Purification and characterization of l,(l/d)-aminopeptidase from Guinea pig serum

Mammalian sera contain enzymes that catalyze the hydrolytic degradation of peptidoglycans and molecules of related structure and are relevant for the metabolism of peptidoglycans. We now report on a novel L,(L/D)-aminopeptidase found in human and mammalian sera. The enzyme hydrolyses the pentapeptide L-Ala-D-iso-Gln-meso-DAP(omegaNH(2))-D-Ala-D-Ala yielding the free L-alanine and the respective tetrapeptide (K(M) 18 mM). L,(L/D)-aminopeptidase from guinea pig serum was highly purified in four chromatographic steps, up to 700-fold. Molecular weight of the enzyme was estimated by HPLC to be approximately 175,000. The configuration of alanine obtained by hydrolysis of the pentapeptide was determined by oxidation with L-amino acid oxidase. The amino acids sequence in the respective tetrapeptide was deduced from the results of mass spectrometry. The novel L,(L/D)-aminopeptidase also hydrolyzed alanine-4-nitroanilide (K(M)=0.6 mM) and several peptides comprising L-amino acids. Peptides containing D-amino acid at the amino end and L-Asp-L-Asp were not the substrates for this enzyme. The purified enzyme also exhibited enkephalin degrading activity, hydrolyzing enkephalins comprising L,L- and L,D-peptide bonds. The enzyme was inhibited strongly by metal chelating agents, bestatin and amastatin.     

Validity of the Force-Velocity Relation for Muscle Contraction in the Length Region, l ≤ l0

Considerable attention has been directed to the characteristic force-velocity relation discovered by A. V. Hill in the study of muscle kinematics. Models of contractile process were tested on the basis of their compatibility with the Hill equation. However, almost all the isotonic data have been restricted to one length, l₀, the maximum length with almost no resting tension; the velocities measured are those initial values when the load begins to move. The force-velocity curve extrapolates to zero velocity for isometric tension, but only for the tension at that one length. Very few efforts have been made to study the profiles of the curves throughout the range of lengths over which shortening takes place. In examining the length region, l ≤ l₀, for an isotonically contracting muscle, not only is the force-velocity relation valid for the initial reference length, l₀, but also for any other length. The analysis in this report indicates that the constants a/P₀ and b/l₀ remain fixed throughout the length change of afterloaded isotonic shortening in the Rana pipiens sartorius muscles.     

Mechanisms of human papillomavirus type 16 neutralization by l2 cross-neutralizing and l1 type-specific antibodies

Pseudovirions of human papillomavirus type 16 (HPV16), the principal etiologic agent in 50% of cervical cancers, were used as a model system to investigate the cell surface interactions involved in the exposure of the broadly cross-neutralizing papillomavirus L2 epitopes. These neutralizing epitopes were exposed only after cell surface binding and a subsequent change in capsid conformation that permitted cleavage by the cellular protease furin at a specific highly conserved site in L2 that is immediately upstream of the cross-neutralizing epitopes. Unexpectedly, binding of L2 antibodies led to the release of the capsid/antibody complexes from the cell surface and their accumulation on the extracellular matrix. Study of the dynamics of exposure of the L2 epitopes further revealed that representatives of the apparently dominant class of L1-specific neutralizing antibodies induced by virus-like particle vaccination prevent infection, not by preventing cell surface binding but rather by preventing the conformation change involved in exposure of the L2 neutralizing epitope. These findings suggest a dynamic model of virion-cell surface interactions that has implications for both evolution of viral serotypes and the efficacy of current and future HPV vaccines.     

Isolation and initial characterization of the mouse Dnmt3l gene

We have isolated and sequenced the mouse zinc finger gene, Dnmt3l (DNA cytosine-5-methyltransferase 3-like), on mouse chromosome 10, showing similarity to members of the DNMT3/Dnmt3 family. The Dnmt3l protein contains an ADD zinc finger, which Dnmt3l shares with other Dnmt3 family members and Atrx. RT-PCR analysis showed Dnmt3l expression in testis, thymus, ovary, and heart, as well as in 7-day, 15-day, and 17-day mouse embryos.     

Bilayer polarity and its thermal dependency in the l(o) and l(d) phases of binary phosphatidylcholine/cholesterol mixtures

Diverse variations in membrane properties are observed in binary phosphatidylcholine/cholesterol mixtures. These mixtures are nonideal, displaying single or phase coexistence, depending on chemical composition and other thermodynamic parameters. When compared with pure phospholipid bilayers, there are changes in water permeability, bilayer thickness and thermomechanical properties, molecular packing and conformational freedom of phospholipid acyl chains, in internal dipolar potential and in lipid lateral diffusion. Based on the phase diagrams for DMPC/cholesterol and DPPC/cholesterol, we compare the equivalent polarity of pure bilayers with specific compositions of these mixtures, by using the Py empirical scale of polarity. Besides the contrast between pure and mixed lipid bilayers, we find that liquid-ordered (l(o)) and liquid-disordered (l(d)) phases display significantly different polarities. Moreover, in the l(o) phase, the polarities of bilayers and their thermal dependences vary with the chemical composition, showing noteworthy differences for cholesterol proportions at 35, 40, and 45 mol%. At 20 degrees C, for DMPC/cholesterol at 35 and 45 mol%, the equivalent dielectric constants are 21.8 and 23.8, respectively. Additionally, we illustrate potential implications of polarity in various membrane-based processes and reactions, proposing that for cholesterol containing bilayers, it may also go along with the occurrence of lateral heterogeneity in biological membranes.     

Diagnostic précoce de l’allergie chez l’enfant

Fibrotest et Actitest sont des alternatives non agressives à la biopsie du foie. Ces deux combinaisons de marqueurs permettent de proposer, chez les sujets contaminés par le virus de l’hépatite C (VHC), en première ligne une prise de sang et de ne réserver la biopsie du foie qu’en deuxième intention. Le Fibrotest permet d’estimer le degré de fibrose (cicatrices) et l’Actitest le degré d’activité (nécrose et inflammation).     

La cytométrie dans l’exploration de l’allergie

Les tests cellulaires pour l’explorationin vitro de l’hypersensibilité immédiate, utilisent le polynucléaire basophile du sang périphérique. Ce leucocyte qui possède la plupart des propriétés du mastocyte, un récepteur membranaire de forte affinité pour les IgE (FceR1) et des granulations contenant des médiateurs préformés de l’anaphylaxie, est un bon témoin de la sensibilisation mastocytaire contre un allergène. Mais comme ce dernier est d’un accès difficile, pour les étudesin vitro chez l’homme on préfère travailler sur le basophile.     

Inhibition of Hill Reaction and Photophosphorylation by l,l,l-Trichloro-2,2-bis(p-chlorophenyl)ethane

The effect of DDT on DCIP and Fe(CN)₆^3– photoreductions,and cyclic and non-cyclic photo-phosphorylations, in some 30varieties of barley from widely different parts of the worldis reported. Whereas resistant barleys were not affected byDDT treatment, chloroplasts from treated susceptible barleysshowed parallel inhibitions of all the investigated aspectsof photosynthesis. However, in a few susceptible varieties inhibitionsof Fe(CN)₆^3– photoreduction or non-cyclic photophosphorylationwere not so pronounced. Possible reasons for these anomaliesare discussed; in particular earlier reports that DDT had noeffect on these latter photosynthetic activities may have beendue to the use of hypotonic media during chloroplast isolation.     

Proceedings towards a natural classification of the worldwide taxa Phellinus s.l. and Inonotus s.l., and phylogenetic relationships of allied genera

The classification of Phellinus s.l., Inonotus s.l. and the phylogenetic relationships of allied genera were studied using nuc-lsu rDNA sequence data. The worldwide taxon sampling comprised 107 species, 99 of them belonging to the Hymenochaetales. The phylogenetic trees were discussed in relation to morphological and anatomical features of the fruit bodies. The Hymenochaetales formed no monophyletic group and several non-Hymenochaetales appeared as intermingled with the Hymenochaetales. Trichaptum abietinum and Oxyporus populinus showed no certain affinities within the Hymenochaetales, whereas Basidioradulum radula was closely related to Phellopilus nigrolimitatus, and Hyphodontia quercina and Schizopora paradoxa were related to Coltricia, Coltriciella and Pyrrhoderma adamantinum. Phellinus s.l. and Inonotus s.l. formed no monophyletic groups, and a subdivision in the following genera is accepted: Phellinus s.s., Inonotus s.s., Inocutis, Fomitiporella, Aurificaria, Phylloporia, Fulvifomes, Mensularia, Pseudoinonotus, Fomitiporia, Porodaedalea, Onnia, Fuscoporia, and Inonotopsis. Coltricia and Coltriciella were confirmed as seperate genera. The taxonomic status of Phellinidium and Pyrrhoderma remained uncertain. 16 new combinations are proposed.     

Phylogeny of the bee subtribe Caenohalictina Michener (Hymenoptera,Apidae s.l., Halictinae s.l.)

Gonçalves, R. B. & Melo, G. A. R. (2009). Phylogeny of the bee subtribe Caenohalictina Michener (Hymenoptera, Apidae s.l., Halictinae s.l.). —Zoologica Scripta, 39, 187–197. Recently, Caenohalictina Michener, 1954 was formally proposed as a group within Halictini based on a phylogenetic analysis using DNA data. Morphological synapomorphies for the subtribe are not stabilized and the relationships among its attributed genera were not completely reconstructed. Also, there are different propositions for the scope and classification of the genera. This study investigates the monophyly of Caenohalictina based on morphological characters and presents a hypothesis for the relationships among its genera and subgenera. For this purpose, a matrix of 36 terminals and 96 characters was coded. The cladistic analysis using character equal weighting resulted in one most parsimonious tree with 189 steps. According to this hypothesis, the subtribe is monophyletic with five synapomorphies supporting it. The relationships among Caenohalictina taxa are: Habralictus(Caenohalictus (((Agapostemon s.s. + Notagapostemon) (Agapostemonoides (Rhinetula (Paragapostemon + Dinagapostemon)))) + ((Ruizanthedella (Ruizantheda + Oragapostemon)) + (Brasilagapostemon (Pseudagapostemon s.s. + Neagapostemon))))). The subgenus Habralictus s.s. is paraphyletic with respect to Habralictus (Zikaniella) crassipes, hence both subgenera are not recognized. Comments on alternative generic classifications and biogeographic patterns are presented.     

Morphogenèse de l’endostructure osseuse de l’ilion humain

Through the human infantile stage, the progressive acquisition of the orthograde posture and bipedal locomotion is responsible for important morphostructural bony changes at the pelvis and the coxofemoral joint. This study uses digital processing techniques applied to a set of radiographic and microtomographic images to characterise the textural properties of the trabecular network and the topographic variation of the cortical shell in an ontogenetic series of 15 ilia from individuals aged between 0 and 40 years. The results suggest that the main structures of the trabecular architecture which are functional to the distribution and dissipation of the postural- and locomotor-related loads, are already recognizable at the age of 1–2 years, in association with the adoption of bipedal walking.