Impaired Surfactant Production by Alveolar Epithelial Cells in a SCID-hu Lung Mouse Model of Congenital Human Cytomegalovirus Infection

Human cytomegalovirus (HCMV) is the leading viral cause of birth defects and life-threatening lung-associated diseases in premature infants and immunocompromised children. Although the fetal lung is a major target organ of the virus, HCMV lung pathogenesis has remained unexplored, possibly as a result of extreme host range restriction. To overcome this hurdle, we generated a SCID-hu lung mouse model that closely recapitulates the discrete stages of human lung development in utero. Human fetal lung tissue was implanted into severe combined immunodeficient (CB17-scid) mice and inoculated by direct injection with the VR1814 clinical isolate of HCMV. Virus replication in the fetal lung was assessed by the quantification of infectious virus titers and HCMV genome copies and the detection of HCMV proteins by immunohistochemistry and Western blotting. We show that HCMV efficiently replicated in the lung implants during a 2-week period, forming large viral lesions. The virus productively infected alveolar epithelial and mesenchymal cells, imitating congenital infection of the fetal lung. HCMV replication triggered apoptosis near and within the viral lesions and impaired the production of surfactant proteins in the alveolar epithelium. Our findings highlight that congenital and neonatal HCMV infection can adversely impact lung development, leading to pneumonia and acute lung injury. We have successfully developed a small-animal model that closely recapitulates fetal and neonatal lung development and provides a valuable, biologically relevant tool for an understanding of the lung pathogenesis of HCMV as well as other human respiratory viruses. Additionally, this model would greatly facilitate the development and testing of new antiviral therapies for HCMV along with select human pulmonary pathogens.     

The Influence of Changes in Magnetic Variations and Light–Dark Cycle on Life-History Traits of Daphnia magna

Day–night cycle is the main zeitgeber (time giver) for biological circadian rhythms. Recently, it was suggested that natural diurnal geomagnetic variation may also be utilized by organisms for the synchronization of these rhythms. In this study, life-history traits in Daphnia magna were evaluated after short-term and multigenerational exposure to 16 h day/8 h night cycle, 32 h day/16 h night cycle, diurnal geomagnetic variation of 24 h, simulated magnetic variation of 48 h, and combinations of these conditions. With short-term exposure, the lighting mode substantially influenced the brood to brood period and the lifespan in daphnids. The brood to brood period, brood size, and body length of crustaceans similarly depended on the lighting mode during the multigenerational exposure. At the same time, an interaction of lighting mode and magnetic variations affected to a lesser extent brood to brood period, brood size, and newborn's body length. The influence of simulated diurnal variation on life-history traits in daphnids appeared distinctly as effects of synchronization between periods of lighting mode and magnetic variations during the multigenerational exposure. Newborn's body length significantly depended on the lighting regime when the periods of both studied zeitgebers were unsynchronized, or on the interaction of light regime with magnetic variations when the periods were synchronized. These results confirm the hypothesis that diurnal geomagnetic variation is an additional zeitgeber for biological circadian rhythms. Possible mechanisms for these observed effects are discussed. Bioelectromagnetics. © 2020 Bioelectromagnetics Society     

Virulence-associated characteristics and phage lysogenicity of two morphologically distinct colonies of Vibrio cholerae O139 serogroup

Abstract The presence of a temperate phage was demonstrated in a strain of Vibrio cholerae O139 isolated from a patient. Spontaneous variants with translucent colonies had lost this phage. The loss of the phage was associated with increased hydrophobicity, indicating the loss of the capsule. These clones were sensitive to serum bactericidal activity, showed decreased expression of such presumed virulence factors as proteases, motility and mannose-sensitive pili. Furthermore, excision of the phage made the strain dependent on purines for growth.     

The systematic approach to describing conformational rearrangements in G-quadruplexes

Conformational changes in DNA G-quadruplex (GQ)-forming regions affect genome function and, thus, compose an interesting research topic. Computer modelling may yield insight into quadruplex folding and rearrangement, particularly molecular dynamics simulations. Here, we show that specific parameters, which are distinct from those commonly used in DNA conformational analyses, must be introduced for adequate interpretation and, most importantly, convenient visual representation of the quadruplex modelling results. We report a set of parameters that comprehensively and systematically describe GQ geometry in dynamics. The parameters include those related to quartet planarity, quadruplex twist, and quartet stacking; they are used to quantitatively characterise various types of quadruplexes and rearrangements, such as quartet distortion/disruption or deviation/bulging of a single nucleotide from the quartet plane. Our approach to describing conformational changes in quadruplexes using the new parameters is exemplified by telomeric quadruplex rearrangement, and the benefits of applying this approach to analyse other structures are discussed.     

The Relationship between the Structure of the Tick-Borne Encephalitis Virus Strains and Their Pathogenic Properties

Tick-borne encephalitis virus (TBEV) is transmitted to vertebrates by taiga or forest ticks through bites, inducing disease of variable severity. The reasons underlying these differences in the severity of the disease are unknown. In order to identify genetic factors affecting the pathogenicity of virus strains, we have sequenced and compared the complete genomes of 34 Far-Eastern subtype (FE) TBEV strains isolated from patients with different disease severity (Primorye, the Russian Far East). We analyzed the complete genomes of 11 human pathogenic strains isolated from the brains of dead patients with the encephalitic form of the disease (Efd), 4 strains from the blood of patients with the febrile form of TBE (Ffd), and 19 strains from patients with the subclinical form of TBE (Sfd). On the phylogenetic tree, pathogenic Efd strains formed two clusters containing the prototype strains, Senzhang and Sofjin, respectively. Sfd strains formed a third separate cluster, including the Oshima strain. The strains that caused the febrile form of the disease did not form a separate cluster. In the viral proteins, we found 198 positions with at least one amino acid residue substitution, of which only 17 amino acid residue substitutions were correlated with the variable pathogenicity of these strains in humans and they authentically differed between the groups. We considered the role of each amino acid substitution and assumed that the deletion of 111 amino acids in the capsid protein in combination with the amino acid substitutions R16K and S45F in the NS3 protease may affect the budding process of viral particles. These changes may be the major reason for the diminished pathogenicity of TBEV strains. We recommend Sfd strains for testing as attenuation vaccine candidates.     

Epidemiological problems of ozena and the means for controlling this infection

Clinical, bacteriological, serological and epidemiological studies of ozena morbidity among the population of Minsk were carried out in 1970-1980. On January 1, 1981, the ozena morbidity rate among the inhabitants of Minsk was 26.72%. Ozena was found to affect mainly children and women. A wide spread of the family foci of this disease (31.68%) was revealed. The results of this study indicate that the source of K. ozaenae is a sick person who begins to excrete the bacteria in the prodromal period of the disease and may continue to excrete them for many years. The transfer of K. ozaenae occurs probably by droplet or contact infection. The droplet infection is less active in the absence of symptoms (coughing, sneezing) facilitating excretion of the infective agent into the air and in cases of the low susceptibility of persons to ozena. The main measures for controlling ozena are the timely detection and sanitation of the sources of ozena, as well as the current disinfection of the infection foci in apartments.     

The efficacy of estrus detection and fertility following synchronization with PGF2a or synchro - mate- B in Zebu cattle

Two experiments were carried out to assess the efficacy of estrus detection and fertility in Zebu cattle after synchronization with prostaglandin F2a or a progestagen. The first experiment compared estrus detection rates and fertility following insemination in 42 cows previously synchronized with either 25 mg of PGF2a or with a 6 mg of Norgestomet implant plus 5 mg i.m. of estradiol valerate (SMB). Differences were observed in the percentage of cows detected in estrus (54 vs 95%, respectively, P < 0.05), but not in fertility at the first synchronized estrus (26 vs 15%), nor in the detection rate and fertility at the subsequent estrous period (38 v 47%). The second experiment evaluated the efficacy of estrus detection at different time intervals in 30 cows, comparing estrus synchronized with PGF2a with the subsequent estrous period. The observation periods were continuous, day and night, for 100 h both after PGF2a treatment and from Day 18 of the treatment cycle (Period 1). In addition, the animals were administered PGF2a again on Day 10 of the second cycle and observed continuously from 0600 to 1800 h, and from Day 18 of the treatment cycle (Period 2). Finally, the same treatment regimen was used except that the observation was between 0600 to 0700 h and 1800 to 1900 h (Period 3). No differences were obtained in the percentage of cows detected in estrus in the synchronized and nonsynchronized groups (average 75%); however, accuracy in the detection of estrus in Period 3 differed in the nonsynchronized and synchronized estrus groups by 40% (P < 0.05) compared with the other two, more intense observation periods.     

Effects of prodigiozan on post-radiation recovery of hemopoietic precursor cells and colony-stimulating factor level in long-term cultures of mouse bone marrow

Prodigiozan injected to long-term cultures of mouse bone marrow 24 h before irradiation increased CFUs and CFU-GM number and colony-stimulating factor (CSF) level by the time of delivery of ionizing radiation. As early as 60 min following irradiation of bone marrow structures with a dose of 2 Gy the number of CFUs and CFU-GM decreased considerably, and from day 3 on after irradiation the indices under study were gradually restored. By day 14 the cultures preinjected with prodigiozan exhibited higher recovery levels. The decrease in the number of precursor cells 60 min after irradiation was accompanied by a drastic increase in the CSF content of cultures; the CSF release in cultures protected with prodigiozan was more moderate than in the irradiated controls.     

Complementary addressed modification of nucleic acids with the alkylating derivatives of oligothymidylate ethyl phosphotriesters. Effect of the phosphotriester fragments' configuration

The alkylating derivatives of four individual diastereomers of the oligonucleotide [dTp(Et)]3dTpU and two individual diastereomers of oligonucleotide [dTp(Et)dTp]4 have been synthesized. The reagents with the phosphorus atoms in the enantiomeric p" configuration are shown to be more efficient in reacting with poly(dA) and with nucleic acids in Krebs-2 ascites carcinoma cells compared to those with the phosphorus atoms in the p' configuration.