[On the problem of the study of the chemical air pollution with chlororganic hydrocarbons at productions of polyvinyl chloride and epichlorohydrin].

Hygienic assessment of working conditions at the chemical productions of polyvinyl chloride (PVC) and epichlorohydrin (EPCH) in East Siberia has shown that the employees are exposed to the chlororganic hydrocarbons of hazard category 1-2, out of them there were found to be more toxical pollutants such as vinyl chloride, 1.2-dicloroethane in the production of polyvinyl chloride; allyl chloride and epichlorohydrin in the production of epichlorohydrin. Multistageness of the technological processes, the absence of the isolation of main stages of the technological processes as well as the heating microclimate contribute to the chemical pollution of the air environment. In spite of the significant improvement of the hygienic situation at the productions mentioned in the recent 10 years according to the chemical factor due to the introduction of the complex of curative measures, the working conditions of the employees still belonged to the harmful category. According to the content of the harmful chemical substances in the air of the working zone and the parameters of microclimate, the working conditions of the employees working at the production of epichlorohydrin and in the shop of vinyl chloride production must be qualified as the harmful ones of the first category of the hazard and danger (Class 3.1), in the production shop for PVC- as the harmful ones which correspond to the second category of the hazard and danger (Class 3.2).

[The specific features of microbiocenoses in children living under conditions of anthropogenic pressing].

The specific features of enteric and nasopharyngeal microbiocenoses and the species composition of bifidobacteria have been studied in children living in the industrial towns of the Irkutsk Region under the existing anthropogenic load. Ambient air pollution is characterized and a presumptive human health risk assessed.

[Biomonitoring of formaldehyde in the urinary samples from the pediatric population in the Irkutsk Region].

Formaldehyde is one of the major pollutants of both ambient and indoors air. Thus, by comparing the concentrations of formaldehyde in ambient air (on study days) and in the air of classes and living rooms, the study has demonstrated that they may be 13.5 and 10.5 times greater than the normal values in the classes and living rooms, respectively. Biomonitoring of formaldehyde in the urinary samples from the pediatric population of the Irkutsk Region as an indicator of its chemical action has revealed the higher average group concentration of the substance in the urinary samples from urban children than that in rural ones (235 children from 6 inhabited localities). A significant correlation has been also found between the levels of formaldehyde in the urine of children (aged 5-10 years) and its concentration in the air of living rooms in the town of Shelekhov.

[Methyl sulphides and their metabolites in biologic materials of workers engaged into sulfate pulping].

The authors studied serum and urine levels of methyl sulphides in workers engaged into sulfate pulping and exposed to methyl sulphides. Blood serum was studied only for dimethyldisulfide. Urine appeared to contain no methyl sulphides, but finding was increased urinary excretion of sulfates which are metabolites of methyl sulphides. Sulfates content of urine correlated with intensity and duration of exposure to methyl sulphides, so this exposure test could be used as a biologic marker in monitoring work conditions and health state of workers exposed to methyl sulphides.

[Evaluation of a chemical factor during fires]. / Otsenka khimicheskogo faktora pri pozharakh.

The working conditions of firemen during active service cannot be assessed without characterizing the degree of ambient gas contamination with toxic substances. Firemen extinguish fires and liquidate accidents under difficult conditions, which presents a threat to their life and health. Fire-induced combustion products can emit as high as 100 types of chemical compounds. Of them, the most common and well-studied compounds are carbon oxide, sulfur dioxide and nitric dioxide, formaldehyde, hydrogen chloride, saturated hydrocarbons (C2-C10), acetone, benzene, toluene, and some chlorohydrocarbons (vinyl chloride, chloroform, and tetrachloromethane). These compounds are hazard classes II and III, they have been detected in the air samples in the foci of a fire in the amenity and industrial premises, during combustion of motor transport, garbage, etc. Toxicological exposure to these compounds is accelerated under elevated temperatures.

Matrix-assisted laser desorption/ionization detection of polymerase chain reaction products by utilizing the 5'-3' exonuclease activity of Thermus aquaticus DNA polymerase.

The 5'-3' exonuclease activity of DNA polymerase was utilized in the polymerase chain reaction system to generate a specific signal concomitant with amplification. These signals were detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). This method obviates the need to perform extensive DNA purification of reaction products that is often necessary for detecting larger DNA molecules by mass spectrometry. Oligonucleotides complementary to the internal region of the amplicon are degraded by the 5'-3' exonuclease activity and the degradation products are analyzed by MALDI mass spectrometry. We refer to this assay as the Exo-taq assay or probe degradation assay. This method should be amenable to automation.

Laser desorption mass spectrometry for microbial DNA analysis.

Recently, we demonstrated that a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS) can be used to determine the molecular weight of polymerase chain reaction (PCR) products of intact 16S rRNA regions and to profile their restriction digests. This is the first time that MALDI-TOF MS with ultraviolet (UV) photoionization has been used to analyze a PCR product of approximately 1600 nucleotides in length.

Nonresonant MALDI of oligonucleotides: mechanism of ion desorption.

Oligonucleotide ions have been detected using matrix-assisted laser desorption/ionization (MALDI) under nonresonant laser irradiation of the sample. When mass resolution was not limited by adduct attachment to the analyte ions, the nonresonant MALDI spectra demonstrated better resolution than the spectra acquired with resonant ultraviolet irradiation. We found that preparation of thin-film samples on absorbing substrate surfaces was critical for the success of NR-MALDI. The possible acoustic mechanisms of ion formation and desorption are discussed.

Regulation of CD34 transcription by Sp1 requires sites upstream and downstream of the transcription start site.

CD34 is a cell surface glycoprotein expressed on hematopoietic stem and progenitor cells, but not on fully differentiated cells in the peripheral blood. To better understand the molecular regulation of early hematopoiesis, we are elucidating the mechanisms of CD34 transcriptional regulation. By deletion analysis we identify a 39-bp element in the proximal region of murine CD34 promoter that is critical for promoter activity. Electromobility shift assays indicate that nuclear proteins of hematopoietic cells bind to this domain; however, the presence of this binding activity does not correlate directly with CD34 expression.Using methylation interference, the DNA binding site for this activity was localized to four guanine residues within the GGGGTCGG sequence from -48 to -54 bp. When the four contact guanines were mutated, both protein binding and promoter activity were abolished. Although this sequence does not contain a standard consensus for Sp1, this transcription factor binds specifically to the 39-bp region and stimulates promoter activity in both hematopoietic cells and in Sp1 null Drosophila S2 cells. In addition, Ku binds to this domain in a sequence-specific manner. Activation of the CD34 promoter by Sp1 requires the presence of a binding domain at -48 bp as well as the 5' untranslated region, which also binds Sp1.A functional interaction between regulatory regions upstream and downstream of the transcription start site is required for CD34 gene expression.

Matrix-assisted laser desorption/ionization time-of-flight analysis of Bacillus spores using a 2.94 microm infrared laser.

The performance of infrared (2.94 microm) and ultraviolet (337 nm) lasers were compared for analysis of purified spores of B. subtilis, B. cereus and B. globigii on a four-inch end-cap reflectron time-of-flight instrument. Infrared matrix-assisted laser desorption/ionization (IR-MALDI) mass spectra of these microorganisms displayed a larger number of biomarker peaks above m/z 4000, compared with UV-MALDI. Biomarker peaks were observed at higher m/z values with the IR laser.

A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene.

Proper regulation of the human CD34 gene requires a combinatorial action of multiple proximal and long-range, cis elements. This report shows that, like the murine CD34 5' untranslated region (UTR), the corresponding region of the human CD34 gene is necessary for optimal promoter activity. We localized the most critical element of this region to base pairs +48/+75. Through oligonucleotide competition and antibody supershift experiments in electrophoretic mobility shift assays, we found that this sequence contains a binding site (CCAAT box) for the transcription factor NFY (nuclear factor Y), a factor mediating cell type-specific and cell-cycle regulated expression of genes. Mutating this site led to a 5-fold decrease in CD34 promoter activity in transient transfection experiments. Interestingly, NFY binds adjacently to the earlier identified c-myb binding site. Here we show that both binding sites are important for CD34 promoter function: mutating either site alone decreased CD34 promoter-driven reporter gene activity 4-fold. We also show that the integrity of the c-myb binding site is necessary for stabilization of NFY binding to its site. Such cooperation between c-myb, which is expressed in early hematopoietic cells, and NFY, which is expressed in many cell types, might contribute to specific activation of CD34 in stem cells. The CCAAT box motif was also noted in the 5' UTR of the murine CD34 gene, however, NFY did not bind to this region. Thus, our results indicate that the functional similarities between the human and murine CD34 5' UTRs are achieved through different molecular mechanism(s).

Gender identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

A rapid, simple, and reliable gender determination of human DNA samples was successfully obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Detection sensitivity reached 0.01 ng or less for DNA samples.

Detection of trinucleotide expansion in neurodegenerative disease by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Genotyping of the dentatorubral-pallidoluysian atrophy (DRPLA) locus in six patient samples, representing four normal individuals and two DRPLA patients, was successfully obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). DRPLA is a dominantly inherited neurodegenerative disorder associated with the expansion of an unstable trinucleotide (CAG) repeat. The accurate determination of repeat length utilizing MALDI supports the use of this methodology for the analysis of genes containing unstable CAG trinucleotide repeats.

Sequencing DNA using mass spectrometry for ladder detection.

Sequencing of DNA fragments of 130 and 200 bp using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for DNA ladder detection was demonstrated. With further improvement in mass resolution and detection sensitivity, mass spectrometry shows great promise for routine DNA sequencing in the future.

Matrix-assisted laser desorption/ionization for short tandem repeat loci.

Matrix-assisted laser desorption/ionization was used for the detection of four base short tandem repeats (STR) for clinical samples using a time-of-flight mass spectrometer. Since STR plays an important role in genetic disease and human identification, this work indicates that laser desorption mass spectrometry has the potential to achieve rapid DNA typing for both forensic applications and genetic disease diagnosis.

[Urinary sulfates as a biologic indicator of exposure to methylsulfate compounds in manufacture of sulfate cellulose]. / Sul'faty v moche kak biologicheskii pokazatel' vozdeistviia metilsernistykh soedinenii v proizvodstve sul'fatnoi tselliulozy.

The authors studied urine excretion of sulfates before and after a shift in workers engaged into brewing and flushing shops of sulfate cellulose production, who are exposed to various amounts of methyl sulfate compounds. Significant increase of urine sulfate appeared by the end of the shift and correlated with intensity of the air pollution with methyl sulfate compounds. Lower urine excretion of sulfates occurred with longer length of service.

Matrix-assisted laser desorption/ionization for sequencing single-stranded and double-stranded DNA.

The DNA sequence of a single-stranded and double-stranded template was determined. The templates were sequenced using the chain termination method and cycle sequencing method and detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The sequencing products were analyzed successfully without the laborious and expensive methods for removal of the template. Direct sequencing of the double-stranded template was achieved with minimal post-reaction purifications, which could be extremely important for mutation analysis and clinical diagnosis. A systematic study of the mechanisms and kinetics of sequencing reactions was also performed. The details of this analysis and directions for future improvements of the quality of sequencing are presented.