Plasma extravasation mediated by lipopolysaccharide-induction of kinin B1 receptors in rat tissues.

The present study was performed to: (a) evaluate the effects of kinin B1 (Sar[D-Phe8]-des-Arg9-BK; 10 nmol/kg) and B2 (bradykinin (BK); 10 nmol/kg) receptor agonists on plasma extravasation in selected rat tissues; (b) determine the contribution of a lipopolysaccharide (LPS) (100 microg/kg) to the effects triggered by B1 and B2 agonists; and (c) characterize the selectivity of B1 ([Leu8]desArg9-BK; 10 nmol/kg) and B2 (HOE 140; 10 nmol/kg) antagonists as inhibitors of this kinin-induced phenomenon. B1 and B2 agonists were shown to increase plasma extravasation in the duodenum, ileum and also in the urinary bladder of the rat. LPS pretreatment enhanced the plasma extravasation mediated only by the B1 agonist in the duodenum, ileum, trachea, main and segmentar bronchi. These effects were prevented by the B1. but not the B2 antagonist. In normal rats, the B2 antagonist inhibited the effect of B2 agonist in all the tissues analyzed. However, in LPS-treated rats, the B2 antagonist was ineffective in the urinary bladder. These results indicate that kinins induce plasma extravasation in selected rat tissues through activation of B1 and B2 receptors, and that LPS selectively enhances the kinin effect on the B1 receptor in the duodenum, ileum, trachea and main and segmentar bronchi, and may increase B1 receptor expression in these tissues.

Effect of quercetin on tachykinin-induced plasma extravasation in rat urinary bladder.

The effect of quercetin on substance P-induced plasma extravasation in rat urinary bladder and its modulation by endogenous peptidases in conscious rats was studied. Plasma protein extravasation (PE) was assayed by measurement of extravasated Evans blue dye (microg/g dry tissue). Intravenous injection of substance P (SP, 10 nmol/kg) significantly increased PE in the urinary bladder. PE evoked by SP was increased significantly by quercetin (20 mg/kg, p.o.) pretreatment in the urinary bladder (73.5 +/- 4.9 to 152.2 +/- 9.9). Pretreatment with captopril, an angiotensin-converting enzyme (ACE) inhibitor (10 nmol/kg, i.v.), or with phosphoramidon, a neutral endopeptidase (NEP) inhibitor (2.5 micromol/kg, i.v.) also potentiated the SP-induced PE in urinary bladder, 286.2 +/- 20.4 and 323.3 +/- 34.0, respectively. Quercetin did not show any effect on neurokinin-A (NKA, 10 nmol/kg, i.v.) -induced plasma extravasation. The present study demonstrates that quercetin potentiates the PE induced by substance P in the urinary bladder. These effects suggest that this flavonoid might cause inhibition of NEP and/or ACE.

Calcium entry through L-type calcium channels causes mitochondrial disruption and chromaffin cell death.

Sustained, mild K+ depolarization caused bovine chromaffin cell death through a Ca(2+)-dependent mechanism. During depolarization, Ca(2+) entered preferentially through L-channels to induce necrotic or apoptotic cell death, depending on the duration of the cytosolic Ca(2+) concentration ([Ca(2+)](c)) signal, as proven by the following. (i) The L-type Ca(2+) channel activators Bay K 8644 and FPL64176, more than doubled the cytotoxic effects of 30 mm K+; (ii) the L-type Ca(2+) channel blocker nimodipine suppressed the cytotoxic effects of K+ alone or K+ plus FPL64176; (iii) the potentiation by FPL64176 of the K+ -evoked [Ca(2+)](c) elevation was totally suppressed by nimodipine. Cell exposure to K+ plus the L-type calcium channel agonist FPL64176 caused an initial peak rise followed by a sustained elevation of the [Ca(2+)](c) that, in turn, increased [Ca(2+)](m) and caused mitochondrial membrane depolarization. Cyclosporin A, a blocker of the mitochondrial transition pore, and superoxide dismutase prevented the apoptotic cell death induced by Ca(2+) overload through L-channels. These results suggest that Ca(2+) entry through L-channels causes both calcium overload and mitochondrial disruption that will lead to the release of mediators responsible for the activation of the apoptotic cascade and cell death. This predominant role of L-type Ca(2+) channels is not shared by other subtypes of high threshold voltage-dependent neuronal Ca(2+) channels (i.e. N, P/Q) expressed by bovine chromaffin cells.

The sea anemone toxin Bc2 induces continuous or transient exocytosis, in the presence of sustained levels of high cytosolic Ca2+ in chromaffin cells.

We have isolated and characterized a new excitatory toxin from the venom of the sea anemone Bunodosoma caissarum, named Bc2. We investigated the mechanism of action of the toxin on Ca(2+)-regulated exocytosis in single bovine adrenal chromaffin cells, monitoring simultaneously fura-2 fluorescence measurements and electrochemical recordings using a carbon fiber microelectrode. Bc2 induced quantal release of catecholamines in a calcium-dependent manner. This release was associated with a sustained rise in cytosolic Ca(2+) and displayed two different patterns of response: a continuous discharge of prolonged duration that changed to a transient burst as the toxin concentration (or incubation time) increased. Continuous secretion was dependent on the activity of native voltage-dependent Ca(2+) channels and showed a pattern similar to that of alpha-latrotoxin; however, its kinetics adjusted better to that of continuous cell depolarization with high K(+) concentration. In contrast, transient secretion was independent of Ca(2+) entry through native voltage-dependent Ca(2+) channels and showed inhibition of late vesicle fusion that was accompanied by "freezing" of F-actin disassembly. These new features make Bc2 a promising new tool for studying the machinery of neurotransmitter release.

Synaptosomal glutamate release induced by the fraction Bc2 from the venom of the sea anemone Bunodosoma caissarum.

The effect of a fraction (Bc2) from the venom of the sea anemone Bunodosoma caissarum on [3H]glutamate release from rat cortical synaptosomes was investigated. Bc2 (2-20 microg/ml) provoked massive glutamate release without causing synaptosome disruption. Glutamate release stimulated by Bc2 was independent of extracellular Ca2+ and of voltage-sensitive Na+ channels, and it was completely abolished by the addition of sphingomyelin. No definitive evidence about the mechanism underlying the stimulatory effect of Bc2 is available as yet. However, a direct interaction with the exocytotic machinery cannot be ruled out.

Valor del isómero dextrógiro del ácido láctico sérico en la isquemia intestinal en ratas / Value of serum d-lactate levels during intestinal isquemia in rats

El D-lactato fue estudiado in vitro como marcador de isquemia intestinal asociada a la presencia de bacterias patógenas en el intestino de ratas. En un estudio in vitro se correlacionó la producción de D-lactato con el crecimiento de dos cepas de E coli. En el estudio in vivo se utilizaron 100 ratas Wistar de 250a 300 gr divididos en ausencia y presencia de isquemia intestinal. Los resultados in vitro demostraron que la producción de D-lactato es acumulativa. Los resultados in vivo demostraron que el nivel sérico de D-lactato dependió de la concentración bacteriana intraluminal y no de la gravedad de la isquemia. Cuando ambas variables fueron asociadas, se produjo un efecto sinérgico en la elevación sérica de D-lactato, mayor en comparación a cada variable aislada. Se observaron resultados similares con la flora natural con o sin isquemia intestinal. Basados en los datos obtenidos, los niveles de D-lactato sérico parecen tener un rol potencial como marcador de isquemia intestinal, cuando se agrega la presencia de bacterias en la luz intestinal

Valor del isómero dextrógiro del ácido láctico sérico en la isquemia intestinal en ratas / Value of serum d-lactate levels during intestinal isquemia in rats

El D-lactato fue estudiado in vitro como marcador de isquemia intestinal asociada a la presencia de bacterias patógenas en el intestino de ratas. En un estudio in vitro se correlacionó la producción de D-lactato con el crecimiento de dos cepas de E coli. En el estudio in vivo se utilizaron 100 ratas Wistar de 250a 300 gr divididos en ausencia y presencia de isquemia intestinal. Los resultados in vitro demostraron que la producción de D-lactato es acumulativa. Los resultados in vivo demostraron que el nivel sérico de D-lactato dependió de la concentración bacteriana intraluminal y no de la gravedad de la isquemia. Cuando ambas variables fueron asociadas, se produjo un efecto sinérgico en la elevación sérica de D-lactato, mayor en comparación a cada variable aislada. Se observaron resultados similares con la flora natural con o sin isquemia intestinal. Basados en los datos obtenidos, los niveles de D-lactato sérico parecen tener un rol potencial como marcador de isquemia intestinal, cuando se agrega la presencia de bacterias en la luz intestinal

In vitro study of enamel erosion caused by soft drinks and lemon juice in deciduous teeth analysed by stereomicroscopy and scanning electron microscopy.

The erosion caused in vitro by cola-type and guaraná-type beverages (the latter is a soft drink sold in Brazil), and a canned lemon juice on the enamel of human deciduous teeth was analyzed. Morphological analysis of affected enamel was done using stereomicroscopy and scanning electron microscopy (SEM). The harmful effect of all test products on deciduous enamel was clearly demonstrated. Stereomicroscopy showed loss of gloss and an alteration in normal color of enamel, with irregular loss of dental tissue in variable degrees. Such a loss became more serious as the time of incubation increased. Different degrees of solubilization of enamel prisms were demonstrated by SEM, affecting initially the sheaths and the heads of prisms and later their tails. Areas of erosion increased in proportion to the time of incubation. All the products showed a great erosive potential on human deciduous dental enamel.