SPC3, an anti-HIV peptide construct derived from the viral envelope,binds and enters HIV target cells

SPC₃ is a peptide construct (eight branches of the GPGRAF motif) derived from the consensus sequence present at the apex of the third variable domain of the human immunodeficiency virus (HIV) envelope (Env). It presents a potent anti-HIV activity and is currently tested in phase II clinical trials (FDA protocol 257A). Its mode of action remains unclear. It was thought that SPC₃ exerts its effect both during HIV interaction with CD4⁺ cells but also through interference either with a post-binding event or with Env processing. Accordingly, SPC₃ was supposed to be able to bind and to enter CD4⁺ cells. In this work, we addressed these points. SPC₃ was found to interact with CD4⁺ cell membrane with a K_0.5 value in the range of 500 nm . The binding of SPC₃ to CD4⁺ cells involves its interaction with a cell membrane associated protein which is pronase sensitive and different from CD4. This interaction was similar from 2 to 37°C. The maximum binding occurred at acidic pH whereas the interaction was inhibited in alkaline conditions. We observed also that SPC₃ was internalized rapidly into the cells—the maximal intracell amount was reached within 30 min—where it remained stable for at least 24 h. Altogether, these data suggest that SPC₃ can exert its antiviral activity via interference with events occurring at the cell surface but also into the target cell. © 1998 European Peptide Society and John Wiley & Sons, Ltd.     

The decrease of catalase or esterase D activity in patients with microdeletions of 11p or 13q does not increase their radiosensitivity

Lymphocyte cultures from patients affected by retinoblastoma (Rb), with or without a microdeletion of chromosome 13, and Wilms tumor (WT), with a microdeletion of chromosome 11p where exposed to gamma-ray radiation during S and G2 phases. Chromatid and chromosome lesions were scored and compared to those observed in controls. No significant differences were detected, neither between patients and controls, nor between patients carrying or not a microdeletion. This lack of difference was unexpected since the genes for catalase and esterase D, also called S-formyl glutathione hydrolase, which are two detoxication enzymes, are deleted in case of microdeletion of 11p and 13q, respectively.     

IMGT standardized criteria for statistical analysis of immunoglobulin V-REGION amino acid properties

IMGT, the international ImMunoGeneTics information system(R) (http://imgt.cines.fr) is a high-quality integrated information system specializing in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of human and other vertebrates. IMGT comprises IMGT/LIGM-DB, the comprehensive database of IG and TR sequences from human and other vertebrates (76 846 sequences in September 2003). In order to define the IMGT criteria necessary for standardized statistical analyses, the sequences of the IG variable regions (V-REGIONs) from productively rearranged human IG heavy (IGH) and IG light kappa (IGK) and lambda (IGL) chains were extracted from IMGT/LIGM-DB. The framework amino acid positions of 2474 V-REGIONs (1360 IGHV, 585 IGKV, 529 IGLV) were numbered according to the IMGT unique numbering. Two statistical methods (correspondence analysis and hierarchic classification) were used to analyze the 237 framework positions (80 for IGHV, 79 for IGKV, 78 for IGLV), for three properties (hydropathy, volume and chemical characteristics) of the 20 common amino acids. Results of the analyses are shown as standardized two-dimensional representations, designated as IMGT Colliers de Perles statistical profiles. They provide a characterization of the amino acid properties at each framework position of the expressed IG V-REGIONs, and a visualization of the resemblances and differences between heavy and light, and between kappa and lambda sequences. The standardized criteria defined in this paper, amino acid positions and property classes, will be useful to study the mutations and allele polymorphisms, to establish correlations between amino acids in the IG and TR protein three-dimensional structures and to extract new knowledge from V-like domains of chains, other than IG and TR, belonging to the immunoglobulin superfamily.     

Synthesis, 1H NMR structure, and activity of a three-disulfide-bridged maurotoxin analog designed to restore the consensus motif of scorpion toxins

Maurotoxin (MTX) is a 34-residue toxin that has been isolated from the venom of the chactidae scorpion Scorpio maurus palmatus. The toxin displays an exceptionally wide range of pharmacological activity since it binds onto small conductance Ca(2+)-activated K(+) channels and also blocks Kv channels (Shaker, Kv1.2 and Kv1.3). MTX possesses 53-68% sequence identity with HsTx1 and Pi1, two other K(+) channel short chain scorpion toxins cross-linked by four disulfide bridges. These three toxins differ from other K(+)/Cl(-)/Na(+) channel scorpion toxins cross-linked by either three or four disulfide bridges by the presence of an extra half-cystine residue in the middle of a consensus sequence generally associated with the formation of an alpha/beta scaffold (an alpha-helix connected to an antiparallel beta-sheet by two disulfide bridges). Because MTX exhibits an uncommon disulfide bridge organization among known scorpion toxins (C1-C5, C2-C6, C3-C4, and C7-C8 instead of C1-C4, C2-C5, and C3-C6 for three-disulfide-bridged toxins or C1-C5, C2-C6, C3-C7, and C4-C8 for four-disulfide-bridged toxins), we designed and chemically synthesized an MTX analog with three instead of four disulfide bridges ([Abu(19),Abu(34)]MTX) and in which the entire consensus motif of scorpion toxins was restored by the substitution of the two half-cystines in positions 19 and 34 (corresponding to C4 and C8) by two isosteric alpha-aminobutyrate (Abu) derivatives. The three-dimensional structure of [Abu(19), Abu(34)]MTX in solution was solved by (1)H NMR. This analog adopts the alpha/beta scaffold with now conventional half-cystine pairings connecting C1-C5, C2-C6, and C3-C7 (with C4 and C8 replaced by Abu derivatives). This novel arrangement in half-cystine pairings that concerns the last disulfide bridge results mainly in a reorientation of the alpha-helix regarding the beta-sheet structure. In vivo, [Abu(19),Abu(34)]MTX remains lethal in mice as assessed by intracerebroventricular injection of the peptide (LD(50) value of 0. 25 microg/mouse). The structural variations are also accompanied by changes in the pharmacological selectivity of the peptide, suggesting that the organization pattern of disulfide bridges should affect the three-dimensional presentation of certain key residues critical to the blockage of K(+) channel subtypes.     

Mechanisms of maurotoxin action on Shaker potassium channels

Maurotoxin (alpha-KTx6.2) is a toxin derived from the Tunisian chactoid scorpion Scorpio maurus palmatus, and it is a member of a new family of toxins that contain four disulfide bridges (, Eur. J. Biochem. 254:468-479). We investigated the mechanism of the maurotoxin action on voltage-gated K(+) channels expressed in Xenopus oocytes. Maurotoxin blocks the channels in a voltage-dependent manner, with its efficacy increasing with greater hyperpolarization. We show that an amino acid residue in the external mouth of the channel pore segment that is known to be involved in the actions of other peptide toxins is also involved in maurotoxin's interaction with the channel. We conclude that, despite the unusual disulfide bridge pattern, the mechanism of the maurotoxin action is similar to those of other K(+) channel toxins with only three disulfide bridges.     

Use of a mathematical model to study the dynamics of Ctenocephalides felis populations in the home environment and the impact of various control measures

The biology of fleas has been studied by a number of authors, as has the impact of various types of control measures. However, there are no mathematical models simulating the dynamics of a population of Ctenocephalides felis felis fleas on their host (the cat) and in their close environment (apartment). The model presented in this paper allows for integration of the numerous biological and behavioural parameters of the parasites and their hosts and for the variation of these same parameters. The various types of control measures can be programmed so that their impact over time can be studied. The model confirms the key role played by adult fleas, or emerged fleas contained in the cocoon. Only regular applications of persistent insecticides to the host animal will enable control of the parasite population. A combination of these insecticides with an IGR (Insect Growth Regulator) will accelerate decontamination of the home environment and see the disappearance of the parasites altogether if they are not reintroduced. The association of additional measures such as vacuum cleaning will accelerate the process of decontamination but will have no impact if carried out in isolation. One-off treatment with insecticide will not enable a reduction in the parasite population, even if carried out frequently. Use of insecticides on the home environment premises alone does not appear to be an adequate means of control. The present model can be used to test various integrated control measures which take into account different factors such as the number of host animals, the frequency of movement outdoors, the impact of the seasons.     

Tree species diversity and forest structure in relation to microtopography in a tropical freshwater swamp forest in French Guiana

Diversity of tree association and forest structure were analysed in relation to microtopography and flooding intensity in a tropical freshwater swamp forest in the Sinnamary river basin, French Guiana. A 530-m-long vegetation transect was established through a hummock-hollow terrain. Nine 10 m× 50 m sample plots, perpendicular to the main transect, were located so that each was as microtopographically uniform as possible. Trees with dbh (diameter at 1.3 m) 10 cm were censused in all plots and trees with 2 cm dbh < 10 cm in three plots. Sixty tree species belonging to 39 genera and 30 families were recorded. The study area was divided into low and high sites according to microtopography and flooding intensity. According to the Czekanowski similarity matrix, the tree association in low, most frequently flooded, sites differed from that in the high sites under intermediate or low flooding intensity. The low sites had higher stand density and lower species richness than the high sites. Trees with dbh 10 cm in low sites were small and stand basal area (SBA) was about the same in low (69.6 m² ha^–1) and high (64.3 m² ha^–1) sites. The low areas were dominated by Pterocarpus officinalis (38% of stems with dbh 10 cm and 36% of SBA) and Malouetia tamaquarina (26 and 15%). Diospyros guianensis (13.4% of stems with dbh 10 cm and 6.1% of SBA), a Caraipa sp. (14.0 and 7.9%), Lecythis corrugata (6.6 and 3.5%) and emergent Caryocar microcarpum (0.9 and 13.9%) were abundant in high sites. Nodulated legume trees, P. officinalis, Hydrochorea corymbosa and Inga disticha, comprised 44% of stems in the low sites. The abundant nodulation suggests that symbiotic dinitrogen fixation may be an adaptation to N-depleted waterlogged soils. Other adaptive responses were litter accumulation between the buttresses of P. officinalis, which formed hummocks above surface water, and clonal growth habit of M. tamaquarina, which resulted in formation of monospecific groves in low sites.