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We report our experience in 66 cases of acute poisoning requiring haemodialysis (HD) in the last 17 years. Barbiturate poisoning was the commonest poisoning (30 cases). Mean blood barbiturate level was 8.9 mg%. Twenty four were in grade IV coma at the time of presentation. Twenty five required one HD and 5 cases needed 2 HD. Four died due to respiratory infection or hypotension. Copper sulphate poisoning was encountered in 19 cases. Common features in this group were: acute renal failure (ARF) (19), haematuria (3), gastrointestinal bleeding (7), intravascular haemolysis (9), jaundice (11), hepatocellular toxicity (8), methaemoglobinuria (8) and circulatory collapse (5). The indication for HD in all these cases was ARF. Seven patients died. There were 9 cases of mercuric chloride poisoning requiring 2-5 HD. Common features in this group were; ARF (9), gastrointestinal bleeding (9), anaemia (8), jaundice (2). Two patients died. Other patients had Mandrax, Naphthalene, Tincture Iodine, Ethylene Bromide and Lithium poisoning. Overall mortality in our study was 24.2%. It is concluded that HD is not the primary mode of therapy for drug intoxication. Adequate supportive management is most important in determining final outcome of these patients.
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PMID:Spectrum of poisoning requiring haemodialysis in a tertiary care hospital in India. 845 67

DNA fragmentation was evaluated in three instances of programmed cell death, interdigital cell death in embryonic mouse limbs, and metamorphic death of both the labial glands and intersegmental muscle in the tobacco hornworm Manduca sexta. In the mouse, we evaluated both developmental cell death and expanded-range cell death induced by retinoic acid. The status of DNA was examined in several ways. Nuclei were examined by electron microscopy and Feulgen staining. Quantitative assessment of total DNA content in Feulgen-stained degenerating nuclei was made for the gland. In the labial gland, DNA content does not drop during the early phases of cell death; nor is an endonucleolytic ladder seen when DNA was examined by ethidium bromide staining or prelabeling with [3H]thymidine. Only by using end labeling of DNA could we detect DNA fragmentation at a very late stage in cell death, day 4 of the collapse of the gland. In contrast, WEHI 7.1 lymphoma cells display an early and extensive ladder after treatment with glucocorticoids. In mouse limb, for which cell death follows a more classic apoptotic morphology, a ladder is likewise not seen. We conclude that activation of an endonuclease is neither a trigger nor a necessary or defining component of the early phases of developmental programmed cell death, and that reported failure by others to find such a ladder may depend on limitations in the system that is under investigation.
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PMID:Delayed internucleosomal DNA fragmentation in programmed cell death. 846 89

This paper examines the accumulation and toxicity mechanism of a cationic detergent, cetyltrimethylammonium (bromide) (CTAB), in energized rat liver mitochondria. The results suggest that: (1) the CTAB ion is accumulated in the mitochondrial matrix by a membrane potential-driven uptake mechanism; (2) accumulation may lead to a toxic effect, since it gives rise to collapse of the mitochondrial membrane potential (delta phi) which is correlated with ATP synthesis and which regulates Ca++ uptake; (3) collapse of delta phi may be due to enhanced permeability of the mitochondrial membrane to the ions (detergent effect); and (4) delta phi collapse and Ca++ and K+ release were also observed in another cationic detergent, NTAB, but not in the presence of anionic detergents.
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PMID:Mitochondrial bioenergetics as affected by cationic detergents. 859 85

The polyene antibiotic amphotericin B (AmB) is known to form two types of ionic channels across sterol-containing liposomes, depending on its concentration and time after mixing (Cohen, 1992). In the present study, it is shown that AmB only kills unicellular Leishmania promastigotes (LPs) when aqueous pores permeable to small cations and anions are formed. Changes of membrane potential across ergosterol-containing liposomes and LPs were followed by fluorescence changes of 3,3' dipropylthiadicarbocyanine (DiSC3(5)). In KCl-loaded liposomes suspended in an iso-osmotic sucrose solution, low AmB concentrations (</=0.1 microM) induced a polarization potential, indicating K+ leakage, but no movement of cations and anions was allowed until AmB concentrations greater than 0.1 microM were added. In agreement with these data, it was found that AmB altered the negative membrane potential held across LPs in a manner consistent with the differential cation/anion selectivity exhibited by the channels formed in liposomes. Thus, LPs suspended in an iso-osmotic sucrose solution did not exhibit any AmB-induced membrane depolarization effect brought about by efflux of anions until 0.1 microM or higher AmB concentrations were added. By contrast, LPs suspended in an iso-osmotic NaCl solution and exposed to 0.05 microM AmB exhibited a nearly total collapse of the negative membrane potential, indicating Na+ entry into the cells. The concentration dependence of the AmB-induced permeability to different salts was also measured across vesicles derived from the plasma membrane of leishmanias (LMVs), by using a rapid mixing technique. At concentrations above 0.1 microM, AmB induced the formation of aqueous pores across LMVs with a positive cooperativity, yielding Hill coefficients between 2 to 3. Measured anion selectivity across such aqueous pores followed the sequence: SCN > NO3 > Cl > I > Br > acetate (SO2-4 being impermeable). Cell killing by AmB was followed by fluorescence changes of the DNA-binding compound ethidium bromide (EB). At low concentrations (</=0.1 microM), AmB was found to be nonlethal against LPs but, above this concentration, leishmanias were rapidly killed. The rate and extent of such an effect were found to be dependent on the type of cation and anion present in the external aqueous solution. For both NH+4 and Na+ salts, the measured rank order of AmB cell killing followed the same sequence that was determined for AmB-induced salt permeation across LMVs. Further, replacement of either extracellular Na+ by choline or Cl- by SO2-4, or its partial substitution by sucrose, in iso-osmotic conditions, led to a complete inhibition of the killing effect exerted by otherwise lethal AmB concentrations. Finally, it was shown that tetraethylammonium (TEA+), an organic cation that is known to block AmB-induced salt permeation across LMVs was able to retard the time lag observed for EB incorporation across LPs, indicating that this parameter can be taken to represent the time taken for salt accumulation inside the parasites. The present results thus indicate clearly that low AmB concentrations (</=0.1 microM) were able to form across LPs, cation channels that collapsed the parasite membrane potential but are not lytic. At high concentrations (>/=0.1 microM), a salt influx via the aqueous pores formed by the antibiotic was followed by osmotic changes leading to cell lysis. This last stage is supported by electron microscopy observations of the changes of parasite morphology immediately upon addition of AmB, which indicated that the typical elongated promastigote cell forms became rounded and the flagella swells and round up. The present work is the first demonstration of the in vitro sensitivity of Leishmania promastigotes to osmotic lysis by AmB.
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PMID:Amphotericin B kills unicellular leishmanias by forming aqueous pores permeable to small cations and anions. 866 Apr 6

Continuous wave EPR spectra of the nitroxyl signals for four spin-labeled high-spin (h.s.) Fe(III) porphyrins showed partially resolved splittings at temperatures near 4 K. Axial ligands were fluoride, chloride, or bromide. As temperature was increased to 20 to 30 K the iron-nitroxyl splitting collapsed due to increasing rates of iron relaxation. Electron spin-echo (ESE) spectroscopy showed that above about 6 K collapse of the iron-nitroxyl spin-spin splitting caused a dramatic increase in the nitroxyl phase memory relaxation rates. Electron spin relaxation rates were determined for Fe(tetratolylporphyrin)X, X = F, Cl, Br, in toluene solution by ESE or inversion recovery at 4.5 to 6 K and by analysis of the temperature-dependent contributions to the continuous wave EPR linewidths between about 10 and 120 K. Above about 10 K iron relaxation rates increase in the order X = F < Cl < Br, which is the order of increasing zero-field splitting. Saturation recovery data for two spin-labeled h.s. iron(III) porphyrins between about 15 and 120 K and for two additional spin-labeled h.s. iron(III) porphyrins between about 85 and 120 K demonstrated that interaction with the h. s. iron enhanced the electron spin relaxation rate of the spin label. The saturation recovery curves for the nitroxyl were analyzed to determine interspin distances using a modified version of the Bloembergen equation and independently determined iron relaxation rates. Interspin distances were between 11.6 and 15.0 A, were independent of axial ligand, and were in good agreement with values obtained previously for low-spin Fe(III) and Cu(II) analogs.
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PMID:Determination of high-spin iron(III)-nitroxyl distances in spin-labeled porphyrins by time-domain EPR. 953 11

The present study was designed to obtain a basic pharmacological profile of venom from the inland taipan (Oxyuranus microlepidotus). Venom (0.05-50 micrograms/ml) produced dose-dependent contractions in guinea-pig ileum, which could not be reproduced upon second administration. The cyclooxygenase inhibitor indomethacin (1 microM), a preceding anaphylactic response induced by egg albumin and inactivation of phospholipase A2 (PLA2) by incubation with 4-bromophenacyl bromide (1.8 mM) all significantly inhibited responses to venom (0.5 micrograms/ml). Venom (0.5 micrograms/ml) caused inhibition of stimulation-induced contractions in the prostatic segment of rat vas deferens which was not significantly affected by the alpha 2-adrenoceptor antagonist idazoxan (0.3 microM). Venom (10 micrograms/ml) caused time-dependent inhibition of the rat electrically stimulated phrenic nerve-diaphragm preparation, positive inotropic and chronotropic responses in rat isolated atria and relaxation in rat endothelium-denuded and -intact isolated aortae. In endothelium-intact aortae, the nitric oxide synthase inhibitor N-nitro-L-arginine (NOLA, 0.1 mM) significantly inhibited the response to venom (10 micrograms/ml). Venom (50 micrograms/kg, i.v.) caused an immediate drop in blood pressure followed by cardiovascular collapse in anaesthetised rats. Venom (10 micrograms/kg, i.v.) caused a gradual fall in blood pressure which was sometimes accompanied by a temporary cessation of respiration. A PLA2 assay detected the presence of PLA2 in the venom. These results suggest that the venom contains a component capable of causing the synthesis of arachidonic acid metabolites and a component capable of relaxing vascular smooth muscle. The inhibitory effect on the phrenic nerve-diaphragm is probably due to the previously identified neurotoxin (paradoxin).
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PMID:Some pharmacological studies of venom from the inland taipan (Oxyuranus microlepidotus). 960 83

We investigated the phase behavior and the phase transitions in aqueous solutions of 100 mM cetyltrimethylammonium hydroxide (CTAOH) with 3-hydroxy-2-naphthoic acid (HNC) and of 100 mM cetyltrimethylammonium bromide (CTAB) with sodium-3-hydroxy-2-naphthoate (SHNC). The naphthoate/surfactant ratio has been varied. As previously observed by the groups of C. Manohar and J. Candau we observed for the second system two viscoelastic gel-like regions, two liquid crystalline regions, and a precipitate region. For the CTAOH/HNC system one finds with increasing concentration of HNC a low viscous solution, a viscoelastic gel, and a viscoelastic liquid crystalline Lalpha-phase. In both surfactant systems the lamellar phase is formed around an equimolar ratio of cationic surfactant and naphthoate. The lamellar phases have been examined by polarization microscopy and freeze-fracture electron microscopy. The Lalpha-phase in the system CTAOH/HNC consists of densely packed multilamellar vesicles while the lamellar phase in the system CTAB/SHNC contains vesicles, as well as stacked bilayers and tubuli. Corresponding to their different microstructures the lamellar phases in the system, CTAOH/HNC and CTAB/SHNC have different rheological properties. The vesicular phase is highly viscoelastic and has a yield stress value while the bilayer phase has a much lower viscosity and no yield stress value. The transition from the micellar to the vesicle phase occurs for CTAOH/HNC over a two-phase region, where micelles and vesicles coexist. In the case of CTAB/SHNC the transition from the micellar to the lamellar phase occurs over a three-phase region, where a surfactant-poor phase coexists with a lamellar and a coacervate phase. In mixtures of CTAB and SHNC a thick precipitate is formed at an equimolar ratio of CTAB and SHNC. This precipitate consists of condensed multilamellar vesicles that contain little water and stick together, as the vesicles collapse due to the shielding of the repulsive forces by NaBr from an unbinding to a binding state. The precipitate can be retransformed to a swollen lamellar phase by charging the vesicles with an excess of ionic surfactant, by adding electrolyte in high concentrations, or by increasing the temperature. As predicted by C. Manohar et al. the vesicle phases show a phase transition at a critical temperature Tc of 46 degreesC. This transition was detected by us for the first time by DSC and by conductivity measurements. It occurs within a narrow temperature range of 2-3 degrees with an enthalpy change of 0.5 kJ/mol. The transition is observed both in the swollen and in the precipitated vesicle phase. It is well separated from the vesicle/rod transition at higher temperatures (>70 degreesC) and the liquid crystalline/crystalline transition at lower temperatures (25-30 degreesC) that has a melting enthalpy of 55 kJ/mol. It is conceivable that the observed transition at 46 degreesC is due to the melting of a two-dimensional solid-like lattice of the HNC-counterions on the vesicle interface. Copyright 1998 Academic Press.
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PMID:Formation and Properties of Lamellar Phases in Systems of Cationic Surfactants and Hydroxy-Naphthoate. 975 56

Aberrant glycosylation of proteins and lipids is a common feature of many tumor cell types, and is often accompanied by alterations in membrane traffic and an anomalous localization of Golgi-resident proteins and glycans. These observations suggest that the Golgi complex is a key organelle for at least some of the functional changes associated with malignant transformation. To gain insight into this possibility, we have analyzed changes in the structure and function of the Golgi complex induced by the conditional expression of the transforming N-Ras(K61) mutant in the NRK cell line. A remarkable and specific effect associated with this N-Ras-induced transformation was a conspicuous rearrangement of the Golgi complex into a collapsed morphology. Ultrastructural and stereological analyses demonstrated that the Golgi complex was extensively fragmented. The collapse of the Golgi complex was also accompanied by a disruption of the actin cytoskeleton. Functionally, N-Ras-transformed KT8 cells showed an increase in the constitutive protein transport from the trans-Golgi network to the cell surface, and did not induce the appearance of aberrant cell surface glycans. The Golgi complex collapse, the actin disassembly, and the increased constitutive secretion were all partially inhibited by the phospholipase A2 inhibitor 4-bromophenylacyl bromide. The results thus suggest the involvement of the actin cytoskeleton in the shape of the Golgi complex, and intracellular phospholipase A2 in its architecture and secretory function.
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PMID:N-Ras induces alterations in Golgi complex architecture and in constitutive protein transport. 991 60

The Papuan taipan (Oxyuranus scutellatus canni) is the third most venomous terrestrial snake in the world, however, little is know about the pharmacology of the venom. In the chick biventer cervicis muscle, venom (10 microg/ml) abolished nerve-mediated twitches (time to 90% inhibition (t90) 44+/-5 min, n = 9). This inhibition was unaffected by prior incubation of the venom with the phospholipase A inhibitor 4-bromophenacyl bromide (4-BPB; 0.72 mM) (t90 48+/-7 min, n = 8). The mouse phrenic nerve diaphragm preparation displayed greater sensitivity to venom (10 microg/ml) (t90 25+/-1 min, n = 6). In the chick biventer muscle, venom (10 microg/ml) significantly inhibited responses to acetylcholine (1 mM) and carbachol (20 microM), but not KCI (40 mM), indicating activity at post-synaptic nicotinic receptors. Venom (10 microg/ ml) did not affect direct muscle stimulation. Venom (3-30 microg/ml) produced dose-dependant contractions of the guinea-pig ileum. Contractile responses were significantly inhibited by indomethacin (1 microM) or prior incubation of the venom with 4-BPB (0.72 mM) indicating involvement of a PLA component. In rat phenylephrine (0.3 microM) precontracted aortae, venom (3-100 microg/ml) produced endothelium-independent relaxation which was unaffected by prior incubation of venom (30 microg/ml) with 4-BPB (0.72 mM). In anaesthetised rats, 10 microg/kg (i.v.) venom produced rapid respiratory and cardiovascular collapse while 5 microg/kg (i.v.) venom produced only a small transient decrease in mean arterial blood pressure. Prior administration of 5 microg/kg (i.v.) venom enabled subsequent administration of 10 and 100 microg/kg (i.v.) venom without respiratory or cardiovascular collapse. Further work is required to identify specific toxins with the above pharmacological activity.
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PMID:A pharmacological examination of venom from the Papuan taipan (Oxyuranus scutellatus canni). 1051 50

Saturated linear fatty acids, derivatized with a near-infrared absorbing fluorescent dye, were separated in 100% methanol with 12.5 mM tetraethylammonium chloride added as a charge carrier. Separation at 380 V/cm was acceptable for acids that differed in length by a single carbon. The labeled linear fatty acids behaved as random coils in the nonaqueous separation medium, as shown in a fit to a simple theoretical expression. However, even in 100% methanol with a trimethylsilylated capillary, significant adsorption to the capillary wall occurred, which reduced resolution and slowed the separation. Addition of water to the methanol medium caused significant differences in separation behavior of high molecular weight acids (>C16). Addition of a cetyltrimethylammonium bromide surfactant to the separation medium dynamically coated the capillary and greatly improved the separation. The surfactant also interacted with the acyl tail, apparently causing it to collapse. Resolution in an optimal separation medium (20 mM surfactant) ranged from 1.6 to 1.1, depending on chain length, and theoretical plate heights were under 4 microm (N > 10(5)). Resolution was more than adequate to separate stearic (C18:0) from oleic (C18:1) acid, as well as other unsaturated C18 homologues.
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PMID:Nonaqueous capillary electrophoresis of fatty acids derivatized with a near-infrared fluorophore. 1081 69


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