The complex analysis of risk factors, influencing on the progression of chronic obstructive ulmonary disease.

AIM: The aim of the study is the analysis of risk factors, influencing on the progression of chronic obstructive pulmonary disease. MATERIALS AND METHODS: 259 patients with chronic obstructive pulmonary disease (COPD) I and III stages, mean age 54,3±11,7 and the duration of the disease 10,1±5,7. Patients were divided into 2 groups: I group (n=119) - with III stage of disease, II group (n=140) - COPD with I stage. 30 healthy were involved in control group. The risk factors of progression of COPD were determined with the use of regressive and correlative analyses. RESULTS: The level of markers of systemic inflammation in serum of patients with COPD: TNF-α (RR 3,25); IL-8 (RR 2,1); hs-CRP (RR 3,52) in combination with traditional risk factors (long course of the disease (RR 2,3), age (RR 1,65), smoking (RR 1,65), and also obesity (RR 2,45), early menopause (RR 3,52), hereditary respiratory predisposition (RR 3,05), and also coexisting cardiovascular pathology increase the risk of COPD progression several times. . CONCLUSION: According to the results of this trial, unmodified risk factors such as hereditary predisposition, early menopause, long course of the diseases and coexisting cardiovascular pathology contribute significantly on progression of COPD. The relative risk increases with increasing of concentration of markers of systemic inflammation (TNF-α, IL-8 and hs-CRP), that correlate with clinical manifestations of COPD: cough, shortness of breath on VAS in mm.

Up-to-date concentrations of long-lived artificial radionuclides in the Tom and Ob rivers in the area influenced by discharges from Siberian chemical combine.

The Siberian Chemical Combine (SCC) is located in Seversk (formerly known as Tomsk-7) in the Tomsk Region of the Russian Federation. The main contribution of radionuclides in the SCC process water discharged into the Tom River was from the single-pass reactors, now removed from service (the last SCC reactor was shutdown on June 5, 2008). The data on the concentrations of (90)Sr, (137)Cs, (239,240)Pu and other artificial radionuclides in water, bottom sediments and flood-plain soils of the Tom and Ob rivers from Tomsk to the confluence of the rivers, are presented and discussed. The results of measurements carried out after shutdown of the last SCC single-pass reactor indicated no radiologically significant consequences of SCC activities for the studied water environment compartments. Contemporary activity concentrations of long-lived artificial radionuclides (3)H, (90)Sr, (137)Cs and (239,240)Pu in river water were below the intervention levels established by current regulations of the Russian Federation for these radionuclides. The results of (3)H analysis in water from the Tom and Samuska rivers demonstrated no inflow of contaminated formation water to surface water from the sites where liquid radioactive wastes of the SCC were injected below the surface. However, the density of flood-plain soil contamination by long-lived (137)Cs in the area influenced by SCC liquid discharges was higher than regional technogenic background. There were local flood-plain areas contaminated not only by (137)Cs, but also other gamma-emitters, such as (60)Co and (152)Eu.

[Mystery and problems of cloning].

The attention of investigators is attracted to the fact that, in spite of great efforts in mammalian cloning, advances that have been made in this area of research are not great, and cloned animals have developmental pathologies often incompatible with life and/or reproduction ability. It is yet not clear what technical or biological factors underlie this, and how they are connected or interact with each other, which is more realistic strategically. There is a great number of articles dealing with the influence of cloning with the nuclear transfer on genetic and epigenetic reprogramming of donor cells. At the same time we can see the practical absence of analytical investigations concerning the technology of cloning as such, its weak points, and possible sources of cellular trauma in the course of microsurgery of nuclear transfer or twinning. This article discusses step by step several nuclear transfer techniques and the methods of dividing early preimplanted embryos for twinning with the aim to reveal possible sources of cell damage during micromanipulation that may have negative influence on the development of cloned organisms. Several new author's technologies based on the study of cell biophysical characteristics are described, which allow one to avoid cellular trauma during manipulation and minimize the possibility of cell damage at any rate.

[Methods of substances and organelles introduction in living cell for cell engineering technologies].

We have presented the classification of more than 40 methods of genetic material, substances and organelles introduction into a living cell. Each of them has its characteristic advantages, disadvantages and limitations with respect to cell viability, transfer efficiency, general applicability, and technical requirements. It this article we have enlarged on the description of our developments of several new and improved approaches, methods and devices of the direct microinjection into a single cell and cell microsurgery with the help of glass micropipettes. The problem of low efficiency of mammalian cloning is discussed with emphasis on the necessity of expertizing of each step of single cell reconstruction to begin with microsurgical manipulations and necessity of the development of such methods of single cell resonstruction that could minimize the possible damage of the cell.

[Biophysical aspects of reconstruction of a single cell by the methods of cell engineering].

The problems of a low efficiency of mammalian cloning are discussed with emphasis on the necessity of the expertise of each step of single cell reconstruction, beginning with microsurgical manipulations. The fact of cell content leakage when the cell is held during microsurgery or microinjections with the help of the conventional method using negative pressure in the holding micropipette was demonstrated in experiments on murine embryos. It was shown that the rate of cell content efflux depends on the value of negative pressure generated in the holding micropipette, and is directly proportional to the dimensions of its orifice and the duration of micromanipulations. An alternative method of cell fixation using the capillary forces of the holding micropipette was proposed. The method optimizes the process of cell fixation, reducing the holding effort by two orders of magnitude. As a result, 92% of embryos remain viable after fixation of embryo, as compared with 39% in the conventional technique. In order to diminish the cell damage produced by the tip of a microinstrument, a new technique of fabricating micropipettes was proposed. The improved method of filling the micropipette with viscous liquids, including DNA, which is described in details in the paper, enabled constant (non-stop) microinjection of more than 1000 cells by hand, without any special automatic device.

[Correlation between the cellular content of mobile water and the viability of lyophilized yeast cells]. / Korreliatsiia velichiny fraktsii podvizhnoi vody i zhiznesposobnosti liofilizirovannykh kletok drozhzhei.

Spin-echo NMR studies showed that lyophilized yeast cells contain isolated mobile water (IMW), whose content varied from 0.25% (of the dry weight of cells) in the lyophilized exponential-phase yeast cells to 3.8% in the lyophilized lag-phase and stationary-phase yeast cells. The viability rate of yeast cells varied from 20% in the lyophilized preparation of exponential-phase cells to 86% in the lyophilized preparation of early-stationary-phase cells. In the lyophilized preparation of yeast cells grown in a chemostat mode at a constant specific rate, the content of IMW depended on the growth-limiting factor, being minimal in the case of growth limitation by carbon source. In the latter case, the viability rate of cells was also minimal. The data obtained show that there is a correlation between the IMW content and the viability rate of yeast cells in lyophilized preparations.

[Fluorescent analysis of cryopreserved totipotent cells of amphibian embryos]. / Fluorestsentnyi analiz kriokonservirovannykh totipotentnykh kletok zarodyshei amfibii.

The cryotolerance of totipotent cells from dissociated embryos of amphibian (grass frog Rana temporaria and grey toad Bufo bufo) was studied. Cell integrity and preservation of the cell barrier function were evaluated by fluorescent analysis. It was shown that the best cryopreservation of the cells was achieved by using the cryoprotective agent 10% dimethyl sulfoxide and 10% saccharose. These cells were successfully used for the homotransplantation of nuclei into enucleated eggs. The development of reconstructed eggs to the blastula stage was noted.

[The reaction of endoplasmic reticulum of Mauthner cells of Xenopus laevis tadpoles in response to the partial denervation]. / Reaktsiia éndoplasmaticheskogo retikuluma na chastichnuiu denervatsiiuMautnerovskikh neironov golovastikov shportsevoi liagushki.

The structure of the Mauthner cells in Xenopus laevis tadpole was investigated by light- and electron microscopy in norm and after early unilateral enucleation. It was found that enucleation at early stages caused a delay in morphological development of the contralateral neurons during embryogenesis. We observed a decrease in size of the soma and nucleus and in the number of dendrites, a marked structural underdevelopment of the majority of cell organoids, as well as proliferation and hypertrophy of transversal cisternae in the contralateral Mauthner cells. The ipsilateral neurostructure remained normal in embryogenesis. The data obtained suggest the availability of some unknown powerful afferent contralateral input to Mauthner cells from the optical analyzer.

Receptor-mediated transport of foreign DNA into preimplantation mammalian embryos.

Mouse and rabbit preimplantation embryos with intact zona pellucida were incubated for 3 hr with DNA-carrying constructs containing insulin as an internalizable ligand: (insulin-polylysine)-DNA and (insulin-polylysine)-DNA-(streptavidin-polylysine)-(biotinylated adenovirus). Video-intensified microscopy demonstrated that the constructs penetrated the zona pellucida and accumulated in the blastomere perinuclear space. The percentage of blastocysts formed was about 70% after incubation of zygotes and two-cell embryos with the constructs. Foreign DNA was detected after 51 hr in 80% of rabbit embryos and after 96 hr in 73% of mouse embryos. Inclusion of various adenoviruses into the construct improved foreign DNA preservation in early embryos. Blot hybridization revealed genome-integrated foreign DNA in 12- and 15-day mouse embryos and in a newborn. Thus, the ligand-mediated mechanism can be employed for introducing foreign genetic material into early mammalian embryos; insulin provides for delivery inside the cell and to the nucleus, while adenoviruses ensure release from endosomes.

[Morphogenesis of Mauthner neurons of Xenopus laevis tadpoles following early unilateral eye enucleation]. / Morfogenez Mauthner neironov golovastikov shportsevoi liagushki v usloviiakh rannei odnostoronnei énukleatsii glaza.

The connection between Mauthner neuron (CMN) and visual analyzer in Xenopus laevis tadpoles was studied using these neurons capacity to change their morphogenesis during the development in case of interruption of their connections with afferent fibres. In phase 31 of development tadpoles were subjected to unilateral enucleation of eye. Morphogenesis of contralateral (experimental) and ipsilateral (control) MN with respect to the operated eye was followed up. Comparison with intact Mauthner neuron of the tadpole of the same age who were not exposed to operation was conducted. Contralateral MN were found to be delayed in development phases 47 and 50. This was associated with 1.5-3.3 fold difference in neuron body central section area and 2-3 fold differences in nuclear section area. Three-dimensional reconstruction revealed lower number of cell processes in contralateral MN as well as decline in their length and diameter. Control and intact MN showed no difference similar to experimental and control ones in phase 41. The data obtained indicate that MN of Xenopus laevis tadpole possess powerful ipsilateral connections originating from the side of visual analyzer and contralateral ones from the side of the eye. This affects MN development and in contrast to octavolateral system (including vestibular apparatus and organs of lateral line) controls the contralateral neuron.

Receptor-mediated transfection of murine and ovine mammary glands in vivo.

Transfection of HC-11 murine epithelial mammary cells as well as murine and sheep mammary glands were carried out using insulin-containing constructs that deliver DNA by receptor-mediated endocytosis to receptor-expressing cells. In vivo transfection of mammary gland tissue with the luciferase gene was carried out by introducing the DNA constructs into the mammary ducts of both mice and sheep. The successful transfection of ewe mammary glands was demonstrated by the detection of luciferase activity in mammary gland biopsy material up to a month after a single administration of the construct. To test whether products of expression of transfected genes could be secreted into the milk in this system, the N-terminal secretory signal sequences of bovine beta-lactoglobulin or the entire coding sequence of human alpha-lactalbumin were fused to the N terminus of the luciferase gene. After transfection with the modified luciferases, both murine and sheep milk could be shown to contain luciferase activity, whereas mice, which had been transfected with the nonmodified luciferase gene, did not secrete any activity in the milk. This approach demonstrates for the first time the possibility of gene transfer in vivo into mammary gland epithelial cells using constructs delivering DNA via receptor-mediated endocytosis.

[Therapeutic endoscopy in combined therapy of gastroduodenal ulcers]. / Lechebnaia éndoskopiia v kompleksnom terapii gastroduodenal'nykh iazv.

The method of therapeutic endoscopy is a very important component in the complex treatment of chronic gastroduodenal ulcers. In 115 of 116 patients with gastric or duodenal ulcers who were treated by the authors the ulcer epithelialized and a cicatrix formed. When therapeutic endoscopy with the application of hippophae oil was added to the basic therapy, the period of the inpatient treatment reduced 1.5-2 times. The late results of the management of chronic gastroduodenal ulcers by this method were studied and confirmed its sufficiently high therapeutic efficacy (the results were positive in 93.7% of cases).

[Effect of the oxidation-reduction potential of the medium on the production of gentamycin]. / Vliianie okislitel'no-vosstanovitel'nogo potentsiala sredy na obrazovanie gentamitsina.

The medium redox potential had an effect on gentamicin production by Micromonospora purpurea v. violacea, strain VNIIA 7R. The Eh influence was shown to be statistically reliable when the results were expressed in relative units against the control. In the laboratory experiments with low volumes of the medium the Eh increase by more than 170 per cent induced inhibition of gentamicin biosynthesis while the Eh increase by 108 to 168 per cent induced stimulation of the activity.

[A method for determining nonspecific body resistance]. / Sposob opredeleniia nespetsificheskoi rezistentnosti organizma.

The general resistance of the organism was studied. This included humoral factors (lysozyme activity, titers of the normal agglutini) and cellular link: total leukocytes and neutrophyls count, the indices of neutrophylic phagocyte response. The index of general resistance of an organism of 110 to 140 indicated its normal level and those below 110 were considered as reduced resistance scores. The technique implies an improved evaluation of an organism's own defenses. This allowed to diagnose the severity or oral mucosa inflammation and so provided grounds for indicating the unspecific stimulating therapy.

[Electrostimulated cell fusion in cell engineering]. / Elektrostimuliruemoe sliianie kletok v kletochnoi inzhenerii.

A survey of studies on reconstructions of animal and plant cells which apply a new physical method--electrostimulated fusion, is presented. Effects of different factors of the medium on the efficiency of electrofusion is discussed. A detailed account is given of the authors' studies on zygotes reconstruction by combined methods of microsurgery and electrostimulated cell fusion. Advantages of the latter as compared to the widely distributed methods of fusion by polyethylenglycol and Sendai virus are considered. This physical method can play an important role in the progress of cellular engineering.