Comparison of antioxidant effects of kolaviron and vitamin C interventions on testicular structures following nevirapine administration in Sprague-Dawley rats / Comparación de los efectos antioxidantes en la intervención con kolaviron y vitamina C sobre las estructuras testiculares después de la administración de nevirapina en ratas Sprague-Dawley

Testicular toxicity has been implicated in highly active anti-retroviral therapy (HAART) treatment. Hence there is need to identify an effective antioxidant product that can alleviate testicular necrosis due to HAART administration. Forty eight adult male Sprague-Dawley rats were used in this study. The animals were divided into eight (8) groups: A-H (n= 6). Group A animals received normal saline as the control; Group B was given Nevirapine (Nv); Group C was given Kolaviron (Kv); Group D was given vitamin C; Group E was given Nv and Kv; Group F was given Nv and Vitamin C; Group G was given Nv for 56 d and Kv for 28 d serving as a withdrawal group; Group H was given corn oil. Nv, Kv and Vit. C were given at 1.54, 200 and 250 (mg·kg)/bw respectively while all administrations were through oral gavage. The body weights were taken every other day. Thereafter, they were anaesthetized with halothane. The testes were excised, weighed, fixed in Bouin's fluid and stained with H&E while the epididymes removed for semen fluid analyses. The results showed a significant (P<0.05) decrease in sperm motility in group E (Nevirapine + kolaviron) when compared with group F (Nevirapine + Vitamin C) while Sperm count was not significantly different (P>0.05) across the groups. The testicular histoarchitectural studies revealed indistinct spermatogonia, necrotic interstititial endocrine cells in the altered interstitial space, fragmented spermatids, atrophy of mature spermatocytes, degenerated germ cells, obliterated seminiferous tubules lumen, undifferentiated spermatogonia and cellular debris in the somniferous tubules lumen of nevirapine administered group but normal across the other groups. In the testis, there were no significant reduction in SOD, Catalase and GPx activities but a significant decrease in GST activity (P<0.001) when group E was compared with group F. In conclusion, vitamin C presents a better remediation in nevirapine induced spermiotoxicity compared to kolaviron in Sprague-Dawley rats.

La toxicidad testicular ha sido implicada en la terapia antirretroviral altamente activa (TARAA). Por lo tanto existe la necesidad de identificar un producto antioxidante eficaz que pueda aliviar la necrosis testicular en la administración de la TARAA. Cuarenta y ocho ratas macho Sprague-Dawley adultas fueron utilizadas. Los animales se dividieron en ocho (8) grupos: AH (n= 6). Grupo A, animales recibieron solución salina normal como el control; Grupo B, recibió Nevirapina (Nv); Grupo C, recibió Kolaviron (Kv); Grupo D, recibió vitamina C; Grupo E, recibió Nv y Kv; Grupo F, recibió Nv y vitamina C; Grupo G, recibió Nv durante 56 d y Kv por 28 d como un grupo de retirada; Grupo H, recibió aceite de maíz. Nv, Kv y Vit. C se administraron en dosis de 1, 54, 200 y 250 (mg · kg) de peso corporal respectivamente; todas las administraciones fueron por sonda oral. Los pesos corporales se tomaron cada dos días. A partir de ese momento los animales fueron anestesiados con halotano. Los testículos fueron extirpados, pesados y fijados en solución de Bouin y teñidos con H&E, mientras que el epidídimo se retiró para analizar el semen. Los resultados mostraron un descenso (p<0,05) en la motilidad de los espermatozoides en el grupo E (Nevirapina + Kolaviron) en comparación con el grupo F (Nevirapina + vitamina C), mientras que el recuento espermático no mostró diferencias significativas (P>0,05) entre los grupos. El estudio de la histoarquitectura testicular reveló espermatogonias indiferenciadas, con células intersticiales necróticas en el espacio intersticial y espermátidas fragmentadas. Además, en el grupo que recibió Nevirapina mostró espermatocitos maduros atrofiados, degeneración de células germinales, lumen de los túbulos seminíferos obliterados, espermatogonias indiferenciadas y restos celulares en el lumen de los tubulos seminíferos. En el resto de los grupos los resultados fueron normales. En el testículo hubo una reducción significativa en las actividades de la superóxido dismutasa, catalasa y glutatión peroxidasa, pero una disminución significativa en la actividad glutatión S-transferasa (P <0,001) al comparar los grupo E y F.

Simultaneous Administration of Dexamethasone and Vitamin E Reversed Experimental Varicocele-induced Impact in testicular tissue in Rats; Correlation with Hsp70-2 Chaperone Expression

ABSTRACTPurpose:This study aimed to investigate the protective effects of isolated and co-administration of vitamin E (VitE) and dexamethasone (DEX) on varicocele (VCL)-induced damages in testicular tissue.Materials and Methods:Wistar rats were divided into five groups (n=6), including; control-sham, non-treated VCL-induced, VitE-treated VCL-induced (VitE, 150 mg/kg, orally), DEX-administrated VCL-induced (DEX, 0.125 mg/kg, i.p.), VitE+DEX-received VCL-induced animals. The antioxidant status analyses, histopathological examinations, hormonal assay and tissue levels of alkaline phosphatase (ALP) were analyzed. The germinal epithelium RNA damage and Leydig cells steroidogenesis were analyzed. Moreover, the Hsp70-2 protein expression was examined based on immunohistochemical and western blot analyses. The sperm parameters, DNA integrity and chromatin condensation were investigated.Results:VitE and DEX in simultaneous form of administration significantly (P<0.05) down-regulated the tissue ALP level and attenuated the VCL-decreased GSH-px, SOD and TAC levels and remarkably (P<0.05) down-regulated the testicular malondialdehyde (MDA) and nitric oxide (NO) contents. The VCL-induced histopathological alterations significantly (P<0.05) improved in VitE and DEX-administrated animals. The VitE and DEX co-administration reduced the VCL-increased RNA damage and elevated the Leydig cells steroidogenic activity. The Hsp70-2 protein level completely (P<0.05) increased in VitE and DEX alone–and-simultaneous-administrated animals. Finally, the VitE and DEX could significantly (P<0.05) improve the VCL-decreased semen quality and improved the sperm DNA integrity and chromatin condensation.Conclusion:Our data suggest that Vit E by up-regulating the antioxidant status and DEX by reducing inflammation-dependent oxidative and nitrosative stresses could improve the VCL-reduced Hsp70-2 chaperone expression and ultimately protected the testicular endocrine activities and promoted the spermatogenesis process.

[Comparison of anti-oxidant enzymes activity and levels of zinc and selenium in sperm and seminal plasma between fertile and idiopathic infertile men]

Reactive oxygen species [ROS]-induced lipidperoxidation can lead to dysfunction of sperm and thereby, infertility may be occurred. So, always there is a balance between amount of ROS and anti-oxidant molecules in semen. Anti-oxidant enzymes of sperm; superoxide dismutase [SOD], glutathione peroxidase [GPX], catalse and zinc and selenium can protect it from destructive effects of ROS. Hence, the present study was designed to compare the activities of these enzymes and trace elements between fertile and idiopathic infertile men. Semen specimens were collected from 30 infertile men with proven infertility by an urologist, and 30 fertile men as control donors, with age range between 20-40 years old. Semen analysis was conducted by CASA method. Atomic absorption method was used for measuring of zinc and selenium concentration. Activity assays of SOD and GPX were performed by Randox Kits. Aebi method also was applied for evaluation of catalase activity. There was no difference between the activities of enzymes in fertile men and infertile ones. Also, it wasn't seen any difference in the selenium and zinc levels of seminal plasma. There was no relationship between evaluated items with sperm parameters. Only, in asthenoteratospermic individuals negative correlations were found between GPX and sperm motility, selenium and sperm morphology. Also, in these individuals, there was a positive correlation between SOD and catalse activity. Measuring activities of SOD, GPx, and catalase and the contents of zinc and selenium of seminal plasma do not appear to be suitable tools for determining the fertility potential of sperm

Protective effect of maternal prenatal melatonin administration on rat pups born to mothers submitted to constant light during gestation

We studied the effects of adverse conditions such as constant light (LL) on the circadian rhythm of malate (MDH, EC 1.1.1.37) and lactate (LDH, EC 1.1.1.27) dehydrogenase activities of the testes of male Wistar rats on postnatal day 28 (PN28), anxiety-like behavior (elevated plus-maze test) at PN60 and sexual behavior at PN120. The rats were assigned to mother groups on day 10 of pregnancy: control (12-h light/dark), LL (light from day 10 to 21 of pregnancy), and LL+Mel (LL and sc injection to the mothers of a daily dose of melatonin, 1 mg/kg body weight at circadian time 12, from day 17 to 21 of pregnancy). LL offspring did not show circadian rhythms of MDH (N = 62) and LDH (N = 63) activities (cosinor and ANOVA-LSD Fisher). They presented a 44.7 percent decrease in open-arm entries and a 67.9 percent decrease in time (plus-maze test, N = 15, P < 0.001, Mann-Whitney U-test and Kruskal-Wallis test), an increase in mounting (94.4 percent), intromission (94.5 percent) and ejaculation (56.6 percent) latencies (N = 12, P < 0.01, Mann-Whitney U-test and Kruskal-Wallis test) and lower numbers of these events (61, 59 and 73 percent, respectively; P < 0.01, N = 12) compared to controls. The offspring of the LL+Mel group presented MDH and LDH circadian rhythms (P < 0.05, N = 50, cosinor and ANOVA-LSD Fisher), anxiety-like and sexual behaviors similar to control. These findings supported the importance of the melatonin signal and provide evidence for the protective effects of hormones on maternal programming during gestation. This protective action of melatonin is probably related to its entrainment capacity, favoring internal coupling of the fetal multioscillatory system.

Arginase status in bull reproductive system

Seven healthy and sexually adult bulls were slaughtered and their reproductive system was isolated. Different tissues including: testes, epididymis, vas deferan, ampulla, accessory sex glands [seminal vesicle, prostate and bulbourethra], muscular and mucosal layer of pelvic and penile urethra were carefully dissected. Total soluble protein and arginase specific activity [ASA] were measured by Lowry and modified paranitrophenylglyoxal [PNPG] method, respectively. The results indicate that the highest arginase specific activity [51.28 +/- 8.79 x10[-3] IU/mg of protein] is present in muscle of pelvic part of urethra. Based on ASA, bulls reproductive system was categorized in three tissue groups: high, medium and low. Muscular layer of penile urethra with the highest ASA level [> 50 X10[-3] IU/mg tissue protein] is in the 1st group, Testes, bulbourethral gland and mucosal layer of penile urethra [30-40 X10[-3] IU/mg tissue protein] are in the2nd group and the rest parts [<25 X10[-3] IU/mg protein] are in 3rd group. Significant differences observed between classified tissues [p<0.05]. The present study indicate that ASA is present at different levels in all parts of bull reproductive system. This condition may be related to different rate of cell proliferation and differentiation or some other unknown physiological and biochemical activities of the enzyme in this system

The expression and cellular localization of phospholipase D isozymes in the developing mouse testis

To examine the involvement of phospholipase D (PLD)isozymes in postnatal testis development, the expression ofPLD1 and PLD2 was examined in the mouse testis atpostnatal weeks 1, 2, 4, and 8 using Western blot analysisand immunohistochemistry. The expression of both PLD1and PLD2 increased gradually with development frompostnatal week 1 to 8. Immunohistochemically, PLDimmunoreactivity was detected in some germ cells in thetestis and interstitial Leydig cells at postnatal week 1.PLD was mainly detected in the spermatocytes andresidual bodies of spermatids in the testis after 8 weeksafter birth. The intense immunostaining of PLD in Leydigcells remained unchanged by postnatal week 8. Thesefindings suggest that PLD isozymes are involved in thespermatogenesis of the mouse testis.

Effect of exogenous selenium on the testicular toxicity induced by ethanol in rats.

The effects of supplementation of selenium at a dose of 10 microg/ kg body weight were investigated on ethanol induced testicular toxicity in rats. In the present study, four groups of male albino rats were maintained for 60 days, as follows: (1) Control group (normal diet) (2) Ethanol group (4g/kg body weight) (3) Selenium (10 microg/kg body weight) (4) Ethanol + Selenium (4g/kg body weight + 10 microg/kg body weight). Results revealed that ethanol intake caused drastic changes in the sperm count, sperm motility and sperm morphology. It also reduced the levels of testosterone and fructose. The activities of 3betaHSD, 17betaHSD in the testis and SDH in the seminal plasma were also reduced. Lipid peroxidation was also enhanced as the lipid peroxidation products were increased and the activities of the scavenging enzymes were reduced. But on coadministration of selenium along with alcohol all the biochemical parameters were altered to near normal levels indicating a protective effect of selenium. These results were reinforced by the histopathological studies.

Utilidad del mapeo testicular en pacientes azooespérmicos no obstructivos / Benefit of the sperm FNA in patients with azoospermia

Objetivos: Ante la presencia de un paciente azoospérmico no obstructivo comienza un verdadero desafío que consiste en localizar y obtener una muestra de tejido testicular donde se haya producido espermatogénesis con el fin de ser utilizada en una técnica de reproducción asistida. Los objetivos del presente trabajo son estimar el peso relativo de la FSH sobre la posibilidad de hallar espermatozoides en una biopsia utilizando la técnica de mapeo testicular con respecto a la biopsia convencional. Material y Métodos: Estudio retrospectivo, comparativo a muestras relacionadas de pacientes a quienes se le realizó mapeo testicular bilateral. Se incluyen 35 pacientes con diagnóstico de azoospermia no obstructiva operados en nuestro Servicio, entre diciembre de 1999 y diciembre de 2000 A todos los pacientes se les realizó mapeo testicular bilateral con seis tomas de cada lado, además se les practicó biopsia convencional con el fin de comparar los hallazgos entre ambos métodos Fueron evaluados también los valores preoperatorios de FSH con el fin de estimar el peso relativo sobre la posibilidad de hallar espermatozoides. Resultados: Los bajos valores de FSH fueron un valor predictivo positivo en la posibilidad de hallar espermatozoides. Con la biopsia convencional se encontraron espermatozoides en un 34,3 por ciento de las muestras mien- tras que el mapeo logró hallarlos en un 48,6 por ciento. Conclusiones: Los valores preoperatorios de FSH podrían desempeñar un valor predictivo sobre la posible obtención de espermatozoides. Se presenta el mapeo testicular como un método que potencialmente podría aumentar el porcen- taje de hallazgos de material espermático en pacientes azoospérmicos no obstructivos evitando tener que cancelar ciclos de fecundación in vitro de no hallarse espermatozoides.

Effect of casein diet on gonadotropin releasing hormone antagonist induced changes in adrenal gonadal functions in male rats.

Adult male rats received daily injections (sc) of gonadotropin releasing hormone antagonist (0.2 mg/kg(-1) x day(-1)) for 21 days when they were sacrificed on day 22, adrenal weight, adrenal A5-3beta (delta 5-3beta) hydroxysteroid dehydrogenase (Delta5-3beta-HSD) activity and serum level of corticosterone were increased significantly while testicular 17beta (17beta) hydroxysteroid dehydrogenase (17beta-HSD) activity and serum level of testosterone and spermatogenesis were decreased in the rats fed on 5% casein diet. GnRH antagonist treated rats fed on 20% casein diet, resulted significant decrease in adrenal weight, serum corticosterone and adrenal A5-3beta-HSD activity while testicular 17beta-HSD activity serum testosterone levels and the weights of sex organs were increased with respect to anti GnRH treated rats fed on 5% casein diet. But the GnRH antagonist treated rats fed on 20% casein diet showed decreased spermatogenesis quantitatively and sperm count appeared similar to anti GnRH treated rats fed on 5% casein diet. These results indicate that high casein diet protects adrenocortical activity and stimulates testosterone synthesis without effecting spermatogenic arrest in GnRH antagonist treated rats. It may be concluded that GnRH antagonist in presence of high milk protein diet may be considered to be a suitable antihormone in the development of an ideal male contraceptive.

Inmunorreactividad de enolasa en el epitelio seminífero senil humano / Immunoreactivity to enolase in the human senil seminiferous epithelium

El metabolismo energético intracelular compartimentalizado, activa la enzimo enolasa para formar ácido pirúvico. Este, substrato energético, debe ingresar a la mitocondria para continuar hacia el ciclo de losácidos tricarboxílicos. Sin embargo, durante la proliferación del epitelio seminífero ocurre una distribución y pérdida citoplasmática progresiva y disposición de las mitocondrias a nivel del flagelo inicial. en el presente estudio se describe la inmunoreactividad de la enzimo enolasa en las diferentes poblaciones celulares del epitelio seminífero, en testiculo humano senil. Se trabajo con un paciente de 70 años, sometido a orquiectomía subcapsular terapéutica. El tejido testicular fue fijado inmediatamente en formalina taponada al 10 por ciento y mantenido por 12 horas. Luego se procesó por técnicas histológicas corrientes e incluyo en parafina para obtener secciones de 5 um. Posteriormente se procedió a la reacción inmunohistoquímica para enolasa y revelación con complejo avidina-biotina. Finalmente se cuantificaron las distintas poblaciones celulares del epitelio seminífero con reacción positivo o negativa. Los resultados preliminares se expresan en porcentajes de células positivas respecto del total de células contadas (40x. se observó que la totalidad de las células de Sértoli presentaron reacción negativa a la enolasa. En el epitelio seminífero se encontró que el 76 por ciento de las gonias (gonias tipo A y B) mostraron reacción negativa a la enolasa, mientras que en citos 1 se redujo al 10 por ciento y ausencia total en espermátidas y espermatozoides. Por lo tanto, las célula somáticas (de origen mesodérmico), del epitelio seminífero presentarían isoenzimas enolasas de reactividad diferente en la relación con la línea germinal (espermatogonias). Adicionalmente, en la línea germinal la inmunoreactividad a enolasa disminuye mientras progresa la espermatogénesis

Impact of linear alkyl benzene sulphonate (LAS) on phosphatase activity in testis of the teleostean fish, Heteropneustes fossilis (Bloch).

Fishes are sensitive indicators of pollutants present in water.These pollutants cause various physical and physiological alterations in fishes. In the present work alteration in the activity of acid and alkaline phosphatase was evaluated in testicular tissue of fresh water fish Heteropneustes fossilis exposed to LC50 value of linear alkyl benzene sulphonate (LAS) for different exposure periods [24 h, 48 h, 72 h and 96 h] With increase in the concentration of chemical LAS, the activity of acid phosphatase (ACP) was reported elevated while a significant fall in the activity of alkaline phosphatase (ACP) was recorded for same exposure period. Elevated activity of ACP, one of the important hydrolases of lysosomes, is quite suggestive of bringing about gross necrosis and dysarchitecture. ALP is involved in various metabolic activities including gonadal maturation and as such decreased activity of this enzyme is definitely one of the important causative factors for reproductive impairment of the fish.

Enzymatic method for branched chain alpha-ketoacid determination: application to rapid analysis of urine and plasma samples from maple syrup urine disease patients

A new method for the determination of branched-chain alpha-ketoacid concentration using lactate dehydrogenase (E C 1.1.1.27) isozyme C4 (LDH) C4) from mouse testes is proposed. The assay is performed on urine and plasma without previous treatment. Alpha-ketoglutarate and pyruvate are determined on the same sample using glutamate dehydrogenase (GDH,EC 1.4.1.2) and lactate dehydrogenase isozyme A4 (LDH5) respectively and subtracted from the total alpha-ketoacid concentration obtained with LDH C4. This value corresponds to the branched chain alpha-ketoacid. Results were linear within the concentration range 8 to 170 mumoles/L. Detection limit was 8 mumoles/L. Analytical recovery was higher than 91 per cent. For microplate assays, recoveries were higher than 84 per cent and the detection limit was 20 mumoles/L. Determinations performed with GDH, LDH A4 and LDH C4 allow differentiation of E3 deficiency from other clinical phenotypes of maple syrup urine disease. The method is simple and fast, and adaptation to microplates would allow screening of newborns.

Effect of amino acids on inhibition of lactate dehydrogenase-X by gossypol

Gossypol acetic acid (GAA) has been shown to have male antifertility effects, but there are pronounced differences among animal species. In the search of endogenous effector molecules, which interfere with the functions of GAA, we have studied the in vitro effect of various amino acids on the inhibition of the purified LDH-X by GAA. Histidine, cysteine and glycine were shown to block the effect of GAA. The effects of these amino acids were concentration dependent. Histidine and glycine protection was found to be complex type in which both the Km and Vmax were decreased compared to control. Arginine, glutamic acid, phenylalanine and valine were found to be ineffective against the inhibitory action of GAA.

Effect of passive immunization with specific anti-FSH on tes-ticular lipids and some lipogenic enzymes of adult bonnet monkeys

The effect of anti-follicular stimulating hormone [FSH] on testicular lipid and the specific activity of testicular enzymes of the isocitrate dehydrogenase [ICDH], pyruvate/malate dehydrogenase [MDH] and malic enzyme involved in lipogenesis were studied in mature bonnet monkeys, Macaca radiata. Immunization of monkey with anti-FSH for 24 days did not produce any significant changes in the body weight, organ weight and pituitary weight. Testicular isocitrate dehydrogenase [ICDH] and malic enzyme activities were decreased significantly but MDH activity was stimulated by anti-FSH treatment. Testicular total lipid, phospholipid and cholesterol, were not altered significantly by the Anti-FSH treatment. Increased level of free cholesterol were also observed after FSH treatment. Among glyceride glycerol sub classes, triacyl glycerol showed a significant increase. Among testicular phospholipid classes, phosphatidyl inositol was markedly decreased by anti-FSH immunization. Data on serum hormonal profile, shows that there were no alteration in serum testosterone, prolactin [PRL] and luteinizing hormones [LH] but FSH was significantly decreased. The present study reveals that immunization with anti-FSH has significant effect on different class of testicular lipids and pyruvate malate enzymes cycle

A rede testicular na cobaia: observaçöes morfológicas e histoquímicas / The rete testis in guinea pig: a morphological and histochemical examination

A rede testicular (RT) da cobaia tem características morfológicas de um tipo intermediário de RT. Esta rede foi caracterizada com cavitária, labiríntica e axial relativamente à disposiçäo de seus canais epiteliais interconectados. Morfologicamente consiste, predominantemente, de câmaras epiteliais interligadas que penetram no parênquima testicular, por uma pequena distância. Os canais e câmaras de todas as partes da rede säo revestidos por epitélio cúbico simples. Reatividades histoenzimáticas fortes foram observadas, predominantemente, na matriz da RT às enzimas fosfatases alcalina e ácida e m-ATPase. Ambas as fosfatases mostraram reatividades médias no epitélio da RT de cobaia, no qual a reatividade da m-ATPase foi fraca. Por outro lado, a NADH-d mostrou reatividade média, porém homogênea, na matriz e no epitélio da RT.

Molecular inactivation of testicular hyaluronidase in solid state after proton irradiation: a study based on target size, substrate binding and thermodynamic analysis of heat denaturation.

Dose response activity curve of testicular hyaluronidase (HDase) following proton irradiation in dry state follows complicated mechanisms which may involve multiple hits and multiple targets of variable sizes giving a constant G value of 1.66. Target analysis appears to be modified by slow recovery of activity when irradiated enzyme is brought to aqueous phase. However, pattern of irradiation at a dose of 1 x 10(5) to 8 x 10(5) Gy reveals that though binding affinity of enzyme to the substrate (hyaluronic acid) increases as shown by declining Km from 500 mg/l to 300-70 mg/l, the reaction rate of catalysis by irradiated HDase is decreased due to decrease in reaction velocity (Vmax: 266 versus 76 units at 8 x 10(5) Gy). Activation analysis of heat denaturation of nonirradiated HDase suggested the involvement of 78 kcal/mole of energy of activation (E*a) which declined to 63-52 k cal/mole after irradiation at 1 x 10(5) to 8 x 10(5) Gy for residual enzyme. The corresponding change in entropy of activation (delta S*) increased from a control value of -291 eu to -236 eu at 8 x 10(5) Gy. From thermodynamic analysis in association with recovery in aqueous phase, it is concluded that HDase is inactivated due to dissipation of proton energy among weak forces including H bonds associated with secondary/tertiary structure of molecules.

Poly ADP ribosylation as a possible mechanism of microwave--biointeraction.

Electromagnetic fields (EMFs) affect the metabolism of the body including the nervous, endocrine, cardiovascular, hematological as well as the reproductive system. EMFs are environmental pollutants, thus posing a health hazard which can cause steric changes in the molecule located at the cell surface. Microwaves are known to cause chromosomal abberations and act as tumor promoters. The process involves a stream of signals from cell membrane to nucleus and other organelles. The present investigations aim to understand the mechanism of biological effects of microwaves (2.45 GHz). The effect was studied on poly ADP-ribosylation, which is a post translational modification of chromatin protein catalysed by the enzyme poly ADPR polymerase using NAD+ as the substrate. Poly ADP-ribosylation has been shown to be involved in several aspects of chromatin structure and function. Twenty-three days old rats weighing 42-48 gms were exposed at a microwave dose level of 1.0 mW/cm2. After exposure for sixty days the animals were sacrificed and an estimation of poly ADPR polymerase activity was undertaken in different organs of these animals. There was an increase of 20% in its activity in liver, 35% in testis, whereas brain showed a 53% decrease in diencephalon and 20% decrease in the cortex in the exposed animals as compared to their respective controls. There was no change in enzyme activity in spleen and kidney. This was accompanied by concomitant changes in NAD+ levels. The above results may be cited as important events in carcinogenesis and tumor promotion related to microwave exposure and the signal transduction mechanism involved. The goal is to shed light on complex ecogenetic interactions leading to cancer modulation of gene expression by epigenetic mechanism.

LDH-x in azoospermia: a new diagnostic alternative to vasography and testicular biopsy.

Activities of different isozymes of lactate dehydrogenase were studied in semen samples from fertile, and different groups of infertile men. No isozyme except the sperm specific LDH-x or C4 showed any difference in activity with changes in sperm density. Activity of LDH-x exhibited a trend positive to changes in sperm density and became absent in obstructive and vasectomized azoospermia. Among other types of azoospermia, the activity became very high in maturation arrest but very low in patients with testicular biopsy showing germ cell aplasia, such as in SCOS and ghostlike tubules. Diagnostic usefulness of the estimation of LDH-x activity in semen is discussed.

Modulation of T cell functions by sperm-specific lactate dehydrogenase: fluorescence analysis of immune competent cells and local graft versus host reaction.

Immune responses to a well-defined sperm-specific isogenic lactate dehydrogenase-C4 (LDH-C4) have been studied in C57Bl/Ks (H-2d) mice after immunization through intra-rectal route. Presence of anti-LDH-C4-antibodies in the sera of females immunized in presence or absence of adjuvant suggested that the immune system of mice becomes exposed to sperm antigens following intrarectal insemination. LDH-C4 primed lymphocytes from both males and females, when transferred in F1 hybrids, suppressed stimulation index of local graft versus host reaction. However, contrary to females, male counterparts which did not elicit measurable anti-LDH-C4-antibody titer, showed the presence of a higher proportion of Ly2+ and Ia+ fluorescence labelled cells in the spleen of LDH-C4 administered mice. Results suggest that males are more susceptible for immune suppression of T cell functions through generation of T suppressor cells. Sex differences in relation to immune deviation by intra-rectal administration of sperm-specific LDH-C4 in mice and their consequences in AIDS and AIDS-related complex diseases are described.

Effect of chlorpromazine and gossypol on Ca(2+)-ATPase activity in the microsomal membranes of rat testes.

The microsomal membranes isolated from rat testes have been found to contain a Mg(2+)-dependent and a Mg(2+)-independent Ca(2+)-ATPase. The enzyme activities were inhibited by two contraceptive drugs--gossypol and chlorpromazine. The inhibition by the former was affected by the presence of ligand(s) and not the substrate in the incubation medium, whereas ligand(s)/substrate did not affect the inhibition by chlorpromazine. This may be explained from the fact that the binding of chlorpromazine and ligand(s)/substrate to the enzyme are independent of each other whereas in case of gossypol the ligand(s) compete with the drug at the binding site of the enzyme.