[Magnesium and vitamin B2 status of children with bronchial asthma and obesity].

Insufficiency or deficiency of some micronutrients may be additional modifying factors that influence the pathogenesis of the disease and the effectiveness of standard pharmacotherapy. The aim of the study - to evaluate the level of magnesium and vitamin B2 in blood serum of patients with bronchial asthma and obesity in order to develop methods for individual correction of deficiency. Material and methods. The study included 51 children aged 12-17 years. The first group included 23 patients (12 girls and 11 boys) with asthma with comorbidities (obesity), and the second group consisted of 28 children (10 girls and 18 boys) with obesity. The concentration of magnesium in blood serum was determined by a colorimetric method without deproteinization, and vitamin B2 - by an immunological microbiological method. Results and discussion. When analyzing the concentration of magnesium in blood serum of the examined children, it was found that in patients with bronchial asthma and obesity, a reduced content of this mineral was observed in 15 (65.2%) patients. The average magnesium concentration was 0.66±0.02 mmol/l at a rate of 0.7-1.2 mmol/l. A statistically significant decrease in the magnesium level in children suffering from asthma and obesity was noted, compared with the level in children with obesity (0.66 [0.57; 0.73] vs 0.71 [0.67; 0.73] mmol/l, р<0.05). Low serum magnesium levels in obese patients were detected more rarely (р<0.05) - only in 6 (21.4%) children, mostly in patients with grade III obesity. The remaining 22 (78.6%) children had magnesium level within the normal range. Patients with low serum magnesium levels showed increased irritability, sleep disturbance, loss of memory and concentration. Vitamin B2 levels in all examined children were within the normal range (137-370 ng/ml). Conclusion. The results indicate a decrease in the concentration of magnesium and normal levels of vitamin B2 in serum in patients with bronchial asthma and obesity. Low serum magnesium levels were observed in children with low bronchial asthma control. To increase the effectiveness of therapy and control the symptoms of bronchial asthma, especially when combined with obesity, correction of the accompanying magnesium deficiency is necessary.

[Two cases of hydrophobia in the Republic of Tatarstan: in vivo and postmortem laboratory diagnosis].

The results of rabies in vivo and postmortem laboratory detection in two cases registered in the Republic of Tatarstan are reported: a victim bitten by a wolf in 2002 and another one bitten by a stray dog on Goa Island, India, in 2013. In the patient bitten by a wolf cornea imprints studies using the method of fluorescent antibodies (MFA) showed rabies-positive result 6 days before the patient's death. The results were confirmed by postmortem examination of different parts of the brain and salivary glands using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. In the patient bitten by a stray dog the rabies virus specific antigen was detected by eye cornea studies using the MFA method and saliva studies using the ELISA. The rabies virus genome was also isolated from saliva and tear fluid using nested reverse-transcription polymerase chain reaction (RT-PCR) 9 days before the patient's death. The in viva studies results were consistent with the postmortem study of different parts of the brain using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. All the infection-positive results of both in viva and postmortem studies were consistent with the clinical studies, i.e. rabies diagnosis was confirmed. The analysis of the rabies virus gene G fragment nucleotide sequence of 238 nd length showed a slight difference between the studied isolates (2 rabies) and the RABV AY9563I9 (1.68%), difference by 10.5% from the Vnukovo-32 vaccine strains and by 10.9% from the SAD B19 rabies strain, respectively (rabies viruses of 1st genotype). It was also significantly different from the lissaviruses of 2,4,5, and 6 genotypes (21 .0-32.7%). The obtained results indicate phylogenetic closeness of the studied isolates (2 rabies) with the RABV AY956319 rabies virus strain belonging to the 1st genotype.

Binding of proteins of HeLa S3 cell extract to oligonucleotides containing the consensus interferon-response sequence (IRS) and to the IRS-containing fragment of the human c-myc gene.

The human c-myc proto-oncogene was recently found to contain a regulatory sequence similar to the consensus interferon-response sequence (IRS) of interferon-activating genes. Binding of regulatory protein(s) to this sequence of cloned fragment of c-myc, lacking the main part of 5'-nontranscribing region, regulates in vitro transcription from I1/I2 initiation sites located in the first intron of the gene. Here, we have shown that HeLa S3 nuclear extract contains different protein factors, at least two, that bind preferentially to the IRS sequence of either the c-myc gene or the interferon-dependent 6-16 gene. Moreover, each of these factors 'cross-binds' to the region of the other gene, although affinity of this interaction is lower. Binding constants of these proteins to oligonucleotide fragments of c-myc and 6-16 genes were determined. In vitro transcription of the human full-length c-myc gene (i.e. the gene containing the complete 5'-noncoding region) initiated from I1/I2 sites, that is controlled by the IRS region, was demonstrated to be blocked. A possible physiological role for the mechanisms described is discussed.

Tryptophanyl-tRNA synthetase in cell lines resistant to tryptophan analogs.

Bovine kidney cell lines resistant to tryptamine and tryptophanol (tryptophan analogs) were selected. The content of tryptophanyl-tRNA synthetase (WRS, EC 6.1.1.2) was assayed by measuring the binding of monospecific polyclonal antibodies to the 35S-labeled enzyme in detergent-soluble and -insoluble forms and measuring the enzyme activity. Both the enzyme content and activity were elevated in the resistant cells. As was found by immunoelectron microscopy, the initial and resistant cells contained WRS in most of their cellular compartments: on free polyribosomes, as large conglomerates in the cytoplasm, on polysomes bound to the rough endoplasmic reticulum membranes and to the outer nuclear membrane, on the cytoskeleton, and in the detergent-insoluble nuclear matrix. Immunochemically stained tangles of filaments were found in the resistant cells, but not in the control cells. WRS was less phosphorylated in the resistant than in the original Madin Darby bovine kidney cells. Karyological and morphometric analysis revealed that, in tryptamine-resistant cells, the marker acrocentric chromosome was longer and the frequency of its duplication rose to 96%. The results of this work indicate that the cultivated cells have become resistant to tryptophan analogs because of an elevated WRS concentration in the cells, possibly due to amplification of the WRS gene.

Human c-myc gene contains a regulatory site similar to consensus of interferon response sequence (IRS).

Expression of c-myc proto oncogene is regulated by multiple mechanisms. Here, we report that the consensus of the regulatory region of interferon-dependent genes, GGAAAN1-3 GAAA, was found after computer search in the 5'-terminal flank of human c-myc gene in position (-76:-67). In vitro transcription of c-myc gene fragments showed that the consensus region competes with oligonucleotide GGGAAAATGAAACT for binding to specific protein(s). This oligonucleotide was shown to bind selectively the interferon-dependent positive transcription factor. Transcription of c-myc fragments lacking 5'-terminal region up to positions -101 or +71 was initiated at two sites located in the first intron. These sites did not coincide with P1 in vivo RNA cap-site. Binding of the protein factor(s) to the regulatory region of c-myc gene -76:-67 blocked the in vitro transcription initiated in the first intron.

Expression of c-myc gene in human ovary carcinoma cells treated with vanadate.

The widely accepted hypothesis of vanadate action on cells postulates that this ion inhibits protein phosphatase(s) that dephosphorylates protein phosphotyrosine residues. This inhibition causes tyrosine hyperphosphorylation of cell proteins followed by changes in physiological action of phosphoproteins resulting in stimulation of cell proliferation, expression of protooncogenes, and transient cell transformation. We have found that treatment of human ovary carcinoma (CaOv) cells with vanadate causes the increase in total protein phosphorylation from 1.5- to 2.0-fold whereas the ratio between phosphoserine, phosphothreonine, and phosphotyrosine content remains unchanged. At the same time, enhancement of c-myc gene expression (not c-fos) was observed. Hence, the increase in the ratio of phosphotyrosine to phosphoserine and phosphothreonine is not an obligatory intermediate stage before vanadate-dependent activation of c-myc expression.

The informosome-like virus-specific ribonucleoprotein (vRNP) may be involved in the transport of tobacco mosaic virus infection.

A new type of informosome-like virus-specific ribonucleoprotein (vRNP) differing from mature tobacco mosaic virus (TMV) particles in buoyant density and structure was found in TMV-infected cells (Yu. L. Dorokhov, N. M. Alexandrova, N. A. Miroshnichenko, and J. G. Atabekov, 1983, Virology 127, 237-252). Two groups of TMV ts mutants were used to discover whether there is a correlation between the vRNP formation and systemic spreading of virus infection (transport) over the infected plant. The first group of mutants (Ni118, flavum) contains a ts mutation in the coat protein gene but are capable of systemic spreading at nonpermissive temperature (tr transport); the second group of mutants (Ni2519, Ls1) cannot spread systemically at restrictive temperature (ts transport). It is shown that vRNP can be produced at restrictive temperature by tr-transport mutants but not by ts-transport mutants. The latter can produce vRNP only at a permissive temperature. The role of vRNP in long-distance transport of the virus infection is supported by two other observations: (a) upper leaves that were maintained at 5 degrees accumulate potentially infective material and material with the properties of vRNP but not virus particles and (b) plants that were simultaneously infected with Lsl and Ni118 at a non-permissive temperature exhibited long-distance transport and vRNP. These results also implicate coat protein in long-distance transport. It is suggested that vRNPs are novel types of virus-specific particles that are involved in both cell-to-cell and long-distance transport of TMV infections.

Stimulation by aurintricarboxylic acid of tobacco mosaic virus-specific RNA synthesis and production of informosome-like infection-specific ribonucleoprotein.

It was shown that aurintricarboxylic acid (ATA), a well-known inhibitor of protein synthesis, markedly stimulates the synthesis of tobacco mosaic virus (TMV)-specific RNA species of the intermediate (I) class (apparent molecular weights 1.1-1.3 x 10(6) and 0.6-0.8 x 10(6)). No stimulation by ATA of full-length genomic TMV RNA or the subgenomic TMV RNA coding for TMV coat protein was detected. Informosome-like infection-specific ribonucleoprotein (vRNP) particles different from mature TMV particles were found in the TMV-infected cells (Yu. L. Dorokhov, N. M. Alexandrova, N. A. Miroshnichenko, and J. G. Atabekov, 1983, Virology 127, 237-252). It is shown here that [3H]uridine incorporation into vRNP RNAs was markedly stimulated in the presence of ATA. vRNP can be released from the TMV-specific polyribosomes by EDTA treatment, which suggests that it is involved in the translation process. The results of the pulse-chase experiments (including those in which TMV RNA synthesis is blocked by 2-thiouracil) suggest that vRNP does not serve as a precursor for mature virion.

Isolation and analysis of virus-specific ribonucleoprotein of tobacco mosaic virus-infected tobacco.

A ribonucleoprotein fraction (vRNP) of a characteristic buoyant density greater than the buoyant density of tobacco mosaic virus (TMV) particles has been isolated from infected tissue by Cs2SO4 density gradient centrifugation. The vRNP particles appear to be TMV specific because they are synthesized in the presence of actinomycin D and have RNAs identified as genomic and I-class subgenomic (apparent Mr 1.1-1.3 x 106 and 0.60.8 x 10(6)) RNAs by their electrophoretic mobility and hybridization to plasmid-bearing RNA sequences. Polypeptides of apparent molecular weights 17,500 (TMV coat), 31,000, 37,000, and 39,000 were major constituents of vRNP. Of the minor polypeptides, those of apparent molecular weights 70,000, 68,000, 55,000, and 25,000 had electrophoretic mobilities similar to mobilities of polypeptides found in a ribonucleoprotein preparation from uninoculated plants. vRNP from common TMV-infected plants, but not from plants infected with a mutant that did not form native coat protein, reacted with immunoglobins against TMV and TMV coat protein. Common TMV and its vRNP differed in the extent of reactivity toward the two immunoglobins, in electron microscopic appearance, and in the higher sensitivity of vRNP to ribonuclease.

Development of systemic TMV infection in upper noninoculated tobacco leaves after differential temperature treatment.

A differential temperature treatment (DTT) of infected plants ensured a partial synchronization of the process of tobacco mosaic virus (TMV) infection. The "infectious entity" enters (and spreads to) basal areas of the upper noninoculated leaves held at low temperature, and then to a distal part of these leaves and basal sites of the lower leaves. The spread of the "infectious entity" over the vascular system of the upper leaves could be followed.