Extra-virgin olive oil ameliorates high-fat diet-induced seminal and testicular disorders by modulating the cholesterol pathway.

BACKGROUND: Rabbits are sensitive to dietary cholesterol and rapidly develop hypercholesterolemia, leading to prominent subfertility. Sterol regulatory element-binding protein isoform 2 drives the intracellular cholesterol pathway in many tissues, including the testicles. Its abnormal regulation could be the mainly responsible for the failure of suppressing cholesterol synthesis in a cholesterol-enriched environment, ultimately leading to testicular and seminal alterations. However, extra-virgin olive oil consumption has beneficial properties that promote lowering of cholesterol levels, including the recovery of seminal parameters altered under a high-fat diet. OBJECTIVES: Our goal was to investigate the effects of high-fat diet supplementation with extra-virgin olive oil at the molecular level on rabbit testes, by analyzing sterol regulatory element-binding protein isoform 2 protein and its corresponding downstream effectors. MATERIALS AND METHODS: During 12 months, male rabbits were fed a control diet, high-fat diet, or 6-month high-fat diet followed by 6-month high-fat diet plus extra-virgin olive oil. Serum lipids, testosterone levels, bodyweight, and seminal parameters were tested. The mRNA and protein levels of sterol regulatory element-binding protein isoform 2, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, and low-density lipoprotein receptor were determined by semi-quantitative polymerase chain reaction and Western blotting techniques. The expression pattern of sterol regulatory element-binding protein isoform 2 protein in the rabbit testicles was studied by indirect immunofluorescence. In addition, testicular cholesterol was detected and quantified by filipin staining and gas chromatography. RESULTS: The data showed that the addition of extra-virgin olive oil to high-fat diet reduced testicular cholesterol levels and recovered the expression of sterol regulatory element-binding protein isoform 2, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, and low-density lipoprotein receptor initially altered by the high-fat diet. DISCUSSION AND CONCLUSIONS: The combination of high-fat diet with extra-virgin olive oil encourages testicular recovery by modifying the expression of the enzymes related to intracellular cholesterol management.

Fibrosis quística: incremento de sobrevida y función pulmonar / Cystic Fibrosis: increased survival and lung function

Objetivo. Se presentan las curvas espirométricas de pacientes que constituyen la población global de un Centro de Fibrosis Quística, en sus valores de VEF1% y FMF25-75%, y se evaluaron distintos parámetros relacionados con ellas: descenso de función pulmonar con la edad y disminución anual porcentual de la misma; también se evalúa el descenso en el tiempo para la adecuada toma de decisiones. Método. La población ya fue estudiada y mostró en sus cohortes del 1999, 2002, 2005 y 2008, un incremento significativo de sobrevida progresivo en cada una de ellas. En el trabajo actual para estudio de VEF1% y FMF25-75% se crean las cohortes 2001 y la cohorte 2010: se seleccionan 11 pacientes con 3 a 5 registros de VEF1% (2001) y 14 pacientes con 3 a 5 registros de VEF1% (2010). Se realiza el análisis de los promedios de los valores medios por año. Se usa el coeficiente de Pearson para establecer la correlación lineal de los valores de VEF1% y FMF25-75% y cuando se comparan curvas se usa el test de student(t). Resultados. Las personas con fibrosis quística en 2010 tienen mejor función pulmonar comparadas con las de la cohorte del 2001. El análisis también muestra que el VEF1% desciende 1,8% por año; el FMF25-75% desciende 2,4% por año. La diferencia entre las rectas que describen la evolución temporal VEF1% y FMF25-75% es estadísticamente significativa para un nivel de significación de 0,0005. Se puede concluir que el FMF25-75% predice el descenso de la función respiratoria con anterioridad al VEF1%. Los valores así determinados permitirán una mejor estimación del caso individual y tomar medidas tales como la revisión de los tratamientos, el adelanto de la solicitud de transplante pulmonar y la evaluación en profundidad del funcionamiento del Centro de Fibrosis Quística.

Objective. Spirometric results for the Cystic Fibrosis Center are presented. FEV1% and FMF25-75% were studied for pulmonary function decrease with age and percent annual decrease. Method. In a previous publication the whole Cystic Fibrosis Center population was divided into the 1999, 2002, 2005 and 2008 cohorts. Increased survival was shown for each cohort. In the present study of FEV1% and FMF25-75% we grouped patients in the 2001 and 2010 cohorts: 11 patients with 3 to 5 values of FEV1% were selected for the 2001 cohort and 14 patients for the 2010 cohort. We calculated the annual mean for these measurements. Pearson coefficient for linear regression was used for values of FEV1% and FMF25-75%. To compare results for these two cohorts we used the student´s test (t). Results. Cystic Fibrosis patients for the 2010 cohort show higher spirometric FEV1% values compared to the 2001 cohort. FEV1% annual percentage decrease was 1,8% and FMF25-75% annual percentage decrease was 2,5%. Difference between regression lines for FEV1% and FMF25-75% is significant for p = 0,0005.We conclude that FMF25-75% shows decreased pulmonary functions before FEV1%. These results will allow for timely consideration of the individual patient´s spirometric course and at the same time evaluate the Cystic Fibrosis Center performance.

Effects of chronic hypoxia on electrogenic transport and transport-related oxygen consumption in rat distal colon.

The distal colon epithelium of rats submitted to chronic hypoxia shows higher short-circuit current (Isc) which, unlike non-hypoxic rat epithelium, has an amiloride-sensitive component despite low serum aldosterone levels. Isc and oxygen consumption (QO2) were simultaneously measured in mucosae from rats submitted to 0.5 atm for 10 days and from control rats in a modified Ussing chamber. Hypoxia increased Isc but not QO2. The slope of the regression line between Isc and QO2 reduction after ouabain addition was decreased in epithelia from hypoxic rats (P = 0.03). Chloride secretion blockade reduced Isc and QO2 in both groups, while sodium channel blockade did so only in the hypoxic group. Dual blockade in hypoxic rat epithelia caused correlated (P = 0.0025) additive decreases in Isc and QO2. Presented results suggest that chronic hypoxia induces an improved coupling between QO2 and electrogenic ion transport, and electrogenic sodium absorption despite low aldosterone levels.

Single-channel response of hamster oocytes to fertilization with homologous spermatozoa.

Electrophysiological events occur early after fertilization, along with changes in intracellular Ca2+ concentration. Passive electrical parameters were determined in golden hamster oocytes by whole cell patch-clamp method. In separate experiments the effect of 4-aminopyridine on resting oocytes was tested. The single-channel patch clamp configuration was employed to assess the electrical response to fertilization with homologous sperm. Structure of oocytes submitted to patch clamp was evaluated with scanning electron microscopy and found to be preserved. Oocyte diameter was 70.2 +/- 2.2 microm; their resting parameters were: membrane potential 23.8 +/- 0.8 mV; total membrane specific resistance 519.1 +/- 94.6 ohms.cm2, and specific capacity 0.99 +/- 0.03 microF.cm(-2). Total membrane current was decreased by 42 % by 4-aminopyridine. Control oocytes and oocytes exposed to sperm differed in their membrane currents in response to a voltage ramp clamping membrane potential from - 100 mV to + 100 mV. In both cases, currents were largest at the most negative potentials, but sperm-exposed oocytes had larger currents. Additionally, while in control oocytes the current was inward at negative potentials but outward at positive potentials, in the presence of spermatozoa oocytes was inward within the whole voltage range tested. This latter current may represent Ca2+ entry.

Single-channel response of hamster oocytes to fertilization with homologous spermatozoa

Electrophysiological events occur early after fertilization, along with changes in intracellular Ca2+ concentration. Passive electrical parameters were determined in golden hamster oocytes by whole cell patch-clamp method. In separate experiments the effect of 4-aminopyridine on resting oocytes was tested. The single-channel patch clamp configuration was employed to assess the electrical response to fertilization with homologous sperm. Structure of oocytes submitted to patch clamp was evaluated with scanning electron microscopy and found to be preserved.Oocyte diameter was 70.2 ± 2.2 µm; their resting parameters were: membrane potential 23.8 ± 0.8 mV; total membrane specific resistance 519.1 ± 94.6 Ù.cm2, and specific capacity 0.99 ± 0.03 µF.cm-2. Total membrane current was decreased by 42 % by 4-aminopyridine.Control oocytes and oocytes exposed to sperm differed in their membrane currents in response to a voltage ramp clamping membrane potential from - 100 mV to + 100 mV. In both cases, currents were largest at the most negative potentials, but sperm-exposed oocytes had larger currents. Additionally, while in controloocytes the current was inward at negative potentials but outward at positive potentials, in the presence of spermatozoa oocytes was inward within the whole voltage range tested. This latter current may represent Ca2+ en try

Single-channel response of hamster oocytes to fertilization with homologous spermatozoa

Electrophysiological events occur early after fertilization, along with changes in intracellular Ca2+ concentration. Passive electrical parameters were determined in golden hamster oocytes by whole cell patch-clamp method. In separate experiments the effect of 4-aminopyridine on resting oocytes was tested. The single-channel patch clamp configuration was employed to assess the electrical response to fertilization with homologous sperm. Structure of oocytes submitted to patch clamp was evaluated with scanning electron microscopy and found to be preserved.Oocyte diameter was 70.2 ± 2.2 Am; their resting parameters were: membrane potential 23.8 ± 0.8 mV; total membrane specific resistance 519.1 ± 94.6 Ù.cm2, and specific capacity 0.99 ± 0.03 AF.cm-2. Total membrane current was decreased by 42 % by 4-aminopyridine.Control oocytes and oocytes exposed to sperm differed in their membrane currents in response to a voltage ramp clamping membrane potential from - 100 mV to + 100 mV. In both cases, currents were largest at the most negative potentials, but sperm-exposed oocytes had larger currents. Additionally, while in controloocytes the current was inward at negative potentials but outward at positive potentials, in the presence of spermatozoa oocytes was inward within the whole voltage range tested. This latter current may represent Ca2+ en try(AU)

Single-channel response of hamster oocytes to fertilization with homologous spermatozoa

Electrophysiological events occur early after fertilization, along with changes in intracellular Ca2+ concentration. Passive electrical parameters were determined in golden hamster oocytes by whole cell patch-clamp method. In separate experiments the effect of 4-aminopyridine on resting oocytes was tested. The single-channel patch clamp configuration was employed to assess the electrical response to fertilization with homologous sperm. Structure of oocytes submitted to patch clamp was evaluated with scanning electron microscopy and found to be preserved.Oocyte diameter was 70.2 ± 2.2 Am; their resting parameters were: membrane potential 23.8 ± 0.8 mV; total membrane specific resistance 519.1 ± 94.6 Ù.cm2, and specific capacity 0.99 ± 0.03 AF.cm-2. Total membrane current was decreased by 42 % by 4-aminopyridine.Control oocytes and oocytes exposed to sperm differed in their membrane currents in response to a voltage ramp clamping membrane potential from - 100 mV to + 100 mV. In both cases, currents were largest at the most negative potentials, but sperm-exposed oocytes had larger currents. Additionally, while in controloocytes the current was inward at negative potentials but outward at positive potentials, in the presence of spermatozoa oocytes was inward within the whole voltage range tested. This latter current may represent Ca2+ en try(AU)

Chronic hypobaric hypoxia effects on rat colon in vitro sensitivity to acute hypoxia and amiloride.

Chronic hypoxia induces many physiological changes, but little is known about its effects on colonic epithelial function. Isolated distal colon mucosa from rats under normobaric conditions and rats submitted to hypobaric hypoxia for either 4 or 10 days was studied in an Ussing chamber. After 4 days of hypoxia, there was only a 15% increase in transepithelial resistivity. However, 10-day hypoxic rats showed higher short circuit current, potential difference, and resistivity. In this group, but not in normal or 4-day hypoxic animals, amiloride dose-dependently depressed short circuit current. The response to acute hypoxia in vitro was unchanged after chronic hypoxia and was not affected by amiloride. Although the amiloride-sensitive increase in short circuit current in 10-day hypoxic rats might resemble mineralocorticoid action, resistivity was increased and serum aldosterone was very low. It is suggested that chronic hypoxia may enhance electrogenic sodium transport by an aldosterone-independent mechanism.