In vivo assembly of genetic constructs in filamentous fungus Talaromyces cellulolyticus.

In the filamentous fungus Talaromyces cellulolyticus, similar to other filamentous fungi, non-homologous recombination predominates over homologous recombination. For instance, to achieve an acceptable integration frequency of a genetic construct into a target site on the intact chromosome, the flanking sequences directing this integration should be approximately 2.5 kb in length.

Acceleration of CRISPR/Cas9-Mediated Editing at Multiple Sites in the Genome.

The application of the CRISPR/Cas9-based genome editing technique to the yeast has made it possible to simultaneously modify several sites, particularly to integrate several expression cassettes. The existing methods provide high efficiency for such modifications; however, common protocols include several preparatory steps, namely, the construction of an intermediate Cas9-expressing strain, the assembly of a plasmid bearing several single guide RNA (sgRNA) expression cassettes, and the surrounding integrated DNA fragments with long flanks for recombination with target loci. Since these preparatory steps are time consuming and may not be desirable in some types of experiments, we explored the possibility of multiple integration without these steps.

Identification of the Talaromyces cellulolyticus Gene Encoding an Extracellular Enzyme with β-galactosidase Activity and Testing it as a Reporter for Gene Expression Assays.

The filamentous fungus Talaromyces cellulolyticus (formerly Acremonium cellulolyticus) is currently being intensively studied as a promising industrial producer of a number of secreted cellulolytic enzymes. In this study, the T. cellulolyticus gene lacA, which encodes a protein orthologous to the fungal extracellular β-galactosidases of family 35, was identified.

Multiple pathways for the formation of the γ-glutamyl peptides γ-glutamyl-valine and γ- glutamyl-valyl-glycine in Saccharomyces cerevisiae.

The role of glutathione (GSH) in eukaryotic cells is well known. The biosynthesis of this γ-glutamine tripeptide is well studied. However, other γ-glutamyl peptides were found in various sources, and the pathways of their formation were not always clear.

Engineering Escherichia coli for autoinducible production of L-valine: An example of an artificial positive feedback loop in amino acid biosynthesis.

Artificial metabolically regulated inducible expression systems are often used for the production of essential compounds. In most cases, the application of such systems enables regulating the expression of an entire group of genes in response to any internal signal such as an aerobic/anaerobic switch, a transition to stationary phase, or the exhausting of essential compounds. In this work, we demonstrate an example of another type of artificial autoinducible module, denoted a positive feedback module.

[Clopidogrel resistance: myth or reality].

The efficacy of clopidogrel, as an established anti-platelet agent for the acute coronary syndrome treatment and for the thrombotic complications prevention after percutaneous coronary angioplasty and coronary artery stenting has been supported by the evidence of several major randomized clinical trials. However, some patients treated with clopidogrel have a minor, but still a risk of coronary artery thrombosis and sudden death. Moreover, clopidogrel was not effective in patients after ischemic stroke, as well as it was not effective for the primary prevention of vascular events.

[Synthesis of xylanolytic enzymes and other carbohydrases by the fungus Penicillium canescens: transformants with an altered copy number of the gene xlnR and an encoding transcriptional activator].

A fragment of Penicillium canescens genomic DNA carrying the xlnR gene coding for a translational activator of xylanolytic genes was isolated. It was demonstrated that a loss of this function in genetically modified transformants resulted in a drastic decrease in the production of P. canescens major xylanases and had a negative effect on the syntheses of several other extracellular xylanases.

[Cloning of an endo-1,4-beta-xylanase gene from Penicillium canescens and construction of multicopy strains].

The complete gene xylA that encodes endo-1,4-beta-xylanase secreted by Penicillium canescens was cloned and sequenced. The coding region of the gene is separated by eight introns. The protein comprises 302 amino acids of the mature protein and 25 amino acids of the signal peptide.

[A morphologic analysis of changes in the lungs and kidneys caused by acetyl glyceryl ether phosphorylcholine and limited by verapamil].

Histological analysis of lung and kidney after multiple PAF i.v. injection into rabbits was performed.

[Features of thrombophilia in nephrotic syndrome in children].

A comprehensive study was made of the hemostatic system in 25 children with the nephrotic syndrome and combined forms of glomerulonephritis. Hypercoagulation and a number of factors preventing thrombosis formation were revealed, namely the presence of so-called thrombin-resistant blocked fibrinogen, a rise of the total antithrombin potential at the expense of proteins differing from antithrombin III, sufficient reserves of plasminogen, and a moderate increase of fibrin degradation products.

[Remote results of the surgical treatment of closed and open injuries of the liver].

Results of examination of 53 patients in remote periods after operation for closed and open traumas of the liver are presented. Two thirds of the patients were found to be practically healthy and one third had complaints of pains in the abdomen (duodenal ulcer was diagnosed in 1 patient, gastroduodenitis in 3 patients, adhesion disease of the abdominal cavity in 14 patients). Biochemical and radionuclide examinations have established disturbances of some liver functions in a number of patients.