Detection and quantification of patulin and griseofulvin by high pressure liquid chromatography in different strains of Penicillium griseofulvum Dierckx

Patulin and griseofulvin production by twelve strains ofPenicillium griseofulvum Dierckx, eleven of which were isolated from pistachio (Pistacia vera) nuts and the other was supplied by the Spanish Collection of Type Culture, was investigated. Six strains of the eleven isolated had ability to produce patulin and griseofulvin in Yes medium. All the strains studied had no ability to produce patulin in Wickerham medium. Griseofulvin production was significant in both media but higher in Wickerham. These metabolites were separated and determined in the chloroform extracts of cultures by high performance liquid chromatography with ultraviolet detection. The best conditions were: acetonitrile — water (45∶55) as mobile phase, a flow rate of 2.0 mL/min and a μ Bondapack C₁₈ column.     

Isolation of highly multidrug-resistant P388 cells from drug-sensitive P388/S cells by flow cytometric cell sorting

To investigate the spontaneous frequency of occurrence of stable multidrug-resistant cells in a population of drug-sensitive cells, we exposed drug sensitive P388/S cells to daunorubicin (dnr) for 1 h, then used fluorescence-activated cell sorting based on intracellular dnr fluorescence to isolate cells within P388/S having different intracellular content of drug. One of the sort windows chosen (low dnr content sort window) isolated only P388/S cells with intracellular drug content equal to or less than that of the known multidrug-resistant subline P388/adr. This sort window constituted approximately 3% of P388/S cells with lowest dnr content. By such a procedure we were able, on one of seven attempts, to isolate and cultivate stable, highly multidrug-resistant cells (comparable to that of P388/adr) from the P388/S cells obtained from the low dnr-content sort window. Net growth of cells in culture was observed 15-20 days after sorting, indicating that of the P388/S cells collected from the low dnr-content sort window, very few were actually highly drug-resistant. On no occasion could resistant cells be cultivated from cells sorted from P388/S with higher dnr content, as would be expected if mutation to a multidrug-resistant phenotype had occurred as a result of exposure to drug. The resistant cells isolated from P388/S by sorting (called P388/LoSort) displayed low intracellular accumulation of dnr that was enhanced by verapamil, were cross-resistant to vincristine and actinomycin-D, and distinct from P388/S, possessed a 150- to 160-kD membrane species identified by Vinca alkaloid photoaffinity labeling.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of prostacyclin analogs on cyclic adenosine monophosphate and superoxide formation in human polymorphonuclear leukocytes stimulated by formyl-methionyl-leucyl-phenylalanine

The modulation of cyclic AMP levels and superoxide release in isolated FMLP-stimulated human PMN by two oxacyclic analogs and one carbacyclic analog of PGI2 and by PGE1 is investigated over a wide range of concentrations of the test compounds. The prostacyclin analogs only marginally increase the cyclic AMP levels in unstimulated PMN but like PGE1 potentiate the FMLP-induced rise in cyclic AMP. The concentration dependency is bell-shaped with a maximum effect at about 10 microM of the prostanoids. In contrast, all prostanoids dose-dependently inhibit FMLP-induced superoxide release almost to completion. The relative inhibitory potency of the prostacyclin analogs corresponds to their prostacyclin-like action in other systems. It is therefore suggested that PMN contain prostacyclin receptors, which, however, have weaker affinities than those in platelets. The lack of correlation between inhibition of superoxide formation and modulation of the cyclic AMP system rules out the possibility that cyclic AMP can simply be considered the second messenger of prostacyclins in PMN. The potential biological relevance of the effects of prostacyclin-like compounds on PMN functions is discussed.     

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.     

Hormonal imprinting and damages caused by fetal steroid treatment. Influence of fetal treatment with pregnancy-protecting steroids (allylestrenol, diethylstilbestrol) on the effect of gonadotropin administered to cockerels perinatally and at the age of six weeks

Single fetal (9th day) treatment with either diethylstilbestrol (DES) or allylestrenol (AE) caused a considerable decrease, both at the age of five days and six weeks, in the weight of the testicles and the diameter of the seminiferous cords, while the ratio of spermatogonia to primary spermatocytes increased. When measured either at the age of five days or six weeks, gonadotropin treatment [a combination of follicle stimulating hormone (FSH) and luteinizing hormone (LH)], administered twice daily for three days after the hatching, led to an increase in the above-mentioned parameter and to a shift in the cell ratio towards the control value. However, the absolute value of the controls treated with FSH-LH was by far not reached. The effect of perinatal treatment could be detected even in adulthood, namely, at the age of five days the response capability was relatively weak in the cockerels treated with DES and AE, while high responsiveness was observed at the age of six weeks. In some cases the relative value of the increment exceeded even that of the control; however in absolute term it was well below the control. On the basis of these experiments it might be concluded that hormonal imprinting evoked by FSH-LH treatment also occurs in the gonad damaged by DES and AE. The setting in of imprinting ameliorates the damages caused by DES and AE and increases the response capability of the cells.     

No effect of isolation on blood pressure and daily electrolyte excretion in rats

The effects of isolation stress on mean blood pressure (BP) and on body weight, water and food intake as well as on urine flow, urinary sodium and potassium excretion were studied in CFY and Long Evans rats. During a 7 day isolation period, food and water intake as well as urine flow, urinary sodium and potassium excretion, as expressed for 100 g body weight, were not changed in either group. Body weight increased similarly in isolated (38 +/- 2 g) and aggregated (41 +/- 5 g) CFY rats. Compared to group housed rats, BP in male CFY animals was not increased after a 7 day isolation (111 +/- 3 vs 111 +/- 3 mmHg, NS). In additional experiments high sodium intake by physiological saline drinking slightly elevated blood pressure but failed to induce arterial hypertension in isolated rats (118 +/- 2 vs 121 +/- 3 mmHg, NS). We conclude that, contrary to some reports from other laboratories, isolation stress has no detectable effect on BP and/or water and electrolyte balance.     

The combined effects of dopamine (DA) and the DA antagonists EGYT-2509, chlorpromazine and haloperidol on the kidney function

In anaesthetized dogs renal function was investigated in four successive 20-min periods in four experimental series. (1) In the first series following the first period (serving as control) 2.5 micrograms/kg/min of dopamine (DA) dissolved in 0.5 ml/min of Ringer's solution was infused into the left renal artery (period 2), than during periods 3 and 4. It was found that first (period 2) and second (period 3) doses of DA induced a significant decrease of about 20-30% in renal vascular resistance, and an increase of about 15-25% in renal blood flow. At the same time, systemic arterial blood pressure fell by 10%. The other investigated parameters of the left kidney (Cinulin, CPAH, sodium, potassium and water excretion) did not differ from the respective parameters of the intact right kidney. (2) In the second experimental series following the first period (prior to period 2) 1.0 mg/kg of the DA antagonist EGYT 2509 was administered intravenously. Prior to the period 3 again 1.0 mg/kg of EGYT 2509 and prior to period 4 2.0 mg/kg of EGYT 2509 was given intravenously. During periods 2 through 4 2.5 micrograms/kg/min of DA was infused into the left renal artery. It could be ascertained that EGYT 2509 abolished the renal effects of DA while not inducing any decrease in arterial blood pressure. (3) In the third experimental series, following the control period, prior to periods 2,3 and 4, 1.0 mg/kg, 1.0 mg/kg and 2.0 mg/kg chlorpromazine respectively, was administered i.v. followed by the infusion of DA into the left renal artery. After the administration of chlorpromazine arterial blood pressure and renal vascular resistance fell concomitantly and DA failed to induce any further changes in these parameters. According to our experiments chlorpromazine abolishes the effect of DA on kidney function. (4) In the fourth series, prior to DA infusion the dogs were given 0.5 mg/kg (period 2) then again 0.5 mg/kg and finally 1.0 mg/kg of haloperidol intravenously. Haloperidol decreased arterial blood pressure as well as renal vascular resistance, thus renal blood flow did not change. Renal blood flow could then be increased by DA infused into the left renal artery. It seems that haloperidol could not abolish the vascular effects of DA in the kidney. Our experiments indicate that substance EGYT 2509 possesses the most marked dopaminergic antagonistic effect, chlorpromazine had also been effective, while haloperidol had proved to be practically ineffective.     

Effect of intraarterial somatostatin infusion on SMA blood flow

Experiments were carried out on anaesthetized cats to study the effect of somatostatin on the mesenteric circulation. Intraarterial infusions of somatostatin were applied into the superior mesenteric artery. The effect of atropine, propranolol and phentolamine were investigated. Catecholamine release or uptake of the mesenteric vascular bed during somatostatin infusions were also measured. We found dose dependent vasodilatory effect of somatostatin on the mesenteric vasculature that was not influenced by atropine and by the adrenoreceptors or by denervation. A direct effect of somatostatin on the vascular smooth muscle membrane is assumed.     

The effect of pentylenetetrazol on the metacerebral neuron of Helix pomatia

The effects of Pentylenetetrazol (PTZ) on the metacerebral giant cell (MCC) of the snail, Helix pomatia were studied. Actions on membrane resistance, time constant, resting and action potentials, outward and inward ionic currents were examined. Superfusion with PTZ in concentrations of 25 to 50 mmol/l, induced a gradually evolving convulsive state, which could be studied by intracellular recording from the MCCs. In the pre-convulsive state an acceleration of the spontaneous activity developed and was followed by paroxysmal depolarization shifts (PDSs), in the convulsive phase. PTZ prolonged the membrane time constant by about 10 percent, but this could not be traced back to alterations in membrane resistance or capacity. The resting membrane potential was not significantly altered; the action potentials were prolonged by slowing down of both the rising and decaying phases. The outward potassium currents were repressed by PTZ in a voltage dependent manner. The decrease of the IA current became more pronounced at increasingly positive command pulses, while IK was relieved from depression especially at longer pulse durations. Inward currents were isolated with the aid of suppression of outward currents by 50 mmol/l TEA. Under these conditions sodium currents, measured in calcium deficient Ringer solution were moderately depressed, while the calcium currents, examined during sodium-free superfusion, were mildly enhanced by PTZ. It is concluded that PTZ effects on ionic conductances, on membrane parameters, on the resting potential and ionic currents explain only modifications of spike potentials occurring in the convulsive state and do not account for the PDS, the central phenomenon of the convulsive electrographic activity, at least in this thoroughly examined type of neuron.     

The effect of prostaglandin F2 alpha and its analogues on progesterone, estradiol and androgen production by the cultured bovine luteal cells

The effect of PGF2 alpha and its analogues on progesterone, estradiol and androgen production in cow corpus luteum in vitro was investigated. The cells derived from cow corpora lutea (CL) and collected in the early and middle luteal phases of the oestrus cycle were cultured as monolayers. The inhibitory effect was not apparent during the first 48 hr of culture, but appeared after this time and persisted through the remainder of the culture period. The direct luteolytic influence of PGF2 alpha was observed in the cultured cells and showed that this compound can act independently of the blood supply.