Age-related expression of TGF beta family receptors in human cumulus oophorus cells.

PURPOSE: During ovarian follicle growth, local cellular interactions are essential for oocyte quality acquisition and successful fertilization. While cumulus cells (CCs) nurture oocytes, they also deliver oocyte-secreted factors (OSFs) that activate receptors on CCs. We hypothesized that disturbance of those interactions contributes to age-related lower reproductive success in women submitted to assisted reproductive technology treatments. METHODS: Women aged 27-48, without recognized personal reproductive disorder, were enrolled in the study and divided in <35- and ≥35-year-old groups. CCs collected upon follicle aspiration were processed for immunocytochemistry and RNA extraction. The expression patterns of OSF receptors BMPR2, ALK 4, ALK5, and activin receptor-like kinase (ALK6) were studied. RESULTS: Independently of age, receptors were found mostly in the cell periphery. The quantitative assay revealed that in older women, BMPR2, ALK 4, and ALK6 were all significantly decreased, whereas ALK5 was slightly increased. CONCLUSIONS: Female age imparts an effect on the expression of OSF receptors in CCs. The findings indicate that reproductive aging affects the local regulation of signaling pathways mediated by BMPR2, ALK6, and ALK4 receptor activation, suggesting their joint involvement.

Superoxide dismutase expression in human cumulus oophorus cells.

Success in assisted reproductive techniques (ART) is influenced by gamete and embryo quality but the assessment of these parameters has been thwarted by the lack of reliable biomarkers. Follicular fluid and cumulus oophorus cells may provide biomarkers due to their close relationship to the oocyte. These cells produce antioxidants and thus protect the oocyte from oxidative damage exerted by reactive oxygen species (ROS). ROS and antioxidants are known to intervene in reproductive physiology and pathology, but their roles are unclear. It is hypothesized that superoxide dismutase (SOD), a first line antioxidant enzyme, is associated with oocyte quality. Cells obtained in the course of ART for the treatment of infertility due to male factor or female pathology were processed for SOD intracellular isoforms (CuZnSOD and MnSOD) immunodetection, total SOD activity and isoforms content. Cells presented strong positive staining for CuZnSOD and MnSOD. SOD activity decreased with increasing female age but was increased in endometriosis and in ovulatory dysfunction. When male factor was the cause for infertility, successful ART was associated with higher SOD activity. Variations in SOD emphasize the relevance of oxidative stress in the oocyte maturation process. These variations also suggest that SOD is a potential biomarker for ART success.

Low-dose oral contraceptives and 24-hour ambulatory blood pressure.

OBJECTIVE: To evaluate in normotensive women the influence of low-dose oral contraceptives (OC, monophasic formulations containing 30 microg of estrogen) on 24-h blood pressure. METHODS: We evaluated prospectively in 15 normotensive healthy women (three smokers) the influence of OC on 24-h ambulatory blood pressure monitoring (ABPM). ABPM was performed (SpaceLabs 90207) before and after 6-9 months of use of OC. We also evaluated ABPM in eight women (two smokers) before and after 6-8 months on an intrauterine device (IUD) as contraceptive method--these were used as control subjects. RESULTS: OC produced a significant increase in 24-h ABPM values (from 120+/-3/75+/-2 to 128+/-4/81+/-2 mmHg, P < 0.04) which was particularly evident for night-time values (from 108+/-2/64+/-2 to 120+/-4/73+/-2 mmHg, P < 0.02). After OC, two normotensive women developed 'hypertensive values'. In OC users there was a slight but significant increase in body weight which did not correlate with the increase of blood pressure. In contrast, in the control group (IUD) neither ABPM values nor weight were modified by the contraceptive maneuver. CONCLUSIONS: In normotensive women, low-dose OC may increase blood pressure to an extent that, at least in some women, may affect blood pressure control towards 'hypertensive values'. This stresses the importance of monitoring blood pressure values during OC treatment.

Inter-observer agreement in analysis of basal body temperature graphs from infertile women.

In this study, we evaluated the reproducibility of analysis of basal body temperature graphs under optimized conditions for agreement. A total of 160 recordings were selected from spontaneous cycles of infertile women and analysed by three experienced clinicians using uniform criteria. Agreement at the various stages of analysis was assessed by the 'proportions of agreement' with 95% confidence intervals. Agreement in identification of the thermal nadir was clearly superior to that reported in previous publications. Reproducibility of 'ovulatory' graph features (i.e. biphasic graphs and adequate thermal shifts) was excellent. Agreement in classification of monophasic graphs and inadequate thermal shifts, although lower, was still good. Thus, with experienced observers and uniform criteria, a good agreement can be achieved in analysis of the most important parameters of the basal body temperature graph. We believe that an effort should be made to generalize the use of uniform analysis criteria, because only then can results from different institutions be compared and the remaining clinical evaluation of the method be performed.

Hypothermia-induced Haemostatic and Biochemical Phenomena. An Experimental Model.

Hypothermia (24°C) was induced in 7 healthy male dogs. Hematological, hemostatic and biochemical parameters were evaluated before the onset and after 3 h of hypothermia. Following hypothermia, there was a significant increase in red cell count, hematocrit and haemoglobin concentration. The white cell and platelet counts were decreased significantly and the mean platelet volume increased significantly. The activated partial thromboplastin time was significantly prolonged whereas the prothrombin time was unchanged. Associated with these changes there was a significant increase in serum glucose and a small increase in aspartate transaminase activity probably due to muscle leakage. No change occurred in the more liver-specific alanine transaminase. The arterial pH decreased and the changes in gasses observed as a result of hypothermia were compatible with a mixed respiratory-metabolic acidosis. The mean aortic blood pressure also dropped markedly. The hematological and hemostatic changes suggest that widespread tissue 'injury' occurs even in short term hypothermia with platelet activation and accumulation in the liver and spleen. This model of hypothermia may be of use in the study of the thrombotic diathesis observed in this condition and serve as a test bed for drugs of potential use in conditions such as Raynaud's syndrome and hypothermic surgery (e.g. coronary artery bypass).

[Primary effect of carotid body stimulation on left ventricular inotropism in swine]. / Efeito primário da estimulação do corpo carotideo sobre o inotropismo ventricular esquerdo no porco.

The cardiovascular response to carotid body stimulation has been a interesting and simultaneously controversial subject. In fact, several authors described different responses depending on the animal species used (cat, dog, rabbit, monkey), time of stimulation (short-term v. long-term) and experimental conditions and procedures. According to our experience, the response to carotid body stimulation with cyanide, in normal breathing dogs, consists in hyperpnoea, bradycardia with arterial blood pressure fall (by means of longer diastolic periods), followed by a marked rise in arterial blood pressure and heart rate, after curarization. Concerning the inotropic response to carotid chemoreceptors stimulation, there exists some controversial and even antagonic opinions. In fact, some authors describe a positive inotropic effect, others a negative inotropic effect, and others still defend no inotropic response at all. In the present work, the authors try to contribute to the solution of this problem, studying a group of animals (pig) anaesthetised with chloralose, in which the major modulators of the inotropic response to carotid body stimulation (ventilation; heart rate; and ventricular load conditions) were controlled. To the evaluation of cardiac contractile state, indexes derived from the systolic isovolumic phase were used (maximum left ventricular pressure, dP/dtmax; dP/dtmin) in all the animals. In some animals, other indexes were also used (Vmax, Vpm, (dP/dt)/DP40, Tau and PB). The authors concluded that, in the present experimental conditions, the primary response to carotid body stimulation, in the pig, includes a positive inotropic effect.

[Effects induced hypothermia on hepatic portal circulation]. / Effets de l'hypothermie provoquée sur la circulation portale hépatique.

Studies of the liver circulation in dogs during hypothermia, showed that portal blood flow is reduced with no significant changes of blood pressure and with important vascular resistance. These effects are probably due to the contraction of pre-capillaries sphincters.

[Effect of induced hypothermia on the systemic circulation]. / Effet de l'hypothermie provoquée dans la circulation systémique.

Studies of the systemic circulation in dogs (n = 5) during hypothermia showed that cardiac output, mean arterial pressure, total peripheral resistance, pulse rate, work L. V. is reduced and the stroke volume is increased. The authors think that these effects are probably due to metabolic alterations during hypothermia.

[Effect of induced hypothermia on the capillary circulation of the liver]. / L'effet de l'hypothermie provoquée dans la circulation capillaire du foie.

Micro-angiographic studies of the liver, in dogs under established hypothermy, showed that the contrast injected in the portal vein does not perfuse the sinusoidal structures. This fact is, probably, due to the contraction of pre- and post-capillaries sphincters. On the contrary, in dogs under normal temperature the injected contrast largely perfused the capillaries and all supra-heptic vascular system.

[Effect of phentolamine on carotid reflexes induced by acetylcholine]. / L'action de la phentolamine sur les réflexes carotidiens déclenchés par l'acétylcholine.

Acetylcholine was injected into the common carotid artery of 15 mongrel dogs. In the animals with tied internal carotid artery, hyperpnoea, bradycardia and hypotension was observed, and in the animals with free internal carotid artery the hyperphoea was followed by apnoea. The administration of phentolamine provokes a temporary decrease or abolition of the reflexes produced by acetylcholine. This action of the alpha-blocker is due to a local effect because when phentolamine is injected intravenously it does not happen and the reflexes produced by acetylcholine with starting place at the carotid sinus of the opposite side are not affected.

[Effect of propranolol and pindolol on carotid reflexes induced by lobeline]. / Action du propranolol et du pindolol sur les réflexes cartidiens déclenchés par la lobéline.

1. On the carotid reflexogenic zone, pindolol provokes a proper action which consists in a respiratory excitation and a diminution of cardiac rate accompanied by decrease of blood pressure. 2. The infiltration of the tissues of the carotid bifurcation with a solution of propranolol annuls temporarily the carotid lobeline reflexes probably by the propranolol local anaesthetic action. 3. The beta blockers propranolol and pindolol, when injected in the vessels of the carotid zone, inhibits temporarily the carotid reflexes of lobeline by a local action on the carotid body. These results suggest that in the excitation of the receptor system of the carotid body by lobeline interferes an adrenergic mechanism.