Biomedical subjects

Vladimir Tsuprun

Publications and source records attributed to Vladimir Tsuprun.

9 recordsLinked to original sources

PubMed2007

Pneumococcal peptidoglycan-polysaccharides induce the expression of interleukin-8 in airway epithelial cells by way of nuclear factor-kappaB, nuclear factor interleukin-6, or activation protein-1 dependent mechanisms.

Cell envelope compounds of bacteria trigger immune and inflammatory reactions by way of chemokines/cytokines. In this study, we demonstrated that pneumococcal peptidoglycan-polysaccharides (PGPS) induced the production of interleukin (IL)-8 by way of nuclear factor (NF)-kappaB, nuclear factor interleukin (NF-IL)6, and activation protein (AP)-1 dependent mechanisms in the human bronchial epithelial cells (NL-20) in a dose- and time-dependent manner in vitro, and the mutation of either the NF-kappaB, NF-IL6, or AP-1 binding sites in the promoter of IL-8 abrogated the IL-8 transcriptional activity. In a similar way, lipopolysaccharides induced the promoter activation of IL-8 in NL-20. However, the PGPS-induced IL-8 promoter activation in rodent middle ear epithelial cells required NF-kappaB and NF-IL6 but not AP-1.

Animals↗

PubMed2006

Age-related functional and histopathological changes of the ear in the MPS I mouse.

OBJECTIVE: Mucopolysaccharidosis type I (MPS I) is an autosomal recessive disorder caused by a mutation in the gene encoding the enzyme alpha-L-iduronidase. This enzyme is responsible for degradation of dermatan and heparan sulfates. Enzyme deficiency results in their accumulation in lysosomes of virtually all organs, resulting in severe somatic and neurological changes. Clinical findings of otitis media with mixed hearing loss are common. Cellular and molecular mechanisms of ear pathology and hearing loss are not understood. The purpose of this study is to describe the age-related audiologic and histopathologic changes of the ear in the mouse model of MPS I. METHODS: Auditory brainstem responses (ABR) were obtained to clicks and tone bursts at 1-32kHz, and pathological changes to middle and inner ears were studied with light and electron microscopy in 53 mice that included: (1) wild type (+/+)-five at 2 months, five at 4-6 months, and five at 13-19 months; (2) heterozygotes (+/-)-four at 2 months, five at 4-6 months, and eight at 13-19 months; and (3) homozygotes (-/-)-five at 2 months, six at 4-6 months, and five at 13-19 months. Histopathology was also done on five newborn -/- mice. RESULTS: In newborns, no lysosomal storage was observed and the ear appeared age appropriately normal. In all other -/- mice, cells with lysosomal storage vacuoles were observed in spiral ligament, spiral prominence, spiral limbus, basilar membrane, epithelial and mesothelial cells of Reissner's membrane, endothelial cells of vessels, and some ganglion cells; their number increased with aging. Hair cell loss was not observed at 2 or 6 months, but there was total loss of the organ of Corti in year-old mice. Hearing of -/- mice was significantly decreased at all ages compared to +/+ and +/-. Hearing loss progressed from mild to moderate loss at 2 months to profound at 6 months and total deafness by 1 year of age. CONCLUSIONS: Progressive age-related changes suggest early therapeutic intervention to prevent sensory cell damage and hearing loss.

Aging↗

PubMed2006

Specificity of aminoacylase III-mediated deacetylation of mercapturic acids.

Trichloroethylene (TCE) and other halogenated alkenes are known environmental contaminants with cytotoxic and nephrotoxic effects, and are potential carcinogens. Their metabolism via the mercapturate metabolic pathway was shown to lead to their detoxification. The final products of this pathway, mercapturic acids or N-acetyl-l-cysteine S-conjugates, are secreted into the lumen in the renal proximal tubule. The proximal tubule may also deacetylate mercapturic acids, and the resulting cysteine S-conjugates are transformed by cysteine S-conjugate beta-lyases to nephrotoxic reactive thiols. The specificity and rate of mercapturic acid deacetylation may determine the toxicity of certain mercapturic acids; however, the exact enzymologic processes involved are not known in detail. In the present study we characterized the kinetics of the recently cloned mouse aminoacylase III (AAIII) toward a wide spectrum of halogenated mercapturic acids and N-acetylated amino acids. In general, the V(max) value of AAIII was significantly larger with chlorinated and brominated mercapturic acids, whereas fluorination significantly decreased it. The enzyme deacetylated mercapturic acids derived from the TCE metabolism including N-acetyl-S-(1,2-dichlorovinyl)-l-cysteine (NA-1,2-DCVC) and N-acetyl-S-(2,2-dichlorovinyl)-l-cysteine (NA-2,2-DCVC). Both mercapturic acids induced cytotoxicity in mouse proximal tubule mPCT cells expressing AAIII, which was decreased by an inhibitor of beta-lyase, aminooxyacetate. The toxic effect of NA-2,2-DCVC was smaller than that of NA-1,2-DCVC, indicating that factors other than the intracellular activity of AAIII mediate the cytotoxicity of these mercapturic acids. Our results indicate that in proximal tubule cells, AAIII plays an important role in deacetylating several halogenated mercapturic acids, and this process may be involved in their cyto- and nephrotoxicity.

Acetylation↗

PubMed2005

Round window membrane and labyrinthine pathological changes: an overview.

The round window membrane is considered the most likely pathway from the middle to the inner ear. Various substances placed in the middle ear have been seen to pass through the round window membrane. Once toxic substances or inflammatory mediators such as cytokines and nitric oxide enter the inner ear, various inner ear sequelae such as labyrinthitis, endolymphatic hydrops, sensorineural hearing loss or more insidious diseases can occur.

Bacterial Infections↗

PubMed2004

The structure of tip links and kinocilial links in avian sensory hair bundles.

Recent studies have indicated that the tip links and kinocilial links of sensory hair bundles in the inner ear have similar properties and share a common epitope, and that cadherin 23 may also be a component of each link type. Transmission electron microscopy was therefore used to study and compare the fine structure of the tip links and kinocilial links in avian sensory hair bundles. Tannic acid treatment revealed a thin strand, 150-200 nm long and 8-11 nm thick, present in both link types. Fourier analysis of link images showed that the strand of both link types is formed from two filaments coiled in a helix-like arrangement with an axial period of 20-25 nm, with each filament composed of globular structures that are approximately 4 nm in diameter. Differences in the radius and period of the helix-like structure may underlie the observed variation in the length of tip and kinocilial links. The similar helix-like structure of the tip links and kinocilial links is in accord with the presence of a common cell-surface antigen (TLA antigen) and similarities in the physical and chemical properties of the two link types. The spacing of the globular structures comprising each filament of the two link types is similar to the 4.3 nm center-to-center spacing reported for the globular cadherin repeat, and is consistent with the suggestion that cadherin 23 is the tip link.

Animals↗

PubMed2003

Structural characterization, tissue distribution, and functional expression of murine aminoacylase III.

Many xenobiotics are detoxified through the mercapturate metabolic pathway. The final product of the pathway, mercapturic acids (N-acetylcysteine S-conjugates), are secreted predominantly by renal proximal tubules. Mercapturic acids may undergo a transformation mediated by aminoacylases and cysteine S-conjugate beta-lyases that leads to nephrotoxic reactive thiol formation. The deacetylation of cysteine S-conjugates of N-acyl aromatic amino acids is thought to be mediated by an aminoacylase whose molecular identity has not been determined. In the present study, we cloned aminoacylase III, which likely mediates this process in vivo, and characterized its function and structure. The enzyme consists of 318 amino acids and has a molecular mass (determined by SDS-PAGE) of approximately 35 kDa. Under nondenaturing conditions, the molecular mass of the enzyme is approximately 140 kDa as determined by size-exclusion chromatography, which suggests that it is a tetramer. In agreement with this hypothesis, transmission electron microscopy and image analysis of aminoacylase III showed that the monomers of the enzyme are arranged with a fourfold rotational symmetry. Northern analysis demonstrated an approximately 1.4-kb transcript that was expressed predominantly in kidney and showed less expression in liver, heart, small intestine, brain, lung, testis, and stomach. In kidney, aminoacylase III was immunolocalized predominantly to the apical domain of S1 proximal tubules and the cytoplasm of S2 and S3 proximal tubules. The data suggest that in kidney proximal tubules, aminoacylase III plays an important role in deacetylating mercapturic acids. The predominant cytoplasmic localization of aminoacylase III may explain the greater sensitivity of the proximal straight tubule to the nephrotoxicity of mercapturic acids.

Acetylcysteine↗

PubMed2003

Structure of the stereocilia side links and morphology of auditory hair bundle in relation to noise exposure in the chinchilla.

Stereocilia side links are directly involved in the maintenance of stereociliary bundle integrity in hair cells. The structure of the stereocilia side links and morphology of the auditory hair bundle in relation to noise exposure in the chinchilla was investigated by transmission electron microscopy. The outer hair cell (OHC) stereocilia side link was suggested to consist of extracellular, juxta-membrane and thin filamentous regions. Two beaded filaments were folded at their distal ends and fastened in one globule in the center between stereocilia. An intracellular, submembraneous layer appeared to form a bridge between the actin core and the extracellular, juxta-membrane region of the side link. In normal physiological conditions, most OHC stereocilia had a regular distribution of side links, forming a 'zipper-like' lattice between stereocilium shafts. Side links of the inner hair cell (IHC) stereocilia had a similar filamentous appearance, but were observed less commonly and had decreased structural organization compared to those of the OHC stereocilia. Ultrastructural analysis of OHC and IHC stereocilia showed that a large number of the side links could survive acoustic stimulation of 114 dB SPL for 2 hrs or 123 dB SPL for 15 min, that resulted in temporarily elevated hearing thresholds in all animals. Disarray, separation, close attachment and fusion of stereocilia were more frequently observed for IHC stereocilia and OHC stereocilia that were poorly connected or that lacked side links. Most disarrayed OHC and IHC stereocilia recovered to a normal erect state with restored orientation of the side links after 14-28 days, which correlated with near-complete recovery of auditory sensitivity. However, direct attachment of plasma membranes, ruptured links, fusion and blebs were seen on some stereocilia even after 28 days and appear to be permanent.

Actin Cytoskeleton↗

PubMed2002

Structure of outer hair cell stereocilia side and attachment links in the chinchilla cochlea.

The structure and symmetry of chinchilla outer hair cell (OHC) stereocilia side and attachment links were investigated by transmission electron microscopy using tannic acid and Cuprolinic blue histochemical procedures. The side links run laterally between and across the rows of the stereocilia and connect the stereocilia together within the bundle. Attachment links form a crown-like array around the tips of only the tallest OHC stereocilia and attach these stereocilia to the Type B fibrils of the tectorial membrane. Computer averaging of the side links from tannic acid-treated tissues showed a central dense region of the link between adjacent stereocilia and a smaller dense portion at the plasma membrane end of the link. Computer averaging of Cuprolinic blue-treated tissues showed low electron density of the central region of the link, and the plasma membrane ends of the link were electron dense. After tannic acid treatment, the attachment links showed a diffused radial distribution around the tips of the tallest OHC stereocilia. After Cuprolinic blue treatment, the attachment links appeared as electron-dense, membrane-bound granular structures arranged with radial symmetry. The central regions of the side links are reactive to tannic acid. These regions appear to contain neutral and basic residues of proteins and participate in side-by-side association of the side links in regular aggregates. Cuprolinic blue-reactive regions of the side and attachment links appear to contain acidic sulfated residues of glycoproteins or proteoglycans, which may be involved in the attachment of these links to the stereocilium membrane.

Animals↗

PubMedDate not supplied

Otosclerosis: etiopathogenesis and histopathology.

Otosclerosis is a disease of the bony labyrinth manifesting clinically as a progressive conductive hearing loss, a mixed-type hearing loss, or a sensorineural hearing loss. The age of onset of the hearing loss caused by otosclerosis is principally between 15 and 40 years. Although histopathological inner ear changes due to otosclerosis have been very well documented, the true etiopathogenesis of the disease has yet to be described despite intensive research. Both genetic and environmental factors have been implicated, however.

Age of Onset↗