Concentrations of Metals in Maxilla and Mandible Deciduous and Permanent Human Teeth

The study shows the results of research on the concentrations of metals (Cd, Pb, Mn, Cu, Cr, Fe, Zn, Na, K, Mg, and Ca) in deciduous and permanent teeth with respect to their location in the oral cavity (maxilla, mandible). It has been found that the concentrations of metals were significantly higher in the deciduous teeth than in permanent ones. Regression analysis and principal component analysis revealed an increased dynamics of the processes of binding the elements by the hydroxyapatite of deciduous teeth. Higher concentrations of the metals were found in the maxilla permanent and deciduous teeth than in the mandible ones.     

Overexpression inEscherichia coli,Folding, Purification, and Characterization of the First Three Short Consensus Repeat Modules of Human Complement Receptor Type 1

We have developed a simple expression, isolation, and folding protocol for an SCR oligomer comprising the first three SCRs of complement receptor Type 1 (C3b/C4b receptor, CD35). A T7 RNA polymerase expression system inEscherichia coliwas used to express the oligomer as inclusion bodies. The oligomer was recovered from solubilized inclusion bodies using batch adsorption on SP–Sepharose. The oligomer was folded by one-step dilution in 20 m ethanolamine/1 m EDTA supplemented with 1 m GSH/0.5 m GSSG. The folded material was processed to a concentrated (>20 mg/ml), usable product of greater than 98% purity using a combination of ultrafiltration, ammonium sulfate treatment, hydrophobic interaction, and size-exclusion chromatography. The yield of folded material varied between 6 and 15 mg/liter culture. The oxidation states of the 12 cysteine residues in SCR(1–3) were identified by HPLC of peptide fragments from a tryptic digest using dual UV/fluorescence detection, collection of selected peaks, and N-terminal sequencing. This methodology confirmed the expected location of disulfide bridges. Equilibrium and velocity sedimentation studies are interpreted in terms of a single sedimenting species with molecular weights of 21,629 and 21,063 by these respective techniques. These values compare to the predicted molecular weight, from amino acid composition, of 21,817. The hydrodynamic properties of the molecule indicate that it is asymmetric with an axial ratio of 1:5.2 or equivalent dimensions of 21 × 110 Å. SCR(1–3) has an unusual CD spectrum exhibiting a broad maximum at 220–230 nm and a minimum at 190 nm. There was little evidence of classical secondary structure. The product exhibited concentration-dependent inhibition of complement-mediated lysis of sensitized sheep red blood cells.     

The influence of the two different light intensities duringthe daytime on the colon motility in patients with irritable bowel syndrome (IBS): a preliminary report

Irritable bowel syndrome (IBS) is the most frequent functional disorder of the gastrointestinal tract, with great economical impact. The etiology and pathogenesis of the disease are unclear. Among patients seeking medical attention for IBS, 70 - 90% have psychiatric co-morbidity, most commonly major depression. In this study we test the influence of bright light on colon motility and subjectively felt symptoms. Eight IBS patients participated in the experiment. The passage rate was evaluated twice for every person: (1) after three days of exposure to bright light -3000 lux (from 8:00 to 18:00), (2) after three days exposure to dim light -100 lux (from 8:00 to 18:00). The comparison of colonic time in IBS patients point to differences in the elimination of markers, depending on experimental light conditions. After bright light conditions a tendency is observed to slower elimination of markers in the IBS-diarrhoea patients, and to a quicker elimination in the IBS-constipated patients. According to subjective feeling, all IBS patients reported an improvement in bowel function and relief of pain after bright but not after dim light application. Results of this preliminary study suggest that phototherapy might be a valuable addition to the conventional treatment of IBS.     

Expression of HvHMA2 in tobacco modifies Zn–Fe–Cd homeostasis

HvHMA2 is a plasma membrane P_1B-ATPase from barley that functions in Zn/Cd root-to-shoot transport. To assess the usefulness of HvHMA2 for modifying the metal content in aerial plant parts, it was expressed in tobacco under the CaMV35S promoter. Transformation with HvHMA2 did not produce one unique pattern of Zn and Cd accumulation; instead it depended on external metal supply. Thus Zn and Cd root-to-shoot translocation was facilitated, but not at all applied Zn/Cd concentrations. Metal uptake was restricted in HvHMA2-transformed plants and the level in the shoot was not enhanced. It was shown that HvHMA2 localizes to the plasma membrane of tobacco cells, and overloads the apoplast with Zn, which could explain the overall decrease in metal uptake observed. Despite the lower levels in the shoot, HvHMA2 transformants showed increased Zn sensitivity. Moreover, introduction of HvHMA2 into tobacco interfered with Fe metabolism and Fe accumulation was modified in HvHMA2-transformants in a Zn- and Cd-concentration dependent manner. The results indicate that ectopic expression of the export protein HvHMA2 in tobacco interferes with tobacco metal Zn–Cd–Fe cross-homeostasis, inducing internal mechanisms regulating metal uptake and tolerance.     

Shigella flexneri 3a Outer Membrane Protein C Epitope Is Recognized by Human Umbilical Cord Sera and Associated with Protective Activity

Shigella flexneri 3a is one of the five major strains of the Shigella genus responsible for dysentery, especially among children, in regions of high poverty and poor sanitation. The outer membrane proteins (OMP) of this bacterium elicit immunological responses and are considered a prime target for vaccine development. When injected into mice they elicit a protective immunological response against a lethal dose of the pathogen. The OMPs from S. flexneri 3a were isolated and resolved by two-dimension-SDS-PAGE. Two 38-kDa spots were of particular interest since in our earlier studies OMPs of such molecular mass were found to interact with umbilical cord sera. These two spots were identified as OmpC by ESI-MS/MS spectrometry. By DNA sequencing, the ompC gene from S. flexneri 3a was identical to ompC from S. flexneri 2a [Gene Bank: 24113600]. A 3D model of OmpC was built and used to predict B-cell type (discontinuous) antigenic epitopes. Six epitopes bearing the highest score were selected and the corresponding peptides were synthesized. Only the peptides representing loop V of OmpC reacted strongly with the umbilical cord serum immunoglobulins. To determine which amino acids are essential for the antigenic activity of the epitope, the loop V was scanned with a series of dodecapeptides. The peptide RYDERY was identified as a minimal sequence for the loop V epitope. Truncation at either the C- or N-terminus rendered this peptide inactive. Apart from C-terminal tyrosine, substitution of each of the remaining five amino acids with glycine, led to a precipitous loss of immunological activity. This peptide may serve as a ligand in affinity chromatography of OmpC-specific antibodies and as a component of a vaccine designed to boost human immune defenses against enterobacterial infections.