Comparative Cytogenetic Variation of the Salivary Gland Polytene Chromosomes in Chironomus riparius Mg., 1804 (Diptera, Chironomidae) from Two Polluted Biotopes of Bulgaria and Russia

The structural–functional variation ofChironomus riparius salivary gland polytene chromosomes was studied in two geographically isolated Palearctic regions, Bulgaria (village Pancharevo) and Russia (St. Petersburg). The two biotopes, where larvae were collected, were polluted with various heavy metals from anthropogenic sources. Hereditary paracentric heterozygous inversions were characteristic of the Russian population, whereas somatic paracentric or pericentric heterozygous inversions were more common in the Bulgarian one. All inversions occurred at low frequencies. Other aberrations found in the two populations included somatic deletions resulting in a pompon structure of chromosome IVG, heterozygous translocation between chromosomes IVG and IIIF, enlargement of individual disks, and the appearance of a heterozygous block close to the centromere of chromosome IVG. In addition, changes in functional activity of the nucleolus organizer and Balbiani rings (BRc/BRb) were observed. Several aberration breakpoints proved to coincide with satellites of the Alu and Hinf families.     

Cytotaxonomy of Chironomidae (Diptera) from Lake Shabla (Bulgaria): cytogenetic evidence for introgressive hydribization

Twenty-two chironomid species were found in Lake Shabla. Five species (Chironomus bonus, Dicrotendipes lobiger, Kiefferulus tendipediformis, Cricotopus suspiciosus and Cricotopus sp.) are new for Bulgaria. Their karytopes are described for the first time.Cytogenetic studies of two closely related chironomid species, Glyptotendipes glaucus and G. pallens, show that in one area of sympatry (Lake Shabla, Bulgaria) about 2.5% of the naturally occuring individuals are hybrids. In most cases, in this hybrid, 1st, IInd and IIIrd chromosomes correspond fully to the chromosomes of G. glaucus. The IVth chromosome is a hybrid, with one homologue of the species G. glaucus; the other homologue is derived from G. pallens. Introgressive hybridization is of particular importance to the production of these combinations. Despite hybridization, natural selection appears to maintain the essential integrity of each separate gene pool.     

Electronmicroscopic observations on the visceral and parietal rat's pleura after contralateral pneumonectomy

The mechanism of compensatory growth and healing of the pleura remains unresolved. Contralateral visceral and parietal (diaphragmatic and costal) pleura were investigated by transmission electron microscopy, following an experimental pneumonectomy (EP). Fifteen young-adult Wistar rats were divided into three groups and with survival times of 1, 5 and 8 days respectively after EP. Three sham-operated (thoracic cavity opened and closed) and three unoperated rats served as controls. One day following EP the superficial mesothelial cells have more microvilli and microvesicles, but a lower number of specialized contacts. Multiplication of extravasal cells leads to an increase of the thickness of the layer over the basal lamina and of the submesothelial layer. Five days after EP the superficial cells show a stratified arrangement in longer sectors of both pleural sheets. Along with typical mesothelial cells there are three new populations of cells: (1) with an abundant granular endoplasmic reticulum and secretory granules, (2) with fibroblast-like characteristics and (3) with a more extensive lysosomal system. The submesothelial layer is thickened due to newly formed blood vessels and collagen bundles. Eight days after EP the mesothelial cells build multi-row arrangement sectors and surround intercellular dilatations covered with microvilli. 'Activated' high mesothelial cells characterize the monolayer sectors. The submesothelial layer remains thicker due to larger collagen bundles and elastic fibers. The changes in the mesothelium and in the connective tissue layer suggest the existence of two periods. The first one is characterized by different mesothelial cell populations, new vasculogenesis and starting of fibrillogenesis. In the second period there are 'activated' mesothelial cells, pleural villi, groups of lymphatic lacunae and significant fibrillogenesis.     

Molecular and Subcellular-Scale Modeling of Nucleotide Diffusion in the Cardiac Myofilament Lattice

Contractile function of cardiac cells is driven by the sliding displacement of myofilaments powered by the cycling myosin crossbridges. Critical to this process is the availability of ATP, which myosin hydrolyzes during the cross-bridge cycle. The diffusion of adenine nucleotides through the myofilament lattice has been shown to be anisotropic, with slower radial diffusion perpendicular to the filament axis relative to parallel, and is attributed to the periodic hexagonal arrangement of the thin (actin) and thick (myosin) filaments. We investigated whether atomistic-resolution details of myofilament proteins can refine coarse-grain estimates of diffusional anisotropy for adenine nucleotides in the cardiac myofibril, using homogenization theory and atomistic thin filament models from the Protein Data Bank. Our results demonstrate considerable anisotropy in ATP and ADP diffusion constants that is consistent with experimental measurements and dependent on lattice spacing and myofilament overlap. A reaction-diffusion model of the half-sarcomere further suggests that diffusional anisotropy may lead to modest adenine nucleotide gradients in the myoplasm under physiological conditions.     

Modeling regulation of cardiac KATP and L-type Ca2+ currents by ATP, ADP, and Mg2+

Changes in cytosolic free Mg(2+) and adenosine nucleotide phosphates affect cardiac excitability and contractility. To investigate how modulation by Mg(2+), ATP, and ADP of K(ATP) and L-type Ca(2+) channels influences excitation-contraction coupling, we incorporated equations for intracellular ATP and MgADP regulation of the K(ATP) current and MgATP regulation of the L-type Ca(2+) current in an ionic-metabolic model of the canine ventricular myocyte. The new model: 1), quantitatively reproduces a dose-response relationship for the effects of changes in ATP on K(ATP) current, 2), simulates effects of ADP in modulating ATP sensitivity of K(ATP) channel, 3), predicts activation of Ca(2+) current during rapid increase in MgATP, and 4), demonstrates that decreased ATP/ADP ratio with normal total Mg(2+) or increased free Mg(2+) with normal ATP and ADP activate K(ATP) current, shorten action potential, and alter ionic currents and intracellular Ca(2+) signals. The model predictions are in agreement with experimental data measured under normal and a variety of pathological conditions.     

Elongation of Mouse Prion Protein Amyloid-Like Fibrils: Effect of Temperature and Denaturant Concentration

Prion protein is known to have the ability to adopt a pathogenic conformation, which seems to be the basis for protein-only infectivity. The infectivity is based on self-replication of this pathogenic prion structure. One of possible mechanisms for such replication is the elongation of amyloid-like fibrils.We measured elongation kinetics and thermodynamics of mouse prion amyloid-like fibrils at different guanidine hydrochloride (GuHCl) concentrations. Our data show that both increases in temperature and GuHCl concentration help unfold monomeric protein and thus accelerate elongation. Once the monomers are unfolded, further increases in temperature raise the rate of elongation, whereas the addition of GuHCl decreases it.We demonstrated a possible way to determine different activation energies of amyloid-like fibril elongation by using folded and unfolded protein molecules. This approach separates thermodynamic data for fibril-assisted monomer unfolding and for refolding and formation of amyloid-like structure.     

Conjugal transfer of streptococcal plasmids to strains of Bacillus sphaericus

Abstract The streptococcal plasmids pIP501, pDC10535 and pSM15346 coding for MLS resistance have been successfully transferred to Bacillus sphaericus strains 1593, 2297, 2362 and local strains after mating on filter. The transfer occurred at high frequency and was demonstrated electrophoretically. Conjugation in liquid media also took place but at lower frequency. The conjugation process was studied by electron microscopy. A kind of a bridge of electron-dense material between the mating cells has been observed. The addition of trypsin did not change significantly the transfer frequency in our experimental conditions.     

Molecular and Subcellular-Scale Modeling of Nucleotide Diffusion in the Cardiac Myofilament Lattice

Contractile function of cardiac cells is driven by the sliding displacement of myofilaments powered by the cycling myosin crossbridges. Critical to this process is the availability of ATP, which myosin hydrolyzes during the cross-bridge cycle. The diffusion of adenine nucleotides through the myofilament lattice has been shown to be anisotropic, with slower radial diffusion perpendicular to the filament axis relative to parallel, and is attributed to the periodic hexagonal arrangement of the thin (actin) and thick (myosin) filaments. We investigated whether atomistic-resolution details of myofilament proteins can refine coarse-grain estimates of diffusional anisotropy for adenine nucleotides in the cardiac myofibril, using homogenization theory and atomistic thin filament models from the Protein Data Bank. Our results demonstrate considerable anisotropy in ATP and ADP diffusion constants that is consistent with experimental measurements and dependent on lattice spacing and myofilament overlap. A reaction-diffusion model of the half-sarcomere further suggests that diffusional anisotropy may lead to modest adenine nucleotide gradients in the myoplasm under physiological conditions.     

Biosynthesis of a thermostable gellan lyase by newly isolated and characterized strain of Geobacillus stearothermophilus 98

The thermophilic strain able to degrade gellan was isolated from Bulgarian hot spring. According to its morphological and biochemical properties and by partial sequencing of its 16S rDNA, it was classified as Geobacillus stearothermophilus. It grew in a synthetic medium with gellan as the only carbon source with a specific growth rate of 0.69 h⁻¹ and generation time of 60 min. The strain produced thermostable gellan lyase extracellularly during exponential phase. Its synthesis was inducible; the enzyme was not registered in culture liquid without gellan. The enzyme activity was increased tenfold in conditions of continuous cultivation compared to data from batch fermentations and enzyme productivity was almost sixfold higher. The enzyme showed optimal activity at 75°C in a very large pH area 4–8.5. This enzyme is the first reported thermostable gellan lyase, its residual activity was 100% after 24 h incubation at 60°C and its half-life was 60 min at 70°C.