A switch to Raltegravir improves antiretroviral associated hepatotoxicity in individuals co-infected with HIV and hepatitis C.

INTRODUCTION: Raltegravir is a switch option for HIV/HCV co-infected individuals due to its hepatic neutral profile. We evaluated the effect of a switch to Raltegravir from other antiretroviral agents in HIV and HCV-co-infected individuals naïve to HCV therapy. METHODS: Observational, single-centre study. Data on alanine aminotransferase levels, HCV-VL, CD4 cell count, HIV viral load levels and hepatic fibrosis score were collated six months pre-switch, at the time of switch and six months post switch to Raltegravir therapy. Results were compared utilizing the Kruskal-Wallis test. RESULTS: Twenty-seven individuals were identified. Median age was 43 years, median duration of HIV infection was 7 years and median documented period of HCV infection at the time of switch was 26 months. A sustained improvement in ALT levels was observed. Median ALT levels were 254 IU/L at the time of switch, decreasing significantly to 176 IU/L, (p = 0.0226) and 90 IU/L (p = 0.0138) 1 month post switch and 6 months post switch respectively. The median Hepatitis C viral load level at the time of the switch was 341,783 copies/mL, which decreased to 224,066 copies/mL 6 months after switch (p = 0.04). DISCUSSION: A switch to Raltegravir in individuals with HIV/HCV co-infection was effective in maintaining HIV virological suppression with improvement in drug-associated hepatotoxicity as measured by ALT.

Laproscopic management of wandering biliary ascariasis.

Ascariasis is one of the most common helminthic diseases in humans, occurring mostly in countries with low standards of public health and hygiene, thereby making ascariasis highly endemic in developing countries. In endemic areas, 30% of adults and 60-70% of children harbour the adult worm. Biliary ascariasis is a rare cause of obstructive jaundice. Conventional management involves endoscopic extraction of worm. We are reporting a rare case of ascaris which induced extrahepatic biliary obstruction in a young male who presented with acute cholangitis. The ascaris was removed by laparoscopic exploration of the common bile duct. Postoperative period was uneventful.

Cyclic gmp is a measure of physiologic stress.

This study was undertaken to determine whether patients who were critically ill evidenced elevated levels of blood cyclic guanosine monophosphate (cGMP). Cyclic guanosine monophosphate levels correlated with severity of illness as measured by the APACHE II severity of illness scoring system (p < 0.01). Cyclic guanosine monophosphate also correlated with the level of carboxyhemoglobin (HbCO) (p < 0.001). The correlation between cGMP and creatinine was p < 0.0001. Patients with end-stage disease (renal or liver) tended to have elevated levels of cGMP (p < 0.0001). We conclude that the induction of these two molecules may be linked in patients with increasing severity of illness.

Development of in vivo bioequivalence methodology for dermatologic corticosteroids based on pharmacodynamic modeling.

BACKGROUND: Dermatologic corticosteroid products produce skin blanching that is related to clinical potency and dose. (For application of the vasoconstrictor assay to bioavailability and bioequivalence assessment, dose is defined in terms of duration of treatment exposure [dose duration], so the terms dose and dose duration have been used interchangeably). The vasoconstrictor assay is the method of choice to assess dermatologic corticosteroid products bioequivalence if dose-response is validated. This article examines dose-response validation to meet objectives of US Food and Drug Administration (FDA) bioequivalence guidance for dermatologic corticosteroid products. METHODS: An exploratory dose-response study was conducted to determine applicability of the empirical maximum effect (Emax) model to the individual subject and population dose-response relationships of six dermatologic corticosteroid product creams that varied from the most to the least potent classes. Products were applied to the skin of 10 healthy subjects in each of two dosing periods for dose durations of 0.5, 1, 2, and 6 hours. Skin blanching was measured by reflectance colorimeter through 24 hours after application. Area under the effect curve (AUEC) was determined for each dose duration. An Emax model was fitted to the AUEC versus dose duration data. A similar analysis was conducted for a bioequivalence study on two formulations of a dermatologic corticosteroid product in 40 healthy subjects. RESULTS: In the exploratory study, the number of individual subject data sets for which the Emax model provided an acceptable fit generally increased with the potency of the dermatologic corticosteroid product. On the basis of population modeling, dose-response data of all products, except the lowest potency cream, were adequately described by the Emax model. Values for population ED50 (the dose duration required to achieve 50% of the fitted AUECmax value) decreased with increase in dermatologic corticosteroid product potency. CONCLUSIONS: Acceptable model fits to all individual subject dose-response data were not achieved for any dermatologic corticosteroid product. However, population dose-responses were adequately described by the Emax model. On the basis of these data, the optimal dose duration used for comparison of multisource dermatologic corticosteroid products is recommended to be equal to the ED50 based on population modeling of pilot dose-response study data.

Oxygenation of crystalloid cardioplegia.

A simple economical apparatus for oxygenation of cold crystalloid cardioplegic solutions is presented. It is sterile, practically feasible for use in open heart surgery, provides a PO2 of 98.7 kPa and sustains it for a period of more than 20 minutes.

Binding and aggregation of the 25-kilodalton toxin of Bacillus thuringiensis subsp. israelensis to cell membranes and alteration by monoclonal antibodies and amino acid modifiers.

The 25-kilodalton toxin of Bacillus thuringiensis subsp. israelensis binds irreversibly to Aedes albopictus cells, Choristoneura fumiferana cells, and erythrocytes. The binding to cells increased with both toxin concentration and time and when the cells were first preincubated with unlabeled toxin. Binding data indicated a two- to threefold increase in the rate of binding after the amount of the membrane-bound toxin reached approximately 3.5 fmol/3 x 10(5) A. albopictus cells or 3.3. fmol/2 x 10(5) C. fumiferana cells. When this level of bound toxin was reached, the toxins also began forming aggregates at the cell membrane. The toxin aggregates were extracted with 10% Triton X-100 and separated from the monomers with a 5 to 20% sucrose density gradient. The toxin aggregates isolated from A. albopictus and C. fumiferana cell membranes were ca. 400 kilodaltons, while those isolated from human erythrocytes were significantly smaller. The proportion of the toxin found in aggregate form increased rapidly with the amount of toxin bound; however, the molecular size of the aggregates remained constant. Eleven monoclonal antibodies raised against the native form of the toxin blocked 80 to 97% of the toxin binding to cells. The epitope of one of these monoclonal antibodies was mapped to a domain which included the cysteine, suggesting the importance of the domain around this amino acid to binding. Toxin binding and cell lysis were also inhibited by treating the toxin with HgCl2, further indicating the importance of the C-terminal hydrophobic cysteine-containing domain in cytolytic activity of the 25-kilodalton protein.

Preliminary X-ray diffraction analysis of crystals of Bacillus thuringiensis toxin, a cell membrane disrupting protein.

Crystals suitable for high resolution X-ray diffraction analysis have been reproducibly grown of the 24,000 Mr protein insect toxin from Bacillus thuringiensis. This protein, which demonstrates substantial insecticidal activity by inserting into phospholipid membranes, crystallizes as long square needles from polyethylene glycol 4000 at neutral pH. The crystals are of space group P4(1) and have cell dimensions of a = b = 33 A and c = 235 A, which suggests to us a predominantly helical motif for the protein's structure.

Cytolytic activity and immunological similarity of the Bacillus thuringiensis subsp. israelensis and Bacillus thuringiensis subsp. morrisoni isolate PG-14 toxins.

The parasporal bodies of the mosquitocidal isolates of Bacillus thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni isolate PG-14 were compared with regard to their hemolytic and cytolytic activities and the immunological relatedness of the 28- and 65-kilodalton (kDa) proteins that occur in both subspecies. The alkali-solubilized parasporal bodies of B. thuringiensis subsp. israelensis caused 50% lysis of human erythrocytes at 1.14 micrograms/ml, whereas those of B. thuringiensis subsp. morrisoni caused similar lysis at 1.84 micrograms/ml. Preincubation of solubilized parasporal bodies with dioleolyl phosphatidylcholine significantly inhibited the hemolytic activity of both supspecies. In cytolytic assays against Aedes albopictus cells, the toxin concentrations causing 50% lysis for B. thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni were 1.87 and 11.98 micrograms/ml, respectively. Polyclonal antibodies raised separately against the 25-kDa protein (a tryptic digest of the 28-kDa protein) or the 65-kDa protein of B. thuringiensis subsp. israelensis cross-reacted, respectively, with the 28- and the 65-kDa proteins of B. thuringiensis subsp. morrisoni. However, neither of these antibodies cross-reacted with the 135-kDa protein of either subspecies. These results indicate that the mosquitocidal and hemolytic properties of B. thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni isolate PG-14 are probably due to the biologically related proteins that are present in the parasporal bodies of both subspecies. The lower hemolytic activity of the B. thuringiensis subsp. morrisoni may be due to the presence of lower levels of the 28-kDa protein in that subspecies.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell membrane interaction of Bacillus thuringiensis subsp. israelensis cytolytic toxins.

Two toxic polypeptides of 24 and 25 kilodaltons (kDa) were purified from parasporal proteinaceous crystals of Bacillus thuringiensis subsp. israelensis. Both of these polypeptides, which are antigenically similar and have identical N terminals, lysed human erythrocytes and cultured mosquito cells. Although the 24-kDa peptide was more toxic than the 25-kDa peptide, both were less toxic than the crude alkali-solubilized crystal toxin. However, a 1:1 mixture of these 24- and 25-kDa proteins was more toxic than either of these polypeptides individually, indicating a possible interaction between these proteins at the cell membrane. Both the 24- and the 25-kDa proteins were inactivated by aqueous suspensions of dioleolylphosphatidylcholine, indicating the involvement of phospholipids in the cytotoxic action of these toxins. Thus the role of cell membrane phospholipids in mediating the toxin action was studied by using phospholipases as probes. Treatment of erythrocytes with high levels of phospholipase D increased their susceptibility to the toxin; however, phospholipase A2-treated erythrocytes were less susceptible to the toxin. These erythrocytes also bound less 125I-labeled 25-kDa toxin. These results support the role of fatty acyl residues at the syn-2 position of membrane phospholipids in toxin action. The cytolytic toxin of B. thuringiensis subsp. israelensis is thought to damage cell membranes in a detergentlike manner. However, there was a difference between the cytolytic action of this toxin and that of a nonionic detergent such as Triton X-100 because phospholipase A2-treated erythrocytes were more susceptible to Triton X-100, whereas such erythrocytes were less sensitive to the toxin. Thus, the cytolytic toxin apparently did not act as a nonspecific detergent, but rather interacted with phospholipid receptors on the cell membrane. Such an interaction of the toxin with phospholipid receptors probably results in the increased cell permeability, thereby causing cell lysis.

Cellular lesions in the central nervous system of Periplaneta americana following insecticide treatment in vitro and in vivo. I. Nerve cell bodies and the neuropile.

Cellular lesions in the metathoracic ganglion of P. americana following insecticide treatment have been examined. Treatment of the isolated ganglia with dieldrin (10 microM) and bioresmethrin (5 microM) induced mitochondrial damage in the neuropile and nerve cell bodies. The mitochondria in treated nerve cells were swollen with broken cristae and devoid of normal morphological appearance. Following in vivo treatment of cockroaches with these insecticides, this type of mitochondrial damage was observed even at the onset of poisoning. In the prostrate cockroaches, however, the mitochondrial swelling was accompanied by the accumulation of electron dense granules. In addition, the neuropiles of insecticide-treated ganglia contained secondary lysosomes which increased in size and number with the progress of poisoning and showed signs of depletion of synaptic vesicles. The action of dieldrin upon the ultrastructure was completely abolished by pretreatment of ganglia with 10 mM Mg2+. On the other hand, pretreatment of ganglia with tetrodotoxin and pentobarbital-sodium had very little effect on the action of dieldrin though these compounds blocked the action of bioresmethrin. The results of this study suggest that cellular lesions in the insect CNS, caused by dieldrin, are due to an enhanced uptake of calcium into nerve terminals which may occur independent of membrane depolarization. The effects of bioresmethrin upon the ultrastructure of the CNS are apparently mediated by nerve excitation and membrane depolarization. It is concluded that treatment of intact cockroaches with dieldrin and bioresmethrin initiates catabolic processes in the nerve cells leading to cellular lesions which are indicative of neuronal degeneration.

Cellular lesions in the central nervous system of Periplaneta americana following insecticide treatment in vitro and in vivo. II. Glial cells.

Glial cell damage in the central nervous system of P. americana following insecticide treatment has been studied. Treatment of isolated metathoracic ganglia with 10 microM dieldrin caused an increase in the electron opacity of the inner and outer glial cells without effecting such a change in the structure of glial cells in the ventral connectives. Glial cells in the ganglia and ventral connectives treated with 5 microM bioresmethrin were found to be swollen and even more electron lucent than the control cells. In the prostrate cockroaches (24 hr after the treatment) the action of bioresmethrin and dieldrin upon peripheral glial cells was identical. Both insecticides caused vacuolation and darkening of outer glial cells; their effects extended to include glial cells in the ventral connectives. At the onset of poisoning, 20-30 min following the application of dieldrin, outer glial cells exhibited slight increase in electron opacity while the inner glial cells showed increase in lysosomal activity. The observed action of dieldrin upon the ultrastructure of glial cells was prevented by pretreatment of the nervous tissue with 10 mM Mg2+. Though tetrodotoxin and sodium-pentobarbital had very little effect upon the action of dieldrin, these drugs blocked bioresmethrin-induced alterations in the fine structure of glial cells. The results of this study suggest that alterations in the ultrastructure of insect neuroglia following treatment with insecticides tested in this study are probably due to perturbations in the neural element of the nervous system.

Pharmacology of insecticide-induced release of hyperlipaemic hormone in the locust, Locusta migratoria.

1. The mechanism of insecticide-induced release of hyperlipaemic hormone from the glandular lobe of the isolated corpora cardiaca (CC) of locusts has been studied using pharmacological agents. 2. Treatment of isolated CC with various insecticides induces the release of hyperlipaemic hormone as judged by bioassay. 3. Reserpinisation of CC (25 micrograms/locust) or treatment of isolated CC with the alpha-adrenergic-receptor blocker phentolamine had no effect on the action of DDT or bioresmethrin, but partially blocked the action of dieldrin and chlorfenvinphos. 4. Treatment of isolated CC with the postsynaptic cholinergic blocker hexamethonium bromide abolished the effect of dieldrin and chlorfenvinphos, but did not block the action of DDT or bioresmethrin. 5. The effects of all the insecticides tested in this study were completely blocked by treatment of the CC with 10(-6) M tetrodotoxin. 6. The results indicate that DDT and bioresmethrin may act directly on the glandular cells via a sodium-dependent mechanism. The results with dieldrin and chlorfenvinphos suggest the presence of two distinct cholinergic pathways, one of which acts via the pre-synaptic aminergic terminals which control the glandular cells, whereas the other acts elsewhere in the CC. Sodium channels are also involved in the ultimate expression of these two insecticides.

Metabolic fate of [14C]dieldrin in Schistocerca gregaria with particular reference to the nervous system.

1. In the metabolism of [14C]dieldrin in Schistocerca gregaria, four metabolites of dieldrin were detected. Three of these have been identified as cis-aldrindiol, trans-aldrindiol and seco-aldrin dicarboxylic acid. 2. Apart from the formation of cis and trans-aldrindiol, another metabolite, M1, was detected in the nervous system. 3. The uptake of the dose of dieldrin was maximal in nerve cords followed by thoracic muscles and gut. Degradation of dieldrin was, however, not detected in thoracic muscles.