Production of monoclonal antibodies to the prion protein and their characterization

Antibodies to the prion protein (PrP), particularly, monoclonal antibodies, are necessary tools in the diagnostics and study of prion diseases and potential means of their immunotherapy. For the production of monoclonal antibodies, BALB/c mice were immunized by a recombinant bovine PrP. Three stable hybridomas producing antibodies of IgM class were prepared. The antibodies were bound to PrP in a solid-phase enzyme immunoassay and immunoblotting. The epitope mapping accomplished with the use of synthetic peptides showed that an epitope located in region 25–36 of PrP corresponds to one antibody, and epitopes located in region 222–229, to the other two. The antibodies to fragment 222–229 purified by affinity chromatography recognized with a high specificity conglomerates of a pathogenic prion in the brain tissue of cows suffering from spongiform encephalopathy. Thus, in nontransgenic mice, PrP-specific monoclonal antibodies were produced, useful in studies and diagnostics of prion diseases.     

Primary structure of the A22 RNA polymerase gene of the foot and mouth disease virus

Complete nucleotide sequence of gene RNA polymerase for the foot-and-mouth disease virus subtype A22 has been determined.     

Cell kinetics in the mouse esophageal epithelium in relation to the time of day

The kinetics of mouse esophageal epithelial cells was investigated throughout 90 h after a single injection of 3H-thymidine at 01 or at 13 h--the times of the peak and minimal magnitudes of the radioisotope index in the circadian rhythm of proliferation. The mitotic cycle parameters in the cells varied but insignificantly. For cells treated with 3H-thymidine at 01 h, T = 24.3 h, ts = 6 h, tG2 min = 1.5 h, tG2+ 1/2 M = 2.9 h and tG1+/2 M = 15.4 h; for those treated with 3H-thymidine at 13 h, T = 25.6 h, ts = 8.4 h, tG2 min = 1 h, tG2+ 1/2 M = 2.2 h, tG1+ 1/2 M = 15 h. Cells labeled at 01 h proliferated more actively for a long period of time as compared to those labeled at 13 h. The synchronism in undergoing several successive mitotic cycles was greater for cells labeled at the peak radioisotope index. The data obtained also suggest that the majority of cells enter the G0 phase after the completion of the first cycle. The duration of the G0 phase varies in different cell populations.     

Mutations in <Emphasis Type="Italic">RAS/BRAF</Emphasis> genes in rectal tumors: From adenomas to early carcinomas

The frequency and spectrum of mutations in RAS/BRAF genes were studied in rectal adenomas, carcinomas in situ, and adenocarcinomas. It is shown that the frequency of KRAS mutations decreases from adenoma to adenocarcinoma; most adenomas and carcinomas in situ are heterogeneous and consist of several subclones. Possible models of colorectal cancer pathogenesis are discussed.     

Features of the circadian rhythm of skin temperature in eight- to nine-year-old children and young adults

The age-related features of the circadian rhythm of skin temperature were studied using the Termochron iButton method in two age groups of subjects: eight-to nine-year-old boys and girls and young men and women aged 20–22 years. For this purpose, the temperature was monitored for 48 h at a recording interval of 10 min. The following authentic chronobiological differences were revealed: the temperature mesor was higher in the girls than in the boys and in the young men it was higher than in the young women. The circadian rhythm’s amplitude was higher in the boys and the young women. Studies of the chronological values in different 24-h periods showed that the average level of temperature at night was lower in all subjects. The amplitude in the adults did not differ in different 24-h periods, whereas it was higher in the children at night.     

Heterogeneity in Colorectal Primary Tumor and Synchronous Liver Metastases

The expression profile of the ZEB1, ZEB2, VIM, CDH1, SFRP2, FOXQ1, TNC, MACC1, PLS3, CFTR, FLNA, MUC2, TFF3, and RARRES3 genes, as well as the mutational status of the KRAS, NRAS, BRAF, and PIK3CA genes, were investigated in 40 patients with colorectal cancer and liver metastases. A comparative analysis of changes in gene expression in primary tumor cells and liver metastases was performed. Statistically significant differences were found between the expression levels of the ZEB2 (p = 0.004), VIM (p < 0.001), FLNA (p = 0.04), and MUC2 (p < 0.001) genes. It was demonstrated that the overall frequency of mutations of the KRAS gene was 18/40 (45%) and the PIK3CA gene was 9/40 (23%). Mutations in the NRAS and BRAF genes were not found. The concordance between the primary tumor and metastases in the liver by mutation status was 100%.     

Study of interneuronal connections by the technic of luminescent demonstration of retrograde transport of primulin

A possibility to study associative and projective connections of the optic cortical fields by the method of luminescent revealing of primuline retrograde transport has been demonstrated in the experiments with cats. Initial neurons of the connective systems have been revealed in various fields of the optic cortex (fields 17, 18, 19) and in the lateral geniculate body of the thalamus. The initial cells have various luminescent intensity.