Die Inzidenz des Diabetes mellitus (DM) nimmt weltweit jährlich zu und erlangt somit große Bedeutung für die Entwicklung der globalen Gesundheit. Die WHO rechnet bis zum Jahr 2030 mit ungefähr 366 Millionen erwachsenen Diabetikern. Es ist bekannt, dass Männer mit Diabetes mellitus Typ 1 (DMI) und 2 (DMII) häufiger an Subfertilität leiden, wobei dies möglicherweise auf erhöhte Apoptoseraten und vermehrte DNA-Fragmentierungen auf zellulärer Ebene zurückzuführen ist. Die Bedeutung der ROS als Regulatoren von physiologischen und pathologischen Signaltransduktionswegen ist bekannt. Demzufolge könnte die Aktivität der Stickstoffmonoxid-Synthetase (NOS) in diesem Zusammenhang eine Bedeutung haben. Das Ziel der vorliegenden Untersuchungen war es, die Auswirkungen von Apoptose und DNA-Fragmentierungen sowie die Bedeutung der NOS im Zusammenhang mit der Fertilitätsfähigkeit von Spermienzellen von DMI und DMII Patienten zu erfassen und damit erste Erklärungsansätze zur Pathophysiologie der diabetesassoziierten Subfertilität zu liefern. Samenproben von Normalspendern und Diabetikern wurden durch Dichtegradientenzentrifugation in Subpopulationen separiert und mittels fluoreszenzbasierten Assays zur Analyse von apoptoseassoziierten Parametern wie dem Zusammenbruch des mitochondrialen Membranpotentials (MMP), Aktivierung von Caspase-3 (CP3), DNA-Fragmentierungen und reaktiven Sauerstoffspezies (ROS) im Flowzytometer (FACS) untersucht. Die Ergebnisse zeigen eine signifikante Erhöhung von Apoptosemarkern (gestörtes MMP, aktivierte CP3), ROS und DNA-Fragmentationsraten in Spermien von DMI und DMII Patienten im Vergleich zu gesunden Normalspendern. Der Effekt ist bei DMII Patienten verstärkt ausgeprägt. Alle gemessenen Parameter korrelieren umgekehrt mit dem Fertilitätspotential der Spermien, gemessen anhand etablierter Spermiogramm-Analysen, womit ein möglicher Erklärungsmechanismus für die Subfertilität bei Diabetikern geliefert werden kann.

No description available.

No / Male-mediated developmental toxicity has been of concern for many years. The public became aware of male-mediated developmental toxicity in the early 1990s when it was reported that men working at Sellafield might be causing leukemia in their children. Human and animal studies have contributed to our current understanding of male-mediated effects. Animal studies in the 1980s and 1990s suggested that genetic damage after radiation and chemical exposure might be transmitted to offspring. With the increasing understanding that there is histone retention and modification, protamine incorporation into the chromatin and DNA methylation in mature sperm and that spermatozoal RNA transcripts can play important roles in the epigenetic state of sperm, heritable studies began to be viewed differently. Recent reports using molecular approaches have demonstrated that DNA damage can be transmitted to babies from smoking fathers, and expanded simple tandem repeats minisatellite mutations were found in the germline of fathers who were exposed to radiation from the Chernobyl nuclear power plant disaster. In epidemiological studies, it is possible to clarify whether damage is transmitted to the sons after exposure of the fathers. Paternally transmitted damage to the offspring is now recognized as a complex issue with genetic as well as epigenetic components.

Thesis (PhD) -- Stellenbosch University, 1995. / ENGLISH ABSTRACT: Graling in the Kruger National Park. south of Phalaborwa Gate, is contaminated with copper, probably from a nearby copper refinery. Cattle in the neighbourhood have died from copper poisoning while the average copper concentration in the livers of impala in the Krgler National Park, was found to be abnormally high. The possibility exists that the ultrastructure of spermatozoa may serve as an indicator of environmental conditions. To examine this statement further, the purpose of this study includes the following: - To describe the ultrastructure of normal impala sperm as well as their abnormalities. - To quantify the sperm abnormalities of each experimental animal and to relate the abnormalities with the copper concentration in the impala liver, in order to determine whether any correlation exists between these two variables. Impala have been terminated monthly, from 1992.06.09 to 1993.05.15. in the vicinity of Phalaborwa Gate and in the Skukuza area for other research projects. Their sperm were made available for this study. The control group consisted of 20 animals which comprised 14 animals from an area, not contaminated with copper, along the Nwaswitshaka spruit near Skukula and 6 animals originally from the same area but which had been kept in captivity at Skukuza on a diet, not contaminated with copper. The experimental group consisted of 44 animals which comprised 32 animals from copper contaminated grazing along the Tshutshi spruit south of Phalaborwa Gate and 12 animals captured along the Nwaswitshaka spruit which were kept on a copper contaminated diet in captivity at Skukuza. Sperm from the epididymis of each animal were collected and fixed in a temporary laboratory. The sperm were processed at the Electron Microscopy Unit of the Faculty of Veterinary Science of the University of Pretoria and studied by means of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The copper concentration in the liver was analyzed and bone marrow collected of each animal in order to determine the condition of the animal. The external morphology of a normal impala sperm was documented by means of SEM and the dimensions of the sperm determined. SEM micrographs of observed sperm abnormalities were taken. Percentages for normal sperm and the percentages of some abnormalities of a control group (Average liver copper concentration - ALCC = 37.9 ppm WB) were calculated. The same percentages were also calculated for an experimental group (ALCC = 104 ppm WB). The control group and the experimental group exhibited 91.6% and 93.6% normal sperm respectively. The difference in percentage sperm abnormalities of the two groups was of the same order. The increased copper in the livers of the experimental group has thus not lead to an increase of the percentage macro-abnormalities observed with SEM. The ultrastructure of normal impala sperm were described and documented with micrographs by means of TEM. The deviations of the normal sperm ultrastructure were also recorded on micrographs. Percentages were calculated for the normal sperm sections, as well as the abnormalities of some sperm sections of a contrl group (ALCC = 33.86 ppm WB). The same percentages were also calculated for an experimental group (ALCC = 122.5 ppm WB). For the control group the average percentage sperm neck sections with vacuoles was : 31.5% and for the experimental group this percentage was : 57.9%. The increased copper concentrations in the livers of the experimental group thus led to an increase of the percentage of sperm necks with vacuoles. There is also reason to believe that the difference of the percentages of sections of the sperm head, in the control and experimental group, should be remarkably greater. A graphic analysis of the TEM-data has also shown that a statistically significant correlation exists between the percentage of sperm with vacuoles in the neck and the copper concentrations in the livers of impalas. The conclusion is that high level copper concentrations exert a detrimental effect on the ultrastructure of the sperm neck. The possibility also exists that copper concentrations in impala liver > 500 ppm WB. will show a correlation with the percentages of some of the other abnormalities observed. / AFRIKAANSE OPSOMMING: Die weiding suid van Phalaborwahek in die Krugerwildtuin word waarskynlik vanaf 'n nabygeleë kopersmeltery met koper besoedel. Van die plaasdiere in die omgewing het reeds van kopervergiftiging gevrek en die gemiddeIde koperkonsentrasie in die lewers van rooibokke is abnormaal hoog. Die moontlikheid bestaan dat die ultrastruktuur van spermatozoa as 'n indikator van omgewingstoestande kan dien. Om bogenoemde moontlikheid verder te ondersoek was die doelwit van hierdie studie die volgende: Om die ultrastruktuur van normaIe rooiboksperme en hulle abnormaliteite te beskryf en die spermabnormaliteite van elke proefdier te kwantifiseer en met die koperkonsentrasie in die lewer in verband te bring om sodoende te bepaal of daar 'n korrelasie tussen die twee veranderlikes bestaan. Rooibokke wat maandeliks vanaf 1992.06.09 tot 1993.05.15 vir ander navorsingsprojekte in die omgewing van Phalaborwahek en Skukuza getermineer is, se sperme is vir die studie beskikbaar gestel. Die kontrolegroep het bestaan uit 20 diere wat saamgestel is uit 14 diere afkomstig van die Nwaswitshakaspruit naby Skukuza, uit 'n gebied wat nie met koper besoedel is nie en 6 diere uit 'n groep wat oorspronklik uit dieselfde gebied gekom het, maar wat te Skukuza in aanhouding geleef het op voedsel wat nie met koper besoedel was nie. Die eksperimentele groep het bestaan uit 44 diere waarvan 32 in die omgewing van die Tshutshispruit suid van Phalaborwahek versamel is en die ander 12 diere is langs die Nwaswitshakaspruit gevang en op 'n koperbesoedelde dieet te Skukuza aangehou. Sperme uit die epididymis van elke dier is in 'n tydelike laboratorium versamel en gefikseer. By die Elektronmikroskopie Eenheid van die Fakutreit van Veeartsenykunde van die Universiteit van Pretoria is die sperme voorberei en met behulp van skandeerelektronmikroskopie (SEM) en transmissie-elektronmikroskopie (TEM) bestudeer. Die koperkonsentrasie in die lewer is bepaal en murgvet is versamel om die kondisie van die diere te bepaaI. Met SEM is die uitwendige morfologie van 'n normale rooiboksperm met mikrograwe gedokumenteer en die mates van die sperm bepaal. Mikrograwe van waargeneemde spermabnormaliteite is met SEM geneem. Persentasies is vir normale sperme en die abnormaliteite van sommige sperme van 'n kontrolegroep (Gemiddelde lewerkoperkonsentrasie - GLKK =37.9 dpm NB) bereken. Dieselfde persentasies is ook vir 'n eksperimentele groep (GLKK = 104 dpm NB) bereken. Van die kontrolegroep het 91.6% en van die eksperimentele groep 93.6% sperme normaal vertoon. Die verskil in persentasies spermafwykings van die twee groepe was ook van dieselfde orde. Die verhoogde koperkonsentrasie in die lewers van die eksperimentele groep het dus nie gelei tot 'n verhoging van die persentasie makro-abnormaliteite wat met SEM waargeneem is nie. Die ultrastruktuur van 'n normale rooiboksperm is met behulp van TEM beskryf en met mikrograwe gedokumenteer. Die afwykings van die spermultrastruktuur is ook op mikrograwe vasgelê. Persentasies is vir die normale spermsneë en die abnormaliteite van sommige spermsneë van 'n kontrolegroep (GLKK = 33.86 dpm NB) bereken. Dieselfde persentasies is ook vir 'n eksperimentele groep (GLKK = 122.5 dpm NB) bereken. Vir die kontrolegroep was die persentasie neksneë met vakuoles = 31.5% en vir die eksperimentele groep was die persentasie = 57.9%. Die verhoogde koperkonsentrasie in die lewer van die eksperimentele groep het dus gelei tot 'n verhoging van die persentasie neksneë met vakuoles. Rede bestaan om te glo dat die verskil van die persentasies kopsneë van die kontrole- en eksperimentele groep ook aansienlik hoër behoort te wees. 'n Grafiese analise van die TEM-data het ook getoon dat daar 'n statisties betekenisvolle korrelasie tussen die persentasie spermnekke met vakuoles en die koperkonsentrasie in die lewer van rooibokke bestaan. Die bevinding is dat hoë lewerkoperkonsentrasies 'n nadelige invloed op die ultrastruktuur van die spermnek uitoefen. Die moontlikheid bestaan ook dat koperkonsentrasies in die lewer > 500 dpm NB ook 'n korrelasie met sommige van die ander abnormaliteite sal toon.

Internal fertilization usually implies that a spermatozoon, with intact attributes for zygote formation, passes all hurdles during its transport through the female genitalia and reaches the oocyte. During this journey, millions to billions of other spermatozoa perish. Spermatozoa are highly differentiated motile cells without synthetic capabilities. They generate energy via glycolysis and oxidative phosphorylation to sustain motility and to maintain the stability and functionality of their plasma membrane. In vivo, they spend their short lifespan bathing in female genital tract fluids of different origins, or are in vitro exposed to defined media during diverse sperm handling i.e. extension, cryopreservation, in vitro fertilization, etc. Being excitable cells, spermatozoa respond in vivo to various stimuli during pre-fertilization (capacitation, hyperactivation, oocyte location) and fertilization (acrosome reaction, interaction with the oocyte) events, mediated via diverse membrane ion-conducting channels and ligand-gated receptors. The present Thesis has mapped the presence and reactivity (sperm intactness and kinematics) of selected receptors, water and ion channels in ejaculated boar spermatozoa. The final aim was to find a relevant alternative cell type for in vitro bioassays that could ease the early scrutiny of candidate drugs as well as decreasing our needs for experimental animals according to the 3R principles. Spermatozoa are often extended, cooled and thawed to warrant their availability as fertile gametes for breeding or in vitro testing. Such manipulations stress the cells via osmotic variations and hence spermatozoa need to maintain membrane intactness by controlling the exchange of water and the common cryoprotectant glycerol, via aquaporins (AQPs). Both AQPs-7 and -9 were studied for membrane domain changes in cauda- and ejaculated spermatozoa (un-processed, extended, chilled or frozen-thawed). While AQP-9 maintained location through source and handling, thawing of ejaculated spermatozoa clearly relocated the labelling of AQP-7, thus appearing as a relevant marker for non-empirical studies of sperm cryopreservation. Alongside water, spermatozoa interact with calcium (Ca2+) via the main Ca2+ sperm channel CatSper. Increments in intracellular Ca2+ initiate motility hyperactivation and the acrosome reaction. The four subunits of the CatSper channel were present in boar spermatozoa, mediating changes in sperm motility under in vitro capacitation-inducing conditions (increased extracellular Ca2+ availability and bicarbonate) or challenge by the CatSper antagonists mibefradil and NNC 55-0396. Uterine and oviduct fluids are richest in endogenous opioids as β-endorphins during mating and ovulation. Both μ- and δ- opioid receptors were present in boar spermatozoa modulating sperm motility, as in vitro challenge with known agonists (μ: morphine; δ: DPDPE and κ: U 50488) and antagonists (μ: naloxone; δ: naltrindole and κ: nor-binaltrorphimine) showed that the μ-opioid receptor maintained or increased motility while the δ-opioid receptor mediated decreased motility over time. Finally, boar spermatozoa depicted dose-response effects on sperm kinematics and mitochondrial potential following in vitro challenge with 130 pharmacological drugs and toxic compounds as well as with eight known mito-toxic compounds. In conclusion, boar spermatozoa expressing functional water (AQPs-7 and -9) and ion (CatSper 1-4) channels as well as μ- and δ-opioid receptors are able to adapt to stressful environmental variations, capacitation and pharmacological compounds and drug components. Ejaculated sperm suspensions are easily and painlessly obtained from breeding boars, and are suitable biosensors for in vitro drug-induced testing, complying with the 3R principles of reduction and replacement of experimental animals, during early toxicology screening.

Des facteurs masculins sont identifiés dans près de la moitié des cas d’infertilité. À ce jour, les tests évaluant la fertilité masculine demeurent peu prédictifs de la survenue d’une grossesse. Dans le but de pallier cette lacune, nous avons mis au point deux nouveaux tests mesurant l’intégrité de l’ADN et le temps de survie des spermatozoïdes. Nous avons effectué une étude prospective portant sur 42 couples infertiles suivis en fécondation in vitro (FIV). Le spermogramme a été effectué selon les critères de l’Organisation Mondiale de la Santé (OMS) et le temps de survie des spermatozoïdes exposés à un détergent cationique a été mesuré en observant la mobilité sous microscope. L’intégrité de l’ADN des spermatozoïdes a été vérifiée par la nouvelle méthode de marquage radioenzymatique et par analyse de la structure de la chromatine (SCSA). Tous les tests ont été réalisés sur la partie des échantillons de sperme non utilisée par la clinique de fertilité. Le projet a été approuvé par le comité d’éthique du Centre Hospitalier Universitaire de Montréal (CHUM) et les patients ont préalablement signé un formulaire de consentement éclairé. L’analyse des paramètres du spermogramme et de l’intégrité de l’ADN n’a montré aucune différence statistiquement significative entre les données chez les couples avec ou sans grossesse. Cependant, le taux de grossesse biochimique était statistiquement plus élevé chez les couples dont le temps de survie des spermatozoïdes était long (>250 s) comparativement à ceux dont ce temps était court (≤250 s): 66% vs 27% respectivement (p<0,05). Les taux de grossesse clinique et d’implantation étaient aussi plus élevés, mais les différences n’atteignaient pas le seuil de signification statistique. Nos résultats confirment que le spermogramme et la mesure de la fragmentation de l’ADN des spermatozoïdes ne sont pas de bons facteurs prédictifs des résultats de la FIV. Par contre, le test de survie des spermatozoïdes serait un meilleur indicateur de la possibilité d’une grossesse en FIV. L’amélioration de sa spécificité et un plus grand nombre de sujets sont nécessaires avant de proposer son application en clinique de fertilité. / Male factors are known to be involved in almost half of the couples consulting for infertility. To date, the tests for evaluating male fertility are poor predictors of pregnancy. We developed two new tests to evaluate sperm function: a sperm survival test and a new method to measure sperm DNA integrity. This prospective study was conducted on 42 infertile couples undergoing in vitro fertilization (IVF). Assessment of sperm parameters was done according to the World Health Organization (WHO) criteria, and sperm survival upon exposure to a cationic detergent was measured by observing motility under the microscope. Sperm DNA integrity was verified by our new radioenzymatic method as well as by the sperm chromatin structure analysis (SCSA) method. All testing was performed on a remainder aliquot of the semen samples. The study was approved by the ethics committee of the Centre Hospitalier Universitaire de Montréal (CHUM), and informed consent was obtained before inclusion. Neither conventional semen analysis, nor sperm DNA fragmentation showed statistically significant difference between conception and non-conception cycles. However, the biochemical pregnancy rate was statistically higher in couples where the sperm survival time was long (>250 s) compared to short (≤250 s): 66% vs. 27% respectively, (p < 0.05). The clinical pregnancy rate and implantation rate were also higher but the differences did not reach statistical significance. Our study confirms that conventional semen analysis and the assay for sperm DNA integrity are not reliable indicators of IVF outcome. In contrast, our new sperm survival test seems to be a better predictor of the pregnancy rate after IVF. Improvement of its specificity and a larger cohort of patients are necessary before proposing its regular application in IVF clinics.

La fitness et a fortiori la survie d'une population dépendent de la stratégie et des performances de reproduction façonnées par son environnement. Par conséquent, les biomarqueurs traduisant une altération de la fonction de reproduction présentent un intérêt particulier. L'atteinte de la fertilité mâle au sein de la faune sauvage a notamment été adressée comme une problématique majeure susceptible de représenter une menace pour le maintien des populations. Toutefois, peu de méthodologies sont aujourd'hui disponibles chez des espèces pertinentes pour aborder cette problématique dans le cadre de programmes de biosurveillance environnementale, notamment chez les crustacés en dépit de leur représentativité au sein du règne animal et de leurs indispensables fonctions au sein des écosystèmes. Dans ce contexte, les présents travaux avaient pour objectif de proposer une ou plusieurs méthodologies basées sur la mesure de marqueurs de fonctionnalité et d'intégrité spermatique chez des crevettes Palaemonidae. Les investigations se sont portées sur deux espèces, une estuarienne, Palaemon longirostris, et une côtière, Palaemon serratus, pour leur complémentarité vis-à-vis du continuum estuaire - littoral. Compte tenu des nombreuses spécificités structurelles et fonctionnelles des spermatozoïdes de crustacés, I nombre de marqueurs transposables vers ces espèces s'est finalement avéré limité. Aussi, après une brève prospection, l'effort de recherche a rapidement été recentré sur la mesure de l'intégrité de l'ADN. Dès lors, la démarche scientifique a été construite de sorte à évaluer, point par point, la pertinence de la méthodologie développée dans une perspective d'application de l'outil dans le cadre de la surveillance environnementale. Une étape préliminaire d'optimisation et de validation méthodologique du test Cornet a démontré que, contrairement à une grande majorité de type spermatique, l'adaptation de ce test sur les spermatozoïdes de Palaemonidae ne nécessite aucune modification particulière du protocole. La dynamique de la réponse biologique en termes d'apparitions, de rémanence et d'effets possibles sur la fitness a été évaluée en conditions contrôlées au laboratoire. Ainsi, des expositions ex vivo et in vivo ont été conduites en utilisant une variété de génotoxiques modèles présentant différents modes d'actions. Les résultats ont attesté de la sensibilité, de la reproductibilité et du caractère intégrateur de la réponse. En revanche, aucun lien entre un ADN spermatique endommagé et une altération du succès de reproduction pré-éclosion n'a pu être établi. Parallèlement, une approche in situ a été conduite en vue de caractériser la valeur basale de la réponse mesurée. Différentes stratégies ont dû être adoptées en fonction des contraintes propres au milieu de vie de chacune des deux espèces. Un référentiel et une valeur seuil, communs aux deux espèces, ont pu être définis, soulignant le potentiel de transférabilité inter-espèces de l'outil. La méthodologie ainsi finalisée, a été éprouvée dans le cadre de plusieurs campagnes de suivi de différentes populations indigènes de l'estuaire et de la baie de Seine en 2015 et 2016. Les résultats se sont révélés très cohérents au regard de la pression de contamination et de la dynamique hydro-sédimentaire de la baie de Seine. En définitive, l'intégrité de l'ADN spermatique chez les Palaemonidae est opérationnelle en l'état pour un déploiement in situ en tant que biomarqueur d'exposition à un stress génotoxique. De futurs études devront néanmoins être conduites (1) pour mieux discerner les implications de ces dommages spermatiques en termes d'impact sur le recrutement des nouvelles cohortes et (2) éprouver la transférabilité de la méthodologie à d'autres espèces de crevettes et sur une plus large échelle géographique. / The environment shapes the reproduction's strategy and performance of a population, influencing its fitness and a fortiori its survival. Therefore, biomarkers that alter reproductive functions represent a great interest in ecotoxicology. The reduction of male fertility in wildlife can represent a threat to the population's survival. Moreover, fcw methodologies are available for species that are relevant to address this issue on envimnmental biomonitoring programs, especially for crustaceans, despite their representativeness in the wildlife and their essential functions within ecosysterns. The present work aimed to propose methodologies based on the measurement of functionality and integrity spermatic biomarkers on Palaemonidae shrimps. We studied two species, an estuarine, Palaemon longirostris, and a coastal species. Palaemon serratus. due to their complementaiity on the continuum estuary-littoral ecosystem. Howevcr, crustaceans' sperm has many structuraI and functional characteristics, the number of transposable markers is limited. Thereafter, the research effort was refocused on the measurement of the DNA integrity and this inethod was evaluated for its adequacy for a biomonitoring study. We optitrtized and validate the Cornet assay for the Palaemonidae species, and the dynamic of the biological response in ternis of appearances, persistence, and possible effects on fitness was evaluated under controlled conditions in the laboratoiy. Furthermore, ex-vivo and in-vivo exposures were conducted using genotoxic models with different modes of action. On the one hand, results attested to the sensibility, the reproducibility and the integrating character of the response, on the other hand, no correlation between damaged sperm DNA and an altération of the pre-hatch stage of development was established. In paralIel, an in-situ approach was conducted to characterize the baseline level of the measured response, taking into consideration the specific constraints of each species' habitat. We were able to define a common baseline level and a threshold value for both species, highlighting the potential of the tool for inter-species transferability. This method was tested with native populations from the estuary and from the Seine Bay in 2015 and 20] 6. And, the results proved to be consistent with the contamination pressure and the hydro-sedimentary dynamics of the Seine Bay. Ultimately, the DNA integrity of sperm in Palaemonidae seems to be functional for in-situ deploytnents as a biomarker of exposure to genotoxic stress. Nevertheless, future studies should be conducted (1) to botter discern the implications of this spermatic damage on the recruitment of new cohorts and, (2) to test the transferability of the methodology to other shrimp species and on a wider geographical scale.

<p>Eurycoma longifolia (Tongkat Ali / TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report&nbsp / regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods This study&nbsp / encompasses two parts (part 1: on spermatozoa / part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups,&nbsp / washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 &mu / g/ml) for 1 hour at 37&deg / C. A sample without addition of TA served as control. After incubation with TA,&nbsp / the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen&nbsp / species (ROS / dihydroethidium test / DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (&Delta / &psi / m) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells&nbsp / incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 &mu / g/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were&nbsp / evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant&nbsp / dose-dependent trends were found&nbsp / for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 &mu / g/ml, yet, no significance could be&nbsp / observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor &Delta / &psi / m.</p>

No description available.

<p>Eurycoma longifolia (Tongkat Ali / TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report&nbsp / regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods This study&nbsp / encompasses two parts (part 1: on spermatozoa / part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups,&nbsp / washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 &mu / g/ml) for 1 hour at 37&deg / C. A sample without addition of TA served as control. After incubation with TA,&nbsp / the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen&nbsp / species (ROS / dihydroethidium test / DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (&Delta / &psi / m) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells&nbsp / incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 &mu / g/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were&nbsp / evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant&nbsp / dose-dependent trends were found&nbsp / for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 &mu / g/ml, yet, no significance could be&nbsp / observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor &Delta / &psi / m.</p>