Cardiopathic effects of dichloroacetate in the fetal Long-Evans rat.

Dichloroacetic acid (DCA) is a by-product of the chlorine disinfection of water and may occur in treated water at levels exceeding 100 micrograms/L. Previous studies revealed teratogenic effects, particularly heart malformations, at high doses (900-2,400 mg/kg given on days 6-15 of pregnancy). In a series of three studies, groups of 7-10 Long-Evans rats were dosed with 1,900 mg/kg of DCA on days 6-8, 9-11, or 12-15; with 2,400 mg/kg on days 10, 11, 12, or 13; and with 3,500 mg/kg on days 9, 10, 11, 12, or 13, in an attempt to determine the most sensitive period and further characterize the heart defect. In a fourth study, six dams were treated with 1,900 mg/kg of DCA days 6-15 of pregnancy, and 56 fetuses were harvested for light microscopy of the heart. Eight control fetuses from four litters were also examined. No heart malformations were seen in the groups treated with 1,900 mg/kg DCA days 6-8 but were present in the group treated on days 9-11 and 12-15, with the higher incidence occurring on days 12-15. Single doses of 2,400 mg/kg DCA given on days 10, 11, 12, or 13 resulted in a much lower incidence of cardiac malformations, which occurred only on days 10 and 12. The high dose of DCA (3,500 mg/kg) did not increase the incidence of heart defects but showed that dosing on day 9 as well as on days 10 and 12 would produce the defect. The defects seen were characterized as high interventricular septal defects (H-IVSD). Light microscopy showed that the defect was caudal to the semilunar valves, with the anterior right wall of the aorta communicating with the right ventricle. Another aspect of the defect is at the level of the semilunar valves, with the right cusp or sinus of Valsalva in communication with the right ventricle. The defects are discussed more fully and methods for further study suggested.     

Sub-compartmentation of the 'cytosolic' glucose 6-phosphate pool in cultured rat hepatocytes

[14C]Glucose release either from endogenous 14C-prelabelled glycogen or from added 14C-labelled glucose 6-phosphate was measured in filipin-treated, permeabilized hepatocytes in 48 h culture. [14C]Glucose output from prelabelled glycogen was not altered by the addition of 5 mM glucose 6-phosphate to the incubation medium. Conversely, [14C]glucose release from 5 mM labelled glucose 6-phosphate was not influenced by different glycogen concentrations in the cells. Moreover, in the permeabilized cells the anion transport inhibitor DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid) inhibited only the liberation of [14C]glucose from labelled glucose 6-phosphate but not from glycogen. It is therefore concluded that there exist at least 2 separate, mutually non-accessible glucose 6-phosphate pools in cultured rat hepatocytes, one linked to glycogenolysis and the other to gluconeogenesis.     

Membrane responses to large hyperpolarizations in trabecles and single cells of frog atrium

Atrial trabeculae (studied in voltage-clamping conditions and in the presence of 0.5 mmol/l BaCl2 to abolish gK1) responded to 1 s hyperpolarizations to beyond approximately E = -140 mV (from HP of about E = -80 mV) with an inwardly directed current increasing with time. Quite similar results were obtained with enzymatically dissociated frog atrial cells studied in whole cell voltage clamp with a patch-clamp pipette. This behaviour could be accounted for by assuming the presence of an "if" current at this quite negative range of potentials or by the fact that the cell membrane may undergo reversible electropermeabilization when its potential is brought to values negative to about -140 mV (St?mpfli 1958). When a brief (1 ms) and large (150 mV) hyperpolarization was applied 1 s before the test pulse, an inwardly directed current increasing with time was elicited by test pulses to beyond approximately E = -120 mV. This current was neither abolished in the presence of 1 mmol/l CsCl nor greatly reduced in the absence of Na+ ions, unlike "if" (Di Francesco 1981). We conclude that this current having a time course similar to that of "if" is of different nature and we argue that it might be accounted for by electropermeabilization of the membrane (reversible within about 2.5 min) due to the electrical shock represented by a brief and large hyperpolarization.     

Monensin inhibits the secretion of α-amylase but not polysaccharide slime from seedling tissues ofZea mays

Summary The effect of monensin on polysaccharide slime secretion by root tips of corn (Zea mays) was studied. Various treatment times and ionophore concentrations were tested: none resulted in inhibition of slime secretion. Because monensin changes the pH of the medium, its effect was also monitored in strongly buffered media and at different pH's. Even in such media, monensin did not inhibit slime secretion. We also measured the effect of the drug after a pulse with [³H]fucose or a pulse followed by a chase. The amount of labeled slimed secreted was not altered by the ionophore. However, 10M monensin affected the development of root tips and drastically reduced their growth. We showed that monensin inhibits the secretion of -amylase by the scutellum of the same plantlet. The importance of the nature of the secretory compound in relation to monensin inhibition of its secretion is discussed.Abbrevations Hepes N-2-hydroxyethylpiperazine-N-2-ethane-sul-fonic acid - Mes 2-(N-morpholino)ethane-sulfonic acid     

Effects of MET-ENK, substance P and SRIF on the behavior of Hemichromis bimaculatus

Administration of 10 and 30 micrograms methionine-enkephalin (MET-ENK)/g bw (n = 10/dose) affected the propensity towards fighting in H. bimaculatus; 10 micrograms increased, while 30 micrograms decreased the aggressive behavior. MET-ENK also affected a number of behavior patterns displayed by the fish. Moreover, the "wet-dog-shakes" observed suggest that MET-ENK acts on opiate-receptors. Treatment with substance P (SP)/g bw (n = 10/dose) induced chafing movements in the fish slightly. It also decreased fighting and increased biting of the air stone, which is evidence that H. bimaculatus is still aggressive, directing its attacks to different objects. When 4, 8, 12 micrograms somatostatin (SRIF)/g bw (n = 10/dose) were injected, H. bimaculatus stopped fighting for several hours after the onset of treatment, depending on the dosage. Somatostatin reduces blood glucose concentration, causing a sudden stop of aggressive behavior, 0.04, 0.1, 0.6, 1.0 and 3.0 IU prolactin (PRL)/g bw (n = 5/dose) eventually decreased fighting and affected a number of behavior patterns displayed by the fish.     

Antagonistic regulation of the glucose/glucose 6-phosphate cycle by insulin and glucagon in cultured hepatocytes.

Flux through the glucose/glucose 6-phosphate cycle in cultured hepatocytes was measured with radiochemical techniques. Utilization of [2-3H]glucose was taken as a measure of glucokinase flux. Liberation of [14C]glucose from [U-14C]glycogen and from [U-14C]lactate, as well as the difference between the utilization of [2-3H]glucose and of [U-14C]glucose, were taken as measures of glucose-6-phosphatase flux. At constant 5 mM-glucose and 2 mM-lactate concentrations insulin increased glucokinase flux by 35%; it decreased glucose-6-phosphatase flux from glycogen by 50%, from lactate by 15% and reverse flux from external glucose by 65%, i.e. overall by 40%. Glucagon had essentially no effect on glucokinase flux; it enhanced glucose-6-phosphatase flux from glycogen by 700%, from lactate by 45% and reverse flux from external glucose by 20%, i.e. overall by 110%. At constant glucose concentrations cellular glucose 6-phosphate concentrations were essentially not altered by insulin, but were increased by glucagon by 230%. In conclusion, under basic conditions without added hormones the glucose/glucose 6-phosphate cycle showed only a minor net glucose uptake, of 0.03 mumol/min per g of hepatocytes; this flux was increased by insulin to a net glucose uptake of 0.21 mumol/min per g and reversed by glucagon to a net glucose release of 0.22 mumol/min per g. Since the glucose 6-phosphate concentrations after hormone treatment did not correlate with the glucose-6-phosphatase flux, it is suggested that the hormones influenced the enzyme activity directly.     

Quantitative investigations on the effect of pentamethylenetetrazol on the supraoptic neurones in rabbit

Summary Quantitative investigations were made on the effect of convulsant doses of pentamethylenetetrazol (PMT) on the neurones of the supraoptic nucleus in rabbit. In contrast to results reported by other workers, the present investigation yielded statistical evidence for an increase in the number of Gomori-positive elements in the supraoptic nucleus following epileptic seizures induced by PMT. The factors that might be responsible for this phenomenon are discussed. It is pointed out that a satisfactory interpretation cannot be given on the basis of histological findings alone. The problem requires further investigation with other techniques, such as electron microscopy and biochemical methods.Dedicated to Professor Dr. Dr. h.c. H. Spatz on the occasion of his 80th birthday.     

Determination of agonist dissociation constants in isolated vasculature: equivalence of estimates obtained by the method of partial irreversible receptor inactivation and a novel application of the operational model of pharmacological agonism

Steady-state contractile responses elicited by activation of the proportional 1-adrenergic and 5-HT2 receptors in isolated rat and rabbit aorta, respectively, were analyzed. Agonist dissociation constants (KA's) obtained by the method of partial irreversible receptor inactivation were compared to KA values determined by fits of the operational model of pharmacological agonism to single concentration response curves (CRCs). The observed nature of the KA estimates obtained with the Furchgott method for phenylephrine (PE) and oxymetazoline (OXY) at the proportional 1-adrenergic receptor and for 5-hydroxytryptamine (5-HT) at the 5-HT2 receptor in isolated rat aorta, and for PE and 5-HT at the proportional 1-adrenergic and 5-HT2 receptors, respectively, in isolated rabbit aorta, was consistent with the hypothesis that the density of membrane receptors is greatly in excess of the density of transducer proteins (i.e., [Ro] much greater than [To]) in these systems. Therefore, KA, efficacy and slope factor estimates were also obtained by computer fits of the operational model to single agonist CRCs in both rat and rabbit aorta, with the empirically determined tissue maximal response (Tmax) substituted for the theoretical parameter Em. In all cases, the mean pKA estimates obtained with the operational model closely approximated and were strongly correlated with the mean pKA estimates determined by the Furchgott method. These studies suggest that, at least in some vascular preparations, Tmax is a good estimate of Em, and moreover, that Em may be not only a specific characteristic of a given receptor-effector system as previously demonstrated by Black and Leff, but that Em may also describe a more general feature of tissue responsiveness that is shared among distinct membrane receptors coupled to similar effector systems. In conclusion, when receptor inactivation studies have indicated that the condition [Ro] much greater than [To] exists, Tmax can be substituted into the operational model to provide valid estimates of agonist KA values at distinct receptor subtypes, in the absence of receptor alkylation.