Liberation of desmosine and isodesmosine as amino acids from insoluble elastin by elastolytic proteases

The development of atherosclerotic lesions and abdominal aortic aneurysms involves degradation and loss of extracellular matrix components, such as collagen and elastin. Releases of the elastin cross-links desmosine (DES) and isodesmosine (IDE) may reflect elastin degradation in cardiovascular diseases. This study investigated the production of soluble elastin cross-linking structures by proteinases implicated in arterial diseases. Recombinant MMP-12 and neutrophil elastase liberated DES and IDE as amino acids from insoluble elastin. DES and IDE were also released from insoluble elastin exposed to monocyte/macrophage cell lines or human primary macrophages derived from peripheral blood monocytes. Elastin oxidized by reactive oxygen species (ROS) liberated more unconjugated DES and IDE than did non-oxidized elastin when incubated with MMP-12 or neutrophil elastase. These results support the exploration of free DES and IDE as biomarkers of elastin degradation.     

Occurrence and characterization of actinobacteria and thermoactinomycetes isolated from pulp and board samples containing recycled fibres

The aim of this study was to characterize the actinobacterial population present in pulps and boards containing recycled fibres. A total of 107 isolates was identified on the basis of their pigmentation, morphological properties, fatty acid profiles and growth temperature. Of the wet pulp and water sample isolates (n=87), 74.7% belonged to the genus Streptomyces, 17.2% to Nocardiopsis and 8.0% to thermoactinomycetes, whereas all the board sample isolates (n=20) were thermoactinomycetes. The identification of 53 isolates was continued by molecular methods. Partial 16S rDNA sequencing and automated ribotyping divided the Streptomyces isolates (n=31) into 14 different taxa. The most common streptomycetes were the mesophilic S. albidoflavus and moderately thermophilic S. thermocarboxydus. The Nocardiopsis isolates (n=11) belonged to six different taxa, whereas the thermoactinomycetes were mainly members of the species Laceyella sacchari (formerly Thermoactinomyces sacchari). The results indicated the probable presence of one or more new species within each of these genera. Obviously, the drying stage used in the board making processes had eliminated all members of the species Streptomyces and Nocardiopsis present in the wet recycled fibre pulp samples. Only the thermotolerant endospores of L. sacchari were still present in the final products. The potential of automated ribotyping for identifying actinobacteria was indicated, as soon as comprehensive identification libraries became available.     

Concanavalin A interactions with asparagine-linked glycopeptides. The mechanisms of binding of oligomannose,bisected hybrid,and complex type carbohydrates

The affinity of concanavalin A (Con A) for simple saccharides has been known for over 50 years. However, the specificity of binding of Con A with cell-surface related carbohydrates has only recently been examined in detail. Brewer and coworkers [J Biol Chem (1986) 261:7306–10; J Biol Chem (1987) 262:1288–93; J Biol Chem (1987) 262:1294–99] have recently studied the binding interactions of a series of oligomannose and bisected hybrid type glycopeptides and complex type glycopeptides and oligosaccharides with Con A. The relative affinities of the carbohydrates were determined using hemagglutination inhibition measurements, and their modes of binding to the lectin examined by nuclear magnetic relaxation dispersion (NMRD) spectroscopy and quantitative precipitation analyses. The equivalence zones (regions of maximum precipitation) of the precipitin curves of Con A and the carbohydrates indicate that certain oligomannose and bisected hybrid type glycopeptides are bivalent for lectin binding. From the NMRD and precipitation data, two protein binding sites on each glycopeptide have been identified and characterized. Certain bisected complex type oligosaccharides also bind and precipitate Con A, while the corresponding nonbisected analogs bind but do not precipitate the protein. The precipitation data indicate that the bisected complex type oligosaccharides are also bivalent for lectin binding, while the nonbisected analogs are univalent. The NMRD and precipitation data are consistent with different mechanisms of binding of nonbisected and bisected complex type carbohydrates to Con A, including different conformations of the bound saccharides.Abbreviations Con A Concanavalin A with unspecified metal ion content - CMPL Con A with Mn²⁺ and Ca²⁺ at the S1 and S2 sites respectively, in the locked conformation [12]; trisaccharide1, 3,6-di-O-(-d-mannopyranosyl)-d-mannose - -MDM methyl -d-mannopyranoside - NMRD nuclear magnetic relaxation dispersion, the magnetic field dependence of nuclear magnetic relaxation rates, in the present case, the longitudinal relaxation rate, 1/T₁, of solvent protons     

Novel alk-1-enyl ether lipids isolated from Clostridium acetobutylicum

Alkenyl ether analogues of phosphatidylglycerol (plasmenylglycerol), bisphosphatidylglycerol (cardiolipin) (plasmenylglycerolphosphatidic acid), monoglycosyldiglyceride and diglycosyldiglyceride were isolated from the polar lipids of Clostridium acetobutylicum and characterized by chemical analyses and degradation. The position of the alkenyl ether bond (at C-1) and of the acyl ester bond (at C-2) as well as the configuration at C-2 of the phospholipids are the same as of the alkenyl ether phospholipids known so far. The alkenyl ether analogue of monoglycosyldiglyceride contains a galactosyl residue, that of diglycosyldiglyceride a glucosyl-galactosyl residue, glucosyl forming the terminal unit.     

Novel facultative anaerobic acidotolerant Telmatospirillum siberiense gen. nov. sp. nov. isolated from mesotrophic fen

Three facultative anaerobic acidotolerant Gram-negative motile spirilla strains designated 26-4b1, 26-2 and K-1 were isolated from mesotrophic Siberian fen as a component of methanogenic consortia. The isolates were found to grow chemoorganotrophically on several organic acids and glucose under anoxic and low oxygen pressure in the dark, tolerant up to 5kPa of oxygen. At low oxygen supply, faint autotrophic growth on the H(2):CO(2) mixture was also observed. All three isolates were able to fix N(2). Major cellular fatty acids were 18:1 omega7c, 17:0 cyclopropane and 16:0. Phylogenetic analyses of the 16S rRNA gene sequences revealed that they formed a deep branch within the family Rhodospirillaceae of the Alphaproteobacteria with the highest similarity of 90.9-92.5% with members of genera Phaeospirillum and Magnetospirillum. Phylogenetic study of nifH (nitrogenase) and cbbL (RuBisCO) amino acid sequence identities confirmed that the new isolates represent a novel group. Based on the phylogenetic analyses and distinct phenotypic characteristics, we are of the opinion that strains 26-4b1, 26-2 and K-1 represent a new species of a novel genus for which the name Telmatospirillum siberiense gen. nov. sp. nov. is proposed.     

Novel phenylanthraquinones,isofuranonaphthoquinones, homoisoflavonoids,and biflavonoids from African plants in the genera Bulbine,Scilla, Ledebouria,and Rhus

The sale of non-cultivated plants poses a real threat to the survival of a large number of plant species. On the other hand, the fact that a given plant has become a traded item is an important lead in the effort to investigate plants for the purpose of discovering novel secondary metabolites that may have marketable properties. This report is largely based on the investigations of plants collected from vendors of medicinal plants. The identification is based on botanical specimens derived from the purchased plant parts either by growing them in a garden or through micropropagation using tissue culture techniques. Bulbine species yielded novel phenyl anthraquinones and isofuranonaphthoquinones, which were found to exhibit antiparasitic and antioxidant properties. Scilla nervosa subsp. rigidifolia and Ledebouria graminifolia yielded twenty homoisoflavonoids and two xanthones. Bulbs derived from tissue culture based specimens of L. graminifolia contained all the compounds isolated from the marketed samples. Some of the homoisoflavonoids were found to possess activity against colon and breast cancer cell lines. Rhus pyroides furnished six novel bichalcones some of which exhibited insect antifeedant, cytotoxic and antiproliferative activities. Structures of the compounds described in this report were elucidated by spectroscopic means including HRMS and 1D-, and 2D-NMR experiments. Interconversion of some of the phenyl anthraquinones was accomplished as part of their structure proof. The total synthesis of three of the biflavonoids was achieved using a synthetic route which involves a microwave irradiation promoted Ulmann synthesis. This revised version was published online in June 2006 with corrections to the Cover Date.     

Vasodilative effect of perillaldehyde on isolated rat aorta

The vasodilative effect of perillaldehyde, one of the major oil components in Perilla frutescens BRITTON, was studied using isolated rat aorta. Perillaldehyde at final concentrations of 0.01 to 1 mM showed dose-dependent relaxation of the aorta contracted by treatment with prostaglandin F2alpha or norepinephrine. Neither the presence of NG-nitro-L-arginine methyl ester nor removal of the aortic endothelium affected the vasodilatation, suggesting that perillaldehyde exerts a direct effect on vascular smooth muscle cells. The vasodilative effect of perillaldehyde was not inhibited by pretreatment with a beta-adrenergic receptor blocker (propranolol), an inhibitor of phosphodiesterase (theophylline), a delayed rectifier K+ channel blocker (tetraethylammonium chloride), or an ATP-sensitive K+ channel blocker (glibenclamide). However, perillaldehyde showed contrasting effects on vasodilatation of the aorta contracted by an influx of extracellular Ca2+ - perillaldehyde caused little vasodilatation on the aorta contracted by the Ca2+ ionophore A23187, while it inhibited the vasoconstriction induced by treatment with high-concentration K+, which dominantly opened the voltage-dependent Ca2+ channel. These results suggest that the vasodilative effect of perillaldehyde is derived from blocking the Ca2+ channels.     

Identification of lower fungi isolated from larvae of mangrove crab,Scylla serrata, in Indonesia

Since 1992, seed production of mangrove crab,Scylla serrata, has been attempted at the Gondol Research Station for Coastal Fisheries, Bali, Indonesia. During the production process, almost all of the larvae have died due to fungal infection. Fungi isolated from the larvae with fungal infection were classified into three species in the order Lagenidiales:Lagenidium callinectes, Haliphthoros milfordensis andHalocrusticida baliensis sp. nov. based on detailed morphological characteristics. The effects of temperature, pH and mineral content of the water on their growth were also examined.     

Characterization and identification of bacteria isolated from micropropagated mint plants

Summary Bacterial isolates from contaminated mint shoot cultures were characterized and identified as a preliminary step in determining an elimination treatment. The 22 bacteria were characterized using biochemical and morphological tests and subjected to sensitivity tests with four antibiotics. The isolates were compared with known organisms and assigned to genera according to similarities in characteristics. Seven isolates were analyzed by fatty acid analysis carried out by a commercial laboratory. Six were classified asAgrobacterium radiobacter; eight asXanthomonas; one each asPseudomonas fluorescens, Micrococcus spp.,Corynebacterium spp., andCurtobacterium spp.; four could not be assigned to genera. Inhibition of growth of the bacteria by most antibiotics was best at pH 7.5. Minimal inhibitory concentration and minimal bactericidal concentrations of gentamicin, rifampicin, streptomycin sulfate, and Timentin varied with genotype.     

Arcobacter ellisii sp. nov., isolated from mussels

As part of a study carried out for detecting Arcobacter spp. in shellfish, three mussel isolates that were Gram-negative slightly curved rods, non-spore forming, showed a new 16S rDNA-RFLP pattern with a specific identification method for the species of this genus. Sequences of the 16S rRNA gene and those of the housekeeping genes rpoB, gyrB and hsp60 provided evidence that these mussel strains belonged to an unknown genetic lineage within the genus Arcobacter. The similarity between the 16S rRNA gene sequence of the representative strain (F79-6^T) and type strains of the other Arcobacter species ranged between 94.1% with A. halophilus and 99.1% with the recently proposed species A. defluvii (CECT 7697^T). DDH results between strain F79-6^T and the type strain of the latter species were below 70% (53 ± 3.0%). Phenotypic characteristics together with MALDITOF mass spectra differentiated the new mussel strains from all other Arcobacter species. All the results indicate that these strains represent a new species, for which the name Arcobacter ellisii sp. nov. with the type strain F79-6^T (=CECT 7837^T = LMG 26155^T) is proposed.     

Morphological aspects ofSaprolegnia diclina Type 1 isolated from pejerrey,Odonthetes bonariensis

Saprolegnia diclina Type 1 (syn.S. parasitica) discovered in pejerrey,Odonthetes bonariensis, is described and illustrated herein.     

Mycotoxin-producing potential of fungi isolated from red kidney beans

The predominant fungi present in samples of reject and retail red kidney beans were Aspergillus glaucus, Penicillium spp. and Alternaria spp. Together with A. ochraceus, A. flavus, Fusarium spp., and Trichoderma, these isolates from the reject beans were screened for numerous mycotoxins by TLC. The most consistently produced mycotoxins were penicillic acid (from A. ochraceus and Penicillium spp.) and Alternaria toxins (tenuazonic acid and alternariol). A. glaucus strains were tested for cytotoxicity in three tissue culture cell lines with positive results.     

Seasonality and toxigenicity ofVibrio cholerae non-01 isolated from different components of pond ecosystems of Dhaka City,Bangladesh

Diarrhoea due toVibrio cholerae non-01 is common in Bangladesh. Four hundred and eighty samples, including plants, water, phytoplankton and sediment, were collected from five ponds in Dhaka every 15 days for one year.V. cholerae non-01 was isolated from 181 (38%) of the samples. Two peaks were evident: one in April and the other in August/September. Forty-three (23%) of the 181 isolates were examined for toxigenicity and 19% were cytotoxic to Y1 adrenal cells. This study provides evidence of the likely infectious nature of some ponds and may have relevance to the epidemiology of diarrhoea caused byV. cholerae non-01 in Bangladesh.     

Structure of a new nonasaccharide isolated from human milk: VI2Fuc,V4Fuc,III3Fuc-p-lacto-N-hexaose

The structure of a new nonasaccharide isolated from human milk has been investigated. By using methylation analysis, FAB-MS and¹H-and¹³C-NMR spectroscopy as basic methods of structural investigation, this oligosaccharide was identified as VI²--Fuc,V⁴-Fuc,III³--Fuc-p-lacto-n-hexaose: Fuc1-2Gal1-3[Fuc1-4]GlcNAc1-3Gal1-4[Fuc1-3]GlcNAc1-3Gal1-4Glc.Abbreviations COSY correlation spectroscope - DP degree of polymerisation - FAB-MS fast atom bombardment-mass spectrometry - HPLC high performance liquid chromatography - NMR nuclear magnetic resonance - GLC gas-liquid chromatography     

Cerrena unicolor isolated from the mycangia of a horntail,Tremex longicollis,in Kochi Prefecture,Japan

A fungus was found to be stored in the mycangia of a horntail,Tremex longicollis, as hyphal fragments. All fungal isolates from the mycangia of 31 adult females of the horntail produced the same colonies on PDA. Basidiocarps ofCerrena unicolor occurred near the emergence hole of the horntail on a dead hackberry tree (Celtis sinensis). The cultures of this fungus were similar to those from the mycangia of the horntail in cultural characteristics. Mating between single-basidiospore mycelia ofC. unicolor and single-arthrospore mycelia from the mycangia of the horntail showed that they were compatible. These results revealed that the fungus isolated from the mycangia ofT. longicollis wasC. unicolor.     

Fungi isolated from skins and pens of healthy animals in Nigeria

The mycoflora of 220 skin scrapings, hair, nail samples and pens’ materials of apparently healthy animals including cows, sheep, goats, rabbits, pigs and dogs were determined. Twenty eight species of fungi belonging to ten genera were recovered. Chrysosporium spp. were the most common and C. keratinophilum was recovered from all animals. Dermatophytes which are known causal agents of dermatophytosis were also isolated indifferent frequencies (Microsporum gypseum, M. canis, Trichophytonmentagrophytes, T. rubrum). This revised version was published online in June 2006 with corrections to the Cover Date.     

The glycopeptides from the human,bovine and porcine stomachs

• 1.1. Glycopeptides were isolated from tryptic digests of human, bovine and porcine gastric mucosae.
• 2.2. Fractionation of the glycopeptide by preparative zone electrophoresis on a Pevikon C-870 block, yielded one glycopeptide fraction from the mucosa of the human stomach, whereas from both mucosae of the bovine fourth stomach and swine stomach it yielded three glycopeptide fractions.
• 3.3. They were characterized by an increase in sialic acid and sulfate as the mobility of Pevikon block electrophoresis increased.