[Combined rehabilitation in the patients presenting with dorsopathies of the lumbar spine and concomitant irritable bowel syndrome based at a therapeutic clinic].

BACKGROUND: Comorbidity constitutes a serious challenge for rehabilitative medicine. The comorbidity of the dorsopathy of the lumbar spine and irritable bowel syndrome mutually complicates the clinical course of both conditions, significantly reduces the patients' quality of life. and increases the costs of diagnostic procedures and restoration of the working capacity. The approaches to the non-pharmacological management of the patients presenting with these diseases remain to be developed. AIMS: The objective of the present study was to evaluate the effectiveness of the proposed combined non-medicinal rehabilitation modality which included a course of therapy with the application of modulated sinusoidal currents, total wrappings with the use of a Rapan saline solution, and sedative inhalations additionally introduced into the basic medicinal therapy of the patients presenting with dorsopathy of the lumbar spine combined with irritable bowel syndrome in the stationary phase. MATERIALS AND METHODS: A total of 59 patients at the age from 20 to 65 years suffering from dorsopathy of the lumbar spine and concomitant irritable bowel syndrome were examined and treated. All the patients were randomized into two groups, the main (n=21) and control (n=38) one, matched for the sex and age. The patients in the control group received the conventional medical treatment in accordance with the adopted medical and economic standards during 2 weeks. The patients of the main group received, in addition to the basal medicinal therapy, the proposed combined rehabilitative physiotherapeutic treatment that included a course of therapy with modulated sinusoidal currents, total wrapping with the use of a Rapan saline solution, and sedative inhalations of the of peony root extract. The effectiveness of these rehabilitation modalities was monitored before and after the course of therapy based on the estimation of bowel function dynamics, the severity of pain syndrome, and the patient's quality of life in terms of the health status with the use of the SF-36 questionnaire. RESULTS: The therapeutic and rehabilitative procedures resulted in a well apparent improvement of the genera condition in the patients of the main group associated with a greater degree of reduction of the incidence of specific clinical symptoms, more rapid relief of the pain syndrome, and a more pronounced improvement of the quality of life indices in comparison with the same variables in the patients comprising the control group. After the course of the treatment and rehabilitation, the incidence of spinal pain in the patients of the main group was significantly reduced by 87% (p=0.001) compared with those of the control group (32%; p=0.005). The frequency of abdominal pain decreased in the main group by 47% (p=0.021) versus the control group (by 27%; p=0.007). The quality of life indices increased 1.2 times in the main group but remained unaltered in the control group. DISCUSSION: The patients of the main group exhibited a more pronounced than in the control group positive dynamics of health conditions characterized by a well apparent reduction in the incidence of the major clinical symptoms of the disease, faster alleviation of the pain syndrome, and the marked improvement of the quality of life indices. CONCLUSIONS: The results of the study with the inclusion of therapy with modulated sinusoidal currents together with total wrapping using the Rapan saline solution and sedative inhalations into the program of the combined treatment of the patients presenting with dorsopathy of the lumbar spine and concomitant irritable bowel syndrome provide a basis for recommending this physiotherapeutic modality for personalized rehabilitation of this group of patients under conditions of a therapeutic clinic.

Influenza A virus M1 protein structure probed by in situ limited proteolysis with bromelain.

Influenza A virus matrix M1 protein is membrane associated and plays a crucial role in virus assembly and budding. The N-terminal two thirds of M1 protein was resolved by X-ray crystallography. The overall 3D structure as well as arrangement of the molecule in relation to the viral membrane remains obscure. Now a proteolytic digestion of virions with bromelain was used as an instrument for the in situ assessment of the M1 protein structure. The lipid bilayer around the subviral particles lacking glycoprotein spikes was partially disrupted as was shown by transmission electron microscopy. A phenomenon of M1 protein fragmentation inside the subviral particles was revealed by SDS-PAGE analysis followed by in-gel trypsin hydrolysis and MALDI-TOF mass spectrometry analysis of the additional bands. Putative bromelain-digestion sites appeared to be located at the surface of the M1 protein globule and could be used as landmarks for 3D molecular modeling.

Trifluralin-induced disorganization of microtubular cytoskeleton alters the development of roots in Hordeum vulgare L.

The extensive use of herbicides in agriculture becomes an important factor in environmental pollution, especially in case of slowly degradable compounds. Some agents act on plants during a long period of time, even if a very low concentration of the herbicide remains in the soil. Here, we investigated the toxicological effect of a low concentration of dinitroaniline herbicide, trifluralin, on growing seedlings of Hordeum vulgare L. Trifluralin in concentration of 1 microg/ml inhibited root growth. The mitotic activity of meristematic cells was suppressed due to the retardation of metaphase progression--alteration that can be caused by cytoskeleton disorder. Using antibodies to alpha-tubulin, we investigated the distribution of microtubules in root meristem cells. During all stages of mitosis, the highly regular system of microtubular cytoskeleton observed in control cells was slightly disorganized. An examination of root structure using light and electron microscopy demonstrated that the cell walls did not form normally during cell division that led to the appearance of large multinucleated cells. Also, the premature (pathological) cell differentiation was induced by trifluralin. A part of differentiating cells showed intracellular structural changes that are consistent with programmed cell death. It seems that the development of alterations in trifluralin-treated roots was due to the microtubular cytoskeleton disorganization.

Cell-to-cell cross-talk between mesenchymal stem cells and cardiomyocytes in co-culture.

The goals of the study were: (1) to explore the communication between human mesenchymal stem cells (MSC) and rat cardiac myocytes resulting in differentiation of the stem cells and, (2) to evaluate the role of mitochondria in it. Light and fluorescence microscopy as well as scanning electron microscopy revealed that after co-cultivation, cells formed intercellular contacts and transient exchange with cytosolic elements could be observed. The transport of cytosolic entity had no specific direction. Noticeably, mitochondria also could be transferred to the recipient cells in a unidirectional fashion (towards cardiomyocytes only). Transmission electron microscopy revealed significant variability in both the diameter of intercellular contacting tubes and their shape. Inside of these nanotubes mitochondria-resembling structures were identified. Moreover, after co-cultivation with cardiomyocytes, expression of human-specific myosin was revealed in MSC. Thus, we speculate that: (1) transport of intracellular elements to MSC possibly can determine the direction of their differentiation and, (2) mitochondria may be involved in the mechanism of the stem cell differentiation. It looks plausible that mitochondrial transfer to recipient cardiomyocytes may be involved in the mechanism of failed myocardium repair after stem cells transplantation.

Dynamics of structural and functional association of nucleolar chromosomes in cells of the hexaploid wheat Triticum aestivum L. at different stages of the cell cycle and during genome polyploidization.

Quantitative analysis of interphase association of the nucleolar chromosomes at different stages of the cell cycle and during genome polyploidization was carried out. Cells of various tissues of hexaploid wheat Triticum aestivum L. (Moskovskaya-35) were used, including diploid root meristematic cells, endopolyploid root cells, triploid endosperm cells and antipodal cells with polytene chromosomes. Interphase nucleoli impregnated with silver or stained with autoimmune antibodies to 53 kDa nucleolar protein served as markers of the nucleolar chromosome association. The following data were obtained: (1) silver-staining revealed two pairs of homologous chromosomes 1B and 6B with active nucleolus-organizing regions in the root meristematic cells; (2) maximal number of nucleoli in diploid meristematic cells reaches four, which corresponds to the number of chromosomes with active organizers; (3) analysis of cells at different stages of the cell cycle has shown that the tendency to the nucleoli association is observed as soon as cells pass individual stages of the cycle; (4) after DNA and chromosome reduplication, the nucleolus-organizing regions in sister chromatids function as a common structure-functional complex; (5) in endopolyploid root cells and antipodal cells with polytene chromosomes, the number of nucleoli does not correlate with ploidy level, and an additional nucleolus revealed in some cells is the result of activation of the latent organizer in one of the nucleolar chromosomes; (6) in the triploid endosperm nucleologenesis, the stage of prenucleolar bodies is missing. Our data suggest that "fusion" of nucleoli and reduction of their number due to the "satellite" association of the nucleolar chromosomes are two independent processes regulated by different mechanisms.

DNA localization in the nucleoli of sea urchin Paracentrotus lividus oocytes.

It is known that in oocytes of P. lividus the nucleus contains a single giant nucleolus of unusual structure where intensive rRNA synthesis occurs. However, the questions of structural and functional relationships between the nucleolar compartments and the sites of active rRNA transcription still remain open. In the present work, we studied the chromatin organization in the nucleoli of P. lividus oocytes using Feulgen's and osmium amine staining procedures. Our results indicate that nucleolar chromatin of small (immature) oocytes differs from that in large mature oocytes. At the early stage of development, the DNA filamentous network 0.2-0.5 microm in diameter is formed in the nucleoli. Profound changes in the structure of the nucleolar DNA are observed in the course of oogenesis. Thus, at the late stage of development, the nucleolar chromatin forms characteristic ring-like structures, which indicates a non-uniform distribution of active ribosomal genes. A model of structural organization of the P. lividus nucleolus is discussed.

Quantitative and ultrastructural analysis of the chondriome in ovogenesis and embryogenesis of the sea urchin Paracentrotus lividus.

The dynamics of chondriome in the ovogenesis of the sea urchin Paracentrotus lividus was studied. Growing oocytes 20-30, 50-60 and 90-100 microm in diameter ("small", "medium-sized" and "large", respectively) and mature eggs were used for the ultrastructural and stereological analysis of mitochondria. Linear parameters of mitochondria (length and thickness) were measured on 3-D reconstructions of serial ultrathin sections using the software developed in the laboratory. The following transformations of chondriome structure were shown to occur during ovogenesis: (1) the number of mitochondria (MT) increases with the growth of cytoplasmic compartment; (2) the modal length of MT increases from 0.5 microm in small oocytes to 1 microm in large ones and decreases again to 0.5 microm in the egg; this process is accompanied by changes in the relative number of spherical MT which decreases in medium-sized oocytes and subsequently rises again in the egg; (3) in medium-sized oocytes, dumbbell-shaped MT appear first, the number of these MT reaching the maximum to the stage of large oocytes. In mature eggs, the dumb-bell-shaped MT are absent; (4) in small and medium-sized oocytes, the orthodox conformation of MT is observed, in contrast to MT with a condensed matrix in large oocytes and eggs; (5) in mature eggs, mitochondrial clusters containing 10 to 20 MT of various size are formed. Based on the data obtained, we suggested that during ovogenesis of the sea urchin, specific differentiation of the chondriome is induced which leads to the increase in the quantity of MT via multiple division acts, while restricting the MT growth and variability of their shape.

Differential decondensation of mitotic chromosomes during hypotonic treatment of living cells as a possible cause of G-banding: an ultrastructural study.

The chromosomal ultrastructure of Chinese hamster cells treated with 0.075 M KCl - a solution ordinarily used for making preparations of spread chromosomes - was studied. The hypotonic treatment was shown to result in differential decondensation of chromosomes which consists in the uneven distribution of deoxyribonucleoprotein (DNP) fibrils along chromatids. Fixation of cells with methanol acetic acid causes an abrupt restructuring of chromosomes. However, the DNP preserves its uneven distribution along chromatids. As seen on ultra-thin sections of marker nucleolus organizer chromosomes, the densely packed regions may correspond to G-bands detected in the selfsame chromosomes by standard methods of differential staining. The results suggest that the capacity of chromosomes for differential staining is based on the different resistance of G- and R-bands to the decondensing action of hypotonic solutions on living cells.

Chromatin-nuclear envelope complex from rat liver: isolation and purification.

We describe a method for the isolation of a fraction of nuclear envelope (NE) from rat liver. The method includes mild treatment of pure nuclei with either endonuclease of DNase I under low ionic strength conditions in the presence of magnesium, which allows the nucleomeric organization of the chromatin (Ch) to be preserved. The NEs were purified by centrifugation in sucrose gradients followed by floatation in sucrose. No more than 3% of the Ch present in the purified Ch-NE complexes was due to the non specific adsorption of Ch to the NE. The main components of the complex (Ch and NE) retained their in situ ultrastructure. The complex consisted of 9--10% DNA, 3--4% RNA, about 63% protein and about 24% phospholipids.