Platelet rich plasma and growth factors cocktails for diabetic foot ulcers treatment: State of art developments and future prospects.

Current advances in diabetic foot ulcers (DFU) treatment are discussed. Normal and pathological wound healing process are observed and the role of growth factors (GFs) is elucidated. Current techniques involving GFs and platelet rich plasma (PRP) are compared. Up-to-date research suggests that treatment with single growth factor (GF) could be insufficient and not encompassing all pathological changes in DFU bed. Efficiency of PRP is rather controversial and lacks evidence. Thus the use of cocktail of particular GFs is suggested. Pro et contra of each approach are discussed.

[Orthotopic bone implants for bone regeneration]. / Organotipichnye kostnye implantaty - perspektiva razvitiia sovremennykh osteoplasticheskikh materialov.

Biotechnology industry is rapidly developing. The elaboration of new biomaterials for bone reconstruction is one of the most perspective directions in tissue engineering. There are millions of surgical operations associated with use of bone graft materials every year. In this article we tried to analyze and systematize data about advanced technologies and modern trends in the preparation of bio-composite bone graft materials. Special attention is given to 3D-prototyping that allows making bone implants with individual form. Introduction of molecular biology technologies such as activating specific cytokines and growth factors at the right time makes it possible to optimize bone regeneration process. The article has also some suggestions on further improvement of the bone engineering technology.

Nanoantibodies for detection and blocking of bioactivity of human vascular endothelial growth factor A(165).

Nanoantibodies (single-domain antibodies, nanobodies) derived from noncanonical single-chain immunoglobulins provide an attractive tool for in vitro and in vivo diagnostics as well as for development of targeted drugs for clinical use. Nanoantibodies against several clinically important targets have been developed and are actively investigated. However, no development of nanoantibodies against vascular endothelial growth factor VEGF-A(165) has been reported. We describe here the generation of nanoantibodies derived from single-chain Bactrian camel immunoglobulins directed against VEGF-A(165). We demonstrate that these nanoantibodies are suitable for enzyme-linked immunoassay to quantify human VEGF-A(165) as well as for blocking its activity. Our results provide a basis for diagnostic kit development for quantification of VEGF-A(165), which emerges as a biomarker useful in various pathological conditions. In addition, the nanoantibodies might be used for development of therapeutic molecules targeting VEGF-A(165)-dependent pathological neoangiogenesis.

[Functional properties of the WNT11 new isoform, expressed in colon carcinoma cell line HT29].

Colon carcinoma is a common type of neoplastic transformation. Mechanisms of its establishment and progression have been studying for several decades. Aberrant activation of the canonical Wnt signaling is frequently observed in colon carcinoma cells. Moreover, expression of the "noncanonical" Wnt ligands is also detected in this type of cancer. However, the implication of the noncanonical Wnt signaling in carcinogenesis and colorectal cancer (CRC) progression is still unclear. Here, to elucidate the characteristic features of the noncanonical Wnt signaling activation in CRC the expression of the "noncanonical" ligand hWnt11 has been studied. It was shown for the first time that expression of the hWnt11 in CRC is accompanied by the alternative splicing. The new hWnt11 isoform (hWnt11sp3) has been identified. Unlike to hWnt11, this isoform is not secreted and lacks the ability to inhibit the canonical Wnt signaling. Considering the canonical Wnt signaling inhibiting activity of hWnt11, different functional properties of the ligand and its isoform may reflect a special role of the alternative splicing in carcinogenesis and tumor progression. Thus, due to the difference in their functional properties an existence of several Wnt isoforms should be taken into account for the investigation of the role of Wnt ligands.

[Nuclear beta-catenin localization is not sufficient for canonical Wnt signaling activation in human meianoma cell lines].

In most cases, advanced stages of melanoma are practically incurable due to high metastatic potential of tumor cells. Multiple observations support the idea that aberrations in Wnt signaling pathway play a significant role in melanoma development and progression. Canonical Wnt signaling activation results in stabilization and accumulation of the major effector molecule called beta-catenin. Mutations promoting beta-catenin stabilization and, thereby, activation of canonical Wnt signaling pathway are frequently found in different cancers, but rarely observed in melanomas. Nevertheless, beta-catenin nuclear and cytoplasmic accumulation is the feature of many human melanoma cell lines and original tumors. That is why, the aim of the investigation was to elucidate the relation between beta-catenin intracellular localization and activity status of Wnt signaling pathway in human melanoma cell lines. Ten human melanoma cell lines were characterized on the basis of the following parameters: canonical Wnt ligand expression, intracellular beta-catenin localization, and activity status of canonical Wnt signaling pathway. Here, it has been demonstrated that nuclear localization of beta-catenin does not always correspond to active status canonical Wnt signaling pathway. Moreover, in the majority of cell lines with nuclear beta-catenin canonical Wnt signaling can't be activated by exogenous expression of an appropriate ligand. Human melanoma cell lines differ in activity of canonical Wnt signaling pathway as well as in mechanisms of its regulation. Therefore, the pathway-targeted potential antineoplastic therapy requires the formation of a "molecular pattern of cancer" for localization of the defect in Wnt signaling cascade in the each case.

[HLA-E molecule induction on the surface of tumor cells protects them from cytotoxic lymphocytes].

Modern immunotherapy has developed powerful tools for mounting antitumor response which nevertheless have had only limited success in clinic. Tumor cells use different mechanisms to escape from immune system. Thus, one of the reasons of unsuccessful immunotherapy might be induction of tolerance of tumor-specific cytotoxic lymphocytes by tumor cells. Previously we have demonstrated expression of HLA-E molecule by the cells of melanoma cell lines. In this paper we have studied HLA-E-dependent mechanism of melanoma cell escape from immune response.

[Functions of protein MTS1 (S100A4) in normal and tumor cells].

To date vast evidence has been accumulated showing the role of protein MTS1 in the metastasis development and cell motility regulation, both in norm and upon pathological change of various tissues. The structure of the protein and its gene, as well as the regulation of the gene expression, are studied in detail. Significant advances have been achieved in understanding molecular mechanisms involving MTS1. This paper reviews the current knowledge of the issue.

[Transcription and mRNA splicing of the human lactoferrin gene controlled by the regulatory region of the bovine alphaS1 casein gene in the mammary gland of transgenic mice and in mouse embryonic stem cells].

The regulatory region of the bovine alphaS1 casein gene was used to obtain two genetic constructs for expression of human lactoferrin in the mammary gland of transgenic animals. Several transfected mouse embryonic stem cell (ESC) lines and primary transgenic mice were generated with these constructs. Recombinant lactoferrin was not detected in milk of transgenic mice by Western blotting. However, a recombinant transcript was found in RNAs isolated from mammary glands of transgenic females during lactation and from transfected ESC lines.

[Use of genetically modified embryonic stem cells for creating transgenic animals]. / Ispol'zovanie genticheski modifitsirovannykh émbrional'nykh stvolovykh kletok dlia polucheniia transgennykh zhivotnykh.

Targeted genetic modification of embryonic stem cells (ESC) was used to obtain nondifferentiated cell clones containing the foreign genetic material in the genome. It was demonstrated that transgenic animals may be obtained by ESC injection in preimplantation embryos and subsequent transplantation of the embryos into a recipient female. Using this method, we constructed chimeric animals with a modified genome.

[Expression analysis of proteins encoded by genes of the tag7/tagL (PGRP-S,L) family in human peripheral blood cells]. / Analiz ékspressii belkov, kodiruemykh semeistvom genov tag7/tagL(PGRP-S,L), v kletkakh perifericheskoi krovi cheloveka.

Various types of human blood cells were tested for expression of the Tag7/PGRP-SA and TagL/PGRP-L proteins, which belong to the family of proteins possessing the lysozyme-like peptidoglycan recognition protein (PGRP) domain. Expression regulation by several factors was demonstrated.

The differentially spliced mouse tagL gene, homolog of tag7/PGRP gene family in mammals and Drosophila, can recognize Gram-positive and Gram-negative bacterial cell wall independently of T phage lysozyme homology domain.

Tag7/PGRP, a recently characterized antimicrobial protein, is conserved from insects to mammals. Recently its involvement in Toll signalling in Drosophila was demonstrated. A number of genes representing a new family homologous to PGRP were identified in Drosophila and human. Here we describe a splicing pattern of the tagL gene, mouse member of tag7/PGRP family. Some of the identified splice variants lacked characteristics for the family T phage lysozyme homology domain (also known as PGRP domain). Accordingly to the predicted transmembrane domains, mouse TagL may be secreted as inducible proteins or retained on intracellular membranes. All detected splice variant isoforms of TagL bound Gram-positive, Gram-negative bacteria and peptidoglycan. This binding did not depend on the presence of T phage lysozyme homology domain but was associated with the C-terminal portion of the polypeptides. Thus, this variety of isoforms of a single gene may play a role in circulating bacteria recognition in mammals.

[Production of recombinant endostatin in milk from transgenic mice]. / Poluchenie rekombinantnogo éndostatina v moloke transgennykh myshei.

Using the bovine alpha S1-casein gene, a genetic construct with an endostatin-coding fragment of the mouse collagen XVIII cDNA was designed to express endostatin in milk of transgenic animals. Several transgenic mice were obtained. The mice secreted endostatin in milk at 70-300 ng/microliter and transmitted this character to their progeny.

[Cloning and study of novel mammalian genes with structural homology with phage lysozyme]. / Klonirovanie i izuchenie novykh genov mlekopitaiushchikh, imeiushchikh oblast' strukturnoi gomologii s fagovym lizotsimom.

The mouse tag7 gene was cloned and characterized in our previous works. This work is devoted to identifying and cloning its homologs. The human homolog of the tag7 gene was cloned. Data of analysis of the primary structure of the human tag7 and results from the study of the production of the corresponding protein in human organs and tissues indicate that this gene plays a major role in the mammalian immune system. The mouse and human tagL gene carrying an extended region of structural homology with the tag7 gene was detected by computer analysis and was subsequently cloned. Sequence analysis suggested the possible membrane localization of the gene, whereas the specific expression pointed to the role of this gene in the immune system. Both genes, tag7 and tagL, were localized in the human chromosome 19.