Development of Pyrosequencing Assay for Evaluation of Genetic Stability of Vaccine Strains of Live Attenuated Influenza Type B Vaccine.

We developed a pyrosequencing protocol for monitoring of stability of attenuating mutations in the genome of vaccine reassortants based on master donor virus of Russian live attenuated influenza vaccine B/USSR/60/69. The developed protocol allows rapid and accurate assessment of mutations and can be used for analysis of genetic stability of reassortants during vaccine strain development and manufacturing, as well as genetic stability of vaccine isolates of influenza B virus during pre-clinical and clinical trials.

[Treating patients with acute myeloid leukemias (AML) according to the protocol of the AML-01.10 Russian multicenter randomized trial: the coordinating center's results].

AIM: To make a randomized comparison of 2 consolidation treatment options (two patient groups): 2 cycles of cytarabine in average (Ig/m2 in Group 2) and standard (100 mg/mi2 in Group 1) doses in combination with idarubicin (8-12 mg/m2) and mitoxantrone (10 mg/m2), after two 7+3 induction cycles of daunorubicin (60 mg/mi2) and subsequent 6 cycles of maintenance therapy. SUBJECTS AND METHODS: In January 2010 to October 2013, a Russian multicenter trial was conducted to treat patients with acute myeloid leukemias (AML) in accordance with the AML-01.10 protocol (ClinicalTrials.gov Identifier: NCT01587430). The trial enrolled 243 AML patients from 21 centers, including 71 patients (median age 38 years) from the State Hematology Center, Ministry of Health of the Russian Federation; 35 and 36 patients were randomized to Groups 1 and 2, respectively. The randomized groups were balanced by basic clinical and laboratory parameters. Favorable, intermediate, and high cytogenetic prognoses were in 14 (21.9%), 40 (62.5%), and 10 (15.6%) patients, respectively. RESULTS: Prior to treatment, 2 patients died; one patient refused treatment. Fifty-eight (85.3%) of the 68 patients achieved complete remission (CR); early deaths was in 2 (2.9%) and resistance in 8 (11.8%). Four (6.9%) patients died during CR. Protocol deviations (doses, intervals, and the number of cycles) were recorded in 12 (20.7%) of the 58 patients. Other 8 (11.8%) patients were switched to low-dose cytarabine because of complications, withdrawn from the protocol and not included into the analysis of randomized comparison. Twenty allogeneic bone marrow transplantations (allo-BMT) (7 related, 12 unrelated, and 1 haploidentical) were performed; of them 15 allo-BMTs were done during first CR. In the 68 patients, 3-year overall survival (OS) was 45.6%; relapse-free survival (RFS) was 41.5%. OS was 64.6% in Group 1 and 58.3% in Group 2; RFS was 62 and 38.8% in Groups 1 and 2, respectively (p>0.5). In the favorable, intermediate, and high prognosis groups, OS was 79.5, 60, and 31.1% and RFS was 81.8, 41.3, and 33.3%, respectively (p=0.1). The consolidation treatment option unchanged survival rates in the above risk groups. Unachieved CR after the first cycle considerably decreased RFS (33.9% versus 60%) and served as an indication for allo-BMT during first CP (RFS without BMT was 0; that with BMT was 78%). CONCLUSION: No differences were found between both consolidation options according to long-term results. Protocol deviations were recorded in one-third of the patients. While implementing the protocol, the efficiency of treatment was high. Allo-BMT during first CR substantially increased RFS if CP was not achieved after the first cycle.

[Seroepidemiology of hepatitis A in Saint-Petersburg in 2009].

AIM: To determine immune structure of different population groups in Saint-Petersburg to hepatitis A virus in 2009 in order to study trends of epidemic process dynamics and planning of prophylactic measures. MATERIALS AND METHODS: Nine hundred and three citizens of Saint-Petersburg aged 3 months to 60 years were tested for anti-HAV by ELISA in 2009. Results were compared with data of previous study conducted in Saint-Petersburg in 1999. Two hundred injection drug users (IDUs) aged 14 - 29 years were tested additionally. RESULTS: In total in 2009, anti-HAV were detected in 32.5 +/- 1.6% citizens of Saint-Petersburg that is 2-fold lower than in 1999 (60.2 +/- 1.5%). Especially sharp decrease of anti-HAV prevalence in 2009 compared to 1999 was observed in age groups 15 - 19 years (by 2.6-fold), 20 - 29 years (by 3.1-fold), and 30 - 39 years (2.8-fold). Anti-HAV were detected in 58.5 +/- 3.5% of tested IDUs that 2.8-fold higher of that value observed in persons 14 - 29 years old from population sample (21.7 +/- 2.4%). CONCLUSION: Decrease of immunity to HA in population of Saint-Petersburg is very unfavorable prognostic factor pointing to strong possibility of HA outbreaks onset during worsening of epidemic situation. In such circumstances, active immunization against HA of wide strata of population should become important prophylactic measure.

Quantitative changes in the fibroarchitectonics of the human cortex from birth to the age of 12 years.

Brains from 74 children aged from birth to 12 years were used to study fibroarchitectonic characteristics in topographically and functional diverse cortical zones (the temporal-parietal-occipital subregion, occipital, precentral, postcentral, and frontal areas) of the cerebral cortex; children aged up to 12 months were studied on the basis of one-month age intervals. Studies were performed by computer analysis of optical images on frontal sections stained with Nissl cresyl violet and silver nitrate impregnation by the modified Peters method. Data on the rate of increase in the thickness of radial bundles of fibers, the distances between bundles, and the age dynamics of the ratios of the specific volumes of neurons and fibers in fields 3, 4p, 6op, 17, 19, 37ac, 44, and 32/10 were obtained. These measurements showed that age-related transformation of fibroarchitectonics in fields of the sensorimotor, somatosensory, occipital, temporal-parietal-occipital, and frontal areas occurred at different times and with different intensities; the most significant changes were seen in the first 2-3 years of life, with changes continuing at a less intense level to age 9-12 years.

[Quantitative changes in fibro-architectonics of the human cerebral cortex from birth to 12 years of age]. / Kolichestvennye izmeneniia fibroarkhitektoniki kory bol'shogo mozga cheloveka ot rozhdeniia do 12 let.

The peculiarities of fibroarchitectonics in topographically and functionally distinct cortical zones (including temporo-parieto-occipital subarea, occipital, pre-, postcentral, and frontal areas) were studied in the brain specimens of 74 children of different ages (from birth to 12 years) taken with yearly intervals, while those in the first year of life--with monthly intervals. Frontal sections stained with Nissl, Peters and Golgi methods were studied using computer analysis of optical images. The data were obtained on the increment rates of radial fiber fascicles' thickness, distances between fascicles and age dynamics of ratios of volume densities of neurons and fibers in areas 3, 4p, 6op, 17, 19, 37ac, 44 and 32/10. It was shown that age-related transformations of fibroarchitectonics in the areas of sensomotor, somatosensory, occipital, temporo-parieto-occipital and frontal cortex were heterochronic and took place with different intensity; most pronounced changes were found to occur during the first 2-3 years, while less intensive ones were observed up to the age of 9-12 years.

[Characteristics of the functional organization of neuronal groups in functionally diverse zones of the human cerebral cortex from birth to 20 years of age]. / Osobennosti strukturnoi organizatsii neironnykh gruppirovok funktsional'no razlichnykh zon kory bol'shogo mozga cheloveka v period ot rozhdeniia do 20 let.

Using computer analysis of histological specimens of motor and posterior associative cortex of the man aged from birth up to 20 yrs age and individual peculiarities of focal groups sublayer III3 of areas 4p, 6op and 37ac, periods of intensive and slow growth of the area of profile fields (PF) of cell groups and total area (TA) of neurons in the group were established. It was shown that greater changes of cell groups PF area and neuron TA occurred in the area 4p during first 3 yrs, while in areas 6op and 37ac from birth up to 7 yrs and 8-9 yrs appropriately. Specific features of the group development in functionally different areas of human brain cortex was demonstrated.

[Characteristics of the ensemble organization of the human cerebral cortex from birth to 20 years of age]. / Osobennosti ansamblevoi organizatsii kory bol'shovogo mozga cheloveka ot rozhdeniia do 20 let.

When studying frontal, somatosensory and visual areas of the human cerebral cortex from birth up to 20 years of age in year-to-year intervals, it has been stated that by birth in neocortex all components of the neuron-glio-vascular ensembles are presented. They are not connected in their composition. During the first year of life the size of all types of neurons increases, long-axonal basket neurons differentiate, fasciculi of radial fibers become thick. By 3 years of life in the ensembles the neurons are definitely grouped as clusters. Sizes of spindle-like and satellite neurons increase; they distribute their axonal collaterals vertically, horizontally and in frontal-posterior direction. By 5-6 years of age the horizontal connection system becomes more complex at the expense of longitudinal growth and ramification of lateral and basal dendrites of the pyramidal neurons. In the section transversal areas occupied with cell groups increase. By 9-10 years of age the pyramidal neurons reach their greatest size. By 12-14 years of age the fibrillar component of the cortex increases considerably, inter- and intraensemble horizontal connections become more complex, the system of local connections becomes more plastic owing to development of short-axonal basket-like neurons. By 16-18 years of age the ensemble cortical organization in its main parameters of architectonics reaches the level specific for mature persons.

[Amino acid sequence of Ondatra myoglobin (Ondatra zibethica). Characteristic features of myoglobins from semiaquatic animals]. / Aminokislotnaia posledovatel'nost' mioglobina ondatry (Ondatra zibethica). Kharakternye osobennosti mioglobinov zhivotnykh, vedushchikh poluvodnyi obraz zhizni.

Based on the amino acid composition of globin, amino acid analysis and N-terminal sequencing of peptides as well as a comparative analysis of the primary structure of beaver, coypu rat and otter myoglobins with the use of the fingerprinting technique, the amino acid sequence of the major component of ondatra myoglobin including 153 amino acid residues was reconstructed. The results of a comparative analysis of the primary structure of myoglobin and the peculiarities of the functional morphology of myoglobins from semi-aquatic animals and sperm whale and the role of amino acid substitutions in the spatial structure of ondatra myoglobin are discussed.

[Chromosomes of 7 transplantable lines of macaque and pavian hematopoietic cells producing herpesvirus type EBV and oncornavirus type C]. / Izuchenie khromosom semi perevivaemykh linii krovetvornykh kletok makak i pavianov, produtsiruiushchikh virus gerpesa tipa EBV i onkornavirus tipa C.

Karyotypes of Su-1, Su-2, KMMA-1, and KMMA-2 (producing oncornavirus C-type), and KMPG-1, SPF-2, and SPG-3 (producing HSV, a virus of EBV type) cell lines were studied in the time course. The lines were derived from hemopoietic cells of brown macaques and baboons bearing lymphomas. Some of the lines had a paradiploid-pseudodiploid karyotype, another part aneuploid type. The selectivity of structural and numerical abnormalities of chromosomes 1, 9, 10, 11, 12, 13, 15, 18, and 20 was detected indicating a certain specificity of karyotype changes upon malignization and transformation. The number of poorly and moderately differentiating chromosomes in aneuploidy was confirmed by greater variability as compared with well-differentiating chromosomes. No parallelism between the karyotype change pattern and the type of virus produced (C or EBV) could be found thus far.