Three new HLA-G alleles and their linkage disequilibria with HLA-A

Three new allelic forms of the HLA-G DNA sequence (HLA-G*II, HLA-G*III, and HLA-G*IV) have been identified. With the HLA-G*I sequence (previously designated HLA 6.0) as a reference, HLA-G*II shows a silent (G A) mutation at the third base of codon 57, HLA-G*III bears a non-synonymous (A T), but conservative, (Thr Ser) substitution at the first base of codon 31, and HLA-G*IV shows two silent substitutions: (A T) at the third base of codon 107 and (G A) at the third base of codon 57. A rapid method of singling out each allele on genomic DNA has been developed by using polymerase chain reaction amplification followed by restriction endonuclease treatment. Also, more or less strong linkage disequilibria has been found between most HLA-A alleles and either HLA-G*I or *II, both being the most prevalent alleles in the population, with a genotypic frequency of 0.55 and 0.38, respectively; HLA-G*III is very rare and HLA-G*IV has a genotypic frequency of 0.07. An evolutive classification of HLA-A alleles results according to their association with either HLA-G*I or HLA-G*II, which does not correlate with the classical serological cross-reacting groups classification. The finding of a strong and selective A/G linkage disequilibria with most HLA-A alleles, together with the existence of less frequent random A/G associations, may suggest that there exist in different haplotypes true and varied A/G genetic distances (and not a recombinational hotspot). It may be inferred from preliminary data that in primates HLA-A/G haplotypes bearing G*II may have appeared later than those bearing G*I.The nucleotide sequence data reported in this paper have been submitted to the GenBank and EMBL nucleotide sequence databases and have been assigned the following accession numbers: EMBL-X60983 (HLA-G*II), GenBank-M99048 (HLA-G*III), and GenBank-L07784 (HLA-G*IV).The contribution to this paper by P. Morales and A. Corell is equal, and the order of authorship is arbitrary. Correspondence to: A. Arnaiz-Villena.     

Diphyllobothriids (Cestoda: Pseudophyllidea) from the long-finned pilot whale Globicephala melas (Traill, 1809) off the Faroe Islands,with comments on the taxonomy of Diphyllobothrium Cobbold, 1858 species from cetaceans

The taxonomy of marine species of the genus Diphyllobothrium, particularly those parasitic in cetaceans, is rather confused. During parasitological investigations of long-finned pilot whales Globicephala melas from waters off the Faroe Islands, five diphyllobothriid species were detected: Diphyllobothrium sp. (possibly D. polyrugosum), D. stemmacephalum, Diphyllobothrium sp. A, Diphyllobothrium sp. B and Diphyllobothriidae sp. D. stemmacephalum is reported for the first time from G. melas. The stituation regarding the taxonomy of Diphylobothrium species from cetaceans is briefly reviewed. It is concluded that the recent development of genetic techniques may be of great value in relation to clarifying the taxonomy of this group.     

Communal Roosting of Wintering Red Kites Milvus milvus (Aves,Accipitridae): Social Feeding Strategies for the Exploitation of Food Resources

Social feeding strategies of wintering red kites are analyzed in relation to age, food, roost-sites and differences from kite residents. Whereas young and adult wintering kites gathered at roost sites almost daily, adult residents did not, and immature residents only occasionally. Kites using roost sites feed more often on prey prelocated by others, while lone roosters also forage and discover food alone. After finding food, kites tend to shift to a new roost site and foraging area. Two details of the ‘information centre’ hypothesis are confirmed in our study: carcasses are unpredictably found patches, divisible between several individuals. But carcasses disappeared fast in the study area, and no increase with time in the number of birds consuming a carcass was observed, so that information transmission was unconfirmed. When kites leave the roost in groups no leader is detectable. It seems that other types of social foraging are operating, and the model best matching our results is network foraging.     

Molecular chaperones and the biogenesis of mitochondria and peroxisomes

Summary— A review of the proteinaceous machinery involved in protein sorting pathways and protein folding and assembly in mitochondria and peroxisomes is presented. After considering the various sorting pathways and targeting signals of mitochondrial and peroxisomal proteins, we make a comparative dissection of the protein factors involved in: i) the stabilization of cytosolic precursor proteins in a translocation competent conformation; ii) the membrane import apparatus of mitochondria and peroxisomes; iii) the processing of mitochondrial precursor proteins, and the eventual processing of certain peroxisomal precursor, in the interior of the organelles; and iv) the requirement of molecular chaperones for appropriate folding and assembly of imported proteins in the matrix of both organelles. Those aspects of mitochondrial biogenesis that have developed rapidly during the last few years, such as the requirement of molecular chaperones, are stressed in order to stimulate further parallel investigations aimed to understand the origin, biochemistry, molecular biology and pathology of peroxisomes. In this regard, a brief review of findings from our group and others is presented in which the role of the F₁-ATPase α-subunit is pointed out as a molecular chaperone of mitochondria and chloroplasts. In addition, data are presented that could question our previous indication that the immunoreactive protein found in the rat liver peroxisomes is due to the presence of the F₁-ATPase α-subunit.     

Ethanol-hypersensitive and ethanol-dependent cdc − mutants in Schizosaccharomyces pombe

Ethanol-hypersensitive strains (ets mutants), unable to grow on media containing 6% ethanol, were isolated from a sample of mutagenized Schizosaccharomyces pombe wild-type cells. Genetic analysis of these ets strains demonstrated that the ets phenotype is associated with mutations in a large set of genes, including cell division cycle (cdc) genes, largely non-overlapping with the set represented by the temperature conditional method; accordingly, we isolated some ets non-ts cdc ^? mutants, which may identify novel essential genes required for regulation of the S. pombe cell cycle. Conversely, seven well characterized ts cdc ^? mutants were tested for their ethanol sensitivity; among them, cdc1–7 and cdc13–117 exhibited a tight ets phenotype. Ethanol sensitivity was also tested in strains bearing different alleles of the cdc2 gene, and we found that some of them were ets, but others were non-ets; thus, ethanol hypersensitivity is an allele-specific phenotype. Based on the single base changes found in each particular allele of the cdc2 gene, it is shown that a single amino acid substitution in the p34^cdc2 gene product can produce this ets phenotype, and that ethanol hypersensitivity is probably due to the influence of this alcohol on the secondary and/or tertiary structure of the target protein. Ethanol-dependent (etd) mutants were also identified as mutants that can only be propagated on ethanol-containing media. This novel type of conditional phenotype also covers many unrelated genes. One of these etd mutants, etd1-1, was further characterized because of the lethal cdc ^? phenotype of the mutant cells under restrictive conditions (absence of ethanol). The isolation of extragenic suppressors of etd1-1, and the complementation cloning of a DNA fragment encompassing the etd1 ⁺ wild-type gene (or an extragenic multicopy suppressor) demonstrate that current genetic techniques may be applied to mutants isolated by using ethanol as a selective agent.     

药敏药片对临床235株细菌实验观察报告

药敏药片经临床对金黄色葡萄球菌、大肠埃希氏菌、铜绿假单胞菌等２３５株考核．表明药片工艺研究先进,药片与培养基结合牢固,无断裂、崩解,不渗出颗粒,抑菌圈呈同心园扩散．边缘清楚。药物含量均匀,释放度好。药片抑菌差仅１～３ｍｍ;而纸片抑菌差为２～１２ｍｍ。药片变黑系数ＣＶ为２．７１～４．２１;而纸片ＣＶ为３．８２～１４．３６。表明纸片片间差大,药片精密度明显好于纸片。     

DEFORMATIVE DISEASE IN IRIDAEA LAMINARIOIDES (RHODOPHYTA): GALL DEVELOMENT ASSOCIATED WITH AN ENDOPHYTIC CYANOBACTERIUM1

This study is the first report of an algal disease, developed in close association with an endophytic organism, documented for the southeastern Pacific. We describe a disease affecting wild populations of the red alga Iridaea laminarioides Bory in central Chile, characterized by gall development on the surface of sporophytic, cystocarpic, and immature thalli. These abnormal growths result in severe morphological alterations of the affected thalli. Diseased fronds display an aggregated spatial distribution and occur throughout the year, with a maximum in summer followed by a decline in winter. The presence of galls was not associated with broken or torn fronds. Although causality has not been unequivocally demonstrated, our field and laboratory observations indicate a strong association of the galls with infections by an endophytic cyanobacterium, probably belonging to the genus Pleurocapsa.     

Chemical structure and conformational features of cell-wall polysaccharides isolated from Aphanoascus mephitalus and related species

The structures of cell-wall mannans isolated from Aphamoascus mephitalus, A. Fulvescens, A. verrucosus, and A. reticulisporus have been investigated by chemical analyses and 1D and 2D ¹H and ¹³C NMR techniques. It was found that all of them consists of a relatively simple comb-like structure of the disaccharide repeating block {→ 6)-[-Man p-(1 → 2)]--Man p-(1 →}. The conformations around the -(1 → 2) and -(1 → 6) linkages in thes kinds of polymers were also studied by using molecular mechanics and dynamics calculations, together with NOE data. The results are similar to those found within the oligosaccharide chains of glycoproteins, with a well-defined conformation for the -(1 → 2) linkage and a certain restriction around the -(1 → 6) bonding imposed by the 2-substitution.     

Influence of liposome charge and composition on their interaction with human blood serum proteins

The roles of sulfhydryl and disulfide groups in the specific binding of synthetic cannabinoid CP-55,940 to the cannabinoid receptor in membrane preparations from the rat cerebral cortex have been examined. Various sulfhydryl blocking reagents including p-chloromercuribenzoic acid (p-CMB), N-ethylmaleimide (NEM), o-iodosobenzoic acid (o-ISB), and methyl methanethiosulfonate (MMTS) inhibited the specific binding of [³H]CP-55,940 to the cannabinoid receptor in a dose-dependent manner. About 80–95% inhibition was obtained at a 0.1 mM concentration of these reagents. Scatchard analysis of saturation experiments indicates that most of these sulfhydryl modifying reagents reduce both the binding affinity (K_d) and capacity (B_max). On the other hand, DL-dithiothreitol (DTT), a disulfide reducing agent, also irreversibly inhibited the specific binding of [³H]CP-55,940 to the receptor and about 50% inhibition was obtained at a 5 mM concentration. Furthermore, 5mM DTT was abelt to dissociate 50% of the bound ligand from the ligand-receptor complex. The marked inhibition of [³H]CP-55,940 binding by sulfhydryl reagents suggests that at least one free sulfhydryl group is essential to the binding of the ligand to the receptor. In addition, the inhibition of the binding by DTT implies that besides free sulfhydryl group(s), the integrity of a disulfide bridge is also important for [³H]CP-55,940 binding to the cannabinoid receptor.     

Immunocytochemical approach to the structure of human blood group B and H glycoconjugate antigens in the three days old rat cochlea

The possible structure of human blood-group antigens, as found in cochlear hair cells of 3-day-old rats, is suggested. Data were obtained from immunocytochemical studies using 77 antibodies against the major human blood group antigens of the ABO, H, I and Lewis genetic systems. Neither the anti-A-related nor the anti-Lewis-related antibodies showed any positive immunoreaction on hair cells. In contrast, anti-B, anti-AB and anti-H antibodies displayed specific positive immunoreactive patterns on the hair cells. The results suggest that, in immature hair cells, two main glycoconjugate structures of the lactoseries are present: H type 2 antigen, which is the precursor of the B type 2 antigen, and the B type 2 antigen itself. Similar H and B carbohydrate structures have been reported in rat olfactory receptors. The type 2 glycoconjugates carrying these H and B antigens of auditive and olfactory receptors are resistant to fixation and paraffin embedding, suggesting that they might be glycoproteins. These auditive and olfactory H and B antigens must be different from the B-related antigens that are expressed by pseudo-unipolar neurons of rat posterior root ganglia, that are built from type 4 core chains, and that are destroyed by routine paraffin embedding procedures.