Prevalence of resistant Helicobacter pylori isolates in Bulgarian children

The aim of this study was to assess the primary and combined resistances of Helicobacter pylori isolates obtained from paediatric patients in 2000-2001 to seven antimicrobial agents. Resistance rates of pre-treatment isolates from 115 children were investigated by the limited agar dilution method alone and by the E-test. The cut-off concentrations for resistance were: metronidazole >8 mg/L, clarithromycin and azithromycin >1 mg/L, clindamycin >4 mg/L, amoxicillin >0.5 mg/L, tetracycline >4 mg/L and ciprofloxacin >1 mg/L. Primary resistance rates were: metronidazole 15.8%, clarithromycin 12.4%, azithromycin 14.6%, clindamycin 20.0%, amoxicillin 0%, metronidazole + clarithromycin 4.5%, ciprofloxacin 6.0%, metronidazole + clarithromycin + ciprofloxacin 1.2%, tetracycline 3.1% and metronidazole + ciprofloxacin 1.2%. There were no significant age (1-9 years versus 10-18 years) or gender differences. Prevalence of both macrolide-resistant and intermediately susceptible strains was 21.9% for azithromycin and 15.9% for clarithromycin. Of 18 metronidazole-resistant isolates, 77.8% exhibited a metronidazole MIC > or = 32 mg/L. H. pylori resistance rates to metronidazole, clarithromycin and both agents were relatively low in Bulgarian children. However, resistance was found to all drugs tested except for amoxicillin. The consumption of newer macrolides and tetracyclines could be related to the prevalence of resistance to the corresponding agents. There were no significant differences in primary resistance rates of H. pylori to antimicrobial agents between children and adults except for metronidazole. Multi-drug resistance to newer macrolides, metronidazole and ciprofloxacin in association with a slightly elevated amoxicillin MIC (0.38 mg/L) was detected in one strain.     

Production of herpes B virus recombinant glycoproteins and evaluation of their diagnostic potential

B virus (cercopithecine herpesvirus 1) is the only deadly alphaherpesvirus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n = 40) and -positive (n = 75) macaque sera identified by a whole antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2.     

Unusual distribution pattern of telomeric repeats in the shrews <Emphasis Type="Italic">Sorex araneus</Emphasis> and <Emphasis Type="Italic">Sorex granarius</Emphasis>

Sorex araneus and Sorex granarius are sibling species within the Sorex araneus group with karyotypes composed of almost identical chromosome arms. S. granarius has a largely acrocentric karyotype, while, in S. araneus, various of these acrocentrics have combined together by Robertsonian (Rb) fusions to form metacentrics, with the numbers and types of metacentrics differing between chromosomal races. Our studies on telomeric sequences in S. araneus and S. granarius revealed differences between chromosomes and between species. In S. araneus (the Novosibirsk race), hybridization signals were present on the telomeres of all the chromosomes after FISH with a PCR-generated telomeric probe. In addition, hybridization signals were observed at high frequencies in the pericentric regions of some but not all metacentrics formed by Rb fusion. There were fewer signals on those metacentrics formed earlier in the evolution of S. araneus. This suggests that S. araneus chromosomes retain at least some telomeric repeats during Rb fusion, but that these repeats are lost or modified over time. These results are critical for the interpretation of the well-studied hybrid zones between chromosomal races of S. araneus, given that Rb fission has been postulated in such hybrid zones and that the likelihood of Rb fission will relate to presence/absence of telomeric sequences at the centromeres of metacentrics. In S. granarius, there were strong signals at the proximal (centromeric) telomeres of the acrocentrics after FISH with a DNA telomeric probe. FISH with a PNA telomeric probe on S. granarius acrocentrics showed that the proximal telomeres were 213 kb on average, while the length of the distal telomeres was 3.8 kb on average. Two-colour FISH, using a telomeric DNA probe and a microdissected probe generated from the pericentric regions of the S. granarius chromosomes a and b, revealed regions on distinct chromatin fibres where telomeric and microdissected probes were colocalized or localized sequentially. The proximal telomeres of S. granarius are highly unusual both in their large size and their heterogeneous structure relative to the telomeres of other mammals.     

Blocking of interleukin-10 receptor--a novel approach to stimulate T-helper cell type 1 responses to hepatitis C virus

Chronic hepatitis C virus (HCV) infection is associated with weak CD4+ T-helper type 1 reactivity and enhanced interleukin-10 production to HCV antigens. Here we demonstrate in vitro that monoclonal antibody-induced blockade of IL-10 receptor (IL-10R) generates a favorable balance of CD4+ T-cell responses to HCV. The addition of anti-IL-10R to mononuclear cells leads to a dose-dependent increase of T-cell proliferative response to HCV core, non-structural proteins 3 and 4. In competition experiments, anti-IL-10R reversed the inhibitory effect of IL-10 on HCV-specific T-cell proliferation. Furthermore, the blockade of IL-10R altered the balance towards type 1 antiviral T-cell reactivity with an increased frequency of HCV-specific IFN-gamma producing T-cells and IFN-gamma secretion. The impact of IL-10R blockade on T-cell reactivity to HCV demonstrates the major role of IL-10 in suppressing antiviral T-cell responses. Blocking IL-10 activity may be a useful immunotherapy approach to enhance the efficacy of antiviral treatment in chronic hepatitis C.     

Suppressive effect of immature erythroid cells on the B-cell proliferation

Immature erythroid cells suppress the proliferative response of preactivated B lymphocytes to lipopolysaccharide. The same effect is observed when recombinant human interleukin-2 or culture medium conditioned by preactivated T or B cells is added to cultured cells. Suppressive nucleate erythroid cells are resistant to leucine methyl ester. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 123, No. 1, pp. 66–70, January, 1997     

The “graft versus leukemia” effect under conditions of mixed chimeric hemopoiesis

The “graft versus leukemia” effect can be induced in (C57B1/6×DBA) F₁ mice by two transplantations of cell suspensions containing bone marrow cells and splenocytes of (CBA×C57B1/6) F₁ mice and an injection of cyclophosphamide. The recipients of cell allotransplants were characterized by mixed chimeric hemopoiesis. These data outline a new approach to induction of the graft versus leukemia effect, requiring no complete elimination of the recipient's hemopoiesis and characterized by a low probability of lethal graft-versus-host reaction. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 123, No. 5, pp. 562–565, May, 1997     

Classical swine fever virus: Antigenic architecture of the E2 glycoprotein elucidated by epitope mapping using synthetic peptides

Detailed epitope characterization of the major envelope glycoprotein E2 of the classical swine fever virus (CSFV) is important for our understanding of interactions between the virus and the host immune system, as well as for the development of CSFV-specific diagnostic assays and epitope- or peptide-based marker vaccines. As was shown previously by competitive binding assay, monoclonal antibodies obtained by our group recognize eight epitopes on the E2 protein. Here, we report mapping of five linear, nonoverlapping B-cell epitopes that use a set of synthetic peptides, which encompass the full sequence of the CSFV E2 protein (Shimen strain). Two of the epitopes are located in the antigenic domain A of the E2, while another three belong to a highly structured region of this glycoprotein. The alignment of the identified gene sequences was performed for 12 CSFV strains, three strains of bovine viral diarrhea virus (BVDV), and two strains of the border disease virus (BDV). The data obtained could be used to improve CSFV diagnostic assays, as well as to investigate the effects of aminoacid substitutions in E2 on its antigenic properties.     

High-field head radiofrequency volume coils using transverse electromagnetic (TEM) and phased array technologies

This article describes technological advances in quadrature transverse electromagnetic (TEM) volume coils and phased arrays reported recently from our laboratory developed for MRI and MRS imaging of the human brain. The first part of this work presents a new method for tuning TEM volume coils based on measurements of the radiofrequency current distribution in the coil elements. This technique facilitates bench adjustment of the coils' homogeneity and is particularly important for tuning double-tuned TEM volume coils. We have also used this method to optimize other TEM configurations such as a quadrature TEM half-volume coil and a split TEM coil. TEM half-volume coils provide greater sensitivity over localized regions than conventional full-volume coils, and the split TEM coil provides greater patient access and ease of use. The second part of this work describes the development of single-tuned and double-tuned transmit TEM volume coils in combination with phased arrays. A variety of different techniques for active detuning of single-tuned and double-tuned TEM volume coils are presented along with the development of phased arrays and transmission line preamplifier decoupling. The final section describes the use of counter rotating current (CRC) surface coils in phased arrays. Because of the intrinsic isolation of CRC coils from transmit volume coils, CRC arrays can be used simultaneously with volume coils for both reception and transmission. Near the center of the human head where both the phased array and the volume coil produce similar sensitivities, simultaneous reception enhances the signal-to-noise ratio. Conversely, simultaneous transmission can be used to boost the transmit field in peripheral brain regions from the volume coil to provide a more homogeneous transmit field.     

Immunomorphological characteristics of animals with different levels of orientation and exploratory behavior

We revealed some relationships between the level of orientation and exploratory behavior, functional activity of the immune system, and structural and functional organization of the CNS in animals. Significant differences in brain morphology and expression of cytokine IL-1, type I IL-1 receptor, and erythropoietin receptor genes in brain cells were detected in (CBA×C57Bl/6)F1 mice with different initial levels of orientation and exploratory behavior. Immunocompetent cells of mice with high and low levels of exploratory behavior differ by spontaneous and mitogen-induced proliferative activity. The formation of humoral and cellular immune response in these animals causes opposite changes in exploratory behavior and the type of these changes depends on the initial level of this behavioral reaction.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 10, pp. 466–469, October, 2004