Application of a fluorometric method to measure glutamate release from single retinal photoreceptors.

Release of the endogenous transmitter, glutamate, was measured from individual cone photoreceptors using a microfluorometric technique. The assay for glutamate was conducted within the lumen of a suction pipette, and was based on the fluorometric measure of the production of NADH from NAD+. This reaction was catalyzed by glutamate dehydrogenase contained in the pipette. Upon introduction of glutamate to the pipette, an increase in the NADH fluorescence was observed, representing the stoichiometric conversion of glutamate to NADH. The fluorescent signal was quantified, allowing an estimate of glutamate release from a single cone upon depolarization. The release observed was elicited upon depolarization of the cell with extrinsic current, and was detectable simultaneous with stimulation of the cell. Depolarization-induced release of endogenous glutamate was from the synaptic pedicle of the cell, and this release decreased with subsequent stimulations. The decrease in the release could be briefly reversed by an increase in the depolarization current used, or by allowing the cell to rest for several minutes.     

Histological, histochemical and electron microscopic changes of the placenta induced by maternal exposure to hyperthermia in the rat.

Both clinical and experimental investigations have shown that maternal hyperthermia during critical stages of embryo development can induce malformations in the offspring. Studies of the effect of heat stress on the placental functions are limited to the ewes, but that on microscopic structure is unknown. In the present study, rats were exposed to 41 or 42 degrees C for 1 h on gestation day (GD) 9. The controls were sham treated. Fetuses and placentas were collected on GD 20. Intrauterine growth retardation (IUGR) and several craniofacial malformations were observed in the fetuses of the heat-treated group. The placentas of the 42 degrees C group were significantly lighter in weight than those of the control. Light microscopy (LM) revealed thickening, hyalinization and occasional lymphocytic infiltration of the decidua basalis. Giant cells were prominent and glycogen cells had degenerated, leaving behind large cysts in the basal (spongy) zone. Best's carmine stain with or without diastase indicated the reduction in number and degeneration of glycogen cells and cyst formation. The labyrinthine zone was relatively thin in comparison to that of the controls. Perivascular fibrosis and paucity of vascularization were other features of the placentas of the hyperthermia group. Electron microscopy (EM) revealed lipid droplet accumulation in the trophoblast, the presence of myelin bodies and an increased production of collagen in the basal zone. Perivascular fibrosis appeared to have contributed to placental barrier thickening. EM also revealed accumulation of glycogen and lipid droplets in the trophoblasts and fibrin secretion into the extracellular space of the labyrinthine zone. These data suggest that placental pathology possibly contributes to fetal growth retardation in maternally heat-stressed rat fetuses.     

Early inhibition of natural and interferon-activated killers in endometrial cancer patients treated with local radiotherapy

The present study was aimed at comparing the effect of clinical staging and radiotherapy on natural killer (NK) and interferon-activated killer (IAK) cell activity in a group of endometrial cancer patients receiving a total dose of 5,000 to 8,000 rads. We report that when compared to age-matched women, a significantly higher number and percentage of patients show low NK and IAK cell activity. At diagnosis, diminished NK activity was seen in about 20% of the patients, while IAK activity was low in 49% of these patients. There was no correlation between these deficiencies and the grade or stage of the disease. In contrast, radiotherapy induced deleterious effects on both populations of NK and IAK cells. These deleterious effects were more pronounced in patients showing a low level of spontaneous NK activity. In an attempt to understand better the mechanism by which the presence of cancer itself and/or radiotherapy affects these activities, we studied in greater detail changes in peripheral blood T-cell numbers and subsets. Before radiotherapy, all lymphocyte counts were within the normal range. In contrast, after radiotherapy the absolute numbers of all T-cell subsets were significantly decreased in the majority of the patients tested, OKT4+ cells being the most radiosensitive and Leu 7+ cells the most radioresistant.     

Eleventh week amniocentesis for prenatal diagnosis of some metabolic diseases.

We report the results of 23 prenatal diagnoses performed at the 11th or 12th week of gestation by the simultaneous analysis of chorionic villi (for direct or indirect enzymatic analysis) and cell-free amniotic fluid (for search of accumulated catabolites). For six cases of citrullinaemia, four cases of argininosuccinic aciduria, seven cases of propionic acidaemia, and six cases of methylmalonic acidaemia, three discrepancies were observed between the two methods used. The amniotic fluid analysis for accumulated catabolites seems to be a safe method and should always be used in conjunction with the enzymatic assays performed for the prenatal diagnosis of these diseases.     

Emergency peripartum hysterectomy.

A total of 54 166 mothers delivered at the Riyadh Armed Forces Hospital between 1990 and 1997, including 6119 (11.3%) caesarean sections. Emergency peripartum hysterectomy for obstetric haemorrhage was carried out in 16 cases (0.3/1000 deliveries). The operation followed major degrees of placenta praevia in 12 (75%) cases and atonic postpartum haemorrhage in four (25%). All patients required blood transfusion. There was one neonatal death and no maternal deaths. Although the operation was straightforward, bladder injury occurred in five (31%) cases which was repaired with no residual damage. Placenta accreta was confirmed histologically in 12 (75%) patients. In conclusion, all obstetricians should be aware of the strong association between a scarred uterus, placenta praevia and placenta accreta which can be very adherent and difficult to remove causing bleeding and necessitating hysterectomy. The operation should be performed by an experienced obstetrician before the patient's condition is extreme.