First chemical synthesis of a scorpion alpha-toxin affecting sodium channels: the Aah I toxin of Androctonus australis hector.

Aah I is a 63-residue alpha-toxin isolated from the venom of the Buthidae scorpion Androctonus australis hector, which is considered to be the most dangerous species. We report here the first chemical synthesis of Aah I by the solid-phase method, using a Fmoc strategy. The synthetic toxin I (sAah I) was renatured in DMSO-Tris buffer, purified and subjected to thorough analysis and comparison with the natural toxin. The sAah I showed physico-chemical (CD spectrum, molecular mass, HPLC elution), biochemical (amino-acid composition, sequence), immunochemical and pharmacological properties similar to those of the natural toxin. The synthetic toxin was recognized by a conformation-dependent monoclonal anti-Aah I antibody, with an IC50 value close to that for the natural toxin. Following intracerebroventricular injection, the synthetic and the natural toxins were similarly lethal to mice. In voltage-clamp experiments, Na(v) 1.2 sodium channel inactivation was inhibited by the application of sAah I or of the natural toxin in a similar way. This work describes a simple protocol for the chemical synthesis of a scorpion alpha-toxin, making it possible to produce structural analogues in time.     

Biological Parameters for Four Coccinellid Species in Central Saudi Arabia

Developmental and reproductive rates ofAdonia variegataGoeze,Coccinella novemnotataHerbst,C. septempunctataL., andC. undecimpunctataL. were studied in wheat and barley in Central Saudi Arabia. Beetles were reared in the laboratory at 25 ± 2°C onBrevicoryne brassicae(L.) andRhopalosiphum padiL.A. variegataandC. undecimpunctatahad significantly higher egg hatching (X = 81.8 and 74.5%), adult longevity (X = 71.8 and 70.0 days), and survival (X = 61.8 and 61.5%), respectively.Coccinella undecimpunctatahad the highest fecundity (X = 370.5 eggs/female) and longest oviposition period (X = 29.8 days).C. septempunctataexhibited the lowest values for these biological parameters. No differences were evident in the total developmental time or egg and pupal duration, although species differences occurred in the larval duration. Larvae ofA. variegatadeveloped faster (X = 11.8 days) than Larvae ofC. septempunctata(X = 12.0 days),C. undecimpunctata(X = 12.2 days), orC. novemnotata(X = 13.1 days). The intrinsic rate of increase (r_m) and predicted population density were higher forC. undecimpunctata(X = 0.093 and 98553) after 60 days than forC. novemnotata(X = 0.088 and 59405),C. septempunctata(X = 0.086 and 35033), orA. variegata(X = 0.082 and 37991), respectively.     

Immunocytochemical identification and localization of lipase in cells of the mycelium of Penicillium cyclopium variety

The localization of lipase in cells of the fungus Penicillium cyclopium was investigated. It was shown by differential centrifugation of a homogenate of mycelial cells that the activity of the enzyme is associated with the cell wall. A study of ultrathin sections of mycelium fixed using the method of Zvyagintseva in an electron microscope showed that the final products of lipolytic activity of the enzyme is localized on the cell wall. Antibodies were raised against the purified A and B lipases from P. cyclopium and their specificity was assessed by enzyme-linked immunosorbent assay. The antibody preparation was used in cytochemical investigation by immunogold labelling. This study permits the localization of cell-bound lipase mainly in the cell wall and in the periplasmic space. The identity of the cell-bound lipase with one of the two extracellular lipases is also demonstrated.     

Precedence of the first male sperm in successive matings in the HymenopteraDiadromus pulchellus

In most insect species where double matings occur, sperm from the second male preferentially fertilize subsequent eggs. However, we demonstrate here that, as already shown for some other hymenopteran species, this is not the case in the ichneumonid waspDiadromus pulchellus (Wesmeal): sperm from the first male usually father all the female progeny. This precedence of the first male sperm is also observed in double matings involving an haploid male and a diploid sterile male, whichever is the first mating male. We discuss the consequences of this phenomenon from an evolutionary point of view.     

Somatic embryogenesis and analysis of peroxidases inPhoenix dactylifera L

To determine some physiological parameters implicated in somatic embryogenesis in date palm (Phoenix dactylifera L.), peroxidases have been studied. Activated charcoal commonly used in date palm tissue culture as an essential antibrowning factor decreased cellular protein contents and peroxidase activities. During the first months of culture, the conventionally used medium (100 mg dm^?3 of 2,4-dichlorophenoxyacetic acid, 3 g dm^?3 charcoal) reduces 2 to 3 and 4 to 6 times protein contents and peroxidase activities, respectively, in comparison with the same one containing only 5 mg dm^?3 of 2,4-D and with or without 150 mg dm^?3 charcoal. In addition, the standard procedure decreased the embryogenic potential which is positively related to the intra- and extracellular (excreted into culture medium) peroxidase activities. In medium with embryogenic calli, extracellular peroxidase activity was three times as high as the activity determined in the same medium with non-embryogenic calli. There were two basic isoforms and four to five acidic bands characterizing the embryogenic calli. It can be suggested that peroxidases play a key role in somatic embryogenesis of date palm and the charcoal used at 3 g dm^?3 constitute a perturbating factor for this process.     

Herbicidal activity of phosphonic,phosphinic, and phosphonous acid analogues of phenylglycine and phenylalanine

A series of phosphonic, phosphinic, and phosphonous acid analogues of phenylglycine and phenylalanine was synthesized and tested as herbicides against Lepidium sativum and Cucumis sativus. Aminobenzylphosphonic acids exhibited notable herbicidal activity and thus represent a group of the most active herbicides found among aminophosphonic acids.     

Structure-lipophilicity relationships of peptides and peptidomimetics

Summary Understanding the physicochemical and structural properties of peptides are important prerequisites for the rational design of bioactive peptides and peptidomimetics. The present contribution reviews methods used for the assessment and prediction of lipophilicity (or hydrophobicity) and their correlation with structural elements of peptides and closely related peptidomimetics.     

Interaction of influenza virus proteins with planar bilayer lipid membranes II. Effects of rimantadine and amantadine

The dependence of the surface potential difference (ΔU), transversal elasticity module (E₁) and membrane conductivity (G₀) on the concentrations of the antiviral drugs, rimantadine and amantadine was studied in the planar bilayer lipid membrane system. The method used was based on independent measurements of the second and third harmonics of the membrane capacitance current. The binding constants of bilayer lipid membranes obtained from the drug adsorption isotherms were 2.1 · 10⁵ M^?1 and 1.3 · 10⁴ M^?1 for rimantadine and amantadine, respectively. The changes in G₀ took place only after drug adsorption saturation had been achieved. The influence of rimantadine and amantadine on the interaction of bilayer lipid membranes with matrix protein from influenza virus was also investigated. The presence of 70 μg/ml rimantadine in the bathing solution resulted in an increase in the concentration of M-protein at which the adsorption and conductance changes were observed. The effects of amantadine were similar to those of rimantadine but required a higher critical concentration of amantadine. The results obtained suggest that the antiviral properties of rimantadine and amantadine may be related to the interaction of these drugs with the cell membrane, which can affect virus penetration into the cell as well as maturation of the viral particle at the cell membrane.     

Evidence for a highly asymmetric arrangement of ether- and diacyl-phospholipid subclasses in the plasma membrane of Krebs II ascites cells

(1) Krebs II ascites cells were taken as a model of the neoplastic cells to investigate the transverse distribution of phospholipids in the plasma membrane. The experimental procedure was based on non-lytic degradation of phospholipids in the intact cell by Naja naja phospholipase A₂ and Staphylococcus aureus sphingomyelinase C and on phopholipid analysis of purified plasma membranes. It was shown that the three major phospholipids, i.e., phosphatidylcholine, phosphatidylethanolamine and sphingomyelin, are randomly distributed between the two halves of the membranes, whereas phosphatidylserine remains located in the inner leaflet. (2) The membrane localization of phosphatidylcholine and phosphatidylethanolamine subclasses (diacyl, alkylacyl and alkenylacyl) was also examined, using a new procedure of ether-phospholipid determination. The method involves a selective removal of diacyl species by guinea pig pancreas phospholipase A₁ and of alkenylacyl species by acidolysis. This analysis revealed a 50% increase of ether phospholipids in the plasma membrane as compared to the whole cell (36.5 and 23.1% of total phospholipid, respectively). Furthermore, a strong membrane asymmetry was demonstrated for the three phosphatidylcholine subclasses, since 1-alkyl-2-acyl-sn-glycerol-3-phosphocholine (alkylacyl-GPC) was entirely found in the inner leaflet, whereas both diacyl- and alkenylacyl-GPC displayed an external localization. The same pattern was observed for phosphatidylethanolamine subclasses, except for 1-alkenyl-2-acyl-sn-glycero-3-phosphoethanolamine, which was found randomly distributed. These results are discussed in relation to the process of cell malignant transformation and to the biosynthesis of platelet-activating factor (PAF-acether or 1-alkyl-2-acetyl-GPC).