Vol 18, No 3 (2018)

In our review, we discuss the inflammatory changes in the pathophysiology of Charcot-Marie-Tooth hereditary polyneuropathy, and this matter is still under further evaluation. Lipid oxidation, inflammatory changes, and blood inflammation agent (e.g., cytokines) concentration changes are important for the pathogenesis of this disorder. Several clinical cases of acute or chronic combination of hereditary and inflammatory polyneuropathies are identified. These clinical cases may be suspected when the typical course of Charcot-Marie-Tooth disease changed to rapid worsening of motor symptoms, inflammatory changes, and pain syndrome. Thus, anti-inflammatory treatment should be considered in these cases.

This study was carried out to explore the methodology of intraportal infusion of autologous bone marrow mononuclear cells in patients in the waiting list for liver transplantation. The study included eight patients. The Model for End-Stage Liver Disease (MELD) score as a predictor of hepatic failure exceeds 10 points, i.e., 12-22 points. According to Child-Turcotte-Pugh (CTP) classification, the severity of cirrhosis ranged from class B to C. The bone marrow volume of 224-320 ml was harvested by bilateral aspiration from the posterior iliac crest. The total volume of the mononuclear cells (MNC) was 47.1-58.3 ml (median, 47 ml). The absolute viable count of isolated MNC was 1.24 · 109-5.84 · 109 and CD34+ cells 2.54 · 106-48.20 · 106. After portal vein catheterization, direct transhepatic portography was performed, and portosystemic collaterals were found in all cases. Portal flowmetry was performed to determine the optimal selective perfusion rate of the segmental branches of the portal vein. Portal vein infusion was performed at 0.3-1.5 ml/sec. Serious side effects were not detected during bone marrow harvesting and MNC infusion. All patients had stable condition (no deterioration was initially noticed) after MNC infusion according to the MELD score and CTP criteria during a 3-month observation. Moreover, improvement in physical health status was observed and obtained using the SF-36 questionnaire. This study confirmed the safety of using intraportal therapy of autologous bone marrow MNC in patients on the waiting list for liver transplantation. Herein, we presented the technique of target delivery of autologous bone marrow MNC to the liver to minimize their loss during intraportal infusion. Prolonged follow-up time and improvement of the present technique are needed for further evaluation of the efficiency of autologous bone marrow MNC.

This study aimed to develop a method of simultaneous immunohistochemical detection of neurons and astrocytes using peroxidase labeling with subsequent analysis by light microscopy. A pair of antigens, each of which specifically identifies either neurons or astrocytes, was chosen (as markers neuronal nuclear antigen and glial fibrillary acidic protein were used, respectively). This method not only identifies the studied cells, but it also assesses their functional state under normal and pathological conditions. The proposed staining protocol allows significant simplification and optimization to simultaneously detect two antigens and to improve the quality of the slices obtained.

The purpose of research is study the features of the flow of reparative regeneration of the cornea in an experiment in vivo with induced bullous keratopathy after the introduction of a suspension of autologous mononuclear leukocytes in the posterior third of the corneal stroma. Materials and methods. The experiment was performed on 27 chinchilla rabbits (27 eyes). At the first stage of the experiment all animals were reproduced the model of corneal bullous keratopathy by mechanical damage and removal of its endothelial layer. At the second stage, 2 weeks after the development of the disease, the animals were divided, depending on the planned treatment, into 2 groups: the main, 12 animals once in the back third of the corneal stroma were administered a suspension of autologous mononuclear leukocytes; comparisons, 12 animals were treated by conservative therapy. Results. Reproduction of in vivo bullous keratopathy of the cornea by mechanical damage and removal of its endothelial layer is accompanied by rapid (within 2 weeks) the development of the disease with diffuse hydropic degeneration of the anterior epithelium, marked edema of the stroma with the bundle of collagen fibers. On the experimental model of the bullous keratopathy of cornea it was found that the introduction of a suspension of autologous mononuclear leukocytes in the posterior third of the corneal stroma in the treatment of the disease provides rapid (1.5 times) dehydration of the stroma, reduces the number of dystrophic cells in the anterior epithelium by 1.2 times compared with conservative treatment. The period of restoration of the normal thickness of the corneal slice in animals of the main group is reduced by 1.4 times. The conclusion. The introduction of autologous mononuclear leukocytes suspension into the posterior third of the corneal stroma in the treatment of experimental bullous keratopathy of cornea induces reparative regeneration, accelerating the restoration of damaged corneal structures and preventing its neovascularization.

The authors provide a genetic construct for obtaining recombinant biologically active wild type streptavidin with high yield. Addition of leader peptide and oligohistidine sequence to the streptavidin sequence makes it possible to isolate soluble streptavidin from the culture medium and from the soluble fraction. Temperature conditions for inducing of protein synthesis were found to have a significant effect on the distribution profile of streptavidin between fractions. The obtained protein product is not contaminated with endogenous biotin. The provided method excludes denaturation and renaturation steps which are necessary for isolation of the desired product from inclusion bodies but substantially reduce the yield. Thus, the provided approach allows to increase the yield of biologically highly active strepatividin which is capable of binding biotin and biotin-containing compounds and can be used for various practical purposes.

Aim. To determine the serotype and virulence factors of the E. coli serogroup O26 isolated from children with diarrheal syndrome. Materials and methods. Fifty-three strains of E. coli O26 isolated in 2014-2016 from the stool of children with clinical manifestations of acute intestinal infection in Saint Petersburg were studied. Phenotypic (enzymatic and antigenic properties), molecular genetic [detection of virulence genes of enteropathogenic (EPEC - eae, bfp, hlyA), and enterohemorrhagic (EHEC - eae, stx1, stx2, ehxA), genes encoding O- and H-antigens (rfb and fliC), genes, defining phylogenetic group (chuA, yjaA и TspE)] methods were used. The phylogenetic group and the production Shiga toxins were determined. Results. All strains were identical to the antigen characteristics of serotype O26:H11 and phylogenetic group B1. Two pathogroups were created based on the set of virulence genes: a-EPEC (64.2%) and EHEC (35.8%). Strains EHEC produced Shiga-toxin 1, encoding gen stx1. No differences in enzymatic activities were found between the strains of E. coli О26:H11 for EPEC and EHEC strains. Conclusion. In the population of E. coli O26:H11, which caused acute intestinal infection in children in Saint Petersburg, more than 30% of the strains belonged to the highly virulent group EHEC. Molecular-genetic methods should be used for reliable detection of pathogens.