Dideoxynucleotide DNA sequencing is one of the principal procedures in molecular biology. Loss of an initial part of nucleotides behind the 3' end of the sequencing primer limits the readability of sequenced amplicons. We present a method which extends the readability by using sequencing primers modified by polyadenylated tails attached to their 5' ends. Performing a polymerase chain reaction, we amplified eight amplicons of six human genes (AMELX, APOE, HFE, MBL2, SERPINA1 and TGFB1) ranging from 106 bp to 680 bp. Polyadenylation of the sequencing primers minimized the loss of bases in all amplicons. Complete sequences of shorter products (AMELX 106 bp, SERPINA1 121 bp, HFE 208 bp, APOE 244 bp, MBL2 317 bp) were obtained. In addition, in the case of TGFB1 products (366 bp, 432 bp, and 680 bp, respectively), the lengths of sequencing readings were significantly longer if adenylated primers were used. Thus, single strand dideoxynucleotide sequencing with adenylated primers enables complete or near complete readability of short PCR amplicons. and M. Beránek, M. Drastíková, J. Petera
Emerging evidence indicates that polychlorinated biphenyls (PCBs) are involved in the development of diabetes mellitus in the obese. The purpose of this study was to determine mechanisms by which PCB 153 (2,2′,4,4′,5,5′-hexachlorobiphenyl) could influence diet-induced obesity and insulin resistance during adipogenesis. Lineage of h-ADMSCs was differentiated either as control (differentiation medium only), or with lipid vehicle modeling high fat nutrition (NuTRIflex) or lipid free vehicle (dimethylsulfoxide) for 28 days with or without PCB 153 daily co-exposure (in three concentrations 0.1, 1, and 10 μM). Gene expression analyses were performed using RT-qPCR at days 4, 10, 21, 24, 28; p rotein l evels A kt a nd phosphorylated Akt (Phospho-Akt) by Western blot at days 4, and 21. PCB 153 treatment of h-ADMSCs only in lipid vehicle was associated with down regulation of key master genes of adipogenesis: PPARγ, SREBP-1, PPARGC1B, and PLIN2 during the whole process of differentiation; and with increased Akt and decreased Phospho-Akt protein level at day 21. We have shown that PCB 153, in concentration 0.1 μM, has a potential in lipid rich environment to modulate differentiation of adipocytes. Because European and U.S. adults have been exposed to PCB 153, this particular nutrient-toxicant interaction potentially impacts human obesity and insulin sensitivity., D. Mullerova, M. Pesta, J. Dvorakova, M. Cedikova, V. Kulda, P. Dvorak, V. Bouchalová, M. Kralickova, V. Babuska, J. Kuncova, J. Langmajerova, L. Muller., and Obsahuje bibliografii
Power generation from biomass is a substantial source of ashes, the extent of which depends on the type of biomass fuel and technology of combustion being used. The current study focuses on comparison of ashes with a special emphasis on hazardous organic compounds, particularly the polycyclic aromatic hydrocarbons (PAHs) fraction, present in ash. The reason is that ashes from combustion of wood or straw are considered as fertilizers in agricultural soils. Ash samples were therefore collected from power plants in the Czech Republic coming from combustion of wood chips, sawdust, bark and straw. The organic fraction was separated by extraction, the final determination of PAHs was performed by GC-UV. The total concentration of PAHs was found to be in the range 15-733 μ g/kg. These compounds are formed during the pyrolysis stage of the combustion process by the secondary ar omatization reactions in char at temperatures above 400 °C. The created PAHs are bound in the porous unburned carbon. For a more detailed qualitative analysis of other organic compounds the GC-MS was used. Higher and branched aliphatic hydrocarbons, ketones, amides and phthalates in the sample with the highest unburned carbon content were found., Pavel Straka and Martina Havelcová., and Obsahuje bibliografické odkazy
In this paper, we study the structure of polycyclic groups admitting an automorphism of order four on the basis of Neumann’s result, and prove that if α is an automorphism of order four of a polycyclic group G and the map φ: G → G defined by gφ = [g,α] is surjective, then G contains a characteristic subgroup H of finite index such that the second derived subgroup H″ is included in the centre of H and CH(α2) is abelian, both CG(α2) and G/[G, α2] are abelian-by-finite. These results extend recent and classical results in the literature., Tao Xu, Fang Zhou, Heguo Liu., and Obsahuje seznam literatury