Influence of endotoxin on the stereoselective pharmacokinetics of oxprenolol,propranolol, and verapamil in the rat

The influence of endotoxin-induced inflammation on the enantioselective pharmacokinetics of propranolol, oxprenolol, and verapamil, which bind to α₁-acid glycoprotein, was studied in the rat. The racemic mixtures were given orally. In the control animals, for propranolol and oxprenolol, the plasma concentrations of the (R)-enantiomer were higher than those of the (S)-enantiomer, while for verapamil the reverse was true. Protein binding and intrinsic clearance are the main factors responsible for this enantioselectivity. After endotoxin treatment, for the three drugs tested the plasma concentrations and the plasma binding of both enantiomers were significantly increased. This effect was more pronounced for (R)-propranolol, (R)-oxprenolol, and (S)-verapamil than for their respective antipodes. The enantioselective effect of endotoxin on the plasma concentrations of the drugs studied seems mainly due to the enantioselective increase in binding to α₁-acid glycoprotein. © 1994 Wiley-Liss, Inc.     

Differential expression of several E2-type ubiquitin carrier protein genes at different developmental stages inArabidopsis thaliana andNicotiana sylvestris

We characterized three genes encoding different E2-type ubiquitin carrier proteins involved in the ubiquitin-mediated proteolytic pathway:UbcAt3 shows homologies to the yeastCDC34 gene andUbcAt4a andUbcAt4b are two different genes homologous to theUbc1/4/5 subfamily in yeast. Their accumulation was analysed and compared with that of the different families encoding polyubiquitins, as well as the monoubiquitin fusion protein, which is considered as a marker for cell division, during various developmental stages including GO/S transition and senescence of higher plant cells. Our results imply that theseUbc genes are under the control of complex mechanisms, and are differentially regulated, but not necessarily co-regulated with ubiquitin genes. Even the closely relatedUbcAt4a andUbcAt4b genes of the same multigene subfamily are controlled by distinct regulatory mechanisms.     

IPG (inositolphosphate glycan) as a cellular signal for TGF-β1 modulation of chondrocyte cell cycle

The knowledge of transforming growth factor (TGF)-β receptors has greatly progressed in the recent years. TGF-β receptors type I and II have been implicated in the modulation of cell proliferation, whereas type III (betaglycan) may act as a component presenting TGF-β to its signaling receptors. In addition, four other proteins that bind TGF-β1 or TGF-β2 have been recently identified in some cell lines, three being anchored to the membrane through a glycosylphosphatidylinositol (GPI). Despite this knowledge, the molecular mechanism of signal transduction through the TGF-β receptors remain an enigma. TGF-β family does not signal via any of the classical pathways. As GPI anchors of membrane proteins have been implicated in the transduction of some hormonal effects, we investigated the putative role of GPI in signaling the TGF-β effects on the proliferation of rabbit articular chondrocytes (RAC). We previously showed that TGF-β1 increased DNA replication rate of RAC, with a recruitment of cells in G2/M followed by a subsequent mitosis wave. Here, we find that the factor causes specific GPI hydrolysis, with correlated increase of inositolphosphate glycan (IPG). This effect was specifically inhibited by antibodies that bind TGF-β1. Using [³H]-inositol labeling and Triton X-114 extraction, we demonstrate that a hydrophobic material from the membrane is cleaved by treatment of cell cultures with phosphatidylinositol specific phospholipase C (PI-PLC) or by exposure to TGF-β, supporting that a PI-anchored molecule gives rise to IPG by TGF-β-induced hydrolysis. The biological relevance of this hydrolysis was demonstrated by the enhancing effect of purified IPG on the DNA synthesis rate, which mimicked the TGF-β action. These results demonstrate that IPG could be an early messenger in the cellular signaling that mediates the effect of TGF-β on RAC growth. © 1993 Wiley-Liss, Inc.     

Management oriented forest map of South India: thematic derivations

The principles and methods of the vegetation mapping undertaken at the French Institute, Pondicherry, are dealt with herein. Particularly, the characterisation of the different types of vegetation and especially the originality of the method: the dynamic interpretation of the vegetation and the depiction of the bioclimatic conditions. The programme of the forest map of South India at scale 1:250 000, undertaken in collaboration with the forest departments of the concerned states, is then described with special attention given to the source and the collection of data. This map has been conceived to serve as a basic document for the sustainable management of the forests. Three examples of its application are given. They concern the detection of anomalies between the existing forest cover and the prevalent climatic environment; the detection of areas for which protection is urgently needed; the selection of regions showing a particular interest in the field of nature conservation or as gene pool reserve. Finally, an example of a thematic map of sensibility of the forests is given, using the vegetation map as a basis.     

Endo's stress analysis of the primate skull and the functional significance of the supraorbital region

A review of Endo's experimental and theoretical procedures and data indicates that the magnitude of the principal strains in the glabella region of both humans and gorillas are low as compared to other parts of the face. Therefore, his data do not provide support for the hypothesis that the glabella region is a highly stressed region during biting. In addition, increased levels of strain in the supraorbital region are directly related to increased levels of masticatory muscle and reaction forces, and not necessarily to anterior tooth loading as opposed to posterior tooth loading. His data also indicate that the supraorbital region in extant humans cannot be accurately modeled as a beam. These conclusions either differ from those of Endo or are not clearly presented or emphasized throughout any of Endo's papers. Therefore, we suggest that a number of investigators have made unsupported or erroneous conclusions based on Endo's work. This is particularly true for those studies that have emphasized the existence of powerful bending stress in the glabella region during incisor biting in both humans and non-human primates.     

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.     

Cloning and characterization of theeapB andeapC genes ofCryphonectria parasitica encoding two new acid proteinases,and disruption ofeapC

Two new proteinases secreted byCryphonectria parasitica, namely EapB and EapC, have been purified. The corresponding structural genes were isolated by screening a cosmid library, and sequenced. Comparison of genomic and cDNA sequences revealed that theeapB andeapC genes contain three and two introns, respectively. The products of theeapB andeapC genes as deduced from the nucleotide sequences, are 268 and 269 residues long, respectively. N-terminal amino acid sequencing data indicates that EapC is synthesized as a zymogen, which yields a mature 206-amino acid enzyme after cleavage of the prepro sequence. Similarly, sequence alignment studies suggest that EapB is secreted as a 203-residue form which shares extensive similarities not only with EapC but also with two other acid fungal proteinases. However, they display distinct structural features; for example, no cysteine residue is found in EapC. TheeapC gene was mutated using a two-step gene replacement strategy which allowed the specific introduction of several stop codons at the beginning of theeapC coding sequence in an endothiapepsin-deficient (EapA⁺)C. parasitica strain. Although the resulting strain did not secrete EapC, it still exhibited residual extracellular proteolytic activity, which could be due to EapB.     

Subsets of sialylated, sulfated mucins of diverse origins are recognized by L-selectin. Lack of evidence for unique oligosaccharide sequences mediating binding

Previous studies have shown that the mucin-type polypeptidesGlyCAM-1, CD34, and MAdCAM-1 can function as ligands for L-selectinonly when they are synthesized by the specialized high-endothelialvenules (HEV) of lymph nodes. Since sialylation, sulfation,and possibly fucosylation are required for generating recognition,we reasoned that other mucins known to have such componentsmight also bind L-selectin. We show here that soluble mucinssecreted by human colon carcinoma cells, as well as those derivedfrom human bronchial mucus can bind to human L-selectin in acalcium-dependent manner. As with GlyCAM-1 synthesized by lymphnode HEY, 2–3 linked sialic acids and sulfation seem toplay a critical role in generating this L-selectin binding.In each case, only a subset of the mucin molecules is recognizedby L-selectin. Binding is not destroyed by boiling, suggestingthat recognition may be based primarily upon carbohydrate structures.Despite this, O-linked oligosaccharide chains released fromthese ligands by beta-elimination do not show any detectablebinding to L-selectin. Following protease treatment of the ligands,binding persists in a subset of the resulting fragments, indicatingthat specific recognition is determined by certain regions ofthe original mucins. How ever, O-linked oligosaccharides releasedfrom the subset of non-binding mucin fragments do not show verydifferent size and charge profiles compared to those that dobind. Furthermore, studies with polylactosamine-degrading endoglycosidasessuggest that the core structures involved in generating bindingcan vary among the different ligands. Taken together, thesedata indicate that a single unique oligosaccharide structuremay not be responsible for high-affinity binding. Rather, diversemucins with sialylated, sulfated, fucosylated lactosamine-typeO-linked oligosaccharides can generate high-affinity L-selectinligands, but only when they present these chains in unique spacingand/or clustered combinations, presumably dictated by the polypeptidebackbone. L-selectin mucins sialic sialic acid sulfate adhesion