Efficacy of chemically characterized Foeniculum vulgare Mill seed essential oil in protection of raw tobacco leaves during storage against fungal and aflatoxin contamination.

AIMS: To report fungal and aflatoxin contamination in stored tobacco leaves and the potential of Foeniculum vulgare (fennel) seed essential oil (EO) as a plant-based preservative in protection of tobacco during storage. METHODS AND RESULTS: Mycological analysis of tobacco samples was done by surface sterilization and serial dilution tests. The Aspergillus flavus isolates were screened for their toxigenicity. Both in vivo and in vitro tests were done to evaluate antifungal and antiaflatoxigenic efficacy of chemically characterized EO. The mycoflora analysis revealed 108 fungal colonies belonging to five genera and nine species. All A. flavus isolates were found aflatoxigenic during screening. Gas chromatography and mass spectrometry analysis of EO identified 19 components (99·66%); estragole being the major component (47·49%). The EO showed broad fungitoxicity at 1·25 µl ml(-1) and 100% inhibition to AFB1 production as well as ergosterol synthesis at 1·0 µl ml(-1) concentration. EO showed 100% protection of stored tobacco samples from aflatoxin B1 contamination. CONCLUSIONS: The fennel EO can thus be formulated as a plant-based preservative for food items. SIGNIFICANCE AND IMPACT OF THE STUDY: The present investigation comprises the first report on antiaflatoxin efficacy of fennel oil and its potency in the protection of tobacco leaves from fungal and aflatoxin contamination during storage.

Effectiveness of a multidimensional approach for prevention of ventilator-associated pneumonia in 21 adult intensive-care units from 10 cities in India: findings of the International Nosocomial Infection Control Consortium (INICC).

We report on the effect of the International Nosocomial Infection Control Consortium's (INICC) multidimensional approach for the reduction of ventilator-associated pneumonia (VAP) in adult patients hospitalized in 21 intensive-care units (ICUs), from 14 hospitals in 10 Indian cities. A quasi-experimental study was conducted, which was divided into baseline and intervention periods. During baseline, prospective surveillance of VAP was performed applying the Centers for Disease Control and Prevention/National Healthcare Safety Network definitions and INICC methods. During intervention, our approach in each ICU included a bundle of interventions, education, outcome and process surveillance, and feedback of VAP rates and performance. Crude stratified rates were calculated, and by using random-effects Poisson regression to allow for clustering by ICU, the incidence rate ratio for each time period compared with the 3-month baseline was determined. The VAP rate was 17.43/1000 mechanical ventilator days during baseline, and 10.81 for intervention, showing a 38% VAP rate reduction (relative risk 0.62, 95% confidence interval 0.5-0.78, P = 0.0001).

Age-related changes underlie switch in netrin-1 responsiveness as growth cones advance along visual pathway.

Retinal axons are led out of the eye by netrin-1, an attractive guidance cue which is secreted at the optic nerve head. In the optic pathway, however, netrin-1 is expressed in areas that exclude retinal axon growth. This suggests that axons may change in their responsiveness to netrin-1 as they advance along the pathway. Indeed, in our 'whole-pathway' preparation in Xenopus, a gradual change from attraction to repulsion occurred as retinal axons emerged from progressively distal points along the pathway. We also found that axons that were aged in culture without pathway experience underwent a similar change, which correlated with a decline in cyclic AMP (cAMP) and netrin-1 receptor expression. Cyclic AMP elevators and adenosine A2b receptor agonists rejuvenated the behavior of old growth cones, causing them to regain attraction to netrin-1, whereas antagonists caused young growth cones to be repelled. These findings show that netrin-1 responsiveness is developmentally regulated and suggest that intrinsic changes that lower cAMP levels underlie this regulation.

Resolution, molecular structure and biological activities of the D- and L-enantiomers of potent anti-implantation agent, DL-2-[4-(2-piperidinoethoxy)phenyl]-3-phenyl-2H-1-benzopyran.

Compound 1 (DL-2-[4-(2-piperidinoethoxy)phenyl]-3-phenyl-2H-1-benzopyran, CDRI 85/287) a potent anti-estrogen and anti-implantation agent has been successfully resolved into its pure D- and L-enantiomers. Biological studies showed L-enantiomer to be the active form, exhibiting a fivefold higher receptor affinity for the rat uterine cytosolic estrogen receptor, 100% contraceptive efficacy at 1.3 mg/kg dose in single day schedule and 89% inhibition of estradiol induced increase of uterine weight at its contraceptive dose. The absolute stereochemistry determined by X-ray crystallographic analysis showed that the L-enantiomer has 2R configuration at its asymmetric centre.

Lecithin-cholesterol acyltransferase activity in normocholesterolaemic and hypercholesterolaemic roosters: modulation by lipid apheresis.

Lipid apheresis, a recently described procedure for the elimination of lipid but not apolipoproteins from plasma, was applied to normocholesterolaemic and hypercholesterolaemic roosters. Lipid apheresis resulted in an immediate reduction in plasma unesterified cholesterol concentration, which was sustained for 150 min. The reduction in unesterified cholesterol concentration was higher in the normocholesterolaemic animals than in the hypercholesterolaemic animals. Lipid apheresis induced changes in the ratio of plasma unesterified to total cholesterol in normocholesterolamic animals but not in hypercholesterolaemic animals. In hypercholesterolaemic animals, lecithin-cholesterol acyltransferase (LCAT) activity was not affected by lipid apheresis, whereas in normocholesterolaemic animals LCAT activity was acutely reduced for 150 min after lipid apheresis. Saturated LCAT kinetics occurred in the hypercholesterolaemic animals but not in the normocholesterolaemic animals. LCAT obeyed Michaelis-Menten kinetics. After lipid apheresis, there was a pool of unesterified cholesterol that was available as substrate for LCAT to a greater extent in hypercholesterolaemic animals than in normocholesterolaemic animals. These observations may have important implications for lipid apheresis as a treatment for atherosclerosis.

Lipid apheresis in an animal model causes in vivo changes in lipoprotein electrophoretic patterns.

Lipid apheresis, a new extracorporeal procedure based on plasma delipidation and showing promise as a possible treatment for atherosclerosis, was recently reported for the first time from this laboratory [Cham et al., J Clin Apheresis 10:61-69, 1995]. In the present study lipid apheresis was applied to hypercholesterolemic and normocholesterolemic roosters to examine its effect on plasma lipoprotein particles. This procedure resulted in conspicuous changes in electrophoretic patterns of plasma lipoproteins. The electrophoretic mobilities of all the lipoprotein fractions had changed considerably. Lipid stainable material was present in at least three bands in the alpha-globulin area. In particular, changes in the electrophoretic region of high-density lipoproteins were observed. Lipid apheresis markedly induced the anti-atherogenic pre- beta-high-density lipoproteins. The observed changes induced by lipid apheresis were more pronounced in the hyperlipidemic animals compared with the normocholesterolemic controls. A novel pre-alpha-lipoprotein band was observed soon after lipid apheresis. This lipoprotein band had a density larger than 1.21. At approximately 150 minutes after lipid apheresis, the electrophoretic pattern had almost returned to its original base pattern. Lipid apheresis results in plasma lipoprotein changes which may induce reverse cholesterol transport and shows promise as a possible treatment of atherosclerosis.

Lipid apheresis: an in vivo application of plasma delipidation with organic solvents resulting in acute transient reduction of circulating plasma lipids in animals.

Despite primary and secondary prevention of coronary disease with lowering plasma cholesterol by diet and drug therapy, coronary heart disease remains the major cause of death in Western countries. Low density lipoprotein apheresis had the potential to make a significant impact as it acutely leads to a marked reduction in plasma cholesterol. However, recent preliminary results suggest that low density lipoprotein apheresis may not be more effective in preventing progression of coronary disease than current drug therapy. We have devised a new technique, termed lipid apheresis, which removes cholesterol and triglycerides from plasma but retains the apolipoproteins. This procedure shows great promise in stimulating regression beyond current therapy. Lipid apheresis, a new extracorporeal procedure based on plasma delipidation with the organic solvent mixture butanol-diisopropyl ether, was applied to hypercholesterolemic and normocholesterolemic roosters. Approximately 25% of the calculated blood volume was removed from the animals. The plasma was separated from the blood cells. The plasma was delipidated for 20 min with the organic solvent mixture. The delipidated plasma containing all proteins, including the apolipoproteins and other ionic constituents, was remixed with the blood cells and infused back into the identical donor animals. Analyses of serial blood samples collected from lipid apheresed and sham treated animals up to 16 h after infusion revealed that lipid apheresis caused acute, marked reductions in plasma lipids. The pattern and extent of the plasma levels of cholesterol were different in the hypercholesterolemic animals when compared with normocholesterolemic animals, indicating that a readily extraplasma cholesterol pool in the hypercholesterolemic animals was rapidly mobilized into the plasma pool.(ABSTRACT TRUNCATED AT 250 WORDS)

Density of sodium channels in insect synaptic nerve endings.

Specific binding of [11-(3)H]saxitoxin (STX) and activity of ouabain sensitive adenosine-triphosphatase (Na(+), K(+)-ATPase) were determined in neuronal membrane fractions using a subcellular preparation from the central nervous system of the cockroach Periplaneta americana. The nerve ending fractions (synaptosomes) contained 90-95% of the total specific activity of ouabain sensitive Na(+), K(+)-ATPase, and 60-70% of specific STX binding of the crude nerve homogenate. Sodium influx induced by veratridine in synaptosomes was inhibited by saxitoxin at a half-maximal concentration of 4 nM, and kinetics were consistent with reversible binding of one molecule of saxitoxin to each sodium channel receptor site, with an equilibrium dissociation constant (K(D)) of approx. 3 nM. The density of saturable binding sites was 2 pmol/mg protein which was estimated to correspond to about 95 binding sites per ?m(2) of synaptic membrane. The results of transport and binding data show that insect synaptosomes possess the capability to conduct inward sodium currents at least comparable to those found in other neuronal membranes, and thus provide a physiologically viable preparation to assess the effect of cation fluxes on the synaptic transmitter release process.

Alkaloid neurotoxins-dependent sodium transport in insect synaptic nerve-ending particles.

1. Sodium uptake associated with the activation of voltage-sensitive sodium channels by alkaloid activators, batrachotoxin, veratridine, and aconitine in presynaptic nerve terminals isolated from the central nervous system of cockroach (Periplaneta americana) was investigated. 2. Batrachotoxin (K0.5, 0.2 microM) was full agonist as for most effective activator of Na+ uptake; veratridine (K0.5, 2.5 microM) and aconitine (K0.5, 7.6 microM) produced a maximal stimulation of 22Na+ uptake that were 71% and 43% respectively of that produced by batrachotoxin. 3. Veratridine-dependent 22Na+ uptake was completely inhibited by tetrodotoxin (I0.5, 11 nM), a specific inhibitor of the nerve membrane sodium channels. 4. The present study describes appropriate conditions for measuring neurotoxins--stimulated sodium transport in insect central nervous system synaptosomes. The data show that voltage-sensitive sodium channels as defined by specific activation by the alkaloid neurotoxins are qualitatively distinct in insect synaptosomes than those previously described for vertebrate brain synaptosomes, cultured neuronal cell, nerve membrane vesicles and neuroblastoma cells.

Effects of phenylalanine and its deaminated metabolites on Na+,K+-ATPase activity in synaptosomes from rat brain.

The effects of phenylalanine (PHE) and its deaminated metabolites phenylpyruvate (PHP), phenyllactate (PHL) and phenylacetate (PHA) on sodium and potassium activated adenosinetriphosphatase (Na+,K+-ATPase) in synaptosomes from rat brain were investigated. At very low concentrations (5-10 microM). PHE, PHL and PHA inhibited the activity, while PHP stimulated the activity. At intermediate concentrations (50-100 microM), all compounds had no effect, but at higher (0.5-1.0 mM) concentrations they inhibited the enzyme activity. Thus all the compounds tested showed a biphasic effect on the enzyme activity. Hydroxylamine inhibited the Na+,K+-ATPase activity when present alone; simultaneous addition of hydroxylamine and PHE, however, eliminated the inhibitory effects of each other. Reversal of mutual inhibition also occurred in the presence of hydroxylamine and very low (5-10 microM) concentrations of PHL or PHA. The inhibitory effects of PHE at aLl concentrations, and of PHL or PHA at low concentrations, were also eliminated in the presence of EGTA. The data indicate that inhibition of brain membrane Na+,K+-ATPase by PHE and by low concentrations of PHL and PHA may involve metal ions, but that the inhibition by high concentrations of these metabolites must occur by a different mechanism. Since Na+,K+-ATPase plays a central role in neuronal function, and the presence of excess PHE and its deaminated metabolites occurs in brain tissue under conditions of experimentally induced hyperphenylalaninemia and genetic phenylketonuria, the neurologic impairment in experimental and genetic PKU may in part be related to the deleterious effects of these compounds on brain ATPase.