Treatment of intestinal dysbacteriosis in children with diabetes mellitus]

The use of the probiotic Acylact in combination with Viferone containing human recombinant interferon-alpha 2 in the treatment of intestinal dysbacteriosis in children with insulin-dependent diabetes mellitus (IDDM) was shown to be highly efficient. Elimination of microecologic disorders in the intestine was accompanied in all the children by immune rearrangements. The quantity of secretory and serum immunoglobulins and complement increased and the white blood cell chemiluminescence proved to be higher. The correction of the immune status of the children with IDDM resulted in a certain decrease of glycemia. The data are indicative of the fact that such a treatment of intestinal dysbacteriosis in children with IDDM is promising.     

Haemophilus influenzae antibiotic resistant strains isolated in Moscow in 2002-2004]

The results of 3-year observation on emergence of H. influenzae antibiotic resistant strains in Moscow are summarized. The study included 566 strains isolated from patients in 2002-2004. The susceptibility was determined by the 2-fold microdilution method on the Haemophilus test medium. The percentage of the resistant strains isolated in 2002, 2003 and 2004 was the following: ampicillin --4.9, 3.2 and 3.6%, tetracycline--3.3, 3.2 and 1.8% and co-trimoxazole--10.9, 20.9 and 20% respectively. The strains isolated in 2003 and 2004 were resistant to azithromycin in 0.6 and 1.8% of the isolates and to clarithromycin in 1.3 and 3.2% of the isolates respectively. Five isolates differed by the minimum resistance to ampicillin whereas producing no beta-lactamase (BLNAR strains). The drugs of choice for the treatment of respiratory tract infections mainly due to H. influenzae, i.e. acute otitis and sinusitis, chronic bronchitis exacerbation and sometimes pneumonia remain betalactam antibiotics. From the microbiological viewpoint the inhibitor-protected aminopenicillins, cefuroxime and cefotaxime have no significant advantages vs. amoxycillin. The use of cotrimoxazole and chloramphenicol should be considered inexpedient.     

Morphocytochemical and electron microscopic research on murine hybridoma cells producing and not producing monoclonal antibodies

Cytochemical and ultrastructural features of mouse hybridomas and also of the parental cells--myeloma P3-X63-Ag8.653 and spleen cells of the Balb/c mice immunized with cell line RPMI-1788 have been studied. Differences in cytomorphological signs and activity of acid phosphatase, acid nonspecific esterase, nonspecific-alpha-naphthyl acetate esterase were shown in hybrid cell lines secreting and not secreting monoclonal antibodies.     

Transient protein–protein complexes in base excision repair

Abstract

Transient protein–protein complexes are of great importance for organizing multiple enzymatic reactions into productive reaction pathways. Base excision repair (BER), a process of critical importance for maintaining genome stability against a plethora of DNA-damaging factors, involves several enzymes, including DNA glycosylases, AP endonucleases, DNA polymerases, DNA ligases and accessory proteins acting sequentially on the same damaged site in DNA. Rather than being assembled into one stable multisubunit complex, these enzymes pass the repair intermediates between them in a highly coordinated manner. In this review, we discuss the nature and the role of transient complexes arising during BER as deduced from structural and kinetic data. Almost all of the transient complexes are DNA-mediated, although some may also exist in solution and strengthen under specific conditions. The best-studied example, the interactions between DNA glycosylases and AP endonucleases, is discussed in more detail to provide a framework for distinguishing between stable and transient complexes based on the kinetic data.

Communicated by Ramaswamy H. Sarma     

Effects of ouabain on proliferation of human endothelial cells correlate with Na<Superscript>+</Superscript>,K<Superscript>+</Superscript>-ATPase activity and intracellular ratio of Na<Superscript>+</Superscript> and K<Superscript>+</Superscript>

Side-by-side with inhibition of the Na⁺,K⁺-ATPase ouabain and other cardiotonic steroids (CTS) can affect cell functions by mechanisms other than regulation of the intracellular Na⁺ and K⁺ ratio ([Na⁺]_i/[K⁺]_i). Thus, we compared the doseand time-dependences of the effect of ouabain on intracellular [Na⁺]_i/[K⁺]_i ratio, Na⁺,K⁺-ATPase activity, and proliferation of human umbilical vein endothelial cells (HUVEC). Treatment of the cells with 1-3 nM ouabain for 24-72 h decreased the [Na⁺]_i/[K⁺]_i ratio and increased cell proliferation by 20-50%. We discovered that the same ouabain concentrations increased Na⁺,K⁺-ATPase activity by 25-30%, as measured by the rate of ⁸⁶Rb⁺ influx. Higher ouabain concentrations inhibited Na⁺,K⁺-ATPase, increased [Na⁺]_i/[K⁺]_i ratio, suppressed cell growth, and caused cell death. When cells were treated with low ouabain concentrations for 48 or 72 h, a negative correlation between [Na⁺]_i/[K⁺]_i ratio and cell growth activation was observed. In cells treated with high ouabain concentrations for 24 h, the [Na⁺]_i/[K⁺]_i ratio correlated positively with proliferation inhibition. These data demonstrate that inhibition of HUVEC proliferation at high CTS concentrations correlates with dissipation of the Na⁺ and K⁺ concentration gradients, whereas cell growth stimulation by low CTS doses results from activation of Na⁺,K⁺-ATPase and decrease in the [Na⁺]_i/[K⁺]_i ratio.     

Analysis of the species composition of the genus <Emphasis Type="Italic">Chrysopilus</Emphasis> (Diptera,Rhagionidae) in the Eastern Palaearctic Region

The species of the genus Chrysopilus, inhabiting Russia and adjacent territories, are analyzed for the first time. The structure of the genitalia is examined, and new diagnostic characters of the species are given. Some species from the boundary areas are considered in detail, since the species composition of the genus has been poorly studied there. The data on 11 species distributed in eastern Russia and Japan are given.     

Mechanism of interaction between human 8-oxoguanine-DNA glycosylase and AP endonuclease

Human 8-oxoguanine-DNA glycosylase (OGG1) is the main human base excision protein that removes a mutagenic lesion 8-oxoguanine (8-oxoG) from DNA. Since OGG1 has DNA glycosylase and weak abasic site (AP) lyase activities and is characterized by slow product release, turnover of the enzyme acting alone is low. Recently it was shown that human AP endonuclease (APE1) enhances the activity of OGG1. This enhancement was proposed to be passive, resulting from APE1 binding to or cleavage of AP sites after OGG1 dissociation. Here we present evidence that APE1 could actively displace OGG1 from its product, directly increasing the turnover of OGG1. We have observed that APE1 forms an electrophoretically detectable complex with OGG1 cross-linked to DNA by sodium borohydride. Using oligonucleotide substrates with a single 8-oxoG residue located in their 5'-terminal, central or 3'-terminal part, we have demonstrated that OGG1 activity does not increase only for the first of these three substrates, indicating that APE1 interacts with the DNA stretch 5' to the bound OGG1 molecule. In kinetic experiments, APE1 enhanced the product release constant but not the rate constant of base excision by OGG1. Moreover, OGG1 bound to a tetrahydrofuran analog of an abasic site stimulated the activity of APE1 on this substrate. Using a concatemeric DNA substrate, we have shown that APE1 likely displaces OGG1 in a processive mode, with OGG1 remaining on DNA but sliding away in search for a new lesion. Altogether, our data support a model in which APE1 specifically recognizes an OGG1/DNA complex, distorts a stretch of DNA 5' to the OGG1 molecule, and actively displaces the glycosylase from the lesion.     

Evaluation of the usefulness of three collections of bacteriophages for typing methicillin-resistant Staphylococcus aureus,isolated in Moscow hospitals

The typing of S. aureus methicillin-resistant strains, isolated in different hospitals of Moscow; was carried out with the use of three collections of phages: the International Set of Phages; the set of phages of the International Center of S. aureus phage typing in London (L); and the experimental collection of phages of the Gamaleya Institute of Epidemiology and Microbiology in Moscow (M). In this study made with the use of both the phages of the International Diagnostic Set and phages L in the standard typing dose of 1 TP about 6% of the cultures under study proved to be sensitive. When the typing dose was increased to 100 TP the phages of the international diagnostic set lyzed 75.5% of the cultures. The typed strains were found to belong to phage types 77 (71.7%), 77/84/85 (19.6%) and 94/96 (6.5%). At a concentration of 100 TP phages L lyzed 83.7% of the cultures, but the dominating phage types could not be determined due to a great variety of phage markers. In contrast to the two preceding collections, the third phage collection M was composed in such a way that in the study of the investigated culture the specificity of its restriction modification was primarily evaluated and only then the presence of antiphage immunity was determined. This latter collection was used in the evaluation of 93.1% of the cultures. By the specificity of their restriction specification system the majority of them were classified with two new groups, heretofore not described. Only this collection M made it possible to differentiate epidemic and sporadic strains and to evaluate the epidemic situation in all 6 hospitals.     

Intra- and interspecies differences in the ISSR-PCR patterns of bumble bee (Hymenoptera: Bombinae)

Variation in the ISSR patterns was studied in queens of five species of bumble bee habitating on the territory of Ukraine and Belarus. Species-specific characteristics were obtained for B. sylvarum, B. lapidarius, B. pascuorum, B. terrestris, and B. hortorum with using (GTG)7A and (AGC)6G as primers. Some ISSR markers were common for the groups of species. From 73 to 100% ISSR markers were invariant within the species. Individual characteristics of B. terrestris from geographically distant populations from Kiev and Minsk Oblasts were similar. The level of intraspecific variation was different among the species. Taking into account intraspecific constancy of the major ISSR-PCR patterns and in the same time their susceptibility to individual differences we proposed these markers could be used to elucidate some complicated questions of bumble bee taxonomy.