Differential Expression of Local Immune Response Genes in the Vagina: Implication for the Diagnosis of Vaginal Infections.

Transcription profiles of genes of local immune response were determined in the vagina of women with bacterial vaginosis, aerobic vaginitis, and vulvovaginal candidosis for detection of the most specific immune markers for these vaginal infections. Laboratory diagnosis of the vaginal infections was performed microscopically; the inflammatory reaction in the vagina (leukorrhea) was defined as the presence of >10 white blood cells per field of view. Transcription profiles of IL1b, IL10, IL18, TNFα, TLR4, GATA3, and CD68 were determined using reverse-transcription quantitative real-time PCR. The strongest predictors of aerobic vaginitis were increased levels of IL1b and IL10 mRNA. Bacterial vaginosis was strongly associated with reduced levels of IL18 and GATA3 mRNA. Increased levels of IL1b and TLR4 transcripts showed significant discriminatory power for vulvovaginal candidosis and leukorrhea. The results of this study suggest differential expression of local immune response genes in the vagina of women with different vaginal infections. Detection of specific immune markers in the vagina using reverse-transcriptase PCR could supplement PCR detection of abnormal vaginal microflora for the diagnosis of vaginal infections.

[Distribution and genetic organization of the pathogenicity island XII among the clinical strains of GBS].

Streptococcus agalactiae or group B streptococci (GBS) is the major cause of various diseases of the newborns and the adults. The genome of GBS contains from 14 to 15 mobile genetic elements (pathogenicity islands, PAI). It is well-known that many GBS virulence determinants are localized not in the core genome, but on the pathogenicity islands. The goal of this work was to determine the distribution and genetic organization of PAI XII containing virulence genes sspB1, scpB, lmb among the clinical strains of GBS. 74 clinical strains of GBS were analyzed using PCR with primers corresponding to the genes of the virulence factors located on PAI XII. The pathogenicity island XII was determined only in 22% of the clinical strains. The genetic organization of the island was different between strains. There was no correlation between the presence of PAI and the serotype of GBS.

[Analysis of point mutations in the ygeD, gyrA and parC genes in fluoroquinolones resistant clinical isolates of Chlamydia trachomatis]. / Analiz tochechnykh mutatsii v genakh ygeD, gyrA i parC klinicheskikh izoliatov Chlamydia trachomatis, ustoichivykh k ftorkhinolonam.

Resistance of 14 clinical isolates of C. trachomatis to fluoroquinolones, i.e. of ciprofloxacin, pefloxaxin and ofloxacin, was assayed. Three isolates with a high resistance degree to all 3 drugs (MIC equal or above 64 microg/ml) were detected. MIC was found to be equal to or below 4 microg/ml for 3 isolates. The remaining isolates had an intermediate resistance level. The nucleotide sequence was established for the Quinolone-Resistance Determining Region (QRDR) genes coding the DNA-gyrase subunit A (gyrA) and DNA-topoisomerase IV subunit C (parC) as well as for the 3'-region of ygeD coding, presumably, the efflux protein. In none of the isolates, the gyrA and gyrC QRDR differed from the corresponding regions in the published C. trachomatis genome sequence. Several silent mutations and mutations resulting in amino acid substitutions were observed in the ygeD 3' region of 2 isolates resistant to high FQ concentrations and in 1 isolate with the intermediate resistance level.

Mutations in a 23S rRNA gene of Chlamydia trachomatis associated with resistance to macrolides.

For six clinical isolates of Chlamydia trachomatis, in vitro susceptibility to erythromycin, azithromycin, and josamycin has been determined. Four isolates were resistant to all the antibiotics and had the mutations A2058C and T2611C (Escherichia coli numbering) in the 23S rRNA gene. All the isolates had mixed populations of bacteria that did and did not carry 23S rRNA gene mutations.

[Experience in wilaprafen treatment of chlamydial prostatitis, complicated by infertility]. / Opyt lecheniia vil'prafenom khlamidii prostitia, oslozhnennogo besplodiem.

The antichlamydial action of wilprafen was studied in patients suffering from urogenital chlamydial prostatitis complicated by infertility. An individual approach to etiotropic and pathogenetic therapy with wilprafen alone and in combination with vobenzim introduced by the authors in men with chlamydial prostatitis complicated by infertility produced a high curative effect.

[Allelic polymorphism of the tem(M) determinant in Mycoplasma hominis and Ureaplasma urealyticum clinical isolates resistant to tetracyclines]. / Allel'nyi polimorfizm tet(M) determinanty v klinicheskikh izoliatakh Mycoplasma hominis i Ureaplasma urealyticum, ustoichivykh k tetratsiklinam.

Of the 130 clinical isolates of Mycoplasma hominis from patients with nonspecific inflammatory diseases of the urogenital tract (UGT), approximately 10% contained the tet(M) gene after the course of treatment with tetracyclines. This gene was found in nine (25%) of the 36 Ureaplasma urealyticum clinical isolates. The nucleotide sequence of 13 tet(M) genes in TcR clinical isolates of M. hominis and five genes in U. urealyticum TcR clinical isolates was determined. A comparison of nucleotide sequences of eight tetM genes of different origin and tet(M) genes of Gardnerella vaginalis and M. hominis and U. urealyticum clinical isolates showed that the mosaic structure of the tet(M) gene is completely identical in 11 of 13 M. hominis TcR isolates but belongs to an unidentified allele different from those described earlier, Another new allelic variant of tet(M) was found in two isolates. In three of five TcR clinical isolates of U. urealyticum, a tet(M) gene, whose mosaic structure was identical to that of tet(M) reported previously for ureaplasmas, and also two new allelic variants, which have not been described so far, were found.

Drift of tetM determinant in urogenital microbiocenosis containing mycoplasmas during treatment with a tetracycline antibiotic.

We studied the correlation between genetic transfer of tetM determinant in Tn916 conjugative transposon by urogenital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) and changes in the bacterial repertoire during treatment with a tetracycline antibiotic. Basic conditions favoring the nonspecific transfer of tetM determinant into mollicute cells are determined and the allele polymorphism of tetM determinant in clinical strains of M. hominis and U. urealyticum is evaluated. The structure of tetM gene in clinical mycoplasma and ureaplasma strains is characterized by a peculiar mosaic pattern and differs from all previously described alleles of this gene. The results suggest that tetracycline resistance in mollicutes is determined by mechanisms alternative to genetic transfer of tetM determinant.

Evaluation of antibiotic sensitivity of Chlamydia trachomatis using RT-PCR.

The sensitivity of 11 clinical strains of Chlamydia trachomatis to azithromycin, ofloxacin, doxycycline, and erythromycin was evaluated. The minimum inhibiting concentrations of all antibiotics for 90% strains, determined by PCR with reverse transcription of omp3B gene RNA (GenBank U68443) corresponded to, and those with reverse transcription of 16S rRNA gene RNA (GenBank X54451) far surpassed the minimum bactericidal concentrations for 90% strains determined by direct immunofluorescence with monoclonal antibodies to the major outer membrane protein.

Genetic heterogeneity of Mycoplasma hominis clinical isolates detected during observation of patients with recurrent urogenital inflammation.

A rapid reproducible effective method for molecular typing of Mycoplasma hominis strains based on random amplified polymorphic DNA (RAPD) technique was developed. RAPD detected genetic heterogeneity of genomes of Mycoplasma hominis clinical isolates and showed changes in the genomes of Mycoplasma hominis clinical isolates from patients with chronic infection.

[Analysis of regions determining resistence to fluoroquinolones in genes gyrA and parC in clinical isolates of Mycoplasma hominis]. / Analiz raionov, opredeliaiushchikh rezistentnost' k ftorkhinolonam, genov gyrA i parC v klinicheskikh izoliatakh Mycoplasma hominis.

Fifteen strains of M. hominis isolated from patients with urogenital inflammations were analyzed. Variations in the quinolone resistance-determining regions (QRDR) have been found in fluoroquinolone-resistant M. hominis clinical isolates in comparison with the reference PG21 strain. In one isolate, parC had Asn substitute at position 91.

[Current problems in the clinical course, diagnosis and treatment of Chlamydia infection in newborns]. / Aktualnye problemy kliniki, diagnostiki i lecheniia khlamidiinoi infektsii u novorozhdennykh detei.

A comprehensive study of the course of the early neonatal period in 120 newborns infected with Chlamydia, analysis of somatic and obstetrical and gynecological anamnesis and the course of gestation, labor, and postpartum period in their mothers, and prospective clinical and microbiological examinations of these infants up to the age of 1 year revealed that the fetus is infected not only during delivery, but antenatally as well. The disease runs an extremely grave course in the neonates, often with generalization of the process. Chlamydial infection in the early neonatal period depends on the time and massiveness of infection of a child, the degree of morphofunctional maturity of the baby and presence of concomitant diseases related to unfavorable conditions of intrauterine development; it may take the following clinical forms: intrauterine sepsis, meningoencephalitis, intrauterine pneumonia, respiratory distress syndrome, gastroenteropathy, conjunctivitis. Problems in the diagnosis and treatment of the disease are discussed.