In vitro effects of two environmental toxicants, beauvericin and glyphosate in Roundup, on cell numbers and steroidogenesis of bovine ovarian cells.

Beauvericin is an emerging Fusariotoxin naturally occurring in cereal grains throughout the world whereas glyphosate (N-phosphonomethyl-glycine) is a non-selective systemic herbicide used worldwide. The purpose of this study is to evaluate a newly developed ovarian cell culture system (that includes both granulosa and theca cells) as an in vitro model for toxicological studies. Specifically, the effects of beauvericin and glyphosate in formulation with Roundup on ovarian cell numbers and steroid production were evaluated. Ovaries collected from cattle without luteal structures were sliced into 30-70 pieces each, and granulosa and theca cells were collected. Harvested cells were cultured for 48 h in 10% fetal bovine serum-containing medium followed by 48 h in serum-free medium containing testosterone (500 ng/mL; as an estrogen precursor) with the following eight treatments: (1) controls, (2) FSH (30 ng/mL) alone, (3) FSH plus insulin-like growth factor-1 (IGF1; 30 ng/mL), (4) FSH plus IGF1 plus beauvericin (3 µM), (5) FSH plus IGF1 plus glyphosate in Roundup (10 µg/mL), (6) FSH plus IGF1 plus fibroblast growth factor 9 (FGF9, 30 ng/mL), (7) a negative control without added testosterone, and (8) IGF1 plus LH (30 ng/mL) with basal medium without added testosterone. In the presence of FSH, IGF1 significantly increased cell numbers, estradiol and progesterone production by severalfold. Glyphosate in Roundup formulation significantly inhibited IGF1-induced cell numbers and estradiol and progesterone production by 89-94%. Beauvericin inhibited IGF1-induced cell numbers and estradiol and progesterone by 50-97% production. LH plus IGF1 significantly increased androstenedione secretion compared with controls without added testosterone indicating the presence of theca cells. In conclusion, the present study demonstrates that toxicological effects of beauvericin and glyphosate in Roundup formulation are observed in a newly developed ovarian cell model system and further confirms that both glyphosate and beauvericin may have the potential to impair reproductive function in cattle.

Effects of N-carbamylglutamate and arginine on steroidogenesis and proliferation of pig granulosa cells in vitro.

Results of in vivo studies indicate dietary N-carbamylglutamate (NCG) and arginine (ARG) can enhance reproductive performance in gilts. It was hypothesized that both NCG and ARG will alter hormone-induced estradiol (E2) production by granulosa cells (GC), explaining why these compounds could improve reproductive performance in pigs. The objective of these studies, therefore, was to evaluate the direct effects of NCG and ARG on porcine GC proliferation and steroidogenesis, using an in vitro cell culture system. The GC from small (SM; 1-5 mm) and large (LG; >5 mm) pig follicles were cultured for 2 days in 5% fetal bovine serum and 5% porcine serum-containing medium followed by 2 days in serum-free medium containing 500 ng/mL of testosterone (as an E2 precursor), and NCG or ARG at various doses in the presence of either follicle-stimulating hormone (FSH; 30 ng/mL), insulin-like growth factor-1 (IGF1; 30 ng/mL), or both. Numbers of GC were determined at the end of the experiment and concentrations of progesterone (P4) and E2 in culture medium were determined. Results indicated that LG-follicle GC were more responsive to NCG and ARG than SM-follicle GC. Specifically, in LG-follicle GC, NCG inhibited (P <  0.05) basal and FSH-induced P4 and E2 production but stimulated cell numbers; whereas ARG inhibited FSH-induced E2 production and cell numbers. In SM-follicle GC, treatment with NCG and ARG decreased IGF1 plus FSH induced P4 production, but E2 production and cell proliferation were not affected. These studies indicate that NCG and ARG may directly affect follicular function in pigs.

Effect of melatonin on bovine theca cells in vitro.

Melatonin affects granulosa cell function in several species but its function in theca cells is less clear, particularly in monotocous animals. Thus, the objectives of this study were to determine the effects of melatonin on theca cell steroidogenesis, gene expression and cell proliferation in a monotocous species, namely cattle. Ovaries were collected from a local bovine abattoir, from which theca cells were isolated from large (8-22mm) follicles and treated with various hormones in serum-free medium for 24h or 48h. Melatonin caused a dose-dependent inhibition (P<0.05) of LH+insulin-like growth factor 1 (IGF1)-induced androstenedione and progesterone production. Also, melatonin inhibited (P<0.05) LH+IGF1-induced expression of steroidogenic acute regulatory protein (StAR) mRNA (via real-time polymerase chain reaction) in theca cells, but it had no effect (P>0.10) on cytochrome P450 11A1 (CYP11A1) and cytochrome P450 17A1 (CYP17A1) mRNA abundance. In LH+IGF1-treated theca cells, melatonin decreased caspase 3 (CASP3) mRNA to levels similar to those observed in LH-treated theca cells. In contrast, melatonin increased (P<0.05) the number of bovine theca cells in both LH- and LH+IGF1-treated cultures. In conclusion, melatonin may act as an endocrine regulator of ovarian function in cattle by stimulating theca cell proliferation and inhibiting differentiation via inhibition of hormone-induced steroidogenesis.

Effects of N-carbamylglutamate and L-arginine on steroidogenesis and gene expression in bovine granulosa cells.

Feeding N-carbamylglutamate (NCG) and arginine (ARG) improves reproductive measures in pigs and reduces systemic steroid levels in pregnant ewes. We hypothesized that the effects of NCG and ARG on reproduction were due to direct effects on the ovary. Thus, the objectives of this study were to investigate the effects of NCG and ARG on granulosa cell (GC) steroidogenesis, gene expression, and cell proliferation in vitro. GC were collected from small (1-5mm) bovine follicles and treated in vitro with NCG or ARG in serum-free medium for 24h to 48h. Both NCG and ARG inhibited (P<0.05) IGF1- and FSH-induced GC estradiol production but only NCG inhibited (P<0.05) progesterone production. In contrast, NCG and ARG increased (P<0.05) GC numbers induced by IGF1 and FSH. NCG inhibited (P<0.05) StAR, CYP11A1 and CYP19A1 mRNA abundance in small-follicle GC, whereas ARG had no effect (P>0.10) on StAR, CYP11A1 or CYP19A1 mRNA abundance. We conclude that NCG and ARG may act directly on GC and therefore may regulate ovarian function by slowing follicular differentiation via inhibiting IGF1 action, and steroid synthesis while stimulating GC proliferation in cattle.

Medición no invasiva de bilirrubina en el recién nacido. Significativo aporte a la práctica clínica en la ictericia neonatal / Non invasive measurement of serum bilirubin in newborns: significant contributions to the clinical practice in the jaundiced newborn

Objetivos del trabajo: 1) comparar la estimación de bilirrubina sérica obtenida por medición transcutánea, BiliCheck, con valores obtenidos espectrofotométricamente y por high performance liquid chromatography (Gold Standard), 2) Determinar puntos de corte de los valores de BiliCheck en los cuales se obtiene la máxima sensibilidad y especificidad, 3) Determinar si el color de la piel, peso de nacimiento, edad gestacional y sexo interfieren con la medición transcutánea de bilirrubina en recién nacidos

Medición no invasiva de bilirrubina en el recién nacido. Significativo aporte a la práctica clínica en la ictericia neonatal / Non invasive measurement of serum bilirubin in newborns: significant contributions to the clinical practice in the jaundiced newborn

Objetivos del trabajo: 1) comparar la estimación de bilirrubina sérica obtenida por medición transcutánea, BiliCheck, con valores obtenidos espectrofotométricamente y por high performance liquid chromatography (Gold Standard), 2) Determinar puntos de corte de los valores de BiliCheck en los cuales se obtiene la máxima sensibilidad y especificidad, 3) Determinar si el color de la piel, peso de nacimiento, edad gestacional y sexo interfieren con la medición transcutánea de bilirrubina en recién nacidos

Beneficios de la preparación industrial de soluciones dextrosadas e hidroeléctricas para uso neonatal / Benefits of the industrial preparations of dextrose and hydroelectrolytic solutions for use neonatal

Con la hipótesis que el uso de soluciones dextrosadas e hidroeléctricas preparadas mediante técnica industrial no utiliza tiempo de enfermería para su preparación, disminuye la incidencia de contaminación microbiana accidental y aumenta la exactitud de su composición química, llevamos a cabo un trabajo de investigación con los siguientes objetivos: 1) determinar si la composición de las fórmulas propuestas para la PI es apropiada para la hidratación en el período neonatal, 2) comparar dos técnicas de preparación : preparación por enfermería (PE) y preparación industrial (PI). Propusimos 4 fórmulas; N§ 1 y N§ 2 conteniendo dextrosa al 5 por ciento y al 10 por ciento respectivamente y 40 mEq/1 de Na y 20 mEq/1 de K en ambas; N§ 3 dextrosa al 10 por ciento, 20 mEq/1 sin K. Los objetivos se cumplieron en tres etapas. Etapa 1: soluciones PE con enfermeras dedicadas al mismo tiempo al cuidado de recien nacidos en terapia intensiva. Etapa 2: soluciones PI. Etapa 3: soluciones PE con enfermeras dedicadas exclusivamente a la preparación de la solución sin tarea asistencial. Las fórmulas propuestas cubrieron el 80 por ciento de las soluciones indicadas en la unidad de cuidado intensivo neonatal. El tiempo utilizado por enfermería para la preparación de las soluciones PE fue mayor en la etapa 1 que en la 3: X 5,22 minutos vs. 3,98 min. (p<.0005). Se detectó contaminación microbiana en 2/57 (3,5 por ciento) de soluciones PE de la etapa 1 vs 0 por ciento en soluciones PE de etapa 3 y soluciones PI. Se encontraron diferencias significativas en la composición química de Na (p<.01) y K (p<.05) entre las soluciones PE 1§ etapa y PI; en las soluciones PE 3§ etapa persistió una mayor variabilidad con respecto a soluciones PI y una diferencia significativa para el K (p<0.05) pero no se observó diferencias significativas en la composición de Na. En conclusión las soluciones PI presentan mayor exactitud y precisión en su composición química, menor riesgo de contaminación microbiana y no se utiliza tiempo de enfermería para la preparación. Persiste la necesidad de las soluciones PE en 20 por ciento; de los casos; dichas soluciones deberían ser preparadas por enfermeras dedicadas exclusivamente a esta tarea dado que se observó menor variabilidad en la composición química, menor tiempo requerido para su preparación y menor riesgo de contaminación microbiana comparándolas con soluciones preparadas por enfemeras dedicadas a la tarea asistencial.

Beneficios de la preparación industrial de soluciones dextrosadas e hidroeléctricas para uso neonatal / Benefits of the industrial preparations of dextrose and hydroelectrolytic solutions for use neonatal

Con la hipótesis que el uso de soluciones dextrosadas e hidroeléctricas preparadas mediante técnica industrial no utiliza tiempo de enfermería para su preparación, disminuye la incidencia de contaminación microbiana accidental y aumenta la exactitud de su composición química, llevamos a cabo un trabajo de investigación con los siguientes objetivos: 1) determinar si la composición de las fórmulas propuestas para la PI es apropiada para la hidratación en el período neonatal, 2) comparar dos técnicas de preparación : preparación por enfermería (PE) y preparación industrial (PI). Propusimos 4 fórmulas; Nº 1 y Nº 2 conteniendo dextrosa al 5 por ciento y al 10 por ciento respectivamente y 40 mEq/1 de Na y 20 mEq/1 de K en ambas; Nº 3 dextrosa al 10 por ciento, 20 mEq/1 sin K. Los objetivos se cumplieron en tres etapas. Etapa 1: soluciones PE con enfermeras