Of monkeys and men: A metabolomic analysis of static and dynamic urinary metabolic phenotypes in two species

Background: Metabolomics has attracted the interest of the medical community for its potential in predicting early derangements from a healthy to a diseased metabolic phenotype. One key issue is the diversity observed in metabolic profiles of different healthy individuals, commonly attributed to the variation of intrinsic (such as (epi)genetic...

Changes at the KinA PAS-A dimerization interface influence histidine kinase function

The Bacillus subtilis KinA protein is a histidine protein kinase that controls the commitment of this organism to sporulate in response to nutrient deprivation and several other conditions. Prior studies indicated that the N-terminal Per-ARNT-Sim domain (PAS-A) plays a critical role in the catalytic activity of this enzyme, as demonstrated by...

Prolonged in vivo gene silencing by electroporation-mediated plasmid delivery of small interfering RNA

For the successful application of RNA interference in vivo, it is desired to achieve (local) delivery of small interfering RNAs (siRNAs) and long-term gene silencing. Nonviral electrodelivery is suitable to obtain local and prolonged expression of transgenes. By intramuscular electrodelivery of a plasmid in which two opposing human polymerase...

Single amino acid residue changes in subsite - 1 of inulosucrase from Lactobacillus reuteri 121 strongly influence the size of products synthesized

Bacterial fructansucrase enzymes belong to glycoside hydrolase family 68 and catalyze transglycosylation reactions with sucrose, resulting in the synthesis of fructooligosaccharides and/or a fructan polymer. Significant differences in fructansucrase enzyme product specificities can be observed, i.e. in the type of polymer (levan or inulin)...

Bioavailability of tryptophan from a single oral dose of a trytophan-enriched peptide mixture in healthy men

The aim of the study was to investigate the bioavailability of tryptophan (Trp) from a Trp-enriched peptide mixture in healthy men. A second objective was to investigate the effect of this Trp-enriched protein hydrolysate on potential parameters of serotonergic activity. serum serotonim melatonin and prolactin and urinary 5-hydroxyindol-acetic...

Allosteric modulation and constitutive activity of fusion proteins between the adenosine A1 receptor and different 351Cys-mutated Gi α-subunits

We studied fusion proteins between the human adenosine A1 receptor and different 351Cys-mutated Gi1 α-subunits (A1-Giα) with respect to two important concepts in receptor pharmacology, i.e. allosteric modulation and constitutive activity/inverse agonism. The aim of our study was twofold. We first analysed whether such fusion products are still...

Phenotypic and molecular characterization of Bruck syndrome (Osteogenesis imperfecta with contractures of the large joints) caused by a recessive mutation in PLOD2

Bruck syndrome (BS) is a recessively-inherited phenotypic disorder featuring the unusual combination of skeletal changes resembling osteogenesis imperfects (OI) with congenital contractures of the large joints. Clinical heterogeneity is apparent in cases reported thus far. While the genes coding for collagen 1 chains are unaffected in BS, there...

Examining the role of glutamic acid 183 in chloroperoxidase catalysis

Site-directed mutagenesis has been used to investigate the role of glutamic acid 183 in chloroperoxidase catalysis. Based on the x-ray crystallographic structure of chloroperoxidase, Glu-183 is postulated to function on distal side of the heme prosthetic group as an acid-base catalyst in facilitating the reaction between the peroxidase and...

Deep-vein thrombosis is not associated with the P/S186 polymorphism of histidine-rich glycoprotein

Background: In several studies, higher plasma levels of histidine-rich glycoprotein (HRG) have been observed in patients with venous thrombosis than in healthy subjects. Apart from environmental factors, such as the use of oral contraceptives, the plasma HRG levels are mainly determined genetically. The most important genetic determinant is P...

Amino acid analysis by reverse-phase high-performance liquid chromatography: Improved derivatization and detection conditions with 9-fluorenylmethyl chloroformate

The radial immunodiffusion assay for plasma Histidine-rich Glycoprotein (HRG) based on a polyclonal antibody, shows a different specificity towards the two variants of a common amino acid polymorphism

Recently, two molecular weight forms of Histidine-rich Glycoprotein have been described which explain 59% of the variation in plasma HRG levels. Here, we demonstrate that this can partly be ascribed to a difference in specificity of the immuno assay for HRG towards the two alleles of the polymorphism.

An amino acid polymorphism in histidine-rich glycoprotein (HRG) explains 59% of the variance in plasma HRG levels

A pedigree-based maximum likelihood method developed by Lange et al. (12) was used to study the contribution of a newly defined di-allelic polymorphism in histidine-rich glycoprotein (HRG) to the plasma levels of HRG. In four families (n = 99) and 20 volunteers we found a heritability of 70%, an age effect of 3% and an effect of individual...

Identification and genetic analysis of a common molecular variant of histidine-rich glycoprotein with a difference of 2KD in apparent molecular weight

Two forms of histidine-rich glycoprotein (HRG) were detected on SDS-PAGE by silver staining and immunoblotting after isolation of the protein from pooled plasma using immune-affinity chromatography followed by chromatography with heparin-Sepharose. Both forms were single-chain molecules and the apparent molecular weights of form 1 and form 2...

Development of a detection system for histidine decarboxylating lactic acid bacteria based on DNA probes, PCR and activity test

On the basis of the comparison of the nucleotide sequences of the histidine decarboxylase genes (hdcA) of Lactobacillus 30A and Clostridium perfringens and the amino acid sequences of these histidine decarboxylases and those of Lactobacillus buchneri and Micrococcus, oligonucleotides unique to the hdcA genes were synthesized and used in PCR. All...

Assignment of the human gene for histidine-rich glycoprotein to chromosome 3

Histidine-rich glycoprotein (HRG) is a monomeric plasma glycoprotein involved in the modulation of coagulation and fibrinolysis. Using Southern analysis of human-rodent somatic cell hybrid DNA with a human HRG-specific cDNA probe, the HRG gene was assigned to chromosome 3. One hybrid that was known to contain only a segment of chromosome 3 also...

Urinary excretion of 3-methylhistidine and creatinine by healthy Dutch children during day and night. The influence of age and sex

The urinary excretion of 3-methylhistidine and creatinine and the urinary 3-methylhistidine to creatinine excretion ratio during day and night were investigated in a group of 103 healthy, normally fed Dutch children (52 boys and 51 girls) aged 2-17 years. The 3-methylhistidine to creatinine ratio of the night urine appeared to be significantly...

Effects of amounts and type of agar on the number of spontaneous revertants in the Ames test

It has been shown that the spontaneous revertants in the Ames test can depend on the amount and type of bottom agar on the plates. The most clear-cut effect was found with his G46 strains, carrying the ??uvrB mutation, and with the new hisG428 strain, TA102. Evidence of mutagenic impurities in agar has been found. Chemicals/CAS: agar, 9002-18-0;...

Prophage induction and mutagenicity of a series of anti-tumour platinum(II) and platinum(IV) co-ordination complexes

Eleven platinum compounds with nitrogen donor ligands, previously tested for anti-tumour activity were studied for induction of prophage lambda and for mutagenicity in the Ames assay, with various strains of Salmonella. The compounds included cis and trans isomers of Pt(II) and Pt(IV) complexes and were tested with and without metabolic...