Screening method for microorganisms producing L-aminoacids from D,L-5-monosubstituted hydantoins

Summary The ability of microorganisms to produce hydantoinase and L-N-carbamoylase could be established by an overlay assay. Enzyme producing strains form clear areas around their colonies caused by the cleavage of D,L-indolylmethylhydantoin. A second overlayer with a tryptophan-auxotroph yeast enables us to detect microorganisms which are able to produce L-tryptophan from D,L-indolylmethylhydantoin.     

Long Term Effects of High CO2 Concentration on Photosynthesis of Water Hyacinth (Eichhornia crassipes (Mart.) Solms)

The photosynthetic response to CO₂ concentration, light intensityand temperature was investigated in water hyacinth plants (Eichhorniacrassipes (Mart.) Solms) grown in summer at ambient CO₂ or at10000 µmol(CO₂) mol^–1 and in winter at 6000 µmol(CO₂)mol^–1 Plants grown and measured at ambient CO₂ had highphotosynthetic rate (35 µmo1(CO₂) m^–2 s^–1),high saturating photon flux density (1500–2000) µmolm^–2 s^–1 and low sensitivity to temperature in therange 20–40 °C. Maximum photosynthetic rate (63 µmol(CO₂)m^–2 s^–1) was reached at an internal CO₂ concentrationof 800 µmol mol^–1. Plants grown at high CO₂ in summerhad photosynthetic capacities at ambient CO₂ which were 15%less than for plants grown at ambient CO₂, but maximum photosyntheticrates were similar. Photosynthesis by plants grown at high CO₂and high light intensity had typical response curves to internalCO₂ concentration with saturation at high CO₂, but for plantsgrown under high CO₂ and low light and plants grown under lowCO₂ and high light intensity photosynthetic rates decreasedsharply at internal CO₂ concentrations above 1000 µmol^–1. Key words: Photosynthesis, CO₂, enrichment, Eichhornia crassipes     

Neodiaptomus intermedius n. sp. (Calanoida: Copepoda) from South India

A new species of Neodiaptomus, N. intermedius n. sp. is described and illustrated. It is compared with related species of the strigilipes — physalipus species group.     

Hypervariability of intronic simple (gt)n(ga)m repeats in HLA-DRB genes

We have investigated the extent of DNA variability in intronic simple (gt)_n(ga)_m repeat sequences and correlated this to sequence polymorphisms in the flanking exon 2 of HLA-DRB genes. The polymerase chain reaction (PCR) was used to amplify a DNA fragment containing exon 2 and the repeat region of intron 2. The PCR products were separated on sequencing gels in order to demonstrate length hypervariability of the (gt)_n(ga)_m repeats. In a parallel experiment, the PCR products were cloned and sequenced (each exon 2 plus adjacent simple repeats) to characterize the simple repeats in relation to the HLA-DRB sequences. In a panel of 25 DRB1, DRB4, and DRB5 alleles new sequences were not detected. Restriction fragment length polymorphism (RFLP) subtyping of serologically defined haplotypes corresponds to translated DNA sequences in 85% of the cases, the exceptions involving unusual DR/DQ combinations. Many identical DRB1 alleles can be distinguished on the basis of their adjacent simple repeats. We found group-specific organization of the repeats: the DRw52 supergroup repeats differ from those of DRB1*0101, DRB4*0101, and DRB5*0101 alleles and from those of pseudogenes. Finally, we amplified baboon DNA and found a DRB allele with extensive similarity to DRB1 sequences of the DRw52 supergroup. The simple repeat of the baboon gene, however, resembles that of human pseudogenes. In addition to further subtyping, the parallel study of polymorphic protein and hypervariable DNA alleles may allow conclusions to be drawn on the relationships between the DRB genes and perhaps also on the theory of trans-species evolution.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number M 34258.     

Tissue specification and intracellular distribution of actin isoforms inVicia faba L.

Summary Two dimensional gel electrophoresis of total cell protein extracts from not expanded, and primary leaves, petioles, and roots ofVicia faba resulted in four actin isoforms at 43 kDa with pI values from 5.9 to 6.05. In contrast to root extracts, in all leaf extracts an additional immunoreactive polypeptide with a molecular mass of 51 kDa and pI 5.75 was detected. This polypeptide was present in high amounts in protein extracts of purified chloroplasts, whereas no actin isoform at 43 kDa could be demonstrated. Compared to the tissue extracts, two actin isoforms at 43 kDa with pI values of 5.9 and 6.0 were enriched, when purified plasma membranes and the membranous fraction of vacuoles were analysed. In contrast, the soluble protein fraction of the plasma membrane preparation contained only two isoactins with pI values of 5.95 and 6.05 and a molecular mass of 43 kDa. These results indicate, that the four actin isoforms at 43 kDa detected in all examined tissues ofV. faba fulfill different functions at specific intracellular compartments, for example, the anchorage of actin microfilaments to membranes.Abbreviations BSA bovine serum albumin - BCIP 5-bromo-4-chloro-3-indolyl phosphate - DDM n-decyl -D-maltopyranoside - EDTA ethylenediamine-tetraacetic acid - HG n-hexyl -D-glucopyranoside - IEF isoelectrical focusing - MES morpholinoethanesulfonic acid - 2-ME 2 mercaptoethanol - NBT nitro blue tetrazolium - pCMB p-chloromercuribenzoic acid - PVP polyvinylpyrrolidone - Tris tris (hydroxymethyl) aminomethane     

Hydrogenosome-Methanogen Assemblages in the Echinoid Endocommensal Plagiopylid Ciliates,Lechriopyla mystax Lynch, 1930 and Plagiopyla minuta Powers, 19331

ABSTRACT. Lechriopyla mystax Lynch, 1930 and Plagiopyla minula Powers, 1933 contain hydrogenosome-methanogen assemblages similar to those reported for other plagiopylid ciliates. These assemblages are stacks of elongate ovoid hydrogenosomes alternating with methanogens; these stacks are surrounded by cisternae of the rough endoplasmic reticulum that are often accompanied by Golgi complexes. The individual methanogens in the larger ciliate, L. mystax, are about four times the volume of those in the smaller ciliate, P. minuta, but both ciliates appear to contain Gram-negative methanococcoid bacteria, possibly Methanoplanus sp. The endoplasmic reticulum-Golgi complex probably plays a significant role in exploitation of the methanogens by the host ciliate.     

Subcellular Distribution of Distinct Nucleolin Subfractions Recognized by Two Monoclonal Antibodies

Monoclonal antibodies binding to different domains of nucleolin have been used to localize nucleolin in tissue culture cells ofXenopus laevis.The monoclonal antibody b6-6E7 binds to an epitope in the N-terminal domain, which contains arrays of phosphorylation consensus sites. This monoclonal antibody binds to nucleolin of oocytes and of eggs with high affinity. In contrast, the monoclonal antibody Nu-1H6 binds poorly to the modified forms of nucleolin arising during meiosis and mitosis. In interphase cells, monoclonal antibody b6-6E7 preferentially stains the periphery of the nucleoli, where most of the rRNA accumulates. Staining by monoclonal antibody Nu-1H6 complements this pattern by staining mainly the center of the nucleoli. The epitope of monoclonal antibody Nu-1H6 is within the central domain of nucleolin, which contains the first two RNA binding domains. RNase treatment of cells results in loss of nucleolin from nucleoli. In mitotic cells, both monoclonal antibodies decorate the surface of condensing chromosomes in prophase. The periphery of the condensed chromosomes in metaphase and anaphase is preferentially stained by monoclonal antibody b6-6E7.     

Alzheimer's disease amyloid-β peptide analogue alters the ps-dynamics of phospholipid membranes

We have investigated the influence of the neurotoxic Alzheimer's disease peptide amyloid-β (25-35) on the dynamics of phospholipid membranes by means of quasi-elastic neutron scattering in the picosecond time-scale. Samples of pure phospholipids (DMPC/DMPS) and samples with amyloid-β (25-35) peptide included have been compared. With two different orientations of the samples the directional dependence of the dynamics was probed. The sample temperature was varied between 290 K and 320 K to cover both the gel phase and the liquid-crystalline phase of the lipid membranes. The model for describing the dynamics combines a long-range translational diffusion of the lipid molecules and a spatially restricted diffusive motion. Amyloid-β (25-35) peptide affects significantly the ps-dynamics of oriented lipid membranes in different ways. It accelerates the lateral diffusion especially in the liquid-crystalline phase. This is very important for all kinds of protein-protein interactions which are enabled and strongly influenced by the lateral diffusion such as signal and energy transducing cascades. Amyloid-β (25-35) peptide also increases the local lipid mobility as probed by variations of the vibrational motions with a larger effect in the out-of-plane direction. Thus, the insertion of amyloid-β (25-35) peptide changes not only the structure of phospholipid membranes as previously demonstrated by us employing neutron diffraction (disordering effect on the mosaicity of the lipid bilayer system) but also the dynamics inside the membranes. The amyloid-β (25-35) peptide induced membrane alteration even at only 3 mol% might be involved in the pathology of Alzheimer's disease as well as be a clue in early diagnosis and therapy.