Confocal-microscopic study of skeletal muscle fibre membrane organelles during Zenker's (spreading) necrosis

The changes of T-system and cellular acidic organelles during spreading (Zenker's) necrosis of frog skeletal muscle fibres have been investigated using laser confocal microscopy and several vital fluorescent dyes acridine orange, RH 414, DiOC6(3), rhodamine 123, fluorescein dextran. The formation of numerous vacuoles as a result of local T-system swelling is most characteristic for initial steps of Zenker's necrosis. Vacuoles can attain tens microns in length. They are located both near nuclear poles and between myofibres. Vacuoles maintain connections with the extracellular space up to the moment of contraction knot rejection, and under definite conditions (glycerol influx to fibre) vacuoles are reversible. They deform nuclei and sarcoplasmic reticulum cisternae. Cellular acidic organelles, accumulating acridine orange (lysosomes, late endosomes, Golgi apparatus cisternae) are situated in direct vicinity with normal and vacuolated T-system. The increase in acidic organelles number and size occur during the pathological process development, and tendency to vacuoles clusterization may be seen. Vacuolation of T-system during necrosis is not followed by vacuole content acidification. The role of cellular acidic organelles and of T-system vacuolation in the development of different muscle pathological changes is discussed.     

Change in localization of cellular vesicular apparatus during differentiation of myoblasts into myotubules in cell culture

The study of changes in the intracellular processes during differentiation of myoblasts into myotubules is of great importance for understanding several fundamental problems of cell biology. At first, this concerns the spatial organization of vacuolar apparatus that reflects the alterations in the properties of cell membranes, cytoskeleton elements and dynamics of vesicular transport in the course of differentiation. The distribution of acidic membrane organelles (lysosomes, late endosomes, Golgi cisternae) during the myotubule formation was revealed. It was shown that perinuclear localization of acidic organelles in myoblasts was replaced by diffuse distribution of these structures in the whole volume of myotubules. Using lipophilic fluorescent dyes, RH 414 and di-8-ANEPPS, the process of formation and dynamics of endocytic vesicles in myoblasts and myotubules was investigated. In the present work, semiconductive nanocrystals, quantum dots (QDs), conjugated with TAT-peptide, which belongs to cell-penetrating peptides, were used to characterize nonspecific endocytosis. It was shown that QDs--TAT complexes penetrate myoblasts but do not penetrate myotubules even after 24 h incubation, which might be connected with plasma membrane changes during the process of skeletal muscle differentiation.     

Pulse electrophoresis of serogroup A streptocci isolated in Moscow

Pulse electrophoresis of group A streptococci (GAS) isolated in 1998 - 2004 in Moscow from patients during outbreaks of tonsillitis in child institutions, military unit and also from sporadic cases in children and adults was performed. During analysis of 48 strains 16 pulse electrotypes were recognized. These data allow to consider that in different child institutions (and other groups) accumulation and spreading of most adapted to such environment variants of GAS are take place and population structure of GAS in such institutions is unique on molecular and other characteristics. This study showed that complex typing of GAS (use of pulse electrophoresis and PCR for detection of erythrogenic toxins A and C) will help to improve molecular-epidemiologic surveillance for streptococcal group A infection, development of recommendations on reduction of morbidity from this infection and also to decrease risk of its severe forms, complications and mortality.     

Molecular epidemiology of infectious diseases

Data on molecular epidemiology of bacterial infections is summarized. The term definitions of "molecular epidemiology", "taxonomic species" are given, the limits and species structure of prokaryotes are described. The basic mechanisms of the prokaryotes variability in the epidemic process, as well as the possibilities and limitations of microbiological, molecular-biological and population--genetic typing methods, are characterized. The tactics of molecular-biological studies in analyzing the population structure on the global, regional and local levels is presented. The economic effectiveness of measures taken with due consideration of information on the clonal structure of causative agents of hospital infections is shown.     

Typing of group A streptococci isolated from urban population of different social status and age

Pulse-electrophoresis, sequencing of emm genes coding protein M and PCR analysis of speA, speB, and speC genes were used for characterization of group A streptococci (GAS) isolated in different years in Moscow and Tuapse mostly from children and military staff. It has been shown that epidemic process of streptococcal infection caused by GAS in Moscow is based on circulation of many independent clones of Streptococcus pyogenes. Obtained data on complex typing of S. pyogenes would be useful for study of molecular epidemiology of diseases caused by GAS and improvement of epidemiologic surveillance.     

Natural foci of leptospirosis in Moscow in 1995 - 2004

The article deals with the results of the 10-year study of the synanthropic urban foci of leptospirosis on the territory of Moscow. Information on the manifestation of the activity of the foci under study, rodents serving as the reservoir of infection in these foci, the etiological structure of the leptospires among these rodents, the state of leptospirosis morbidity among humans is presented.     

Complex typing of Salmonella typhimurium hospital strains

A salmonellosis outbreak, caused by S. typhimurium, was investigated with the use of some microbiological and molecular-biological methods of typing. This investigation revealed that the outbreak was caused by the "outbreak" electrotype of the multi-resistant variant of the infective agent, found to have several plasmidovars. The possibilities and limitations of typing by sensitivity to antibiotics and plasmid DNA profile were shown. These methods of intraspecific typing were regarded as methods making it possible to establish the heterogeneity of S. typhimurium with the use of intraclonal markers.     

Birdsong Overlapping: Random Coincidence or Acoustic Interactions

Doklady Biological Sciences - Birdsong overlapping may either indicate communicative interaction between birds or be a simple coincidence. We developed a program that allowed us to mathematically...     

Confocal microscopy study of membrane organelles of the skeletal muscle fiber in the process of Zenker’s (spreading) necrosis

Using laser confocal microscopy and some vital fluorescent dyes (acridine orange, RH 414, DiOC₆(3), rhodamine 123, fluorescein dextran), changes of the T-system and cellular acidic organelles were studied during spreading (Zenker’s) necrosis of isolated frog skeletal muscle fibers. The most characteristic of the initial stages of development of Zenker’s necrosis is the formation of numerous vacuoles as a result of local T-system swellings. The vacuole length can reach tens of micrometers. They are located both near nuclear poles and between myofibrils. Until the moment of contraction knot separation, the vacuoles preserve their connections with normal T-tubules and under certain conditions (glycerol influx to the fiber) are reversible. The vacuoles deform nuclei and cisternae of the sarcoplasmic reticulum. Acidic cell organelles accumulating acridine orange (lysosomes, late endosomes, trans-Golgi cisternae) are located in the immediate vicinity both of normal and of vacuolated T-tubules. In the course of the development of the pathological process, the size and number of acidic organelles increases and they tend to be clustered. Vacuolation of the T-system during necrosis was not accompanied by vacuole content acidification. At late stages of necrosis, alterations of nuclei and sarcoplasmic reticulum were observed. The role of cellular acidic organelles and of the T-system vacuolation in development of various muscle pathologies is discussed.