[Influence of connective tissue differentiation on scar tissue formation in children]. / Vliyanie differentsiatsii soedinitel'noi tkani na formirovanie rubtsovoi tkani v detskom vozraste.

OBJECTIVE: To study the immunohistochemical features of various scar tissues in children without connective tissue pathology and with undifferentiated connective tissue dysplasia syndrome. MATERIAL AND METHODS: Tissue biopsy was performed in 217 children who underwent surgical treatment for various lesions, such as injuries, burns, as well as other procedures. There were 127 boys (58.5%) and 90 (41.5%) girls. The main group consisted of 98 (48.2%) children with scar tissue; group of UCTD syndrome - 65 (30.0%) children; control group - 43 (24.8%) patients without pathological scars. Histological examination of scar tissue and intact skin was carried out during primary or redo reconstructive surgery. Immunohistochemical study of antibodies against CD34, CD105, CD140b, PDGFs, COL types I, III and IV was performed. RESULTS: The study showed a quantitative characteristic of expression of COL type I in hypertrophic scar with predominance in the main group (77.5±5.4%; p<0.05), and decrease in COL type IV. Keloid form was associated with predominance of granulation tissue in all layers of dermis and high levels of all types of collagen. In the group of UCTD, COL type III prevailed in all pathological forms of the scar. We determined quantitative indicators of expression of vascularization factors (CD34; CD105) and fibroblastic activity (CD140b; PDGFs). CONCLUSION: Understanding the process of fibrinogenesis and analysis of stages of triggering mechanisms are essential for development of preventive algorithms. Individualized approach should be considered in the treatment. These studies are especially important in children with UCTD syndrome as high-risk group for pathological scarring. Thus, further research is required.

[Multicenter study of the effectiveness of antiscar therapy in patients at different age periods]. / Mul'titsentrovoe issledovanie effektivnosti protivorubtsovoi terapii u patsientov razlichnykh vozrastnykh grupp.

OBJECTIVE: Was to evaluate the effectiveness of anti-scar treatment with Contractubex gel in children and adults. MATERIAL AND METHODS: A group of researchers based on clinical hospitals and university medical clinics carry out the multicenter study to evaluate the effectiveness of anti-scar treatment with Contractubex gel containing cepalin, allantoin and heparin, with its early appointment in groups of children from 12 to 18 years old and adults from 21 to 35 years old. The study included data from 216 patients. Patients of both age groups were initially divided into two: the main and control ones with an equal distribution according to the type of surgical intervention (hernia repair and appendectomy), age, gender, and anamnestic data. The dynamic observation was carried out using two rating scales - filled out by a doctor (Vancouver scale) and a patient (author's rating scale in the Scar Diary mobile application). RESULTS: Based on the results of the analysis of the data obtained, a high efficiency of the use of Contractubex gel at the early stages of scar formation among patients of the main group in comparison with the control was revealed. By 90 days, the treatment result according to the Vancouver scale was 0,16±0,1 points in the main group and 0,39±0,2 points in the control group. At the same time, with a dynamic scale for assessing the cicatricial process, there was a significant (p<0.05) improvement in the main group (0,2±0,06 points) compared with the control group (0,6±0,17 points). In addition, was noticed the strong commitment to anti-scar treatment in pediatric patients. CONCLUSIONS: The work confirms the undoubted need for anti-scar treatment in the early stages of scar formation after surgical interventions, which accelerates the psychophysical rehabilitation of patients after surgery and improves the quality of life.

[Comparison of different methos of compression of scars with prolonged release plate]. / Sravnenie razlichnykh metodik kompressii rubtsov plastyrem prolongirovannogo deistviia.

The purpose of the study was to compare the effectiveness of various compression techniques with a prolonged-action patch on postoperative scars. MATERIAL AND METHODS: The study included 21 patients aged 3.6±1.7 years who had scars after surgery. The patients were divided into two groups: group No 1 (n=11), in which they applied longitudinal patch of prolonged-action Contractubex according to the standard technique; group No 2 (n=10) - transverse imposition of the patch on the postoperative scar (according to the author's method). RESULTS: In 20 (95.5%) patients, good compliance was noted. By the 90th day after surgery, the width of the scar in the group No. 1 was significantly larger than in the group No. 2, being 2.5±0.15 and 2.0±0.1 (p<0.05). CONCLUSION: Thus, the effectiveness of the proposed technique of transverse imposition of the patch Contractubex on the postoperative scar after surgical intervention was confirmed by a good cosmetic and functional result of scar formation.

Regulation of c-met expression by transcription repressor Daxx.

The protooncogene c-met encodes the tyrosine kinase receptor for the hepatocyte growth factor/scatter factor (HGF/SF). While overexpression of c-met is documented in many types of tumors, the mechanism of c-met regulation remains elusive. Here, we demonstrate Daxx as a repressor of c-met transcription. The expression of c-met is elevated in Daxx knockout mouse cells and is reversed by Daxx reconstitution. C-met promoter analysis of Daxx-/- cells reveled changes in chromatin acetylation, but not in DNA methylation. Daxx binds to the mouse c-met promoter and Daxx-binding region is sufficient for transcription repression, while HDAC2 is associated with c-met promoter mostly in Daxx+/+ cells, pointing to Daxx-dependent HDAC2 recruitment as a potential mechanism of c-met repression. HGF-induced cell mobility and invasion confirmed augmented activity of c-Met/HGF pathway in Daxx-/- cells. Finally, inverse correlation between Daxx and c-Met in cancer cell lines and in metastatic breast cancer specimens suggests potential function of Daxx as a c-met repressor during cancer progression.

PML is critical for ND10 formation and recruits the PML-interacting protein daxx to this nuclear structure when modified by SUMO-1.

Nuclear domain 10 (ND10), also referred to as nuclear bodies, are discrete interchromosomal accumulations of several proteins including promyelocytic leukemia protein (PML) and Sp100. In this study, we investigated the mechanism of ND10 assembly by identifying proteins that are essential for this process using cells lines that lack individual ND10-associated proteins. We identified the adapter protein Daxx and BML, the RecQ helicase missing in Bloom syndrome, as new ND10-associated proteins. PML, but not BLM or Sp100, was found to be responsible for the proper localization of all other ND10-associated proteins since they are dispersed in PML-/- cells. Introducing PML into this cell line by transient expression or fusion with PML-producing cells recruited ND10-associated proteins into de novo formed ND10 attesting to PMLs essential nature in ND10 formation. In the absence of PML, Daxx is highly enriched in condensed chromatin. Its recruitment to ND10 from condensed chromatin requires a small ubiquitin-related modifier (SUMO-1) modification of PML and reflects the interaction between the COOH-terminal domain of Daxx and PML. The segregation of Daxx from condensed chromatin in the absence of PML to ND10 by increased accumulation of SUMO-1-modified PML suggests the presence of a variable equilibrium between these two nuclear sites. Our findings identify the basic requirements for ND10 formation and suggest a dynamic mechanism for protein recruitment to these nuclear domains controlled by the SUMO-1 modification state of PML.

The activation of protein kinase C induces higher production of reactive oxygen species by mononuclear cells in patients with multiple sclerosis than in controls.

OBJECTIVE: Recent findings have increasingly shown the importance of reactive oxygen species (ROS) in causing oxidative damage to macromolecules and in contributing to tissue degeneration in target organs of autoimmune diseases. This study was aimed at comparing the base line and induced production of ROS by peripheral blood mononuclear cells (PB MNCs) of patients with multiple sclerosis (MS) in remission and relapse, of patients with other neurological diseases (OND) and of healthy controls. In addition, we analyzed the underlying mechanism of ROS production. METHODS: PB MNCs were separated from 28 MS patients in remission and 13 in relapse, and from 29 healthy controls and 10 OND. ROS was measured by spectrofluorometry. Expression of proinflammatory cytokines was assessed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Mitochondrial (mt) DNA haplotypes were determined by using restriction site polymorphism analysis. RESULTS: The base line and tumor necrosis factor (TNF)-alpha or interferon (IFN)-gamma induced ROS values were similar in the four groups, and the individual measures did not show a correlation with MS associated mtDNA haplotypes. Phorbol ester activation of protein kinase C (PKC) induced higher ROS production in all groups, however, with significantly greater values in the MS remission group. Calphostine C, a PKC inhibitor decreased or eliminated ROS production in a dose-dependent manner, suggesting further that it was predominantly or exclusively generated by PKC activated NADPH oxidase. A trend of increased TNF-alpha and IFN-gamma expression was noted in the MS relapse group, in contrast to the high ROS release in the MS remission group. CONCLUSION: The detected phase difference between the highest ROS production vs TNF-alpha expression is compatible with the hypothesis that different subpopulations of monocytes/macrophages are involved. We suggest that the ROS producing subpopulation preferentially migrates into the central nervous system (CNS) during a relapse. The present study together with our previous observation on oxidative damage to DNA in active plaques delineates a molecular pathway likely involved in the histologic evolution of inflammatory demyelination.

Human placenta DNA primase: purification of enzyme and analysis of RNA primer synthesis.

The immunoaffinity purification of human placenta DNA primase devoid of DNA polymerase alpha activity is described. Primase consists of 52 and 59 kDa polypeptides. They form a single protein of 330 kDa under native conditions. The polypeptide structure of primase is believed to be (52 + 59)3. Primase synthesizes the oligoribonucleotides 2-9 monomers long and multimeric oligoribonucleotides of a modal length of about 10 monomers. The following model of RNA primer synthesis is proposed: 1) primase, being in free state or in complex with Pol alpha, forms a protein trimer or another structure that includes several primases; 2) primase synthesizes de novo only the oligonucleotides 2-10 monomers in length; 3) the newly synthesized oligonucleotides dissociate in solution or translocate to either Pol alpha or a neighbouring primase unit to be further elongated with the next 7-10 mononucleotides.

Eukaryotic DNA primase. Abortive synthesis of oligoadenylates.

Calf thymus DNA polymerase alpha-primase, human placenta DNA polymerase alpha-primase and human placenta DNA primase synthesized oligoriboadenylates of a preferred length of 2-10 nucleotides and multimeric oligoribonucleotides of a modal length of about 10 monomers on a poly(dT) template. The dimer and trimer were the prevalent products of the polymerization reaction. However, only the oligonucleotides from heptamers to decamers were elongated efficiently by DNA polymerase alpha.