ATPases, ion exchangers and human sperm motility.

Human sperm has several mechanisms to control its ionic milieu, such as the Na,K-ATPase (NKA), the Ca-ATPase of the plasma membrane (PMCA), the Na(+)/Ca(2) (+)-exchanger (NCX) and the Na(+)/H(+)-exchanger (NHE). On the other hand, the dynein-ATPase is the intracellular motor for sperm motility. In this work, we evaluated NKA, PMCA, NHE, NCX and dynein-ATPase activities in human sperm and investigated their correlation with sperm motility. Sperm motility was measured by Computer Assisted Semen Analysis. It was found that the NKA activity is inhibited by ouabain with two Ki (7.9 × 10(-9) and 9.8 × 10(-5) M), which is consistent with the presence of two isoforms of α subunit of the NKA in the sperm plasma membranes (α1 and α4), being α4 more sensitive to ouabain. The decrease in NKA activity is associated with a reduction in sperm motility. In addition, sperm motility was evaluated in the presence of known inhibitors of NHE, PMCA and NCX, such as amiloride, eosin, and KB-R7943, respectively, as well as in the presence of nigericin after incubation with ouabain. Amiloride, eosin and KB-R7943 significantly reduced sperm motility. Nigericin reversed the effect of ouabain and amiloride on sperm motility. Dynein-ATPase activity was inhibited by acidic pH and micromolar concentrations of Ca(2) (+). We explain our results in terms of inhibition of the dynein-ATPase in the presence of higher cytosolic H(+) and Ca(2) (+), and therefore inhibition of sperm motility.

TNF-alpha in bulls experimentally infected with Trypanosoma vivax: a pilot study.

There are few studies about the immune response during trypanosomosis in cattle. The objective of this research was to evaluate the effect of experimental infection with Trypanosoma vivax (T. vivax) on serum levels of TNF-alpha in bulls and its relationship to hematocrit, body temperature and parasitemia. Two adult crossbred bulls were infected experimentally with T. vivax and two were used as controls. The bulls were evaluated during a 64 day period in terms of temperature, hematocrit, and parasitemia. Serum TNF-alpha levels were determined by ELISA, using an antibody specific for bovine. TNF-alpha in serum began rising on the seventh day after infection and reached a peak on day 40 of post-infection, then dropped. The lowest hematocrit levels corresponded to the upper levels of TNF-alpha, for each animal. In conclusion, the experimental infection of cattle with T. vivax promotes the release of TNF-alpha, demonstrating a pro-inflammatory immune response to this hemotropic parasite. Moreover, the lowest hematocrit levels coincide with high concentrations of TNF-alpha, suggesting that this cytokine can be linked to the observed anemia during the course of infection by T. vivax in cattle.

Effect of tumor necrosis factor-α on the intracellular Ca(2+) homeostasis in human sperm.

PROBLEM: Inflammation and genital infections promote the increase in leukocytes, pro-inflammatory cytokines, and oxygen reactive species, impairing sperm functions such as motility, capacitation, and acrosome reaction. All these functions are primarily regulated by cytoplasmic concentration of Ca(2+) ([Ca(2+) ]cyto ). This study evaluated the effect of tumor necrosis factor (TNF)-α on the [Ca(2+) ]cyto and its regulation in human sperm. METHOD OF STUDY: Sperm loaded with fura-2 were incubated with or without TNF-α (0-500 pg/mL) from 0 to 120 min. After incubation, the basal [Ca(2+) ]cyto and membrane permeability to Ca(2+) were evaluated by spectrofluorometry, before and after Ca(2+) addition to the extracellular medium. RESULTS: Without TNF-α, the addition of Ca(2+) promotes an transitory increase in [Ca(2+) ]cyto in the spermatozoa, that returns in a few minutes to a basal level, indicating calcium regulation activation. TNF-α decreases the Ca(2+) permeation and increases the basal level of [Ca(2+) ]cyto after a Ca(2+) pulse (P < 0.04); affecting calcium regulation in a way that is time and concentration dependent. TNF-α effect was partially prevented by the addition of an antioxidant (butylated hydroxytoluene) (P < 0.03). CONCLUSION: Tumor necrosis factor-α decreases membrane permeability to Ca(2+) and affects Ca(2+) regulation in sperm cells in vitro, probably via lipid peroxidation, which may explain the decrease in sperm fertilizing capacity during inflammatory and infectious processes.

Markers of accessory sex glands function in men with varicocele, relationship with seminal parameters.

INTRODUCTION: Varicocele has been associated with decreased semen quality, not much is known about the effect of varicocele on the accessory sex glands function. The purpose of this study was to evaluate the relationship among varicocele, seminal parameters and biochemical markers of accessory sex glands: neutral alpha glucosidase (NAG, epididymis), fructose (seminal vesicles), prostatic acid phosphatase (PAP) and zinc (prostate). MATERIALS AND METHODS: A clinical study was performed in 190 men with varicocele and 100 men normozoospermic as control group. Semen analysis, hypoosmotic swelling test (HOST), polymorphonuclear (PMN), fructose, zinc, PAP and NAG were determinate. Differences were evaluated by, t test, ANOVA and a Pearson's coefficient correlation. RESULTS: Varicocele group showed a decrease in sperm motility, normal morphology, HOST and vitality. No differences were observed in fructose, PAP and zinc levels between control and varicocele group. The NAG was significantly decreased in varicocele group. A positive correlation was observed between both fructose and PAP with semen volume, sperm concentration, PMN, and zinc levels. Additionally, a decrease of NAG was correlated with a decrease of normal sperm morphology, motility, vitality and HOST. CONCLUSIONS: Varicocele does not alter fructose secretion by seminal vesicles and PAP and zinc by prostate. Varicocele is associated with a decrease of NAG activity in seminal fluid, suggesting epididymal dysfunction possibly associated with a detrimental in sperm quality.

Serum levels of interleukin-6, interleukin-1beta and human chorionic gonadotropin in pre-eclamptic and normal pregnancy.

Studies in placentas from the first trimester and in vitro models indicate that interleukin (IL)-1beta and IL-6 induce the release of human chorionic gonadotropin (hCG). During pre-eclampsia there is an increase of pro-inflammatory cytokines; however, its relationship with hCG levels during the third trimester of pregnancy has not been determined. The aim of the present study was to evaluate the relationship between blood levels of IL-6, IL-1beta and hCG in normal pregnancy and pre-eclampsia. Blood samples during the third trimester of pregnancy from women with severe pre-eclampsia (n = 20) or normal pregnancy (n = 20) were assayed for hCG by immunoassay, IL-6 and IL-1beta by enzyme-linked immunosorbent assay. Serum level of IL-6 was significantly higher in pre-eclamptic than in normal women (16.5 +/- 2.1 vs. 4.9 +/- 1.1 pg/ml); however, IL-1beta was similar in both groups. Although hCG was higher in pre-eclampsia than normal pregnancy, the difference was not statistically significant. Furthermore, IL-1beta in normal pregnancy was correlated negatively with hCG (r = -0.69, p < 0.001). In conclusion, serum levels of IL-6 were increased in pre-eclampsia but were not correlated with hCG or IL-1beta; however, in normal pregnancy there was a negative correlation between IL-1beta and hCG. The interaction between IL-1beta and hCG at the third trimester needs to be investigated.

Sperm lipid peroxidation and pro-inflammatory cytokines.

AIM: To investigate if interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interferon-gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF-alpha) are able to stimulate the level of lipid peroxidation of sperm membranes, either alone or in the presence of leukocytes. METHODS: Semen samples from normozoospermic donors were prepared by density gradient. The sperms were exposed to the indicated cytokines, at physiological and infection-inflammation concentrations, in the absence or presence of leukocytes. Lipid peroxidation of the sperm membranes was determined by measuring malondialdehyde (MDA) and 4-hydroxialkenals (HAE) formation. RESULTS: TNF-alpha, IL-8 and IFN-gamma increased the level of sperm membrane lipid peroxidation when tested at physiological concentrations. At infection-inflammation concentrations, only IL-8 was able to produce a higher effect. When assayed in the presence of leucocytes, IL-8 and TNF-alpha showed a higher effect at infection-inflammation concentrations than at physiological concentrations. Finally, IL-8 showed a higher effect in the presence of leukocytes than in their absence at both physiological and infection-inflammation concentrations. TNF-alpha also showed a higher effect when assayed in the presence of leukocytes than in their absence, but only at infection-inflammation concentrations. There was no effect of IL-6 or IL-10 in any of the tested conditions. CONCLUSION: Several pro-inflammatory cytokines at physiological concentrations increase the level of lipid peroxidation of sperm membranes, which could be important for the sperm fecundation process. However, infection-inflammation concentrations of some cytokines, such as IL-8 and TNF-alpha, either alone or in the presence of leukocytes, could drive the lipid peroxidation of the spermatozoa plasma membrane to levels that can affect the sperm fertility capacity.

Alteraciones en la citología cervical y respuesta inmune contra Chlamydia trachomatis en trabajadoras sexuales

Resumen. Existen controversias en cuanto a la asociación entre infección por Chlamydia trachomatis (C. trachomatis) y el desarrollo de neoplasia cervical. El objetivo del presente estudio fue evaluar la relación entre la presencia de anticuerpos contra la C. trachomatis de tipo IgG en suero e IgA en moco cervical y la prevalencia de modificaciones en citología cervical. Se evaluaron 166 mujeres trabajadoras sexuales, controladas en el servicio de Infecciones de Transmisión Sexual de la Unidad Sanitaria de los Teques, Estado Miranda, Venezuela. Fueron entrevistadas y evaluadas para cervicitis y se tomó muestras de moco cervical y sangre para la determinación de anticuerpos contra C. trachomatis de tipo IgA e IgG (ELISA). Se tomaron muestras cervicales y se estudiaron por coloración de Pap. Los resultados fueron evaluados estadísticamente (Chi cuadrado, Keldal Tau). La prevalencia de IgA contra C. trachomatis en moco cervical fue de 45,2% y la de IgG en suero sanguíneo fue de 69,3%. El 38,0% de las citologías cervicales fue reportada normal o con cambios inflamatorios leves, el 35,5% inflamación moderada, el 25,3% inflamación severa y un 1,2% presentó lesiones intraepiteliales escamosas de bajo grado (LIEBG). No se encontró correlación estadísticamente significativa entre la presencia de anticuerpos contra C. trachomatis (IgA e IgG) y la de cervicitis, ni con los resultados de la citología. En conclusión, no se observó asociación entre respuesta inmune contra C. trachomatis y resultados de la citología cervical ni la presencia de cervicitis. Sin embargo, la existencia de otras infecciones simultáneas pudiera haber interferido en la observación de una posible asociación entre estas variables.

Abstract. There are controversies about the relationship between Chlamydia trachomatis (C. trachomatis) infections and development of cervical carcinoma. The objective of this investigation was to evaluate the relation between the presence of IgG against C. trachomatis in serum and IgA in cervical mucus with the prevalence of abnormalities in the Papanicolaou smears. One hundred sixty-six sexual workers were evaluated. They were checked up for sexual transmitted diseases (STD) at the "Unidad Sanitaria de Los Teques, Edo. Miranda". Sexual workers were interviewed and had a gynecological evaluation. Cervical mucus and peripheral blood samples were obtained to determine antibodies IgA and IgG against C. trachomatis. Cervical samples were taken to be evaluated by Papanicolaou staining. The association among the different variables was statistically evaluated (Chi squared and Kendal Tau). The prevalence of antibodies isotype IgA against C. trachomatis in cervical mucus was 45.2% and IgG in serum was 69.3%. Papanicolaou smear showed that 38% of women had normal or low inflammation grade, 35.5% moderate inflammation, 25.3% severe inflammation and 1.2% presented Low Grade Squamous Intraepithelial Lesion (LGSIL). There was no statistical correlation among presence of antibodies against C. trachomatis, presence of cervicitis and Papanicoloau smears. In conclusion we could not observe any relationships between immune response against C. trachomatis, cervicitis and Papanicolaou smears abnormalities. However, it is important to consider that other infections, that could be present simultaneously, could interfere in the interpretation of the results.

Bioactive to immunoreactive ratio of circulating human chorionic gonadotropin as possible evaluation for the prognosis of threatened abortion.

OBJECTIVE: In an effort to evaluate the prognosis of threatened abortion, we established the ratio of serum human chorionic gonadotropin (hCG), as measured by bioassay or radioimmunoassay techniques, of samples from patients with threatened abortion or normal pregnancy. STUDY DESIGN: Peripheral blood samples from patients in their first trimester of pregnancy with threatened abortion (n=24), or normal pregnancy (n=12), were assayed for progesterone (RIA), and for immunoactive (DELFIA) and bioactive (mouse Leydig cell testosterone production assay) hCG. RESULTS: Serum progesterone was not statistically different between the threatened-continuing and the threatened-miscarried groups. The ratio of hCG bioactive/hCG immunoactive (B/I) was significantly lower for the patients of the threatened group that experienced abortion. The B/I ratio for the control and threatened-continuing patients was similar. CONCLUSION: The hCG bioactive/hCG immunoactive ratio could be a good indicator of the prognosis of threatened abortion.

Prevalencia de hepatitis B, hepatitis C y sífilis en trabajadoras sexuales de Venezuela / Prevalence of hepatitis B, hepatitis C and syphilis in female sex workers in Venezuela

OBJETIVO: En Venezuela las trabajadoras sexuales reciben un control sanitario para la sífilis y el virus de inmunodeficiencia humana (VIH). Sin embargo, otras importantes infecciones de transmisión sexual no son evaluadas. Así, se realizó este estudio con el objetivo de determinar el nivel socio-cultural de un grupo de trabajadores sexuales y su relación con la sero-presencia de marcadores de Hepatitis C y Hepatitis B, en adición a la evaluación de rutina. MÉTODOS: Se evaluaron 212 trabajadoras sexuales, que acudieron al control sanitario en el servicio de infecciones de transmisión sexual, de la ciudad de Los Teques, Venezuela. Fueron entrevistadas en cuanto a edad, nivel educativo, uso de anticonceptivos y del condón. Se les tomó una muestra de sangre para determinar sífilis, antígeno de superficie de hepatitis B (HBsAg) y la presencia de anticuerpos contra el core de hepatitis B (anti-HBc), virus de hepatitis C (anti-HC) y VIH. Los datos fueron evaluados estadísticamente por Chi-cuadrado y correlación de Pearson. RESULTADOS: La prevalencia fue de 2,4por cento para sífilis, 0,5por cento para anti-HC, 3,8por cento para HBsAg y 13,8por cento para anti-HBc. Un aumento en la prevalencia de marcadores de hepatitis B se correlacionó con un bajo nivel educativo (p<0,05) e incremento en la edad (p<0,05). No se encontró ningún caso positivo de VIH. La encuesta reveló que el 38,5por cento de las trabajadoras sexuales nunca utilizan el condón y un 25,6por cento de ellas no utiliza ningún tipo de método anticonceptivo. CONCLUSIONES: Es necesario implementar planes de inmunización de hepatitis B en este grupo de mujeres, así como campañas de educación sobre la importancia del uso del condón para disminuir la probabilidad de contraer infecciones de transmisión sexual

[Changes in cervical cytology and immune response against Chlamydia trachomatis in sexual workers]. / Alteraciones en la citología cervical y respuesta inmune contra Chlamydia trachomatis en trabajadoras sexuales.

There are controversies about the relationship between Chlamydia trachomatis (C. trachomatis) infections and development of cervical carcinoma. The objective of this investigation was to evaluate the relation between the presence of IgG against C. trachomatis in serum and IgA in cervical mucus with the prevalence of abnormalities in the Papanicolaou smears. One hundred sixty-six sexual workers were evaluated. They were checked up for sexual transmitted diseases (STD) at the "Unidad Sanitaria de Los Teques, Edo. Miranda". Sexual workers were interviewed and had a gynecological evaluation. Cervical mucus and peripheral blood samples were obtained to determine antibodies IgA and IgG against C. trachomatis. Cervical samples were taken to be evaluated by Papanicolaou staining. The association among the different variables was statistically evaluated (Chi squared and Kendal Tau). The prevalence of antibodies isotype IgA against C. trachomatis in cervical mucus was 45.2% and IgG in serum was 69.3%. Papanicolaou smear showed that 38% of women had normal or low inflammation grade, 35.5% moderate inflammation, 25.3% severe inflammation and 1.2% presented Low Grade Squamous Intraepithelial Lesion (LGSIL). There was no statistical correlation among presence of antibodies against C. trachomatis, presence of cervicitis and Papanicoloau smears. In conclusion we could not observe any relationships between immune response against C. trachomatis, cervicitis and Papanicolaou smears abnormalities. However, it is important to consider that other infections, that could be present simultaneously, could interfere in the interpretation of the results.