Oviductal secretion and gamete interaction

Experimental evidence from the last 30 years supports the fact that the oviduct is involved in the modulation of the reproductive process in eutherian mammals. Oviductal secretion contains molecules that contribute to regulation of gamete function, gamete interaction, and the early stages of embryo development. The oviductal environment would act as a sperm reservoir, maintaining sperm viability, and modulating the subpopulation of spermatozoa that initiates the capacitation process. It could also contribute to prevent the premature acrosome reaction and to reduce polyspermy. Many studies have reported the beneficial effects of the oviductal environment on fertilization and on the first stages of embryo development. Some oviductal factors have been identified in different mammalian species. The effects of oviductal secretion on the reproductive process could be thought to result from the dynamic combined action (inhibitory or stimulatory) of multiple factors present in the oviductal lumen at different stages of the ovulatory cycle and in the presence of gametes or embryos. It could be hypothesized that the absence of a given molecule would not affect fertility as its action could be compensated by another factor with similar functions. However, any alteration in this balance could affect certain events of the reproductive process and could perhaps impair fertility. Thus, the complexity of the reproductive process warrants a continuous research effort to unveil the mechanisms and factors behind its regulation in the oviductal microenvironment.Fil: Ghersevich, Sergio Albino. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaFil: Massa, Estefanía María Alejandra. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Zumoffen, Carlos María. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; Argentin

Mammaglobin A: overview and clinical utility

Mammaglobin A is a protein that belongs to the secretoglobin superfamily. It has highly specific expression in cells from most breast cancers and may be used to detect circulating or disseminated tumor cells. In addition, mammaglobin A is currently under investigation as a potential therapeutic target for immune therapies that target breast cancer. The present review will highlight our current understanding of mammaglobin A at the genetic and protein level and its potential clinical applications. Characteristics of breast cancer and methods used to isolate and detect circulating tumor cells will also be presented.Fil: Ceballos Mancini, María Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Fisiología Experimental (i); ArgentinaFil: Ghersevich, Sergio Albino. Universidad Nacional de Rosario; Argentin

Assessing endocrine and immune parameters in human immunodeficiency virus-infected patients before and after the immune reconstitution inflammatory syndrome

ABSTRACT Objective The present study compares immune and endocrine parameters between HIV-infected patients who underwent the Immune Reconstitution Inflammatory Syndrome (IRIS-P) during antiretroviral therapy (ART) and HIV-patients who did not undergo the syndrome (non-IRIS-P). Materials and methods Blood samples were obtained from 31 HIV-infected patients (15 IRIS-P and 16 non-IRIS-P) before ART (BT) and 48 ± 2 weeks after treatment initiation (AT). Plasma Interleukin-6 (IL-6) and Interleukin-18 (IL-18) were determined by ELISA. Cortisol, dehydroepiandrosterone sulfate (DHEA-S) and thyroxin concentrations were measured using chemiluminescence immune methods. Results Concentrations of IL-6 (7.9 ± 1.9 pg/mL) and IL-18 (951.5 ± 233.0 pg/mL) were significantly higher (p 0.05). Levels of DHEA-S in IRIS-P decreased AT (1080.5 ± 124.2 vs. 782.5 ± 123.8 ng/mL, p < 0.05) and they were significantly lower than in non-IRIS-P (782.5 ± 123.8 vs. 1203.7 ± 144.0 ng/mL, p < 0.05). IRIS-P showed higher values of IL-6 and IL-18 BT and lower levels of DHEA-S AT than in non-IRIS-P. Conclusion These parameters could contribute to differentiate IRIS-P from non-IRIS-P. The significant decrease in DHEA-S levels in IRIS-P after ART might suggest a different adrenal response in these patients, which may reflect the severity of the disease

Assessing endocrine and immune parameters in human immunodeficiency virus-infected patients before and after the immune reconstitution inflammatory syndrome

Objective: The present study compares immune and endocrine parameters between HIV-infected patients who underwent the Immune Reconstitution Inflammatory Syndrome (IRIS-P) during antiretroviral therapy (ART) and HIV-patients who did not undergo the syndrome (non-IRIS-P). Materials and methods: Blood samples were obtained from 31 HIV-infected patients (15 IRIS-P and 16 non-IRIS-P) before ART (BT) and 48 ± 2 weeks after treatment initiation (AT). Plasma Interleukin-6 (IL-6) and Interleukin-18 (IL-18) were determined by ELISA. Cortisol, dehydroepiandrosterone sulfate (DHEA-S) and thyroxin concentrations were measured using chemiluminescence immune methods. Results: Concentrations of IL-6 (7.9 ± 1.9 pg/mL) and IL-18 (951.5 ± 233.0 pg/mL) were significantly higher (p 0.05). Levels of DHEA-S in IRIS-P decreased AT (1080.5 ± 124.2 vs. 782.5 ± 123.8 ng/mL, p < 0.05) and they were significantly lower than in non-IRIS-P (782.5 ± 123.8 vs. 1203.7 ± 144.0 ng/mL, p < 0.05). IRIS-P showed higher values of IL-6 and IL-18 BT and lower levels of DHEA-S AT than in non-IRIS-P. Conclusion: These parameters could contribute to differentiate IRIS-P from non-IRIS-P. The significant decrease in DHEA-S levels in IRIS-P after ART might suggest a different adrenal responseFil: Rateni, Liliana Beatriz. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina.Fil: Lupo, Sergio. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentina.Fil: Lupo, Sergio. Center for Assistance and Comprehensive Clinical Research (CAICI); Argentina.Fil: Racca, Liliana. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina.Fil: Palazzi, Jorge. Center for Assistance and Comprehensive Clinical Research (CAICI); Argentina.Fil: Ghersevich, Sergio. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina

Relationship between genotoxic effects of breast cancer treatments and patient basal DNA integrity

It is accepted that radiotherapy and chemotherapy cause genotoxic effects as part of their therapy side effects, which are highly variable among different patients. This study evaluated DNA integrity using the comet assay in peripheral blood lymphocytes from breast cancer patients before (“pre-treatment patients”; n = 47) and after (“post-treatment patients”; n = 24) radiotherapy and/or chemotherapy treatment and from healthy donors (n = 15). Comet evaluation was made by visual (types 0-4) and digital (percentage of DNA remaining in the comet head = % head DNA) analysis. The association between the level of DNA damage and cancer prognostic factors was assessed. The treatments caused a significant increase in DNA damage registered by both visual (p < 0.001) and digital (p < 0.001) analysis. No significant associations between the level of DNA damage in pre-treatment patients and cancer prognostic factors were found. Results showed a significant correlation between the comet results from each patient before and after treatment (r = 0.64, p = 0.001). The % head DNA in post-treatment samples from patients with high level of DNA damage before treatment (30.3 ± 3.1%, p < 0.01) was lower than in post-treatment samples from patient with low to medium level of DNA damage before therapy (49.2 ± 4.4%). Results support the usefulness of the comet assay as a sensitive technique to evaluate the basal DNA status and the damage caused by cancer treatments. The assay could contribute to the treatment decisions, taking into account the patients’ basal DNA damage before therapy.Fil: Capitaine Funes, Juan. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Ceballos, María Paula. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental (UNR-CONICET); ArgentinaFil: Capitaine Funes, Juan. Provincia de Santa Fe. Hospital Provincial del Centenario. Servicio de Mastología; ArgentinaFil: Massa, Estefania. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Cipulli, Germán. Provincia de Santa Fe. Hospital Provincial del Centenario. Servicio de Mastología; ArgentinaFil: Benitez Gil, Alfonso. Provincia de Santa Fe. Hospital Provincial. Servicio de Mastología; ArgentinaFil: Capitaine Funes, Carlos. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Capitaine Funes, Carlos. Provincia de Santa Fe. Hospital Provincial del Centenario. Servicio de Mastología; ArgentinaFil: Tozzini, Roberto. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Tozzini, Roberto. Provincia de Santa Fe. Hospital Provincial del Centenario. Servicio de Mastología; ArgentinaFil: Ghersevich, Sergio. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin

Assessing endocrine and immune parameters in human immunodeficiency virus-infected patients before and after the immune reconstitution inflammatory syndrome

<div><p>ABSTRACT Objective The present study compares immune and endocrine parameters between HIV-infected patients who underwent the Immune Reconstitution Inflammatory Syndrome (IRIS-P) during antiretroviral therapy (ART) and HIV-patients who did not undergo the syndrome (non-IRIS-P). Materials and methods Blood samples were obtained from 31 HIV-infected patients (15 IRIS-P and 16 non-IRIS-P) before ART (BT) and 48 ± 2 weeks after treatment initiation (AT). Plasma Interleukin-6 (IL-6) and Interleukin-18 (IL-18) were determined by ELISA. Cortisol, dehydroepiandrosterone sulfate (DHEA-S) and thyroxin concentrations were measured using chemiluminescence immune methods. Results Concentrations of IL-6 (7.9 ± 1.9 pg/mL) and IL-18 (951.5 ± 233.0 pg/mL) were significantly higher (p < 0.05) in IRIS-P than in non-IRIS-P (3.9 ± 1.0 pg/mL and 461.0 ± 84.4 pg/mL, respectively) BT. Mean T4 plasma level significantly decreased in both groups of patients after treatment (p < 0.05). In both groups cortisol levels were similar before and after ART (p > 0.05). Levels of DHEA-S in IRIS-P decreased AT (1080.5 ± 124.2 vs. 782.5 ± 123.8 ng/mL, p < 0.05) and they were significantly lower than in non-IRIS-P (782.5 ± 123.8 vs. 1203.7 ± 144.0 ng/mL, p < 0.05). IRIS-P showed higher values of IL-6 and IL-18 BT and lower levels of DHEA-S AT than in non-IRIS-P. Conclusion These parameters could contribute to differentiate IRIS-P from non-IRIS-P. The significant decrease in DHEA-S levels in IRIS-P after ART might suggest a different adrenal response in these patients, which may reflect the severity of the disease.</p></div

Effect of exposure to ulipristal acetate on sperm function

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Objective A pill containing ulipristal acetate (UPA) is used for emergency contraception (EC). Considering that, following its intake, spermatozoa may be exposed to UPA in the female genital tract we intended to evaluate sperm functions after incubation with this compound. Methods Motile spermatozoa were selected by swim-up and were incubated under capacitating conditions with UPA (at concentrations of 1, 10, 100, 1,000, and 10,000 ng/ml) or control medium. The main outcome measures were sperm vitality, sperm protein tyrosine phosphorylation (TyrP), spontaneous acrosomal reaction (AR), and human follicular fluid (hFF)-induced AR. Results Sperm vitality and TyrP pattern were similar between spermatozoa exposed to UPA or control. In addition, spontaneous AR ranged from 14.0. +/- 1.5% to 18.0. +/- 1.9% after exposure to UPA or control medium without significant differences, and UPA did not prevent hFF-induced AR. Conclusions Incubation of sperm with UPA at concentrations around the expected plasma levels after ingestion of this EC pill (similar to 100-200 ng/ml) did not modify the signal transduction of TyrP involved in sperm capacitation. Moreover, UPA showed no agonist effect on progesterone receptors because it did not induce AR. Considering that progesterone in hFF is essential for AR induction, and UPA did not prevent the hFF-induced AR, an antagonist action of UPA on the AR is unlikely.176428437Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

S100 A9 is expressed and secreted by the oviduct epithelium, interacts with gametes and affects parameters of human sperm capacitation in vitro

Our previous findings demonstrate that some oviductal secretion proteins bind to gametes and affect sperm physiology and gamete interaction. One of these proteins possesses an estimated molecular weight of 14 kDa. The objective of this study was to isolate and identify this 14 kDa protein, to localize it in the human oviduct, to detect gamete binding sites for the protein, and to evaluate its effects on sperm capacitation parameters and gamete interaction. Explants from the human oviductal tissues of premenopausal women were cultured in the presence of [35S]-Methionine-proteins ([35S]-Met-proteins). De novo synthesized secreted [35S]-Met-proteins were isolated from the culture media by affinity chromatography using their sperm membrane binding ability and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Using liquid chromatography-tandem mass spectrometry peptide sequencing, human S100 A9 was identified as one of the isolated proteins from the 14 kDa protein band. S100 A9 was detected in oviduct epithelium and oviduct secretion using immunohistochemistry and a Western blot. S100 A9 binding to human oocytes and spermatozoa was assessed by indirect immunofluorescence. The acrosome reaction (AR) affected S100 A9 ability to bind sperm cells. The presence of S100 A9 significantly increased both the induced AR and the sperm protein tyrosine phosphorylation, with respect to controls. However, the protein did not affect sperm-zona pellucida interaction. Results indicate that S100 A9 is present in the human oviduct and that it modulates parameters of sperm capacitation in vitro. Hence, the protein might contribute to the regulation of the reproductive process in the oviductal microenvironment.Fil: Massa, Estefanía María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Bioquímica Clínica; ArgentinaFil: Prez, Gastón Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Bioquímica Clínica. Bioquímica Clínica; ArgentinaFil: Zumoffen, Carlos María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Bioquímica Clínica. Bioquímica Clínica; ArgentinaFil: Morente, Carlos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Bioquímica Clínica. Bioquímica Clínica; ArgentinaFil: Ghersevich, Sergio Albino. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Bioquímica Clínica. Bioquímica Clínica; Argentin

Effects of lactoferrin, a protein present in the female reproductive tract, on parameters of human sperm capacitation and gamete interaction

In a recent study, lactoferrin (LF) was detected in human oviductal secretion. The protein was able to bind to oocytes and sperm, and modulated gamete interaction. The aim of the present study was to investigate the effect of LF on parameters related to human sperm capacitation and sperm-zona pellucida interaction. Semen samples were obtained from healthy normozoospermic donors (n = 7). Human follicular fluids and oocytes were collected from patients undergoing in vitro fertilization. Motile sperm obtained by swim-up were incubated for 6 or 22 h under capacitating conditions with LF (0-100 μg/mL). After incubations, viability, motility, presence of α-d-mannose receptors (using a fluorescent probe on mannose coupled to bovine serum albumin), spontaneous and induced acrosome reaction (assessed with Pisum sativum agglutinin conjugated to fluorescein isothiocyanate), and tyrosine phosphorylation of sperm proteins were evaluated. Sperm-zona pellucida interaction in the presence of LF was investigated using the hemizone assay. The presence of LF did not affect sperm viability or motility, but caused a dose-dependent significant decrease in sperm α-d-mannose-binding sites, and the effect was already significant with the lowest concentration of the protein used after 22 h incubation. Dose-dependent significant increases in both induced acrosome reaction and tyrosine phosphorylation of sperm proteins were observed in the presence of LF. The present data indicate that LF modulates parameters of sperm function. The inhibition of gamete interaction by LF could be partially explained by the decrease in sperm d-mannose-binding sites. The presence of the LF promoted sperm capacitation in vitro.Fil: Zumoffen, Carlos María. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Massa, Estefanía María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Caille, A. M.. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Munuce, María José. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Ghersevich, Sergio Albino. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin

Efectos de lactoferrina in vivo e in vitro sobre parámetros reproductivos en ratas

Lactoferrina (LF) está presente en el tracto reproductor femenino. La proteína es expresada y secretada por el tejido oviductal humano y puede unirse a las gametas. La presencia de LF fue capaz de reducir la interacción de gametas humanas y modular la función espermática in vitro. El objetivo de este estudio fue investigar el efecto de LF sobre parámetros reproductivos in vivo (tasas de nacimiento y preñez) e in vitro (tasa de fertilización in vitro -FIV-) en ratas Wistar. Ratas hembra (80 días) fueron asignadas aleatoriamente a uno de cuatro grupos de tratamiento y recibieron una inyección diaria durante el ciclo estral de 100 mg LF/kg (n=5), 200 mg LF/kg (n=5), 400 mg LF/kg (n=5) o 0,9% w/v cloruro de sodio (controles, n=7). En el día del proestro, a las ratas hembras tratadas se les permitió aparearse con un macho y se documentó el número de hembras preñadas y de crías nacidas de cada grupo. Se obtuvieron espermatozoides móviles de cauda epididimario de ratas macho (100-140 días). Se recuperaron ovocitos del oviducto de ratas (n=9) luego de la estimulación hormonal ovárica y los mismos fueron colocados en grupos de a 3 o 4 en gotas de 100 µl de medio HTF, inseminados con espermatozoides e incubados en ausencia (controles) o presencia de LF (100 μg/ml) a 37 ºC y 5% PCO2 por 24 hs. Después de la incubación, se evaluó el número de ovocitos fertilizados por tinción con Hoechst 33258 en un microscopio de fluorescencia y se estimó la tasa de fecundación in vitro. Los resultados indicaron que el 64% de los animales controles quedaron preñados. Ninguna de las ratas tratadas in vivo con 100 mg LF/kg resultó preñada mientras que los tratamientos con 200 mg y 400 mg LF/kg resultaron en 63% y 38% de animales preñados, respectivamente. La dosis de 100 mg LF/kg se asoció significativamente con la ausencia de preñez (p<0,05). La dosis de 200 mg LF/kg redujo significativamente el número de crías respecto de los controles (7,5 2,0 vs. 11,6 0,5, respectivamente p < 0,01). El número de crías observado con la dosis de 400 mg LF/kg (13,0 0,4) no difirió de los controles. La recuperación promedio de ovocitos de cada rata estimulada hormonalmente fue de: 17,2 ± 3,3 ovocitos/rata. La tinción fluorescente demostró que la presencia de LF disminuyó significativamente el promedio de la tasa de FIV respecto de controles (33,3 ± 8,5)% vs. (60,7 ± 6,6) %, respectivamente (n=9, p<0,05). Los datos sugieren que LF podría afectar el proceso de fecundación y la preñez en ratas, tal vez en forma dosis dependiente, dado que distintas dosis administradas in vivo causaron efectos distintos.Fil: Massa, Estefanía María Alejandra. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Lo Celso, Agustina Laura. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaFil: Armesto, Rubina. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaFil: Madariaga, Maria Jose. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaFil: Pelusa, Fabián. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaFil: Ghersevich, Sergio Albino. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Departamento de Bioquímica Clinica. Bioquímica Clinica; ArgentinaXXI Congreso y XXXIX Reunión Anual de la Sociedad de Biología de RosarioRosarioArgentinaSociedad de Biología de Rosari