Hafnia alvei lipopolysaccharides: Isolation, sugar composition and SDS-PAGE analysis

Abstract Lipopolysaccharides (LPS) of 33 strains of Hafnia alvei were isolated and purified. LPS content of the dry bacterial mass ranged from 1.2 to 4.5%. All examined lipopolysaccharides contained glucose, glucosamine, heptose, 3-deoxy-octulosonic acid and often galactose. Rhamnose, mannose, galactosamine, mannosamine and unidentified amino sugars were found in some H. alvei strains. Sialic acid was present in LPS of one strain. d -3-Hydroxybutyryl groups also were identified in lipopolysaccharides of 5 strains of this genus.
SDS-PAGE of the lipopolysaccharides was presented in the paper. According to these results two core types exist in H. alvei .     

Calcium Regulates the Activity and Structural Stability of Tpr,a Bacterial Calpain-like Peptidase

Porphyromonas gingivalis is a peptide-fermenting asaccharolytic periodontal pathogen. Its genome contains several genes encoding cysteine peptidases other than gingipains. One of these genes (PG1055) encodes a protein called Tpr (thiol protease) that has sequence similarity to cysteine peptidases of the papain and calpain families. In this study we biochemically characterize Tpr. We found that the 55-kDa Tpr inactive zymogen proteolytically processes itself into active forms of 48, 37, and 33 kDa via sequential truncations at the N terminus. These processed molecular forms of Tpr are associated with the bacterial outer membrane where they are likely responsible for the generation of metabolic peptides required for survival of the pathogen. Both autoprocessing and activity were dependent on calcium concentrations >1 mm, consistent with the protein''s activity within the intestinal and inflammatory milieus. Calcium also stabilized the Tpr structure and rendered the protein fully resistant to proteolytic degradation by gingipains. Together, our findings suggest that Tpr is an example of a bacterial calpain, a calcium-responsive peptidase that may generate substrates required for the peptide-fermenting metabolism of P. gingivalis. Aside from nutrient generation, Tpr may also be involved in evasion of host immune response through degradation of the antimicrobial peptide LL-37 and complement proteins C3, C4, and C5. Taken together, these results indicate that Tpr likely represents an important pathogenesis factor for P. gingivalis.     

Excitons in Metal‐Halide Perovskites

The unprecedented increase of the power conversion efficiency of metal‐halide perovskite solar cells has significantly outpaced the understanding of their fundamental properties. One of the biggest puzzles of perovskites has been the exciton binding energy, which has proved to be difficult to determine experimentally. Many contradictory reports can be found in the literature with values of the exciton binding energy from a few meV to a few tens of meV. In this review the results of the last few years of intense investigation of the exciton physic in perovskite materials are summarized. In particular a critical overview of the different experimental approaches used to determine exciton binding energy is provided. The problem of exciton binding energy in the context of the polar nature of perovskite crystals and related polaron effects which have been neglected to date in most of work is discussed. It is shown that polaron effects can reconcile at least some of the experimental observations and controversy present in the literature. Finally, the current status of the exciton fine structure in perovskite materials is summarized. The peculiar carrier–phonon coupling can help to understand the intriguing efficiency of light emission from metal‐halide perovskites.     

Arsenic content in the femur head of the residents of southern and central Poland

Arsenic content was assayed in the samples of the femur head of the people living in southern and central Poland (Kraków, n=13; Silesian region, n=13; Łódź, n=12). The average age being 68.7±8.7 yr. Arsenic content in the femur head was determined applying the hydride generation atomic absorption spectrometry (HG-AAS) method after microwave mineralization. The average arsenic contents in the femur head of the residents of the Łódź, Kraków, and Silesian regions were 0.41 μg/g, 0.37 μg/g, and 0.18 μg/g, respectively. No correlation has been found between arsenic content in the femur head and the content of other metals. Neither the age nor sex of the people tested affected the arsenic content in the femur head.     

Synthesis and antiprotozoal activity of 2,5-bis[amidinoaryl]thiazoles

Seven novel diamidino 2,5-bis(aryl)thiazoles (5a–g) were synthesized and evaluated against Trypanosoma brucei rhodensiense (T. b. r.) and Plasmodium falciparum (P. f.). The diamidines were obtained directly from the corresponding bis-nitriles (4a–g) by the action of lithium bis(trimethylsilyl)amide. The bis-nitriles 4a–f were synthesized in four steps starting with the Stille coupling of 2-tributyltinthiazole with the appropriate cyanoaryl halide. The bis-nitrile 5g was obtained by the palladium facilitated coupling of the mixed tin-silyl reagent 2-trimethylsilyl-5-trimethyltinthiazole with 2-bromo-5-cyanopyridine. The amidoxime potential prodrugs 6a–e, 6g were obtained by the reaction of hydroxylamine with the bis-nitriles. O-Methylation of the amidoximes gave the corresponding N-methoxyamidines 7a–c, 7e, 7g. The diamidines showed strong DNA binding affinity as reflected by ΔT_m measurements. Four of the diamidines 5a, 5b, 5d and 5e were highly active in vitro against P. f. giving IC₅₀ values between 1.1 and 2.5 nM. The same four diamidines showed IC₅₀ values between 4 and 6 nM against T. b. r. The selectivity indices ranged from 233 to 9175. One diamidine 5a produced one of four cures at an ip dose of 4 × 5 mg/kg in the STIB900 mouse model for acute African trypanosomiasis. The amidoxime and N-methoxyamidine of 5a were the only produgs to provide cures (1/4 cures) in the same mouse model on oral dosage at 4 × 25 mg/kg.     

In vitro study of AMP-deaminase from fish (Cyprinus carpio) treated with hydrolyzable tannins

AMP-deaminase (EC 3.5.4.6) is an enzyme responsible for stabilising adenylate energy changes. The properties of this enzyme are controlled by various ligands of hydrophobic nature. An investigation of enzyme activity alterations under the influence of natural phenolic acids (tannic, ellagic and gallic) which are soluble in water, could evidence the biological toxicity of these compounds. In our study purified AMP-deaminase isolated from white muscle of Cyprinus carpio was exposed to phenolic acids in the concentration range of 1 to 50 microM as well as to tannic acid in the presence of Cu2+ ions (5 microM). On the basis of the obtained results we can conclude that among the tested acids, gallic acid did not contribute to the change in AMP-deaminase activity, whereas ellagic acid diminished its activity at the highest concentration (50 microM). Tannic acid caused a significant decrease in the enzyme activity in comparison to control for all used concentrations. Cu2+ ions alone reduced the activity of AMP-deaminase for all studied concentrations. A combined action of a chosen Cu2+ ions concentration (5 microM) with tannic acid at the concentration higher than 2 microM resulted in a decrease in the enzyme activity, but for lower tannic acid concentration of 1 microM the activity of AMP-deaminase was stimulated. These experiments showed that tannic acid may stop free radical chain reactions only at low concentrations (1 microM) in the presence of Cu2+ ions (5 microM).     

Quantitative immunogold study of increased expression of metallothionein-I/II in the brain perivascular areas of diabetic scrapie-infected mice

Quantitative immunogold procedure was used to study the distribution of metallothionein I/II (MT-I/II) at the ultrastructural level in the perivascular areas, including microvascular endothelial cells (ECs) and astrocytes with their perivascular end-feet, in brains of scrapie-infected hyperglycemic (diabetic) and normoglycemic (non-diabetic) mice. Samples of the fronto-parietal cortex obtained from diabetic and non-diabetic scrapie-infected, as well as from non-infected (control) SJL/J mice, were processed for immunocytochemical examination. In control mice, the labelling of the ECs was of low intensity, restricted to few immunogold particles in the cytoplasm. More intense labelling was present in the cytoplasm of astrocytic perivascular processes and perikarya, where it was associated with endoplasmic reticulum and fibrils. A few immunosignals were also present inside the nuclei of astrocytes. In diabetic mice the labelling of the EC cytoplasm was slightly increased, whereas in the cytoplasm of perivascular processes and pericarya of astrocytes, including their nuclei, there was significant enhancement of labelling. In these cells the density of immunosignals was highest in the areas of cytoplasm containing bundles of fibrils. In non-diabetic, scrapie-infected mice the intensity of immunolabelling was higher than in control mice but slightly lower than in diabetic mice. These results are similar to those in Alzheimer’s disease reported by other authors, and suggest that neurodegenerative diseases as well as metabolic stress enhance the metallothionein expression in perivascular regions of brain cerebral cortex, predominantly in astrocytes.     

Assessment of various diagnostic methods of ureaplasma respiratory tract infections in newborns

We compared three methods used microbial culturing for detection of ureaplasmas in endotracheal aspirate from 500 prematurely born neonates with respiratory disturbances: BioMerieux test, PCR and microbial culturing. Ureaplasmas were detected in respiratory tracts of 79 (16%) newborns. Correlation of the results of culture with those obtained with the BioMerieux kit, culture with PCR and BioMerieux kit with PCR was 97%, 89% and 90%, respectively. Sensitivity and specificity of PCR in comparison with culture was 86% and 98%, respectively, and of the BioMerieux kit 96% and 98%. PCR can be recommended in rapid diagnostics of respiratory infections in newborns suffering from respiratory disorders. It allows the detection of ureaplasmas in case of parallel infections and identification of their species.