 13-cis retinoic acid and poor sperm quality is warranted to determine if this association is present in infertile men.

HLA-E polymorphisms in hematopoietic stem cell transplantation.

Human leukocyte antigen (HLA)-E is an inhibitory ligand of natural killer cells and γ/δ T-cells. Differential expression of HLA-E alleles on the cell surface has been reported to influence outcome of hematopoietic stem cell transplantation (HSCT). We performed HLA-E genotyping in 116 HSCT patients and their HLA-matched unrelated donors. The impact of HLA-E genotypes on patient's overall survival (OS), disease free survival (DFS), cumulative incidences for relapse, transplant-related mortality (TRM) and acute graft vs host disease (aGvHD) was assessed. Neither univariate nor multivariate analysis showed any influence of HLA-E polymorphisms on the investigated endpoints of HSCT in our cohort. We could not confirm any of the previous observations in our cohort and consider it unlikely that HLA-E polymorphisms affect outcome of HSCT.

Respiration activity monitoring system (RAMOS), an efficient tool to study the influence of the oxygen transfer rate on the synthesis of lipopeptide by Bacillus subtilis ATCC6633.

The effect of oxygen transfer rate (OTR) on the synthesis of mycosubtilin, a non ribosomal lipopeptide antifungal biosurfactant, was investigated in the respiration activity monitoring system (RAMOS) for two Bacillus subtilis strains. These cultures were performed under definite oxygen-limited conditions without the adding of any anti-foam in the culture medium. By using four different filling volumes (FV) in the shaken bioreactors, different levels (20, 14, 9 and 7 mmol O(2)l(-1)h(-1)) of oxygen-limited growth could be obtained. A 25-fold increase of the specific productivity of mycosubtilin was observed for B. subtilis ATCC6633 in the case of the most severe oxygen limitation. But nearly no effect could be found with strain BBG100 carrying the constitutive P(repU) promoter instead of the natural P(myc) promoter. Transcript analysis of the fenF gene belonging to the myc operon indicated that the P(myc) promoter regulation could be slightly oxygen sensitive. Additionally, different patterns of the synthetised mycosubtilin homologues were obtained for different level of oxygen-limited growths. At the present state of investigation, oxygen regulation was thus shown to act at different levels suggesting the existence of a complex regulatory system of NRPS lipopeptide synthesis in the natural B. subtilis ATCC6633 strain.

Miglustat has no apparent effect on spermatogenesis in normal men.

BACKGROUND: In mice, administration of the glycosphingolipid biosynthesis inhibitor miglustat results in reversible infertility, characterized by impaired sperm motility and markedly abnormal sperm morphology. This observation suggested that miglustat might have utility for fertility control in man. To ascertain the impact of miglustat on human spermatogenesis, we conducted a pilot study of miglustat administration in normal men. METHODS: After a 2-week baseline period, seven normal men were administered miglustat 100 mg, orally, twice daily for 6 weeks. During treatment, subjects had frequent seminal fluid analyses to assess the impact of treatment on sperm concentration, motility and morphology and the ability to undergo the acrosome reaction by in vitro assays. RESULTS: Five subjects completed all aspects of the study. In these subjects, there was no apparent effect of miglustat on sperm concentration, motility or sperm morphology after 6 weeks of therapy. In addition, no changes in acrosome structure or function were observed with treatment, despite therapeutic concentrations of miglustat in the serum and seminal plasma. All subjects experienced gastrointestinal upset, diarrhoea and mild weight loss during treatment. No other abnormalities in blood counts, serum chemistries, vision or overall health were observed. CONCLUSION: In contrast to the observations in mice, the oral administration of miglustat does not appear to affect human spermatogenesis. Further elucidation of the mechanism underlying the species specificity of miglustat may improve our understanding of the role of glycosphingolipids in spermatogenesis and result in alternative approaches to male fertility control.

[Acute pancreatitis]. / Akute Pankreatitis.

While interstitial acute pancreatitis usually takes a benign course, necrotizing acute pancreatitis takes a severe course, mainly because of severe local and systemic complications. After a quick diagnosis it is necessary to rapidly assess a degree of severity of the disease and thus the prognosis. The clinical picture and the result of imaging procedures do not always correspond. The management basically includes to treat pain as well as to administer fluid, electrolyte, protein and calories. In addition, systemic treatment of complications such as shock or respiratory and renal insufficiency--if occurring--is necessary. In case of pancreatic necrosis, prophylactic administration of pancreas-penetrable antibiotics is recommended to avoid infection. In the severely ill with infected pancreatic necrosis, surgery is the treatment of choice. In approximately 10% of all patients with alcohol-induced pancreatitis, there is a gradual transition to chronic pancreatitis.

Comparison of microscopic methods for detecting inflammation in expressed prostatic secretions.

PURPOSE: We evaluated microscopic methods of detecting inflammation in expressed prostatic secretions. MATERIALS AND METHODS: Methods of counting expressed prostatic secretion leukocytes were compared in 251 samples from 159 patients with chronic prostatitis/chronic pelvic pain syndrome, including traditional wet mounts, hemocytometer derived concentrations and expressed prostatic secretion smears stained with Gram's method or DiffQuick stain (Dade International, Inc., Miami, Florida). RESULTS: Of 159 initial patient evaluations 84 (53%) showed inflammation by hemocytometer concentration at 500 leukocytes per mm.3 or greater but only 37 (23%) were considered inflammation by the traditional wet mount method (p <0.001). Inflammation was identified in 149 of 251 specimens (59%) by hemocytometer but in only 82 (33%) by wet mount (p <0.001). When inflammation was defined as 1,000 leukocytes per mm.3 or greater the hemocytometer still identified significantly more patients (41%) and specimens (48%) with inflammation than the wet mount. The hemocytometer method had a substantially lower interassay and intra-assay coefficient of variation than the wet mount method. Polymorphonuclear neutrophils and macrophages were the most common cells observed on stained smears, which detected inflammation in 147 specimens (59%) by DiffQuick but in only 98 (39%) by Gram's method. CONCLUSIONS: Detecting inflammation in expressed prostatic secretions is method dependent. Significantly more cases of inflammation were detected by hemocytometer than by the traditional wet mount technique. Because the wet mount method also proved more variable than the hemocytometer and highly sensitive to volume, its use is not recommended. These findings support the adoption of hemocytometer and staining methods for accurate evaluation of expressed prostatic secretion inflammation in men with chronic prostatitis/chronic pelvic pain syndrome.

Incidence, risk factors, and clinical outcome of stroke after acute myocardial infarction in clinical practice. MIR and MITRA Study Groups. Myocardial Infarction Registry. Maximal Individual Therapy in Acute Myocardial Infarction .

In this analysis of ischemic and hemorrhagic strokes after acute myocardial infarction (AMI) in 21,330 consecutively included patients with AMI, we found an incidence of stroke after AMI of 1.2% and a very poor prognosis. Previous stroke, atrial fibrillation, and older age were the strongest predictors of stroke after AMI; thrombolysis was a borderline risk factor and early therapy with aspirin was associated with a reduction in stroke after AMI.

Does gastroscopy induce myocardial ischemia in patients with coronary heart disease?

BACKGROUND AND STUDY AIMS: Gastroscopy has been reported to be dangerous for unstable patients with coronary heart disease (CHD). The aims of this study were to find out whether endoscopy is equally liable to cause myocardial ischemia in stable CHD patients, and whether this can be predicted prior to endoscopy, and to find out the frequency of abnormal findings in patients for whom a secondary prophylaxis with acetylsalicylic acid (ASA) is indicated. PATIENTS AND METHODS: Electrocardiograph recording using a Holter monitor was performed during gastroscopy in 71 patients with stable CHD, to check for silent ischemia. To predict potential ischemia during gastroscopy, the Holter monitoring ECG was applied prior to a treadmill test, and withdrawn after gastroscopy 16-22 hours later. RESULTS: During gastroscopy, 30 patients (42%) had silent ischemia, but only 1 patient (1%) became symptomatic. Ischemia was dependent on heart-rate (median heart rate with ischemia 124 beats/min, without 104 beats/min). Abnormal findings on gastroscopy were found in 53 patients (75%). They implied a potential bleeding risk in 30 patients (42%) and prevented the indication for ASA in 6 of them (8%). CONCLUSIONS: Gastroscopy is potentially a harmful procedure for CHD patients, but the incidence of ischemic periods may be reduced by conscious sedation and, if the patient is receiving beta-blocking agent therapy, by applying this medication prior to gastroscopy.

Massive intraperitoneal bleeding from tryptic erosions of the splenic vein. Another cause of sudden deterioration during recovery from acute pancreatitis.

Acute bleeding is a rare, but frequently fatal complication of pancreatitis. Bleeding into the gastrointestinal tract may occur owing to gastric or duodenal erosions, peptic ulcers, or varices in the esophagus, stomach, or colon following splenic vein thrombosis, or intraperitoneally from eroded vessels in pancreatic pseudocysts or expanding pseudoaneurysms. We report a novel case of massive intraperitoneal bleeding owing to tryptic erosions of the splenic vein in a patient recovering from acute pancreatitis. Diagnosis of the bleeding was made by ultrasound and ultrasound-guided blood aspiration. The source of the bleeding was identified intraoperatively, and a left-sided pancreatectomy and a splenectomy were performed.

Inhibition of human sperm-zona-free hamster oocyte binding and penetration by protein C inhibitor.

Protein C inhibitor is a heparin-dependent serine protease inhibitor present in plasma at about 0.08 mumol l-1. Protein C inhibitor inhibits activated protein C and other coagulation factors. Previously, we described the presence of high protein C inhibitor levels in human semen (3.1 mumol l-1) and showed potential roles of the inhibitor in human reproduction. Here, we show that protein C inhibitor is present in an active form in follicular fluid at about 0.1 mumol l-1 and that purified, functionally active human plasma-derived and inactive, semen-derived protein C inhibitor and a synthetic peptide derived from its sequence inhibited both binding and penetration of zona-free hamster oocytes by human sperm. The binding inhibition by protein C inhibitor was dose dependent, with 50% inhibition at 0.037 mumol l-1 inhibitor (45 +/- 17 sperm per egg versus 90 +/- 23 in control experiments). The inhibitor also blocked in a dose-dependent manner the penetration of zona-free hamster eggs by human sperm (20 +/- 7% fertilized eggs at 0.1 mumol l-1 protein C inhibitor versus 55 +/- 10% in control experiments). Polyclonal antiprotein C inhibitor or antipeptide antibodies partially abolished the effect of protein C inhibitor and peptide on the inhibition of the binding and penetration of zona-free hamster oocytes by human sperm. The effect of the protein C inhibitor was not dependent on its antiprotease activity since purified semen-derived protein C inhibitor which did not have antiprotease activity gave comparable results. We conclude that protein C inhibitor may be involved in human reproduction at several steps, including the fertilization process.

Deletion of type IIalpha regulatory subunit delocalizes protein kinase A in mouse sperm without affecting motility or fertilization.

Cyclic AMP stimulates sperm motility in a variety of mammalian species, but the molecular details of the intracellular signaling pathway responsible for this effect are unclear. The type IIalpha isoform of protein kinase A (PKA) is induced late in spermatogenesis and is thought to localize PKA to the flagellar apparatus where it binds cAMP and stimulates motility. A targeted disruption of the type IIalpha regulatory subunit (RIIalpha) gene allowed us to examine the role of PKA localization in sperm motility and fertility. In wild type sperm, PKA is found primarily in the detergent-resistant particulate fraction and localizes to the mitochondrial-containing midpiece and the principal piece. In mutant sperm, there is a compensatory increase in RIalpha protein and a dramatic relocalization of PKA such that the majority of the holoenzyme now appears in the soluble fraction and colocalizes with the cytoplasmic droplet. Unexpectedly the RIIalpha mutant mice are fertile and have no significant changes in sperm motility. Our results demonstrate that the highly localized pattern of PKA seen in mature sperm is not essential for motility or fertilization.

Advanced semen analysis: a simple screening test to predict intrauterine insemination success.

OBJECTIVE: To determine if a simple screening test of sperm recovery through a density gradient preparation and sperm survival after a 24-hour incubation is predictive of IUI success. DESIGN: Prospective nonrandomized descriptive study. SETTING: Tertiary assisted reproductive technology center. PATIENT(S): Four hundred fourteen couples undergoing IUI for male factor and unexplained infertility. INTERVENTION(S): An advanced semen analysis, which consisted of a basic semen analysis, sperm processing through a density gradient preparation, and a 24-hour sperm incubation, was performed on all couples before beginning IUIs. MAIN OUTCOME MEASURE(S): Cumulative and per cycle pregnancy rates (PRs) were calculated for routine semen parameters, number of sperm processed through density gradient, and percent motile sperm after a 24-hour incubation. RESULT(S): None of the basic semen analysis parameters accurately predicted IUI success. When the processed total motile sperm available for insemination was > or = 10 x 10(6) and their 24-hour survival was > or = 70%, 89% (162 of 182) of couples achieved a pregnancy with a 21.4% (162 of 757) per cycle PR compared to a 2.8% (11 of 403) per cycle PR and 4.7% total PR when survival was <70%. With use of these cutoff values for the advanced semen analysis, the test had a sensitivity of 94% and specificity of 86%. CONCLUSION(S): The number of motile sperm available for insemination and especially their 24-hour survival are highly predictive of IUI success. This advanced semen analysis is an excellent screening test to evaluate couples considering IUI.

Assessment of interlaboratory and intralaboratory sperm morphology readings with the use of a Hamilton Thorne Research integrated visual optical system semen analyzer.

OBJECTIVE: To evaluate the level of variance produced in a multicenter study with the use of a computer-assisted sperm morphology analyzer. DESIGN: A multicenter, prospective, blinded study. SETTING: Assisted reproduction research laboratories. PATIENT(S): Semen samples produced for assisted reproductive procedures. INTERVENTION(S): Hamilton Thorne Research (Beverly, MA) integrated visual optical system semen analyzers were used at five different centers to evaluate the same set of 30 slides that were prepared and numerically coded at Tygerberg Hospital in Tygerberg, South Africa. MAIN OUTCOME MEASURE(S): The percentage of normal sperm. RESULT(S): Interlaboratory coefficients of variation (CVs) ranged between 16.31% and 23.09%. One of the participating laboratories produced an approximately 14% (-6.5-7.7) limits of agreement analysis, with a CV of 11.36%, for its duplicate readings. The use of a 10% normal sperm morphology cutoff point to determine discordance levels produced rates ranging between 10% and 23.3% for the interlaboratory and intralaboratory readings. This level of discordance equates with < or = 7 of the corresponding readings from two laboratories falling into a different normal sperm morphology group (< or = 10% or >10%). CONCLUSION(S): The magnitudes of variation produced by the readings performed in our study reached the same level as for the manual evaluation of sperm morphology. A < 10% CV can be obtained if the correct quality control measures are implemented.

Contrasting associations of blood and semen lead concentrations with semen quality among lead smelter workers.

While blood lead concentration has been inversely associated with indicators of reproductive health in occupationally exposed male workers, the utility of lead concentration in semen as an indicator of lead exposure to the male reproductive system has not been fully explored. Blood and semen lead concentrations from 81 lead smelter workers were examined in relation to semen quality and endocrine function parameters. Mean blood and semen lead concentrations were 22.8 micrograms/dl (range 5-58) and 1.9 micrograms/dl (range 0.1-17.6), respectively. Total sperm count and concentration decreased with increasing blood lead concentration; p for trend was 0.003 and 0.009, respectively. Semen lead concentration was inversely related to total sperm count (p = 0.05), ejaculate volume (p = 0.001), and serum testosterone (p = 0.004), but not to sperm concentration. The association between semen lead concentration and total sperm count was eliminated when volume was included in the model. Blood lead concentration was more consistently associated with indicators of sperm production than was semen lead. In contrast, semen lead concentration was negatively associated with circulating testosterone concentrations. Our findings indicate that measurement of semen lead may not be a valuable adjunct to conventional blood lead monitoring for investigations of male reproductive system toxicity.

Vasectomy reversal associated with increased reactive oxygen species production by seminal fluid leukocytes and sperm.

PURPOSE: Reactive oxygen species, which are primarily produced by leukocytes, are generally detrimental to sperm. High reactive oxygen species levels are found in men with abnormal sperm function. Since men often have poor sperm characteristics and infertility after vasectomy reversal, we compared reactive oxygen species in seminal cells of men after vasovasostomy to those of fertile men to determine if reactive oxygen species were elevated in the former group. MATERIALS AND METHODS: We studied semen samples of men with proved fertility (39) and those with previously proved fertility who had undergone vasectomy reversal (45). The presence of leukocytes was determined by Bryan-Leishman staining. Reactive oxygen species endogenous activity was monitored by luminol dependent chemiluminescence in washed cells, including all cells in the semen, and Percoll density gradient purified sperm. RESULTS: After vasovasostomy men had significantly lower sperm concentration, motility and computerized motility measurements than fertile men. Mean reactive oxygen species in washed seminal cells after vasovasostomy was 684 relative light units per second compared to 49 for fertile controls (p < 0.0001). Density gradient purified sperm had 53 and 0.64 relative light units per second, respectively (p < 0.0001). When men with leukocytospermia were excluded from analysis, differences between the groups remained, although 9 times more reactive oxygen species were detected in men after vasectomy reversal with than those without leukocytes in semen. CONCLUSIONS: Higher levels of reactive oxygen species are found in washed seminal cells and purified sperm after vasectomy reversal than in those of fertile men. Although leukocytes are probably a significant source of reactive oxygen species in these groups, they may not account for all of the increased reactive oxygen species after vasovasostomy. Low motility after vasectomy reversal may be related to the detrimental effects of reactive oxygen species produced by leukocytes or sperm, even in men without clinical leukocytospermia.

Effects of clinical stage and immunological status on semen analysis results in human immunodeficiency virus type 1-seropositive men.

Complete semen analyses including computer-assisted sperm motility and morphology assessments were performed to determine if semen and sperm differed between HIV-seropositive men and fertile controls, or differed with symptoms, or CD4+ peripheral cell count categories. Previous studies included small numbers of men and presented conflicting conclusions. Two hundred and fifty non-vasectomized HIV-seropositive men and 38 fertile controls each provided one semen sample. Non-parameteric statistics were used to analyse both continuous and nominal data. Fertile men had significantly greater semen volume, sperm concentration, percent motility, percent rapid and linear motility and total strictly normal spermatozoa than HIV seropositive men. Neither total number nor subtypes of leukocytes in semen differed between the two groups. Among the HIV seropositive men, significant differences in semen analyses were found between CD4+ cell count, clinical, and AIDS categories. Lower CD4+ cell counts (< 200 mm-3) were associated with significantly lower percent motility, percent normal sperm morphology by strict criteria, significantly more spermatids in semen, and higher percentages of teratozoospermia, oligoasthenoteratozoospermia and leukocytospermia. Healthier men, based on clinical categories, had significantly more normal shaped spermatozoa and fewer had azoospermia, oligoasthenoteratozoospermia or leukocytospermia. Many HIV-seropositive men have normal semen analyses, but as the disease progresses more defects are found, particularly in strict criteria sperm morphology.

Liver biopsy is a useful predictor of response to interferon therapy in chronic hepatitis C.

AIM: To evaluate the usefulness of easily assessable morphological parameters in liver biopsies in order to predict efficacy of interferon-alpha (IFN) treatment in patients with chronic hepatitis C. METHODS AND RESULTS: Inflammatory activity and fibrosis (according to Scheuer), and the hepatic iron content (according to Rowe and DiBisceglie) were assessed in pre-treatment liver biopsies of 73 de novo patients with chronic hepatitis C. Furthermore the presence of fat, lymphoid aggregates, and bile duct lesions was evaluated. With respect to IFN therapy patients were classified as responders alanine aminotransferase (ALT) normal and negative hepatitis C virus (HCV) RNA in serum at the end of treatment, n = 33) or non-responders (n = 40). Non-responders had more advanced fibrosis (P = 0.0001) and more extensive iron storage (P = 0.0008) than responders. In contrast absence of stainable iron was frequently (46%) associated with sustained response. Absence of fat droplets in hepatocytes was associated with response (P = 0.0001). Stepwise logistic regression analysis indicated that the stage of fibrosis, the hepatic iron grade, and the presence or absence of fat were independent predictors of response. CONCLUSIONS: Liver biopsy provides useful information for selection of patients with hepatitis C for IFN therapy.

Bladder petechiae after cystoscopy and hydrodistension in men diagnosed with prostate pain.

PURPOSE: We determined if men with prostate pain syndromes have petechiae in the bladder after hydrodistension. MATERIALS AND METHODS: A total of 60 men with the diagnosis of prostate pain and without bacteriuria underwent cystoscopy and hydrodistension under a general or regional anesthetic. RESULTS: Of the 60 men 35 (58%) had moderate to severe petechiae similar in appearance to women with interstitial cystitis after hydrodistension. Men with moderate to severe bladder petechiae had fewer leukocytes in expressed prostatic secretions, smaller bladder capacities and less often testicular pain than men with more normal appearing bladders after hydrodistension. Symptomatic improvement 2 to 6 weeks after hydrodistension was more common in men with moderate to severe petechiae than in those with fewer petechiae. Absence of rectal pain predicted symptomatic improvement after hydrodistension. CONCLUSIONS: We suggest that bladder petechiae, and possibly interstitial cystitis or a related condition, may be more frequently associated with prostate pain syndromes in men than previously appreciated.