Age-related changes in seminal polymorphonuclear elastase in men with asymptomatic inflammation of the genital tract.

AIM: To investigate age-related inflammatory events in the male genital tract. METHODS: In a total of 4265 randomly collected patients attending the andrological outpatient clinic of the Center for Dermatology and Andrology, University of Giessen, Germany, ejaculate volume, pH-value, sperm concentration, total and progressive sperm motility, concentration of polymorphonuclear (PMN) elastase, number of peroxidase-positive cells and fructose were measured and correlated with patient's age. RESULTS: While ejaculate volume, motility and fructose all correlated negatively with age, sperm concentration, PMN elastase and the pH-value showed a positive correlation. The prevalence of male genital tract inflammation (as defined by PMN elastase > 250 ng/mL) and its severity increased significantly. PMN elastase did not correlate with sperm motility. Fructose as a marker of seminal vesicle function showed a significant negative relationship with the PMN elastase levels, the number of peroxidase-positive cells and sperm motility. CONCLUSION: The significant increases of PMN-elastase levels as marker of male genital tract inflammation in older men appear to be indicative of age-related changes in local immunoregulatory mechanisms. Because there is no association of PMN elastase with sperm motility, a direct inhibitory effect of this enzyme can be excluded.

Chronic pelvic pain syndrome/chronic prostatitis affect the acrosome reaction in human spermatozoa.

Male genital tract inflammations constitute an important factor of male subfertility in which different mediators are discussed damaging sperm membranes. As acrosome reaction (AR) is ultimately a membrane function, this study aimed at investigating the effects of the chronic pelvic pain syndrome on AR. In a total of 56 patients with chronic pelvic pain syndrome NIH IIIA and NIH IIIB, AR, concentration of leucocytes in ejaculate and urine after prostate massage, sperm concentration, motility, vitality and morphology were determined. The control group (n = 95) consisted of patients with normozoospermia and proven absence of signs of inflammation. Compared to the control, both patients groups showed poorer sperm morphology (P < 0.0001) and sperm concentration (P < 0.002). In addition, while maximum AR was not affected, inducibility of AR was significantly lower (P < 0.004) and spontaneous AR increased (P < 0.02). Our data clearly demonstrate a significant influence of the inflammation on acrosomal functionality, which could be mediated by inflammatory mediators like ROS.

Seasonal changes of neutral alpha-glucosidase activity in human semen.

Although there are contradictory reports, the biochemical evaluation of the seminal activity of neutral alpha-glucosidase (NAG) has repeatedly been described as an important parameter to test epididymal patency and is recommended by the WHO. Because, for a number of diagnostic parameters, seasonal variations have been described even in the human, it was the aim of this study to investigate possible circannual changes of NAG. This is an important aspect of andrological diagnosis, as seasonal changes of specific diagnostic parameters might have an impact on the accuracy and predictive power of these parameters, which in turn might have an effect on the therapeutic concept for the patients. In a total of 473 patients, sperm concentration, volume of the ejaculate, total motility, progressive motility, pH value, number of peroxidase-positive cells, concentration of fructose, and NAG as functional markers of the seminal vesicles and the epididymis, respectively, were analyzed according to standard procedures. Seminal activity of NAG was significantly correlated with the sperm concentration (P < .0001), ejaculate volume (P < .0001), and the pH (P = .0025). Moreover, significant (P = .0008) seasonal changes in the activity of seminal NAG with the maximum in spring (76.87 mU/ejaculate) and the minimum in autumn (58.55 mU/ejaculate) were found. The incidence of low-ranged activity of the enzyme was 9.2% in spring, while it was 20.3% in autumn. Thus, our data clearly demonstrate circannual changes of the seminal activity of neutral alpha-glucosidase. This in turn has clinical impact as the predictive power of the test system changes throughout the year.

Effects of single non-ortho, mono-ortho, and di-ortho chlorinated biphenyls on human sperm functions in vitro.

Polychlorinated biphenyls (PCBs) are ubiquitous pollutants in the environment. They are present in body fluids associated with reproduction such as follicular fluid, seminal fluid and cervical mucus. Most PCB effects are mediated through the aryl hydrocarbon receptor, which is present in human spermatozoa. Additionally, PCBs may alter various biochemical reactions, such as calcium homeostasis. Therefore we investigated the effects of single non-ortho PCB 126, mono-ortho PCB 118, and di-ortho PCB 153 on human sperm motility, vitality, and calcium-dependent acrosome reaction (AR) in vitro. Human spermatozoa were either treated with different single PCB congeners or their combinations for 5 h at 37 degrees C (spontaneous AR), or for 16 h at room temperature and 4 degrees C (induced AR). Motility was measured after 5 h of incubation. Compared with the controls, PCB exposure had no effects on the percentage of living acrosome reacted spermatozoa, vitality, and motility. There was no difference in the inducibility of the AR between treatment groups and the respective controls after long term incubation. The PCB concentrations used were far higher than those found in cervical mucus or seminal fluid. In vivo effects of PCB congeners on human ejaculated spermatozoa seem to be unlikely. However these results cannot be easily transferred to the in vivo situation, because individual susceptibility has to be considered, and there is no information about synergistic or additive effects with other chemicals present in the male and female reproductive tract.

Molecular aspects of declining sperm motility in older men.

OBJECTIVE: To investigate the molecular mechanism for declining sperm motility in older men. DESIGN: Retrospective and prospective study. SETTING: University-based andrology unit. PATIENT(S): Semen was collected from 2,111 patients and sperm donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Age, sperm concentration, ejaculate volume, sperm motility (including velocity average path, velocity straight line, velocity curvilinear, linearity, and lateral head displacement), normal sperm morphology, percentage of abnormally blue-stained flagella, and total and free T, FSH, and LH concentrations were analyzed. In addition, the zinc concentration in the seminal fluid, flagella, and the whole spermatozoa was measured by means of atomic absorption spectrometry. RESULT(S): Patient age correlated negatively with T concentration and sperm motility, including velocity, but positively with the percentage of abnormally stained flagella and the flagellar zinc. Whereas the percentage of abnormally stained flagella correlated negatively with motility and sperm velocity, there was a positive relationship with the flagellar zinc content. Flagellar zinc content was negatively correlated with motility. Testosterone showed a positive relationship with motility and sperm concentration. CONCLUSION(S): Because the epididymis is functionally T-dependent, our data suggest that the removal of zinc from the outer dense fibers during epididymal sperm maturation is affected in aging men, which in turn will result in decreased sperm motility.

Effects of Basella alba and Hibiscus macranthus extracts on testosterone production of adult rat and bull Leydig cells.

AIM: To determine the androgenic effects of Basella alba and Hibiscus macranthus extracts in the rat and the bull, and to develop a novel in vitro test system using Leydig cells from bull testes. METHODS: The effect of methanol extracts from both plants on testosterone production in isolated Leydig cells from the rat and the bull was analyzed using 125I-radioimmunoassay (125I-RIA). Rat Leydig cells were obtained by common methods, whereas a novel technique was used to purify Leydig cells from bull testes. RESULTS: Bull testes from the slaughter house were a cheap source of pure Leydig cells. In culture, these cells produced testosterone for 5-6 days, which can be stimulated by human chorionic gonadotrophin (hCG). Basella alba extracts significantly enhanced testosterone production in bull and rat Leydig cells in a concentration-dependent manner. Hibiscus macranthus showed no androgenic effect but was shown to inhibit testosterone production at higher concentrations. CONCLUSION: Leydig cells purified from bull testes can be used as an alternative tool in experimental animal research. Certain fractions of Basella alba extract demonstrated androgenic potential whereas Hibiscus macranthus extracts did not.

Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients.

OBJECTIVE: To investigate whether there is an impact of different sources of reactive oxygen species (ROS) on sperm functions. DESIGN: Prospective study. SETTING: Patients at the Center for Dermatology and Andrology, Giessen, Germany. PATIENT(S): Semen collected from 63 randomly collected patients attending the IVF unit of the University of Giessen, Germany. INTERVENTION(S): Only patients with nonleukocytospermia were included in this study. MAIN OUTCOME MEASURE(S): Sperm count and motility before and after sperm separation by swim-up, morphology, DNA fragmentation, and extrinsic (by leukocytes) and intrinsic ROS production (by spermatozoa) were evaluated. RESULT(S): Leukocytes correlated significantly with extrinsic ROS production (r = 0.576), but markedly less with intrinsic ROS production (r = 0.296). Sperm count, morphology, and motility in the ejaculate were markedly more affected by extrinsic than by intrinsic ROS. The DNA fragmentation was strongly positively correlated with intrinsic ROS production, whereas this correlation was weaker for extrinsic ROS production. No correlation was found between DNA fragmentation and the number of leukocytes, whereas the correlations with motility in the ejaculate and the motile sperm count after swim-up were highly significant. Moreover, significant differences were observed for extrinsic and intrinsic ROS production between groups of patients having a high (> or = 1 x 10(6)/mL) and a low number (<1 x 10(6)/mL) of leukocytes in the ejaculate. CONCLUSION(S): The origin of ROS seems to have an influence on the site of the damage. Because leukocyte counts <1 x 10(6)/mL caused a significant decrease of motility and DNA integrity, the threshold given by the World Health Organization (WHO) should be re-evaluated.

[Stress and psoriasis -- a psychoneuroimmunological study]. / Stress und Psoriasis -- eine psychoneuroimmunologische Studie.

UNLABELLED: Psoriasis is a polygenetic hereditary multifactorial disease which may be influenced by a number of environmental factors. To date only a few studies experimentally investigated the influence of stress on psoriasis. One problem of these studies is that it remains unclear whether the experimental findings are relevant for the entire group of patients, or whether there are subgroups who are particularly susceptible to stress. Therefore our main objective is to examine whether experimental stressors can identify subgroups of patients who are particularly susceptible to stress and if these differ in immunological parameters. METHOD: The Trier Social Stress Test (TSST) was used as stressor. The severity was recorded both objectively using the PASI (Psoriasis Area and Severity Index) as well as subjectively by the patient. Somatic parameters for which stress reactivity is known but no direct relationship to psoriasis is assumed were selected and exploratively examined within the present study (salivary cortisol). The second set of parameters for which the stress reactivity was unclear included variables which are linked to the pathophysiology and/or the severity of psoriasis. In addition to salivary cortisol, eosinophils, ICAM-3, and sIL-2R were determined in serum. 38 psoriasis patients and 38 control subjects were examined (21 male and 17 female participants within each group). RESULTS: The PASI correlated very inconsistently with the subjective severity parameters. The relationships between severity and blood parameters tested showed a systematic relationship for eosinophils only. The TSST is suitable for eliciting stress in psoriasis patients. In one subgroup, there was an increase in skin affliction, while skin affliction in the second group remained constant or decreased. A classification into stress-reactive or non-reactive patients cannot, however, be supported by the immunological parameters tested.

Sperm function and assisted reproduction technology.

The evaluation of different functional sperm parameters has become a tool in andrological diagnosis. These assays determine the sperm's capability to fertilize an oocyte. It also appears that sperm functions and semen parameters are interrelated and interdependent. Therefore, the question arose whether a given laboratory test or a battery of tests can predict the outcome in in vitro fertilization (IVF). One-hundred and sixty-one patients who underwent an IVF treatment were selected from a database of 4178 patients who had been examined for male infertility 3 months before or after IVF. Sperm concentration, motility, acrosin activity, acrosome reaction, sperm morphology, maternal age, number of transferred embryos, embryo score, fertilization rate and pregnancy rate were determined. In addition, logistic regression models to describe fertilization rate and pregnancy were developed. All the parameters in the models were dichotomized and intra- and interindividual variability of the parameters were assessed. Although the sperm parameters showed good correlations with IVF when correlated separately, the only essential parameter in the multivariate model was morphology. The enormous intra- and interindividual variability of the values was striking. In conclusion, our data indicate that the andrological status at the end of the respective treatment does not necessarily represent the status at the time of IVF. Despite a relatively low correlation coefficient in the logistic regression model, it appears that among the parameters tested, the most reliable parameter to predict fertilization is normal sperm morphology. (Reprod Med Biol 2005; 4: 7-30).

Diseases on hair follicles leading to hair loss part I: nonscarring alopecias.

Hair follicles are unique structures with exceptional regenerative potential. They are believed to be crucial for epidermal homeostasis and reepithelialization after damage to human skin. Like other, more active and quickly proliferating organ systems, hair follicles may be easily disturbed in their normal growth cycle by systemic and local influences, including specific skin diseases. This may lead to hair loss, a very common complaint in men and women. The difficulties in reviewing the diseases of hair follicles lay in the long list of different etiologic factors (infectious, autoimmune, inflammatory, neoplastic, physical, chemical, congenital) and a still missing classification system according to etiopathogenetic principles. In this article (Part I of II), the structure and function of hair follicles, the diagnostic approach to diseases causing hair loss, and the most common nonscarring alopecias (telogen effluvium, anagen effluvium, androgenetic alopecia, and alopecia areata) are reviewed in regard to pathogenesis, clinical findings, and current options of treatment. Part II will focus on scarring alopecias.

Male sexuality with advancing age.

People aged 60 years or older are commonly given the image of grandparents who are sexually rather inactive. However, a high percentage of men over 60 years have sexual desires and are sexually active. Aging men need more time and more direct stimulation of the penis to achieve full erection. The risk of developing erectile dysfunction increases with age. Almost one man in 22 aged 60-69 years without previous erection problems develops erectile dysfunction within 1 year. Hormone replacement therapy is only indicated if testosterone deficiency is combined with the syndrome of the "aging male" and lack of other disorders that are possibly causative and could be treated. For hormone replacement therapy testosterone gel preparations are preferable because they have a short half-time and can be immediately discontinued in the event that a pre-existing occult prostate carcinoma is stimulated.

Influence of deoxyribonucleic acid damage on fertilization and pregnancy.

OBJECTIVE: To investigate sperm DNA damage in relation to fertilization and pregnancy. DESIGN: Prospective study. SETTING: The Institute of Reproductive Medicine, Giessen, Germany. PATIENT(S): Semen collected from 249 patients attending the IVF program. MAIN OUTCOME MEASURE(S): The percentage of terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling- (TUNEL-), Fas-, and annexin-V-positive sperm and the proportion of green-fluorescing sperm in the acridine orange stain was determined and correlated with sperm concentration, motility, fertilization, and pregnancy. RESULT(S): Significant correlations with the concentration of motile sperm were only found for the acridine orange stain (before and after sperm separation) and for the TUNEL assay (after sperm separation). Moreover, patients whose sperm had a high percentage of DNA fragmentations showed significantly lower pregnancy rates (TUNEL assay: 19.05% vs. 34.65%; acridine orange stain: 24.58% vs. 37.93%). The apoptosis parameters (annexin V binding and Fas expression) showed no statistically significant differences. CONCLUSIONS: Our data clearly demonstrate that DNA fragmentation, as determined by the TUNEL assay, is predictive for pregnancy in IVF. This implies that spermatozoa with DNA fragmentation can still fertilize an oocyte but that when paternal genes are "switched on," further embryonic development stops, resulting in failed pregnancy. It seems that, at least in the patients we analyzed, apoptosis in the sperm does not play a role for fertilization. This would imply that DNA fragmentation in human spermatozoa is caused by external factors, such as reactive oxygen species, rather than by apoptosis.

Interdisciplinary diagnostics in environmental medicine--findings and follow-up in patients with chronic medically unexplained health complaints.

PROBLEM: In patients attributing their chronic, medically unexplained complaints to environmental factors the greatest challenge is to overcome their disabling belief in toxicogenic explanations. METHOD: Patients presenting with health complaints that they attributed to environmental causes in an environmental outpatient department (EOPD) within a university medical center in Germany were studied. An interdisciplinary review of previously diagnosed medical conditions, current clinical consultations, personal risk communication and therapeutic advice is presented. Additionally, patient contentedness, complaint development, and belief in environmental attribution in a follow-up interview are given. RESULTS: The open, prospective study comprises 51 patients reporting more than one complaint. Symptoms had lasted for more than 3 years in 63% of the cases. Seventy percent attributed their complaints to more than one environmental cause. The clinical diagnostic procedure reduced the number of prediagnosed clinical conditions by 50%. Numerous foregoing environmental laboratory analyses had overestimated toxicologically relevant findings. These were not confirmed in 80% (8/10) of the cases. In 8% (n = 4) of the patients a relevant environmental or occupational medical condition was found. A mental or behavioral condition was not considered to have first priority in explaining all complaints in 43% (22/51) of the patients. Among these, mostly respiratory or skin-related diseases were found. All patients contacted participated in a follow-up study after a minimum of 21 months. Sixty-seven percent reported having felt that they were taken seriously, 38% felt better after the beginning of the study, and 45% were no longer certain about the importance of the environmental attribution. Since 83% of the patients with a preceding residential diagnosis of MCS or SBS still believed in environmental causes of their complaints in the follow-up study, we conclude that these prediagnoses appear to be a risk for persisting attribution of the environmental factor. About one third (37%) of these patients with complaints that had not been medically explained by an organic condition during interdisciplinary diagnostics had meanwhile consulted a psychotherapist. CONCLUSIONS: Interdisciplinary diagnostics and scientifically based risk assessment in a specialized clinical center were effective and mostly well accepted by the patients and resulted in reduced attribution of complaints to environmental conditions. No indication was found that patients with complaints not medically explained by organic conditions were managed less successfully by this approach. Considering the high costs that these patients have previously caused, it appears valuable to apply an interdisciplinary diagnostic strategy.

DNA fragmentation of spermatozoa and assisted reproduction technology.

Despite the ever-increasing knowledge of the fertilization process, there is still a need for better understanding of the causes of sperm DNA fragmentation and its impact on fertilization and pregnancy. For this reason, human sperm DNA fragmentation was investigated by means of the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) assay and the production of reactive oxygen species (ROS) in the ejaculate and in the spermatozoa themselves. These data were correlated with fertilization and pregnancy data from IVF and intracytoplasmic sperm injection (ICSI) patients. Sperm DNA fragmentation did not correlate with fertilization rate, but there was a significantly reduced pregnancy rate in IVF patients inseminated with TUNEL-positive spermatozoa. ICSI patients exhibited the same tendency. This implies that spermatozoa with damaged DNA are able to fertilize an oocyte, but at the time the paternal genome is switched on, further development stops. The determination of ROS in the ejaculate and the percentage of ROS-producing spermatozoa revealed markedly stronger correlations between sperm functions (i.e. motility) and the percentage of ROS-producing spermatozoa. The influence of seminal leukocytes, known to produce large amounts of oxidants, on sperm DNA fragmentation should not be neglected.