Thermostable Recombinant Polypeptides as the Source of L-Amino Acids for Culture Media.

Mutant homologues of small chemotactic and DNA-binding proteins from thermophilic bacteria Thermotoga petrophila RKU-1 and Thermotoga naphthophila were obtained. These proteins can be expressed in the recombinant form in E. coli cells.

[PEGylated recombinant L-asparaginase from Erwinia carotovora: Production, properties, and potential applications].

N-hydroxysuccinimide ester of monomethoxy polyethylene glycol hemisuccinate was synthesized. It acylated amino groups in a molecule of recombinant L-asparaginase from Erwinia carotovora. A method of L-asparaginase modification by the obtained activated polyethylene glycol derivative was developed.

Induction of Alternative Splicing and Inhibition of Activity of Telomerase Catalytic Subunit by Apoptotic Endonuclease EndoG in Human T, B, and NK Cells.

The effect of apoptotic endonuclease EndoG on alternative splicing of mRNA of human telomerase catalytic subunit hTERT (human telomerase reverse transcriptase) and telomerase activity in normal human lymphocytes were studied. Human CD4, CD8, B, and NK cells were transfected with a plasmid pEndoG-GFP containing EndoG gene or control plasmid pGFP. The levels of mRNA of EndoG or hTERT splicing variants were analyzed by real-time PCR.

[Construction and characterization of a recombinant mutant homolog of the CheW protein from Thermotoga petrophila RKU-1].

In the work a recombinant chemotaxis protein CheW from Thermotoga petrophila RKU-1 (TpeCheW) and its mutant homolog (TpeCheW-mut) were created. It was shown that, despite the low homology with CheW prototypes from intestinal bacteria, these proteins didn't cause metabolic overload and were well expressed by cells of E. coli laboratory strains.

Inhibition of telomerase activity and induction of apoptosis by Rhodospirillum rubrum L-asparaginase in cancer Jurkat cell line and normal human CD4+ T lymphocytes.

Rhodospirillum rubrum L-asparaginase mutant (RrA) down-regulates telomerase activity due to its ability to inhibit the expression of telomerase catalytic subunit hTERT. The aim of this study was to define the effect of short-term and long-term RrA exposure on proliferation of cancer Jurkat cell line and normal human CD4 T lymphocytes. RrA could inhibit telomerase activity in dose- and time-dependent manner in both Jurkat and normal CD4 T cells.

Rhodospirillum rubruml-asparaginase targets tumor growth by a dual mechanism involving telomerase inhibition.

Rhodospirillum rubruml-asparaginase mutant RrA E149R, V150P, F151T (RrA) was previously identified to down-regulate telomerase activity along with catalyzing the hydrolysis of l-asparagine. The aim of this study was to define the effect of prolonged RrA exposure on telomerase activity, maintenance of telomeres and proliferation of cancer cells in vitro and in vivo. RrA could inhibit telomerase activity in SCOV-3, SkBr-3 and A549 human cancer cell lines due to its ability to down-regulate the expression of telomerase catalytic subunit hTERT.

[Prediction of protein thermostability from their primary structure: the current state and development factors].

The construction of proteins and peptides with desired properties, including resistance to high temperatures, as well as optimization of their amino acid composition, is an important and complex task, which attracts much attention in various branches of the basic sciences, and also in biomedicine and biotechnology. This raises the question: what method is more relevant for the at the pilot stage of research in order to estimate the influence of the planned amino acid substitutions on the thermostability of the resultant protein construct? In this brief review we have classified existing basic practical and theoretical approaches used in studies and predicting the thermal stability of native and recombinant polypeptides. Particular attention has been paid to the predictive potential of statistical methods for studying the thermodynamic parameters of the primary protein structure and prospects of their use.

[Bioactive proteins and peptides: current state and new trends of practical application in the food industry and feed production].

Proteins and polypeptides play a key role in the life of a human body. Scientific and practical interest to the natural proteinaceous substances could be explained by the diversity of their functions in metabolic processes. Biologically active substances of the protein origin have a rich history of applications in different sectors of the economy.

[The development of nomenclature of complex medical services on specialty «pathologic anatomy»].

The article presents definitions of concepts and analysis of classification schemes of medical services of specialty «pathologic anatomy" for last decade and directions of their development. The listing of services of such kind is to include both intravital and postmortem pathomorphologic analyses and organizational legal activity related to control and expertise of quality of medical care on the basis of collegiate clinicalpathologicoanatomic comparisons. It is appropriate to distinguish among pathologicoanatomic services the primary and secondary analyses with their reflection in statistical reports of institutions/subdivisions ofpathologicoanatomic service.

[Bacterial recombinant L-asparaginases: properties, structure and anti-proliferative activity].

For more than 40 years L-asparaginases are used in combined therapy of acute lymphoblastic leukemia in children and the range of tumors sensitive to these enzymes constantly extends. This review summarizes results of studies aimed at creation of new systems for heterological expression of bacterial L-asparaginases as Erwinia carotovora (EwA), Helicobacter pylori (HpA), Yersinia pseudotuberculosis (YpA) and Rhodospirillum rubrum (RrA); special attention is paid to isolation of purified enzymes and their crystallization, modification by chitosan/polyethylene, physicochemical, kinetic and structural properties characterization, and the study of the cytotoxic or anti-proliferative activity of new recombinant L-asparaginases on cell cultures in vitro. The resultant recombinant L-asparaginases (EwA, YpA, HpA и RrA) exhibit reasonable cytotoxic action on the human leukemia cells comparable to the pharmacologically available L-asparaginase EcA and represent practical interest in respect to creation, on their basis, new effective antineoplastic remedies.

Identification of functional regions in the Rhodospirillum rubrum L-asparaginase by site-directed mutagenesis.

Site-directed mutagenesis of Rhodospirillum rubrum L-asparaginase (RrA) was performed in order to identify sites of the protein molecule important for its therapeutic and physico-chemical properties. Ten multipoint mutant genes were obtained, and five recombinant RrA variants were expressed in E. coli BL21(DE3) cells and isolated as functionally active highly purified proteins.

[Antitumor activity of L-asparaginase from Erwinia carotovora from against different leukemic and solid tumours cell lines].

We have studied dose- and time-dependent antitumor and cytotoxic effects of Erwinia carotovora L-asparaginase (ECAR LANS) and Escherichia coli L-asparaginase (MEDAC) on human leukemic cells and human and animal solid tumor cells. We determined the sensitivity of tumor cells to L-asparaginases, as well the effect L-asparaginases on cell growth rate, protein and DNA synthesis per se and with addition of different cytostatics. The data obtained demonstrated that ECAR LANS L-asparaginase suppressed growth of all tested solid tumor cells.

[Anticancer activity of oxovanadium compounds].

Cytotoxic and antitumor activity of the biligand vanadyl derivative of L-malic acid (bis(L-malato)oxovanadium(IV) (VO(mal)2) was investigated in comparison with inorganic vanadium(IV) compound--vanadyl sulfate (VOSO4) and also with oxovanadium monocomplex with L-malic acid (VO(mal)) and vanadyl biscomplex with acetylacetonate. In this purpose the effect of vanadyl compounds on growth of normal human skin fibroblasts and tumor cells of different lines: mouse fibrosarcoma (L929), rat pheochromocytome (PC12), human liver carcinoma (HepG2), virus transformated mouse fibroblast (NIN 3T3), virus transformated cells of human kidney (293) were investigated. The results showed that VO(mal)2 was not toxic for normal human skin fibroblasts but considerably inhibited growth of cancer cells in culture.

[Diagnosis and causes of death in diabetes mellitus].

In the pandemic of diabetes mellitus (DM), there is no unified approach to making the final clinical and postmortem diagnoses of deaths caused by coronary heart disease (CHD), cerebrovascular diseases (CVD), and some other vascular catastrophes. The acute and chronic forms of CHD and CVD and acute and chronic ischemic lesions in other vascular basins, which result in death in DM, should be regard as its complications. Moreover, DM is a peripheral vascular disease.

[Antitumor activity of L-asparaginase from Yersinia pseudotuberculosis].

The cytotoxic activity of L-asparaginases from Yersinia pseudotuberculosis and from Erwinia carotovora were investigated in vitro using several tumor cells lines: Jurkat and Molt-4 (human T-lymphoblastic leukemia), MCF-7 (human breast adenocarcinoma), LnCap (human prostate carcinoma), NGUK1 (rat Gasser node neurinoma). E. coli L-asparaginase produced by "Medak" (Germany) was used as a reference.

[Uterine arteriovenous anomaly in a young woman].

The paper presents a case of uterine arteriovenous anomaly as mixed hamartoma with the signs of racemose and intramuscular angiomas, which was a morphological finding in an 18-year-old woman when the uterus removed for misdiagnosed postpartum septic endometritis was examined.

Role of matrix metalloproteinases and their inhibitors in tumor invasion and metastasis.

The role of various matrix metalloproteinases (MMP)--such as gelatinases, stromelysins, matrilysin, collagenase-3, and membrane-bound MMP (MB-MMP)--in tumor invasion and metastasis is discussed. Data suggesting significance for malignant growth of the expression level of these enzymes and also of their activators and inhibitors are presented. It is concluded that at different stages of tumor progression the activity of different MMPs is displayed, which is regulated by various growth factors and oncogenes.

Cationic lipid-DNA complexes-lipoplexes-for gene transfer and therapy.

Cationic lipid-mediated gene transfer and delivery still attract great attention of many gene therapy laboratories. From the point of view of the most important characteristics of lipoplex particles, e.g.

Effects of isatin on atrial natriuretic peptide-mediated accumulation of cGMP and guanylyl cyclase activity of PC12 cells.

We have previously demonstrated that isatin (indole-2,3 dione), an endogenous compound widely distributed in mammalian tissues and body fluids, effectively inhibits atrial natriuretic peptide (ANP) receptor binding and ANP-stimulated guanylyl cyclase activity of rat membrane preparations. In the present study the effects of isatin on ANP-mediated accumulation of cGMP and guanylyl cyclase (GC) activity of PC12 cells were studied. Isatin (0.

The lacZ gene transfer into L929 cells and [14C]-DNA tissue distribution following intraperitoneal administration of new pH-sensitive lipoplexes in mice.

The efficacy of lacZ gene transfer into the L929 cell line and a local [l4C]-DNA delivery in male NMRI mice (10-12 weeks old), were studied using new pH-sensitive liposomes, containing phosphatidylcholine/glycyrrhizin (PC/GL) or alpha-tocopherol ester of succinic acid (PC/TSA). The reporter gene (pQE-LacZ plasmid) was transferred into L929 cells using corresponding lipoplexes, 0.5% of cells being transfected.

Matrix metalloproteinases of normal human tissues.

This review considers biochemical properties of the family of matrix metalloproteinases (MMPs) of normal human tissues and the involvement of these enzymes in morphogenesis. Four main MMP subfamilies are characterized, and a group of other MMPs is described. Data on mechanisms of activation and inhibition of MMPs in certain tissues during various physiological processes (embryogenesis, angiogenesis, tissue growth and involution) are considered.