hTERT-immortalized adipose-derived stem cell line ASC52Telo demonstrates limited potential for adipose biology research.

In the modern world obesity and insulin resistance contribute to a high impact on the structure of mortality. Basic research and pharmacological screenings for the search of new targets and insulin sensitizers require relevant cell models of adipocytes. Today the 3T3-L1 preadipocytes cell line is a widely used mouse-based model for investigation of adipocyte biology. Nonetheless, animal studies cannot be transferred directly in human research and nowadays the search for relevant and renewable cell models of human adipocyte is of undeniable importance. In the present study, we have compared pooled culture of human adipose-derived stem cells (ADSC) with immortalized ADSC cell line ASC52Telo. Both cell types had mesenchymal stem cell phenotype verified by flow cytometry. However, the efficacy of adipogenic differentiation, stimulation of FABP4 and PPARg protein expressions, and glucose uptake stimulation by insulin were reduced for ASC52Telo-derived adipocytes in comparison with ADSC-derived adipocytes. In addition, the analysis of insulin signaling has shown impaired phosphorylation of IRS1 and AS160 in ASC52Telo-derived cells. In summary, we have shown that immortalized cell line of human ADSC ASC52Telo have mesenchymal stem cell phenotype. Nevertheless, ASC52Telo-derived adipocytes demonstrate impaired adipogenesis and insulin sensitivity that are the main properties of healthy adipocytes.

Identification of novel inhibitors for the tyrosyl-DNA-phosphodiesterase 1 (Tdp1) mutant SCAN1 using virtual screening.

Spinocerebellar ataxia syndrome with axonal neuropathy (SCAN1) is a debilitating neurological disease that is caused by the mutation the Tyrosyl-DNA phosphodiesterase 1 (TDP1) DNA repair enzyme. The crucial His493 in TDP1's binding site is replaced with an arginine amino acid residue rendering the enzyme dysfunctional. A virtual screen was performed against the homology model of SCAN1 and seventeen compounds were identified and tested in a novel SCAN1 specific biochemical assay. Six compounds showed activity with IC50 values between 3.5 and 25.1 µM. The most active ligand 5 (3.5 µM) is a dicoumarin followed by a close structural analogue 6 at 6.0 µM. A less potent series of ß-carbolines (14 and 15) was found with potency in the mid-teens. According to molecular modelling an excellent fit for the active ligands into the binding pocket is predicted. To the best of our knowledge, data on inhibitors of the mutant form of TDP1 has not been reported previously. The virtual hits were also tested for wild type TDP1 activity and all six SCAN1 inhibitors are potent for the former, e.g., ligand 5 has a measured IC50 at 99 nM. In the last decade, TDP1 is considered as a promising target for adjuvant therapy against cancer in combination with Topoisomerase 1 poisons. The active ligands are mostly non-toxic to cancer cell lines A-549, T98G and MCF-7 as well as the immortalized WI-38 human fetal lung cells. Furthermore, ligands 5 and 7, show promising synergy in conjunction with topotecan, a clinically used topoisomerase 1 anticancer drug. The active ligands 5, 7, 14 and 15 have a good balance of the physicochemical properties required for oral bioavailability making the excellent candidates for further development.

[Urgent therapeutic penetrating autokeratoplasty for a purulent corneal ulcer in the only functional eye].

The article presents a case of severe purulent corneal ulcer in the only functional eye of a young patient with Frank-Kamenetsky glaucoma and high-degree myopia. Combination therapy was ineffective and the patient developed a descemetocele. Due to the absence of cadaver corneas and small size of the only available conserved Alloplant cornea, the contralateral cornea (of the blind eye, which was enucleated under general anesthesia) was used for urgent therapeutic penetrating autokeratoplasty. Postoperative period was complicated with ocular hypertension, which was surgically resolved. To overcome the critical situation around keratoplasty, with results from Russian regulation gaps, the authors suggest creating interregional eye banks in large cities that would provide sampling, conservation and storage control of donor material in accordance with the existing legislation.

[Agrobacterium-mediated in Planta transformation of maize germ cells].

Abstract-A transfer DNA (T-DNA) carrying the marker gene nptII was detected in the genomes of diploid and haploid maize plants obtained after the treatment of pistil filaments with a suspension of Agrobacterium during artificial pollination. PCR analysis of total DNA isolated from 155 canamycin-resistant diploid F1 seedlings revealed T-DNA insertions in the genomes of 111 plants (32.7% of the total number of analyzed seeds). The example of matroclinal haploids was used to demonstrate that T-DNA may be transported to the egg cell by the growing pollen tube (PT). Twelve out of 16 analyzed haploid plants contained the T-DNA insertion. The possible mechanism of the transfer of the Agrobacterium T-DNA to the maize genome during pollination is discussed.

[The levels of stable organic pollutants in the breast milk of women living in the Irkutsk region].

The concentrations of stable organic chlorine pollutants (SOCP) in the breast milk of women living in Irkutsk, Baikalsk, and the settlement of Kachug are lower than those in the increased SOCP-burdened areas of the Irkutsk Region (the town of Usolye-Sibirskoye, settlements on the shore of the Baikal Lake) and comparable with those in Russia and industrially developed countries of the world. The content of SOPC is much lower than those in the developing countries where this pesticide continues to be applied. The breast milk levels of OCP, TEQ (polychlorinated dibento-para-dioxines and polychlorinated dibenzofurans) in all towns and individual indicator polychlorinated biphenyls (PCB) in Irkutsk significantly decreased as compared with those in the 1980s. However, in Kachug and Baikalsk, the concentrations of PCB practically unchanged and the level of some congeners increased. The absence of changes in the content of PCB in Kachug and Baikalsk may be associated with no decrease in environmental pollution in the Irkutsk Region. Altered breast concentrations of SOCPs caused a reduction in their intake in babies.

The influence of soil contamination on the concentrations of PCBs in milk in Siberia.

Although atmospheric deposition is generally the dominant pathway of PCBs into agricultural food chains, soil ingestion by livestock can be important in some cases. The relationship between PCB levels in cow's milk and in pasture soil was studied in the Irkutsk region in Siberia where an historical atmospheric source(s) of PCBs has led to widespread contamination of soil. Milk samples were collected in spring and again in autumn from 18 different farms and analyzed for PCBs. Pasture soil samples were also collected and analyzed. The PCB concentrations in both milk and soil ranged over more than an order of magnitude between the farms. A good correlation was obtained between PCB levels in autumn milk and in soil. This together with a range of other evidence suggested that ingestion of pasture soil was the dominant source of the PCB contamination in the milk. The average soil ingestion rate was estimated to be 1700 g/d, which is at the upper end of values reported in the literature. This may be due to the arid summer climate or the animal husbandry practices in Siberia.

Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in Baikal seal.

Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) were determined in blubber tissue of one 3 month and two adult female seals from Lake Baikal. High PCDD and PCDF concentrations were found in the Baikal seal. The levels are comparable with those reported for ringed seals (phoca hispida) living in the Baltic sea, and Barrow Strait and Admiralty Inlet in the Canadian Arctic.

[Peculiarities of diagnosis and treatment for diabetic feet]. / Osobennosti diagnostiki i lecheniia diabeticheskikh porazhenii stop.

Multifactorial pathogenesis of the "diabetic foot" syndrome suggests advisability to single out some clinical pathogenetic forms, depending on basic causes of the lesions: due to neuropathy, osteoarthropathy, neuro-ischemic factors. The authors has developed the algorithm which enables to diagnose various forms of the syndrome and differentiate the treatment. The most important treatment modalities include unloading and podiatric measures as well as surgery, systemic antibacterial and glucolytic therapy. Conservative or surgical antiischemic measures are indicated only in demonstration of the degree of the ischemic damage in the lower extremities by Doppler ultrasonography and measurement of local oxygenation of the skin. The combined differentiated treatment in 142 patients with diabetic lesions of the feet has resulted 97.2% favourable outcomes in neuropathy and 86.5% successful outcomes in neuroischemic damages. Local oxygenation of dorsal skin of the foot (< 20 mm Hg) indicates that amputation above the ankle is highly probable.