A Prostaglandin D2 system in the human testis

As shown recently, cyclooxygenase 2 (COX2), the inducible key enzyme for the prostaglandin (PG) biosynthetic pathway, is abundantly present in interstitial cells of testes of men suffering from different forms of impaired spermatogenesis and sub- or infertility, but it is absent in human testes with normal spermatogenesis. Although the spectrum of the downstream products of COX2 action in testis, namely PGs, and their effects are not known, our results show that Prostaglandin D2 (PGD2) likely plays a role. We describe (a) PGD2 synthetases, as well as receptors for PGD2 (DP) in testicular interstitial cells of men suffering from spermatogenic damage and infertility, and report that (b) PGD2 is produced by and can affect Leydig cells of an animal model, which expresses testicular COX2 and DP.Fil: Schell, Cristoph. Ludwig Maximilians Universitat; AlemaniaFil: Frungieri, Monica Beatriz. Ludwig Maximilians Universitat; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Albrecht, Martín. Ludwig Maximilians Universitat; AlemaniaFil: Gonzalez de Calvar, Silvia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Köhn, Frank M.. Andrologicum Munich; AlemaniaFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Mayerhofer, Artur. Ludwig Maximilians Universitat; Alemani

Medical Yoga for Patients with Stress-Related Symptoms and Diagnoses in Primary Health Care: A Randomized Controlled Trial

An increasing number of patients are suffering from stress-related symptoms and diagnoses. The purpose of this study was to evaluate the medical yoga treatment in patients with stress-related symptoms and diagnoses in primary health care. A randomized controlled study was performed at a primary health care centre in Sweden from March to June, 2011. Patients were randomly allocated to a control group receiving standard care or a yoga group treated with medical yoga for 1 hour, once a week, over a 12-week period in addition to the standard care. A total of 37 men and women, mean age of 53±12 years were included. General stress level (measured using Perceived Stress Scale (PSS)), burnout (Shirom-Melamed Burnout Questionnaire (SMBQ)), anxiety and depression (Hospital Anxiety and Depression Scale (HADS)), insomnia severity (Insomnia Severity Index (ISI)), pain (visual analogue scale (VAS)), and overall health status (Euro Quality of Life VAS (EQ-VAS)) were measured before and after 12 weeks. Patients assigned to the Yoga group showed significantly greater improvements on measures of general stress level (P<0.000), anxiety (P<0.019), and overall health status (P<0.018) compared to controls. Treatment with medical yoga is effective in reducing levels of stress and anxiety in patients with stress-related symptoms in primary health care

Testicular mast cells and male infertility.

Die Fibrose der Wand der Samenkanälchen ist eine seit langem bekannte typische Veränderung bei Männern mit Spermatogenesedefekten, deren Ursache und deren direkte Konsequenz aber nicht bekannt sind. Da im Hoden von Männern mit Spermatogenesestörungen deutlich vermehrt Mastzellen vorkommen, von denen viele aktiviert und degranuliert sind, ist zu erwarten, daß Produkte der Mastzellen freigesetzt werden. Wir stellen die Hypothese auf, daß sie an der Entstehung der Tubulusfibrose und der Spermatogenesestörung beteiligt sind. Untersuchungen in menschlichen Zellkulturen zeigten, daß Tryptase, das Hauptprodukt der Mastzellen, die Bildung von COX-2 und von Prostaglandinen stimuliert. Bestimmte Prostaglandine (J2/15dJ2) stimulieren dann über ihren Rezeptor (PPARgamma) das Fibroblastenwachstum, eine Voraussetzung für die Fibrose. Diese Ergebnisse sind für krankhafte Bindegewebsumbauvorgänge und speziell für die Tubulusfibrose bei Spermatogenesedefekten von Bedeutung: Bei Spermatogenesedefekten fanden wir, daß Mastzellvermehrung und Aktivierung mit der Expression von COX-2 in interstitiellen Zellen einhergehen. Bei Hoden mit normaler Spermatogenese (und nur wenigen Mastzellen) war COX-2 dagegen nicht nachweisbar. Rezeptoren für Tryptase sind auf interstitiellen Zellen und relevante Prostaglandinrezeptoren auf peritubulären Zellen nachweisbar. Es ist demnach möglich, daß Mastzelltryptase COX-2- und Prostaglandinbildung im infertilen Hoden anregt und dies letztlich zur Fibrose in der Tubuluswand führt, eine Veränderung, die sicherlich zur Spermatogenesestörung beiträgt. Die Aufklärung des Signalweges der Tryptase erlaubt es erstmals auch, gezielt über eine Beeinflussung des Fibroseprozesses nachzudenken: Mastzellstabilisatoren, Antagonisten von Tryptase, von COX-2 oder PPARgamma bieten sich an.Fibrotic thickening of the wall of the seminiferous tubules is a hallmark of male infertility, but neither causes nor the direct consequences of tubular fibrosis are known. Since male infertility is also associated with increased numbers of activated mast cells, we propose that secreted mast cell products may be involved in the pathogenesis of tubular fibrosis and possibly male infertility. Evidence for this hypothesis is derived from cell culture and ex-vivo experiments: We found that actions of the major mast cell product, tryptase, includes formation of COX2 and synthesis of prostaglandins in target cells. Prostaglandin J2/15dJ2, which act via their receptor (PPARgamma), are responsible for fibroblast proliferation, a prerequisite of fibrosis. This may be of relevance for fibrosis in general and for testicular tubular fibrosis in particular: We found COX2 in testes of infertile men, but not of men with normal spermatogenesis, implying formation and action of prostaglandins. Since peritubular cells bear PPARgamma and since activated mast cells are at least tripled in infertile men, we propose a chain of events initiated by mast cell tryptase eventually leading to tubular fibrosis. Since tubular fibrosis is likely to be involved in the process leading to the damage of spermatogenesis, these results may pinpoint new targets for possible therapeutic interventions: mast cells, tryptase, COX2 and prostaglandins, as well as PPARgamma.Fil: Mayerhofer, Artur. Universitat Technical Zu Munich; AlemaniaFil: Meineke, V.. Sanitätsakademie der Bundeswehr; AlemaniaFil: Weidinger, S.. Universitat Technical Zu Munich; AlemaniaFil: Köhn, F. M.. Universitat Technical Zu Munich; AlemaniaFil: Frungieri, Monica Beatriz. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Mast cell-sperm interaction: evidence for tryptase and proteinase-activated receptors in the regulation of sperm motility

The detection of significant levels of tryptase in human seminal plasma and follicular fluid and of tryptase-positive mast cells (MCs) in the wall of human Fallopian tubes lead us to hypothesize that tryptase may exert regulatory actions on human spermatozoa. Immunoelectronmicroscopy revealed proteinase-activated receptor 2 (PAR-2) in the membranes of the acrosomal region and midpiece of human spermatozoa. These PAR-2 were functional, as exposure of spermatozoa from healthy men (n = 12) with regular standard semen parameters to human recombinant tryptase significantly decreased motility in a dose- and time-dependent fashion. Motile spermatozoa (WHO a + b) were significantly decreased within 10 min of incubation with 1.000 ng/ml tryptase (P = 0.045). After 30 and 60 min, significant reduction of motility was also observed in the presence of lower tryptase concentrations (100 ng/ml, P = 0.037; 10 ng/ml, P = 0.046). The inhibitory effects of tryptase progressed throughout an observation period of 180 min. Furthermore, tryptase effects were reversible after washing procedures and could be inhibited by pretreatment with anti-tryptase antibody or anti-PAR-2 antiserum.Fil: Weidinger, S.. Technische Universität München; AlemaniaFil: Mayerhofer, Artur. Universität München; AlemaniaFil: Frungieri, Monica Beatriz. Universität München; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Meineke, V.. Military Academy Munich; AlemaniaFil: Ring, J.. Technische Universität München; AlemaniaFil: Köhn, F. M.. Technische Universität München; Alemani

Divergent effects of the major mast cell products histamine, tryptase and TNF-alpha on human fibroblast behaviour

Fibroblast proliferation is a key process in tissue remodeling and mast cells (MCs) are thought to play a crucial role. Having established that the three major MC products, tryptase, histamine and TNF-alpha (TNF) are normally present in human skin MCs, which are in close proximity to dermal fibroblasts, we studied their individual effects on cell cycle-controlled human dermal fibroblasts (HFFF2). These cells express receptors (H1, PAR2, TNFR1/2) for the major MC mediators, but only tryptase or a PAR2 agonist peptide stimulated proliferation and gene expression. TNF was antimitotic, and histamine, while elevating intracellular Ca2+ levels at high concentrations, did not affect proliferation. We conclude that MC products but also composition and numbers of respective receptors on fibroblasts are crucially responsible for fibroproliferative events. © Birkhäuser Verlag, 2005.Fil: Albrecht, Martín. Ludwig Maximilians Universitat; AlemaniaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Ludwig Maximilians Universitat; AlemaniaFil: Kunz, L.. Ludwig Maximilians Universitat; AlemaniaFil: Rämsch, R.. Ludwig Maximilians Universitat; AlemaniaFil: Meineke, V.. Sanitätsakademie der Bundeswehr; AlemaniaFil: Köhn, F.M.. Technical University; AlemaniaFil: Mayerhofer, Artur. Ludwig Maximilians Universitat; Alemani

Evidence for a histaminergic system in the human testis

The complete lack of information about mast cells as a source of histamine and potential target cells for histamine in human testes prompted us to investigate these issues in testes of fertile and infertile patients using a combination of laser microdissection, reverse transcription-polymerase chain reaction (RT-PCR), and immunohistochemistry. We show for the first time the expression of the rate-limiting enzyme in histamine synthesis - histidine decarboxylase - by human testicular mast cells and the expression of the histamine (H) receptors 1 (H1) and 2 (H2) by germinal, interstitial, and peritubular cells in the testes of fertile and infertile patients. ©2005 by American Society for Reproductive Medicine.Fil: Albrecht, Martín. Ludwig Maximilians Universitat; AlemaniaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Ludwig Maximilians Universitat; AlemaniaFil: Gonzalez Calvar, Silvia I.. Fundación de Instituto de Biología y Medicina Experimental; ArgentinaFil: Meineke, Viktor. Sanitätsakademie der Bundeswehr; AlemaniaFil: Köhn, Frank M.. Universitat Technical Zu Munich; AlemaniaFil: Mayerhofer, Artur. Ludwig Maximilians Universitat; Alemani

Prostaglandin E 2 (PGE 2 ) is a testicular peritubular cell-derived factor involved in human testicular homeostasis

In man, blockage of prostaglandin (PG)-production e.g. by non-steroidal anti-inflammatory drug (NSAIDs) may have negative testicular side effects, implying beneficial actions of PGs in the testis. We examined human testicular samples and isolated human testicular peritubular cells (HTPCs) to explore sites of PG-synthesis and targets. HTPCs express cyclooxygenase 1 (COX1) and secrete PGE 2 . Receptors (EP1, 2, 4) were specifically identified in peritubular cells. In HTPCs PGE 2 significantly increased mRNA levels of the contractility protein calponin, but did not induce contractions. PGE 2 , as well as EP1 and EP4 receptor agonists, significantly increased glia cell line derived neurotrophic factor (GDNF) mRNA and/or protein levels. Importantly, the NSAID ibuprofen reduced PGE 2 and this action also lowered SMA and calponin mRNA levels and levels of secreted GDNF protein. The results reveal an unknown PGE 2 system in the human testis, in involving peritubular cells, which may be prone to interference by NSAIDs.Fil: Rey Ares, Veronica. Ludwig Maximilians Universitat; AlemaniaFil: Rossi, Soledad Paola. Ludwig Maximilians Universitat; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Dietrich, Kim Gwendolyn. Ludwig Maximilians Universitat; AlemaniaFil: Köhn, Frank Michael. Ludwig Maximilians Universitat; AlemaniaFil: Ullrich Schwarzer, J. Andrologicum; Múnich, Alemania; AlemaniaFil: Welter, Hared. Ludwig Maximilians Universitat; AlemaniaFil: Frungieri, Monica Beatriz. Ludwig Maximilians Universitat; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Mayerhofer, Artur. Ludwig Maximilians Universitat; Alemani

The Ca2+-activated, large conductance K+ channel (BKCa) is a player in the LH/hCG signaling cascade in testicular Leydig cells.

In Leydig cells, hormonal stimulation by LH/hCG entails increased intracellular Ca(2+) levels and steroid production, as well as hyperpolarization of the cell membrane. The large-conductance Ca(2+)-activated K(+)-channel (BK(Ca)) is activated by raised intracellular Ca(2+) and voltage and typically hyperpolarizes the cell membrane. Whether BK(Ca) is functionally involved in steroid production of Leydig cells is not known. In order to explore this point we first investigated the localization of BK(Ca) in human and hamster testes and then used a highly specific toxin, the BK(Ca) blocker iberiotoxin (IbTx), to experimentally dissect a role of BK(Ca). Immunohistochemistry and RT-PCR revealed that adult Leydig cells of both species are endowed with these channels. Ontogeny studies in hamsters indicated that BK(Ca) becomes strongly detectable in Leydig cells only after they acquire the ability to produce androgens. Using purified Leydig cells from adult hamsters, membrane potential changes in response to hCG were monitored. HCG hyperpolarized the cell membrane, which was prevented by the selective BK(Ca) blocker IbTx. Steroidogenic acute regulatory (StAR) mRNA expression and testosterone production were not affected by IbTx under basal conditions but markedly increased when hCG, in submaximal and maximal concentration or when db-cAMP was added to the incubation media. A blocker of K(V)4-channels, expressed by Leydig cells, namely phrixotoxin-2 (PhTx-2) was not effective. In summary, the data reveal BK(Ca) as a crucial part of the signaling cascade of LH/hCG in Leydig cells. The hyperpolarizing effect of BK(Ca) in the Leydig cell membrane appears to set in motion events limiting the production of testosterone evoked by stimulatory endocrine mechanisms.Fil: Matzkin, Maria Eugenia. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Lauf, S.. University of Munich. Anatomy III. Cell Biology; AlemaniaFil: Spinnler, K.. University of Munich. Anatomy III. Cell Biology; AlemaniaFil: Rossi, Soledad Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Köhn, F.. Andrologicum; AlemaniaFil: Kunz, L.. University of Munich. Department Biology II. Neurobiology; AlemaniaFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Mayerhofer, A.. University of Munich. Cell Biology; Alemani

Alpha 1 adrenergic receptor-mediated inflammatory responses in human testicular peritubular cells

Stress activates the sympathetic nervous system and is linked to impaired fertility in man. We hypothesized that catecholamines by acting on testicular cells have a role in these events, possibly by fostering an inflammatory environment. The cells of the wall of seminiferous tubules, human testicular peritubular cells (HTPCs), express adrenergic receptors (ADRs) α1B, α1D, β1 and β2. A selective α1-ADR agonist, phenylephrine, increased intracellular Ca 2+ -levels in cultured HTPCs and induced COX-2, IL-6 and MCP-1 mRNA expression without affecting IL-1β mRNA. These changes were paralleled by a significant increase in the secretion of IL-6 and MCP-1. Epinephrine was also effective, but salbutamol, a selective β2-ADR agonist was not. Our results suggest that stress-associated elevation of catecholamines may be able to promote inflammatory events by targeting peritubular cells in the human testis. Blockage of α1-ADRs may therefore be a novel way to interfere with stress-related impairment of male reproductive functions.Fil: Rossi, Soledad Paola. Ludwig Maximilians Universitat; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Walenta, Lena. Ludwig Maximilians Universitat; AlemaniaFil: Rey Ares, Veronica. Ludwig Maximilians Universitat; AlemaniaFil: Köhn, Frank-Michael. Andrologicum; AlemaniaFil: Ulrich Schwarzer, J.. Andrology Center Munich; AlemaniaFil: Welter, Harald. Ludwig Maximilians Universitat; AlemaniaFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Mayerhofer, Artur. Ludwig Maximilians Universitat; Alemani