Antioxidant enzyme profile and lipid peroxidation products in semen samples of testicular germ cell tumor patients submitted to orchiectomy

ABSTRACT Purpose To determine enzymatic antioxidant and lipid peroxidation levels in seminal plasma of patients orchiectomized for testicular tumors. Materials and Methods The study included 52 patients: 26 control men and 26 orchiectomized patients for testicular tumor, of which 12 men had seminoma tumor and 14 men non-seminoma tumor. After semen analysis performed according to the WHO guidelines, an aliquot of semen was centrifuged and the seminal plasma was collected. Lipid peroxidation was performed by thiobarbituric acid reactive substances (TBARS) assay and antioxidant profile was assessed by analyzing catalase, glutathione peroxidase (GPx) and superoxide anion (SOD) activities using colorimetric assays with a standard spectrophotometer. Data were tested for normality and compared using one-way ANOVA (p<0.05). Results Seminoma and non-seminoma groups presented lower sperm concentration and morphology when compared to control group (p=0.0001). Both study groups (seminoma and non-seminoma) presented higher TBARS levels when compared to control group (p=0.0000013). No differences were observed for SOD (p=0.646) andGPx (p=0.328). It was not possible to access the enzymatic activity of catalase in any group. Conclusion Patients with testicular tumor present increased semen oxidative stress, but no differences were observed in antioxidant levels, even after orchiectomy. This indicates that most likely an increased generation of oxidative products takes place in these patients.

Glutathione and male fertility

Considerable efforts are now focused on the identification of ultrastructural and/or molecular defects in the seminal fluid from infertile men. The tripeptide glutathione is the most abundant non-thiol protein in mammalian cells. It is a part of the protein identified as the major constituent that embeds the helix of mitochondria in the midpiece of spermatozoa. The objective of this work is to clarify the role played by seminal plasma glutathione on male fertility. A randomly selected group of 25 nonazoospermic infertile patients were included in this study, who attended the Institute of Embryo Research and Infertility Treatment at Al-Kadhimiya City/ Baghdad, with a history of infertility of at least 1 year duration between March and June 2009. Controls consisted of samples obtained from healthy donors of proven fertility [n = 10]. The seminal plasma glutathione concentration and percentage of spermatozoa with low mitochondrial membrane potential [MMP] were studied in the ejaculate from infertile patients against control donors. Results were compared with the standard semen characteristics [concentration, motility and morphology]. Infertile patients had significantly lower amount of GSH in the seminal plasma [P = 0,00001], lower progressive motility [grades a and b] [P = 0,002] and poorer sperm morphology [P = 0.00001]; while they had higher percentage of spermatozoa with low MMP [P = 0,00001] as compared to controls. Most of infertile patients were non oligozoospermic therefore; no significant concentration difference was found compared to controls. Seminal glutathione concentration was significantly inversely correlated with low MMP [r = - 0.665, P = 0.001], yet it was positively related to progressive motility [r = 0.51, P = 0.02] and normal morphology [r = 0.611, P = 0.004]. Comparison between patients with asthenoteratozoospermia [low percentage of progressive motility and normal morphology] [n = 18] against other patients [n = 7] revealed significantly lower seminal GSH concentration [P = 0.02] and higher percentage of low MMP [P = 0.00001]. This further support the relation between GSH and mitochondrial status, motility and normal morphology of spermatozoa. Seminal plasma GSH levels may play an important role in male fertility through protection against oxidative damage and preserving the mitochondrial integrity and function of the spermatozoa as the main energy source

Sesame radiatum phytoestrogens stimulate spermatogenic activity and improve sperm quality in adult male Sprague Dawley rat testis / Fitoestrógenos sesame radiatum estimulan la actividad espermatogénica y mejoran la calidad del esperma en el testículo de ratas Sprague Dawley adulto de sexo masculino

Increasing concern has been expressed about the declining sperm count of humans and the potential environmental effects of both synthetic and natural estrogenic endocrine disruptors (EEDs) on human reproductive health in the last few decades. However, due to paucity of knowledge, we evaluate the chronic reproductive toxicity of sesame phytoestrogenic lignans on the male Sprague Dawley (SD) rats' testis. Thirty adult male SD rats weighing 150-200g were divided into three groups. Two treated groups received a daily dose of aqueous leaves extract of Sesamum radiatum at 14.0 mg/kg bw and 28.0mg /kg bw respectively via gastric gavage, while equal volume of normal saline was administered to the control group for six weeks. Seminal analysis and hormonal assay study were analyzed using SPSS software and P< 0.05 was considered statistically significant. The results showed significant (P< 0.05) body weight gains observed in all the animals with significant (P< 0.05) weight increase in their raw testicular weights compared to control. The relative testicular weight per 100g bw was significantly (P< 0.05) higher in control than treated. However, the weight gain was dose related with a reversal in their relative testicular weight. The cauda sperm count including the motility and morphology of the treated were significantly (P< 0.05) higher than control in a dose related manner. In addition, significant (P > 0.05) increases in testosterone and a significant decrease in FSH in the high dose (treated) compared to control. Sesame phytoestrogenic lignans improves spermatozoa quality in a dose related manner.

Una mayor preocupación se ha expresado en las últimas décadas por la disminución en la cantidad de espermatozoides de los seres humanos y los posibles efectos ambientales de disruptores endocrinos estrogénicos (DES) sintéticos y naturales sobre la salud reproductiva humana. Sin embargo, debido a la escasez de conocimientos, evaluamos la toxicidad crónica para la reproducción de fitoestrógenos lignanos del sésamo en el testículo de ratas Sprague Dawley (SD) macho. Treinta ratas macho adultas, de un peso de 150-200g se dividieron en tres grupos. Dos grupos de tratamiento recibieron una dosis diaria de extracto acuoso de las hojas Sesamum radiatum de 14,0 mg/kg de peso corporal y 28,0mg / kg de peso corporal, respectivamente, a través de una sonda gástrica, mientras que igual volumen de solución salina normal se administró al grupo control durante seis semanas. El análisis seminal y estudio de ensayo hormonal fueron analizados mediante el software SPSS y P <0,05 fue considerado estadísticamente significativo. Los resultados mostraron significativas (P <0,05) ganancias de peso corporal observados en todos los animales con un peso significativo (P <0.05), aumento de peso en su peso testicular bruto en comparación con el control. El relativo peso testicular por 100 g de peso corporal fue significativamente mayor (P <0,05) en control que en tratamiento. Sin embargo, el aumento de peso dosis se ha relacionado con un retroceso en su peso relativo testicular. La cantidad de espermatozoides, incluyendo la motilidad y morfología de los que recibieron tratamiento fue significativamente más alto que el control (P <0,05) en una dosis relacionados. Además, significativo aumento en los niveles de testosterona (P> 0,05) y una disminución significativa de FSH en la dosis alta (tratados), en comparación con el control. Fitoestrógenos lignanos del sésamo mejoran la calidad de los espermatozoides en una dosis adecuada.

Activity of antioxidant enzymes in seminal plasma and their relationship with lipid peroxidation of spermatozoa

PURPOSE: To determine the activity of seminal plasma catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) and their relationship with malondialdehyde (MDA), as a marker of lipid peroxidation, content of spermatozoa and seminal plasma in normozoospermic and asthenozoospermic males. MATERIALS AND METHODS: Semen samples were obtained from 15 normozoospermic and 30 asthenozoospermic men. RESULTS: We observed inverse correlations between activities of CAT (k/mL) and SOD (U/mL) in seminal plasma with MDA content of spermatozoa from normozoospermic samples (r =- 0.43, p < 0.05 and r =- 0.5, p < 0.05, respectively). Significant correlations were observed between total activity CAT (k/total seminal plasma) with total SOD (U/total seminal plasma) and GPX activity (mU/total seminal plasma) in seminal plasma from normozoospermic samples (r = 0.67, p = 0.008 and r = 0.455, p = 0.047, respectively). Furthermore, we found positive correlations between total activities of CAT, SOD and GPX with total content of MDA in seminal plasma (nmoL/total seminal plasma) from normozoospermic samples (r = 0.67, p = 0.003; r = 0.73, p = 0.003; r = 0.74, p = 0.004, respectively). In asthenozoospermic samples, there were no significant correlations observed between activities of CAT (k/mL), SOD (U/mL) and GPX (mU/mL) of seminal plasma with MDA content of spermatozoa. However, we found significant correlations between total activities of CAT (k/total seminal plasma) and SOD (U/total seminal plasma) with total content of MDA in seminal plasma (r = 0.4, p = 0.018 and r = 0.34, p = 0.03, respectively). CONCLUSION: These findings indicate a protective role for antioxidant enzymes of seminal plasma against lipid peroxidation of spermatozoa in normozoospermic samples.

Human seminal proteinase and prostate-specific antigen are the same protein.

Human seminal proteinase and prostate-specific antigen (PSA) were each isolated from human seminal fluid and compared. Both are glycoproteins of 32-34 kDa with protease activities. Based on some physicochemical,enzymatic and immunological properties,it is concluded that these proteins are in fact identical.The protein exhibits properties similar to kallikrein-like serine protease, trypsin,chymotrypsin and thiol acid protease.Tests of the activity of the enzyme against some potential natural and synthetic substrates showed that bovine serum albumin was more readily hydrolysed than casein.The results of this study should be useful in purifying and assaying this protein.Based on published studies and the present results,the broad proteolytic specificity of human seminal proteinase suggests a role for this protein in several physiological functions.

Evaluación del sistema antioxidante en el semen normal / Evaluation of antioxidant system in normal semen

Background. Reactive oxygen species (ROS) formation have the ability to alter reversibly or irreversibly the cellular function in humans. It has been proposed that the ROS alters the biochemistry and the physiology of the sperm. On the other hand, the antioxidative mechanisms could protect the sperms from the damage produced by free radicals. Aim. To determine the normal values for superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA) and nitric oxide (NOx) in the seminal liquid of healthy humans. Procedures. Semen samples from 45 healthy men (22 to 47 years of age) were studied. The samples were obtained by masturbation and were collected in conical sterile tubes. Once centrifuged at 4 °C they were divided in aliquots to measure the concentration of SOD, GPx, MDA, and NOx. The analysis of the samples was realized in conformity with biochemical widely accepted methods. Results. The concentrations of SOD and MDA both in the seminal liquid and in the spermatozoids were similar, SOD 0.43 ± 0.09 U/mg prot. in the seminal liquid and 0.45 ± 0.07 U/ mg prot. in spermatozoids, and MDA 0.33 ± 0.07 nmoles/mg prot. and 0.37 ± 0.10 nmoles/mg prot. in the seminal liquid and spermatozoids respectively. With regard to GPx it increased almost 13 times more in the spermatozoids (2547.77 ± 48.59 U/mg prot.) than in the seminal liquid (197.54 ± 25.21 U/mg prot.). The NOx also increased lightly in the spermatozoids (4.45 ± 0.43 /imol) when compared with the seminal liquid (3.91 ± 0.16 /imol). Conclusions. The measurement of the antioxidative and oxidative agents could serve to evaluate human infertility in those cases where the result of the spematobioscopy appears normal.

Antecedentes. Las especies reactivas del oxígeno (ERO), tienen la capacidad de alterar reversible o irreversiblemente la función celular. Se ha propuesto que las ERO modifican la bioquímica y la fisiología del espermatozoide. Por otro lado, los mecanismos antioxidativos pudieran proteger a los espermatozoides del daño producido por las ERO. Objetivo. Determinar los valores normales para el superóxido dismutasa (SOD), glutatión peroxidasa (GPx), malondialdehído (MDA) y óxido nítrico (NOx) en el líquido seminal y espermatozoides de humanos sanos. Procedimientos. Se estudiaron 45 muestras de semen de sujetos aparentemente sanos. Las muestras se obtuvieron por masturbación y se colectaron en tubos estériles. Una vez centrifugadas, se fraccionaron en alícuotas para medir la concentración de SOD, GPx, MDA y NOx. El análisis de las muestras se realizó conforme a métodos bioquímicos ampliamente aceptados. Resultados. Las concentraciones de SOD y MDA en el líquido seminal como en los espermatozoides fueron similares (SOD 0.43 ± 0.09 en semen y 0.45 ± .07 U/mg prot. en espermatozoides, y MDA 0.33 ± .07 y 0.37 ± 0.10 nmoles/mg prot. en líquido seminal y espermatozoides, respectivamente. Con respecto a la GPx, está aumentada casi 13 veces más en los espermatozoides (2547.77 ± 48.59 U/mg prot.) que en el líquido seminal (197.54 ± 25.21 U/mg prot.), el NOx también se incrementa ligeramente en los espermatozoides (4.45 ± 0.43 µmol) cuando se compara con el líquido seminal (3.91 ± 0.16 µmol). Conclusiones. La medición de los antioxidantes y oxidantes pudieran servir para evaluar la infertilidad humana en aquellos casos donde los resultados de la espermatobioscopia aparezcan como normales.

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.

Effect of psychological stress on the L-arginine-nitric oxide pathway and semen quality

It has been reported that mental stress causes abnormality of spermiogram parameters. We investigated the effect of psychological stress on the L-arginine-nitric oxide (NO) pathway. Semen samples were collected from 29 healthy fourth semester medical students just before (stress) and 3 months after (non-stress) the final examinations. Psychological stress was measured by the State Anxiety Inventory questionnaire. After standard semen analysis, arginase activity and NO concentration were measured spectrophotometrically in the seminal plasma. Measurements were made in duplicate. During the stress period, sperm concentration (41.28 ± 3.70 vs 77.62 ± 7.13 x 10(6)/mL), rapid progressive motility of spermatozoa (8.79 ± 1.66 vs 20.86 ± 1.63 percent) and seminal plasma arginase activity (0.12 ± 0.01 vs 0.22 ± 0.01 U/mL) were significantly lower than in the non-stress situation, whereas seminal plasma NO (17.28 ± 0.56 vs 10.02 ± 0.49 æmol/L) was higher compared to the non-stress period (P < 0.001 for all). During stress there was a negative correlation between NO concentration and sperm concentration, the percentage of rapid progressive motility and arginase activity (r = -0.622, P < 0.01; r = -0.425, P < 0.05 and r = -0.445, P < 0.05, respectively). These results indicate that psychological stress causes an increase of NO level and a decrease of arginase activity in the L-arginine-NO pathway. Furthermore, poor sperm quality may be due to excessive production of NO under psychological stress. In the light of these results, we suggest that the arginine-NO pathway, together with arginase and NO synthase, are involved in semen quality under stress conditions.

Association between mental stress & some antioxidant enzymes of seminal plasma.

BACKGROUND & OBJECTIVE: Mental stress, which is responsible for various disorders, is one of the most important medical and social problems. It is reported that mental stress causes abnormality in sperm quality. Most of the previous investigations done to study the association between mental stress and infertility were carried out with infertile men. Infertility itself and/or its therapy may lead to stress. Further, most studies investigating the association between psychological stress and semen quality have lacked information on biochemical parameters. In the present study, we investigated the effect of mental stress due to final exams on two important antioxidant enzymes of the seminal plasma, superoxide dismutase (SOD) and catalase in normal healthy medical students. METHODS: Semen samples were collected from 27 healthy male volunteers, who were third semester students of a medical school, just before (stress period) and 10.19+/-0.83 wk after (non-stress period) the final examinations. Psychological stress of participants was measured by the State Trait Anxiety Inventory. After standard semen analysis, semen samples were centrifuged at 10,000 x g for 15 min. Superoxide dismutase (SOD) and catalase activities were measured in the seminal plasma. RESULTS: During stress period, stress scores and SOD activities increased significantly compared to the non-stress period. Catalase activities showed no change. Spermatozoa concentrations, motility index and percentage of rapid progressive motility decreased under stress. INTERPRETATION & CONCLUSION: Our results indicated that mental stress negatively affected semen quality. Increase in SOD activities led to poor quality of semen parameters.

Determinación de células peroxidasa positivas en líquido seminal: es un parámetro confiable para el diagnóstico de infección genital asintomática? / Determination of peroxidase positive cells in semen: is it a secure parameter for the diagnosis of silent genital infections?

BACKGROUND: The presence of leukocytes, detected by peroxidase test in semen, can be a good indicator of infections in the male genital tract. Peroxidase positive cells have been positively correlated with elevated values of elastase, one of the major proteases liberated by granulocytes at the inflammation place. However, seminal granulocytes may not be adequately detected by the peroxidase test in comparison with immunological methods. AIM: To correlate the determination of peroxidase positive cells with the elastase level in the seminal plasma. MATERIAL AND METHODS: Seminal plasma from 64 patients with a high number of round cells (> 106/ml) in semen, was studied. Correlation analysis was done using the Pearson correlation coefficient. RESULTS: No correlation between the level of granulocyte elastase and the number of peroxidase positive cells (r = 0.2237, p > 0.05), or even the number of round cells (r = 0.03934, p > 0.05) was observed. CONCLUSIONS: Our results suggest that the determination of peroxidase positive cells is not a reliable indicator of leukocytes in the seminal plasma and their absence do not discard a silent genital tract infection.

Efecto de enzimas proteolíticas y de la abstinencia sexual en la cuantificación bioquímica de alfa glucosidasa en plasma seminal humano / Effects of proteolytic enzymes and sexual abstinence on alfa glucosidase levels in human seminal plasma

Background: a-glucosidase is found in human seminal plasma as an acid form, located in accessory glands, and as a neutral form secreted almost exclusively by the epididymis. Quantification of a-glucosidase activity is a marker of the secretory function of the epididymis and indemnity of the sperm transport pathway Aim: To obtain reference values for a-glucosidase in normal samples of seminal plasma, to evaluate its behavior in serial samples and to determine the effect of proteolytic enzymes. Material and methods: Fifty donors, with normal semen analysis according to the criteria of the World Health Organization, were evaluated. For the study with alpha-quimotrypsin, 0.1 to 10 mg/ml of the enzyme was added to the seminal plasma from a group of donors. a-glucosidase was also measured in semen obtained from nine patients at different time intervals. Results: Normal a-glucosidase values ranged from 14.52 to 25.69 µU/ml. Concentrations up to 10 mg/ml of alpha-quimotrypsin (10 times of that usually used in the liquefaction of the semen) did not alter the quantification of a-glucosidase. Serial determinations revealed oscillations in their magnitude, which stayed in each patient's characteristic range. However a subgroup presented a marked reduction of the activity of a-glucosidase as the abstinence diminished (40 percent). Conclusions: Evaluation of a-glucosidase in seminal plasma gives reliable information of the secretor state of the epididymis and especially replaces invasive methods used to evaluate the indemnity of the spermatic transport from the epididymis to the anterior urethra

Acrosome intactness and seminal hyaluronidase activity: relationship with conventional seminal parameters.

Seminal hyaluronidase activity was estimated after liquefaction in semen samples of 100 male partners of infertile couples including 16 azoospermic (no spermatozoon) men and 48 fertility proven men by a method based on measurement of the area of digestion of substrate (hyaluronic acid) in agar plate. Semen samples were also evaluated for Acrosomal Intactness (AI) test except the azoospermics of the studied samples. Seminal hyaluronidase activity was completely absent in azoospermic specimens confirming its cellular origin. Seminal hyaluronidase activity was found to be significantly correlated, statistically, with sperm density (r = 0.708, p < 0.001), % motility (r = 0.6478, p < 0.001) and % normal sperm morphology (r = 0.5724, p < 0.001). Acrosomal Intactness (AI) test scores were also well correlated with sperm density (r = 0.6477, p < 0.001), % motility (r = 0.5965, p < 0.001) and % normal morphology (r = 0.6237, p < 0.001). Both values were higher in semen samples with normal routine parameters (proven fertility and normozoospermic infertile groups) than those compared with abnormal routine parameters (oligozoospermic). We also found very highly significant correlation (r = 0.8442) between seminal hyaluronidase activity and Acrosomal Intactness scores, statistically (p < 0.001). This could be because; normal germinal semineferous epithelium generates abundant number of sperms with normal motility and morphology that are also having intact acrosome. Intact acrosome prevents loss of acrosomal enzymatic activity (e.g. hyaluronidase) until released after liquefaction during seminal analysis and during acrosomal reaction in female genital tract prior to fertilization. Seminal hyaluronidase activity, thus determined, is primarily dependent upon the intact status of acrosome. As each sperm contributes to the seminal hyaluronidase activity, it is directly correlated with sperm density; but at the same time it exhibits goods correlation with % motility and % normal morphology. Therefore AI score and seminal hyaluronidase activity can be considered as good indicators of sperm function.

Cigarette smoking and spermatozoa I. histochemical differences between spermatozoa of cigarette smokers and non-smokers

Thirty semen specimens were taken from non-smokers and another thirty specimens from heavy smokers. Sperms of both groups were examined histochemically for some enzymes concerned with energy production [adenosine triphosphatase, lactate dehydrogenase and succinic dehydrogenase] and penetrating power [non-specific esterase and acrosin]. The histochemical reactions for these enzymes were lower in the sperms of the smokers than those of non-smokers revealing more aspects of the detrimental effects of cigarette smoking on spermatozoa

diagnostic value of seminal - glucosidose enzyme index for sperm motility and fertilizing capacity

To find out the diagnostic value of seminal a-glucosidase enzyme index in the ejaculated semen samples of proven-fertility and primary infertility patients, for the sperm motility and fertilizing capacity. The study which lasted approximately 2 years was carried out in the Andrology Department of the Arab Fertility Center, Jeddah, Kingdom of Saudi Arabia. Eighty-five healthy male patients [age 25-38 years] were admitted into the study. They were divided into 2 main groups as proven-fertility group and primary infertility group. Each group was again subdivided into 4 classes based on the sperm concentrations. The sperm motility assessment was carried out quantitatively by Semi automated Computer assisted Semen analysis using Autosperm analyzer. Simultaneously, a-glucosidase enzyme index was determined in all the semen samples. The results were tabulated as per the standard sperm motility grades of the World Health Organisation, under the classified sperm concentrations of the study. The progressive motility grades were compared with the enzyme index of a-glucosidase in both the groups. The total progressive motility was found higher in the high-density [classes I and II] than the low-density [classes III and IV] semen samples. The enzyme index readings of the various classes of sperm concentrations were significant [p<0.05] and showed a stepwise fall from 89% +/- 9% to 52% +/- 3% in the proven fertility group. Although, a similar trend has been observed [48% +/- 5% to 20% +/- 2%] in the primary infertility group, the enzyme index class II sperm concentration [44% +/- 6%] was not found significant with class 1 [48% +/- 5%] sperm concentration, but the significance was achieved with the other 2 classes. The readings of enzyme index between the groups were found highly significant [p, 0.001]. The enzyme index of a-glucosidase estimations in semen samples may indicate a positive correlation with progressive motility and fertilizing capacity of spermatozoa and may serve as diagnostic value in individuals showing normal levels of sperm concentration and serum androgen

concentration of semen N-acety1-3-D glucosaminidase [NAG] and plasma reproductive hormones in infertile men

Semen analysis is considered to be the single most useful procedure of assessing subfertility. Thus, the level of fructose, antibodies, Zinc and reproductive hormones have been studied in different types of subfertile males. The level of N-acetyl[3-D glucosominidase [NAG] activity, an enzyme in the aerosomal cap, has also been studied but its levels have not been compared with asthenospermic and necrospermic males. The current study was conducted to evaluate the role of semen NAG activity and plasma reproductive hormone [LH, FSH and Testosterone] that correlate with the different types of subfertility in males. Thirty eight normal healthy, 24 oligospermic, 14 azoospermic, 5 asthenospermic and 2 necrospermic males between 24-45 years of age with no physical or endocrinal disorder were included in the study. Semen samples were collected with the standard procedure in sterile glass tubes and sperm count was measured by using Horwell Fertility Counting Chember. The NAG activity in seminal plasma was estimated by the fluoremetic method. Semen protein content was determined by Lowry's method. Plasma LH, FSH and testosterone in all subtypes of sub fertile men were determined by radioimmunoassay method. NAG activity in normospermics was found to be 0.349'0.0031 ug/mg protein, whereas, in oligospermic and in azoospermic males, the activity significantly [P<0.05] raised as compared to values in normospermic males. Plasma Testosterone concentration was found significantly lower in oligospermic and higher in asthenospermic whilst there was no difference between azoospermic and normosperic count. Whereas, plasma LH and FSH were significantly elevated in oligospermic and azoospermic groups. The current data indicate a direct relationship between plasma testosterone level and semen NAG activity and an inverse relationship with plasma LH/FSH. Semen NAG activity in males could be used to derive a male fertility prediction value in selected couples with no identifiable female factor of infertility

LDH isozymes with anomalous bands in semen of leprosy patients.

Activity of LDH isozymes was evaluated electrophoretically on 7% acrylamide gel in semen of 37 leprosy patients (15 with borderline, 12 with borderline tuberculoid and ten with lepromatous leprosy) and ten fertile men of 30-45 years of age. Significantly lower activities were recorded of LDH1 in all categories of leprosy patients. Similarly, lowering of LDH2 activity was noticed in borderline and lepromatous cases only, lowering of LDH4 activity in lepromatous cases only and LDH5 activity was lowered in borderline leprosy patients. Lowest activity of LDH3 and absence of LDHx were found in lepromatous leprosy. However, in borderline tuberculoid patients, LDH3 and LDHx were significantly higher. This exceptional increase in activity was found to be due to presence of additional (anomalous) isozymes bands of LDH3, LDHx and LDH4 in 25% of borderline tuberculoid patients. Additional bands of LDH3 have also been located in 40% of the borderline leprosy patients.

Efeito da insulaçäo escrotal sobre os constituintes do plasma seminal de bodes / Effect of scrotal insulation on seminal plasma of bucks

No plasma seminal de bodes da raça Moxotó e ½ sangue Moxotó-Pardo Alpina foram quantificadas a concentraçäo de frutose, ácido cítrico e proteína total, além das enzimas aspartato aminotransferase (AST) e alanina aminotransferase (ALT), em espectrofotometria, com comprimento de onda de 505 mm, antes e após aquecimento do saco escrotal. A insulaçäo do saco escrotal com bolsa plástica, de parede dupla, durante, 6,5 dias, provocou aumento expressivo na concentraçäo da frutose seminal, inversamente proporcional à concentraçäo espermática, só retornando à normalidade com a regeneraçäo do quadro espermático. A concentraçäo do ácido cítrico e da proteína total näo apresentou diferenças estatísticas significativas entre grupos genéticos, diante do desafio calórico. As concentraçöes da AST e ALT reduziram-se pela metade após o desafio térmico até o nono período (semana) e só retornaram à normalidade a partir do 10º período experimental. Dez semanas após o início do desafio escroto-testicular todos os animais mostraram regeneraçäo dos níveis de frutose, ácido cítrico, proteína total e das enzimas AST e ALT no plasma seminal

Características escroto-testiculares e do ejaculado em bodes mestiços submetidos à insulaçäo escrotal / Scrotal-testicular and ejaculation characteristics in crossbred bucks submitted to scrotal insulation

Estudaram-se as características do saco escrotal e do ejaculado de seis bodes ½ sangue Moxotó-Pardo Alpina, adultos, mantidos em confinamento, antes e após a insulaçäo do saco escrotal com bolsa plástica, de parede dupla, durante 6,5 dias. O volume apresentou-se com oscilaçöes atípicas e o aumento da temperatura no escroto afetou as características escroto-testiculares e do ejaculado de modo consistente. Os defeitos espermáticos começaram e elevar-se aos sete dias após o início da insulaçäo. A motilidade individual progressiva atingiu os menores valores à terceira semana após o início da insulaçäo, retornando aos valores normais entre a oitava e nona semanas. Entretanto, a degeneraçäo seminal ocorreu em todos os animais na quarta semana após iniciado o desafio térmico, evidenciando a diminuiçäo na concentraçäo, o aumento da patologia espermática e a reduçäo do vigor celular, culminando com acinesia e necrospermia. Dez semanas após o início do desafio escroto-testicular todos os animais mostraram regeneraçäo do perímetro escrotal, da consistência testicular e do quadro espermático

Seminal profiles of lysosomal enzymes in normal and infertile men.

Human seminal plasma is known to possess considerable proteolytic activity, much of which is associated with lysosomes. The activities of lysosomal hydrolases like alkaline proteinase, cathepsin-D, aryl-sulfatase and N-acetyl-beta-D-glucosaminidase in seminal plasma from randomly chosen infertile and vasectomised men have been compared. These enzymes have been implicated in the coagulation and liquefaction processes. The role of fructose and proteins in these processes has also been studied. The results indicate that cathepsin-D and aryl-sulfatase activity in infertile men were significantly lower than normo-spermic subjects. N-acetyl-beta-D-glucosaminidase was lowest in azoospermia suggesting that it could be used as a biochemical marker for azoospermia. Conversely, alkaline proteinase showed increased levels in all the infertile cases.

study of some enzymes in the seminal plasma of infertile males

Quantitation of enzymatic activity of alpha-glucosidase, N- acetyl- beta-glucosaminidase [NAG], beta-glucuronidase, lactate dehydrogenase and phospholipase-A2 was performed for 18 oligozoospermic, 28 azoospermic and 15 normospermic males. The study revealed significant decrement of alpha-glucosidase and phospholipase-A2 in both oligo- and azoospermic subjects compared with controls. The decreased activity was significantly marked in azoospermic subjects. NAG activity levels were insignificantly decreased in oligo- and azoospermic males in comparison with control normospermic subjects. Lactate dehydrogenase was decreased in both oligo- and azoospermic males, but the difference was significant only in azoospermic patients. Beta- glucuronidase appears to be secreted by the epididymis and the activity present in azoospermic semen can provide information concerning the etiology of the azoospermia and the functional state of the epididymis. A2 activity correlated significantly with sperm concentration [r = 0.5 and P <0.01]. The enzyme in seminal plasma seems not only to be derived from spermatozoa but also from the prostate gland and/or seminal vesicle gland. In a similar manner lactate dehydrogenase total activity in seminal plasma is derived from testicles and spermatozoa together with prostate gland and the seminal vesicles. Therefore, enzymatic measurements in seminal plasma are valuable in studying male infertility. The enzymes of special value are alpha- glucosidase, lactate dehydrogenase and phospholipase-A2