Five Kunitz protease inhibitor group B genes were isolated from the genome of the diploid non-tuber-forming potato species Solanum palustre. Three of five new genes share 99% identity to the published KPI-B genes from various cultivated potato accessions, while others exhibit 96% identity. Spls-KPI-B2 and Spls-KPI-B4 proteins contain unique substitutions of the most conserved residues usually involved to trypsin and chymotrypsin-specific binding sites of Kunitz-type protease inhibitor (KPI)-B, respectively. To test the inhibition of trypsin and chymotrypsin by Spls-KPI proteins, five of them were produced in E. coli purified using a Ni-sepharose resin and ion-exchange chromatography. All recombinant Spls-KPI-B inhibited trypsin; K(i) values ranged from 84.8 (Spls-KPI-B4), 345.5 (Spls-KPI-B1), and 1310.6 nM (Spls-KPI-B2) to 3883.5 (Spls-KPI-B5) and 8370 nM (Spls-KPI-B3). In addition, Spls-KPI-B1 and Spls-KPI-B4 inhibited chymotrypsin. These data suggest that regardless of substitutions of key active-center residues both Spls-KPI-B4 and Spls-KPI-B1 are functional trypsin-chymotrypsin inhibitors.

No abstract

Background We suggest that differential gene expression in patients with AS includes the immune system, intracellular or extracellular signaling pathway, and bone matrix biosynthesis pathway, which might play important roles in hip joint ligament ossification. The development of inflammation in the AS pathogenesis (sacroileitis, spondylodiscitis, arthritides, etc.) is associated with an increase of C-reacting protein, CRP and BASDAI rates and ESR, often complexed with HLA-B27. Objectives We set a task to study the biomarkers indicative of the cell death processes and their possible patterns (apoptosis, necrosis), as well as methylation levels, through an investigation of an associations with inflammation intensity (ESR, CRP) biomarkers, exDNA level and the proteins complexed with exDNA. Methods Phenol-based technique was used to extract exDNA from blood serum of 37 patients with AS and 18 healthy donors. 85% patients were positive for HLA-B27, BASDAI 5.31±3.28. The exDNA concentration was determined using fluorescent technique with Hoechst 33258. The protein component bound to exDNA, was determined using fluorescence of the protein-dye complex (coumarin-benzotiametincyanine, 580 nm). The level of methylation by exDNA restriction with methyl-specific nucleases HpaII and MspI. The statistical processing - Statistica 8.0. Results 71% of AS patients showed an increase in exDNA concentration, and 50% demonstrated high methylation levels (1.56±0.34 a.u.), which correlated with the exDNA concentrations and increased leukocyte count (p=0.008, t=0.7467). Raised concentration of monomethine proteins, complexed with DNA, as compared with the normal, was found in 74% of AS patients. A direct correlation was detected between the levels of monomethine proteins and the level of DNA methylation (p=0.03, t=0.867). 84% of patients with CRP concentration >5 mg/l, demonstrated a rise in exDNA concentration (501.5±238.5 ng/ml). Direct correlation between ESR and CRP (p=0.04, t=3.85) was observed, and an increased leukocyte count (10.26±4.8) was found. In a group where CRP level were <5 mg/l, only 50% of patients had elevated exDNA (314.3±254.6 ng/ml) and methylation levels. One can suppose that hypermethylated DNA is released in the process of apoptosis. The statistically significant association between methylation level and elevated leukocyte count suggests a role of hypermethylated DNA in the increased leukocyte resistance to death, pathogenesis, and inflammation at AS. Conclusions Correlation between elevated monomethine protein and leukocyte count in the association with high DNA methylation levels can be considered as a marker of neutrophil resistance to apoptosis and participation of neutrophils in the processes of destruction and inflammation. Hence, the data we obtained helps us to develop a concept of AS pathogenesis as cause-effect relationships of the inflammation processes, destruction, and proliferation with an increase in cell death rate and in hypermethylated DNA. Presence of hypermethylated DNA is associated ...

The paper analyses development of academic science in the Urals during the Great Patriotic war. The paper shows process of its transition from the stage of formation reached in the prewar period to the stage of final formation, which made it possible to implement innovative discoveries and move forward scientific and technical progress. The paper pays special attention to the creative interaction of central management structures, institutes and laboratories of Academy of Sciences of the USSR, evacuated to the Urals, with local scientific organizations. The paper also analyses issues of solving everyday problems, training and placement of scientific personnel, and describes material and technical support of research works, scientific results and their practical use. The paper concludes that Ural scientists coped with the requirements of wartime, successfully solving many issues of defence, and finally ensured promotion of Ural academic science to a new qualitative level.