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Biomedical subjects

G A Shipulin

Publications and source records attributed to G A Shipulin.

At least 19 recordsLinked to original sources

[Esomeprasol and rabeprasol effects on esophageal acidification in patients with gastroesophageal reflux disease, intensively metabolizing inhibitors of proton pump].

AIM: To compare action of different esomeprasol and rabeprasol doses on esophageal acidification in patients with gastroesophageal reflux disease (GERD). MATERIAL AND METHODS: A cross-over study was made in 27 GERD patients (12 males and 15 females). It included esophagoduodenoscopy with biopsy for detection of H-pylori and 24-h pH-metry of esophageal content. Rabeprasol and esomeprasol efficacy was compared for daily doses 20 mg of each (stage I) and rabeprasol 20 mg and esomeprasol 40 mg (stage II). RESULTS: All the patients proved H-pylori positive. Before the treatment mean daily time of marked esophageal acidification was 29-36%. At stage I significant differences between the drugs were absent. At stage If esomeprasol was more effective from the first day of therapy both in relation to time to effect and its intensity. CONCLUSION: Esomeprasol provided a fast and noticeable correction of esophageal acidification. This relieves symptoms of GERD and accelerates esophageal mucosa epithelization.

2-Pyridinylmethylsulfinylbenzimidazoles↗

[Alcohol-induced arterial hypertension and genetic polymorphism of alcohol-metabolizing enzymes].

AIM: To study arterial pressure (AP) and heart rate (HR) in patients suffering from alcohol withdrawal syndrome (AWS) with relation to genetic polymorphism of alcohol dehydrogenase-2 (ADG2) and aldehyde dehydrogenase-2 (AlDG2). MATERIAL AND METHODS: AP and HR were analysed for 36 alcoholics (83 hospitalizations, including repeated ones) with genotypes ADG2 and AIDG2 at admission to and discharge from narcological hospital. RESULTS: ADG2 genotypes distribution was the following: ADG2-1/1--61.1% (n = 22); ADG2-1/2--36.1% (n = 13); ADG2-2/2--2.8% 9n = 1). All the patients had a genotype AIDG2-1/1. A hypertensive reaction occurred in 75.9% inpatients with AWS. At admission, patients with genotype ADG2-1/1 had significantly higher systolic AP and HR vs those with allele ADG2-2: 146.6 +/- 17.0 mmHg against 141.2 +/- 14.9 mmHg, 95.6 +/- 13.9 b/min vs 88.5 +/- 12.2 b/min, respectively. In homozygous genotype ADG2-1/1 AP and HR were higher than in heterozygotes: SAP 152.3 +/- 12.9 mm Hg vs 145.2 +/- 16.2 mm Hg, pulse pressure 57.2 +/- 11.9 vs 50.0 +/- 15.1 mm Hg, HR 96.8 +/- 13.4 b/min vs 87.7 +/- 12.0 b/min. The groups had similar mean diastolic pressure. At discharge, AWS standard therapy resulted in a significant lowering of AP and HR in the study group. Mean values of the parameters in groups with different genotypes did not differe at discharge. CONCLUSION: Population of alcoholics from the Moscow Region had allele polymorphism ADG2. Genetic polymorphism AlDG2 is not typical for this group. Hypertensive reaction was registered in the majority of alcoholics in AWS. Higher systolic, pulse pressure and heart rate were significantly higher in the AWS group with genotype ADG2-1/1. Controlled alcohol withdrawal entails a significant reduction of AP and HR.

Adult↗

[Development and evaluation of the kit for detection of SARS-associated Coronavirus RNA].

AIM: To develop a diagnostic kit for detection of SARS (severe acute respiratory syndrome)-related coronavirus RNA based on reverse transcription and polymerase chain reaction and to estimate its specificity and sensitivity. MATERIAL AND METHODS: 68 virus and bacterial cultures, 240 clinical samples from people without SARS symptoms and also 22 RNA samples from patients with SARS symptoms received during the epidemic in Beijing were used. RESULTS: The specificity of the kit was determined using animal coronaviruses and other bacterial and viral strains, causing acute respiratory and intestinal infections, and was shown to be 100%. The sensitivity of the kit in different clinical samples was 2.2 x 10(3) genome equivalents of recombinant SARS RNA in 1 ml of the specimen. The kit was evaluated in the Institute of Microbiology and Epidemiology of Beijing (China) using SARS-cov viral suspension and clinical samples from patients with suspected SARS. It was shown that kit was able to detect 10 TCID/50 ml of SARS-Cov virus. Testing of clinical samples from patients with suspected SARS showed that diagnostic sensitivity of the kit was 95%. Detection of the SARS-Cov RNA was more effective in feces compared to sputum 990 and 40%, respectively). CONCLUSION: The kit "AmpliSens SARS" for qualitative detection of SARS-related coronavirus RNA by reverse transcription and polymerase chain reaction (PCR) in nasopharyngeal wash/aspirates, naso/oropharyngeal swabs, plasma, and extract from feces has been developed in the Central Research Institute for Epidemiology of the RF Ministry of Health. The kit contains reagents for RNA isolation and purification, cDNA synthesis by reverse transcription of RNA, for PCR and for electrophoretic analysis of amplified products. The kit also contains recombinant positive and internal control samples allowing to control efficiency of analysis and showed good analytical and diagnostic characteristics.

Evaluation Studies as Topic↗

[Multilocus sequence-typing for characterization of Moscow strains of Haemophilus influenzae type b].

Haemophilius influenzae, type b (Hib) bacteria, were genotyped by multilocus sequence typing (MLST) using 5 loci (adk, fucK, mdh, pgi, recA). 42 Moscow Hib strains (including 38 isolates form cerebrospinal fluid of children, who had purulent meningitis in 1999-2001, and 4 strains isolated from healthy carriers of Hib), as well as 2 strains from Yekaterinburg were studied. In MLST a strain is characterized, by alleles and their combinations (an allele profile) referred to also as sequence-type (ST). 9 Sts were identified within the Russian Hib bacteria: ST-1 was found in 25 strains (57%), ST-12 was found in 8 strains (18%), ST-11 was found in 4 strains (9%) and ST-15 was found in 2 strains (4.5%); all other STs strains (13, 14, 16, 17, 51) were found in isolated cases (2.3%). A comparison of allelic profiles and of nucleotide sequences showed that 93% of Russian isolates, i.e. strain with ST-1, 11, 12, 13, 15 and 17, belong to one and the same clonal complex. 2 isolates from Norway and Sweden from among 7 foreign Hib strains studied up to now can be described as belonging to the same clonal complex; 5 Hib strains were different from the Russian ones.

Adenylate Kinase↗

[Laboratory diagnostics of Crimean hemorrhagic fever by polymerase chain reaction].

Our group developed, within the present case study, two techniques' variations, i.e. a single-step RT-PCR and nested RT-PCR assays, for the purpose of detecting the Crimean-Congo hemorrhagic fever RNA virus in human samples. The above assays as well as those previously recommended by the Ministry of Health of the Russian Federation were simultaneously used in 14 clinical samples obtained from patients with Crimean hemorrhagic fever. After assessing the detection accuracy, it was found that the developed-by-us test system displayed the same or even better diagnostic values versus the previously recommended nested RT-PCR and a 1000-fold advantage over the previously recommended single-step RT-PCA. The single-step RT-PCR assay variation is always more preferable in sense of technical and economic motivations. Finally, the test system developed by us has every reason to become the method of choice in routine PCR diagnosis of Crimean hemorrhagic fever after all official trials and approvals are duly complied with.

DNA Primers↗

[Comparative effectiveness of the antisecretory action of rabeprazole and esomeprazole in people with rapid metabolism of proton pump inhibitors].

AIM: To compare antisecretory effects of Rabeprazole and Esomeprazole in proton pump inhibitors extensive metabolizers in an open, randomized, two-way crossover study. METHODS: Sixteen GERD H. pylori-positive patients (8 men, mean age 49.6 and 8 women, mean age 49.3) with the homozygous extensive metabolizer genotype of CYP2C19 determined by polymerase chain reaction-restriction fragment length polymorphism analysis received Rabeprazole 20 mg or Esomeprazole 20 mg daily on days 1-6, with a 14-day wash-out period. Intragastric pH was recorded continuously on days 0, 1, 5 and 7. RESULTS: On days 1 and 5 no differences were found between Rabeprazole 20 mg and Esomeprazole 20 mg in 24-hours median pH (day 1: 5.9 versus 5.0; day 5: 6.45 versus 6.3) or in percent of time with pH 4 (day 1: 57.8% versus 50.5%; day 5: 81.4% versus 81.2%). On day 1, mean percent of time pH 4 were significantly greater after Esomeprazole 20 mg 52.6% (95% CI: 23.6-68.2) than Rabeprazole 20 mg 33.0% (95% CI: 15.3-48.2) during 0-6 h (p = 0.02). On day 7 (24 later the last dose), 24-hours median pH was higher after Esomeprazole 20 mg than Rabeprazole 20 mg (2.7 versus 5.05; p = 0.02). CONCLUSIONS: Rabeprazole 20 mg and Esomeprazole 20 mg are equally effective in increasing gastric pH in H. pylori-positive PPI extensive metabolizers on days 1 and 5. Esomeprazole 20 mg is more effective than Rabeprazole 20 mg in maintaining pH 4 during the first 6 hours on the first day and increasing of intragastric pH on the day 24 hours later the last dose.

2-Pyridinylmethylsulfinylbenzimidazoles↗

[Determination of HIV RNA concentration for evaluation of the efficacy of combined therapy of HIV infection].

Plasma HIV RNA (viral load) and count of CD4+ T cells were evaluated in 23 patients with HIV infection treated with invirase (1800 mg/day), zidovudine (600 mg/day), and zalcitabine (2.25 mg/day) for 6 months in order to evaluate the efficiency of antiretroviral therapy. Viral load was measured by AMPLICOR HIV-1 Monitor test. The reproducibility of HIV RNA measurements was in line with reported data (CV 15-41%), allowing highly accurate (15%) evaluation of RNA in a standard control sample provided by National Institute for Biological Standards and Control, Great Britain. Plasma HIV RNA concentration decreased to an undetectable level (below 400 RNA copies/ml plasma) after 6 months of treatment in 52.2% patients. In 17.4% the therapy failed, and in 30.4% it resulted in a reduction of viral load to > 1 lg, although HIV RNA was still detected in the plasma after 6-month therapy. The count of CD4+ T cells increased by 9.5%. Changes in the viral load outstripped changes in CD4+ cells. Viral load was in high correlation with the count of CD4+ lymphocytes: -0.53, p = 0.01 before treatment and -0.61, p = 0.002 after 6-month treatment.

Acquired Immunodeficiency Syndrome↗

[Multilocus sequencing--a new method of genotyping bacteria and first results of its use].

Comparative characterization (molecular typing) of isolates within a bacterial species is one of the major problems in microbiology and epidemiology. However, it is rather difficult to correlate data obtained in various laboratories, because traditional, including molecular, methods employed in typing pathogenic microorganisms cannot be standardized. In 1998, Maiden et al. proposed multilocus sequence typing (MLST); through which alleles of several housekeeping genes are directly assessed by nucleotide sequencing, each unique allele combination determining a sequence type of a strain. The advantages of this approach are that the culturing of pathogenic microorganisms is avoided, as their gene fragments are amplified directly from biological samples, and that the sequencing data are unambiguous, easy to standardize, and electronically portable. The latter makes it possible to generate an expandable global database for each species at an Internet site, in order to use it for the purposes of genotyping pathogenic bacteria (and other infectious agents). MLST protocols have been elaborated for Neisseria meningitidis, Streptococcus pneumoniae, and Helicobacter pylori; those for Streptococcus pyogenes, Staphylococcus aureus, and Haemophilus influenzae are now being developed. Basic principles and the first results of MLST have been reviewed, including data on the distribution and microevolution of N. meningitidis clones causing epidemic meningococcal infection, the relative recombination and mutation rates in the N. meningitidis genome, the identification of antibiotic-resistant S. pneumoniae clones causing severe generalized infection, the grouping of H. pylori isolates from various geographic regions, etc.

Bacteria↗

Meningococcal disease and polymorphism of FcgammaRIIa (CD32) in late complement component-deficient individuals.

Late complement component-deficient (LCCD) individuals lack plasma bactericidal activity and are highly susceptible to meningococcal disease. Phagocytosis plays a significant role in immune defence against meningococci and involves FcgammaRIIa (CD32) on leucocytes. Two allotypic forms are currently recognized: FcgammaRIIa-R131 and RIIa-H131. Neutrophils with the IIa-H/H131 allotype are more effective in phagocytosis than IIa-R/R131. We studied the distributions of IIa-R131 and IIa-H131 allotypes among 29 Russian LCCD patients who had suffered from recurrent episodes of meningococcal disease. The distribution of IIa-R/R131 to heterozygous IIa-R/H131 to homozygous IIa-H/H131 genotypes was 0.14:0.29:0.57 for LCCD patients who developed the first episode of disease before 10 years of age. The distribution was 0.21:0.64:0.14 for patients who experienced meningococcal disease above the age of 10 years (chi2 = 6, P < 0.05, odds ratio for IIa H/H131 versus R/R131 = 8). Meningococcal disease had a 'grave' course in 14 of 31 disease episodes in patients with IIa-R/R131 and IIa-R/H131 allotypes, in contrast to 1 of 18 episodes in patients with IIa-H/H131 allotype (chi2 = 7, P < 0.01, odds ratio = 14). We conclude that IIa-H/H131 individuals appear to have a higher acquired antibody-mediated phagocytosis-dependent resistance to meningococcal disease above the age of 10 years. Additionally, effective CD32-mediated phagocytosis may restrict the severity of meningococcal disease in LCCD patients with IIa-H/H131 phenotype.

Adolescent↗

Association of human Fc gamma RIIa (CD32) polymorphism with susceptibility to and severity of meningococcal disease.

Phagocytosis of bacteria constitutes an important defense mechanism against invasive bacterial diseases. Efficacy of phagocytosis by polymorphonuclear neutrophils is known to vary between allotypes of Fc gamma RIIa (a class of Fc receptors for immunoglobulins that is constitutively expressed on neutrophils). We compared the distribution of Fc gamma RIIa-R131 and Fc gamma RIIa-H131 allotypes in 98 Slavic complement-sufficient patients with meningococcal disease with that of the allotypes in 107 healthy controls. A strong association was found between the IIa-R/R131 allotype and the development of meningococcal disease after the age of 5 years, compared with IIa-R/H131 and IIa-H/H131 allotypes (P < .03; odds ratio [OR], 2.9). A severe course of meningococcal disease was observed in 21 (68%) of 31 episodes in patients with IIa-R/R131 genotype and in 22 (54%) of 41 episodes in patients with IIa-R/H131 genotype, in contrast to eight (31%) of 26 episodes in patients with IIa-H/H131 genotype (P < .02; OR, 4.7). Our data show that individuals older than 5 years of age who have the IIa-H/H131 allotype are less susceptible to severe meningococcal disease than are individuals with the IIa-R/R131 or IIa-R/H131 genotype.

Adolescent↗

Heterozygous C8beta complement deficiency does not predispose to meningococcal disease.

We have identified 42 Russian patients with homozygous C8beta complement component deficiency, all of whom had experienced at least one episode of systemic meningococcal disease. About 90% of these individuals have a C --> T exchange in exon 9, leading to a premature stop codon. If, like the homozygous-deficient state, heterozygous C8beta deficiency constitutes a risk factor for meningococcal disease, it would be expected to be detected with increased frequency among individuals suffering from this disease. Using allele-specific polymerase chain reaction (PCR), we studied 153 consecutive patients with meningococcal disease admitted to the Moscow Hospital for Infectious Diseases to determine the frequency of C8 null allele. No individuals with heterozygous C --> T exchange were identified among these 153 patients, despite the fact that seven persons were detected who had homozygous C8beta deficiency, caused by the same C --> T exchange in exon 9, and one patient who had C7 component deficiency. Thus, heterozygous deficiency, although more frequent than homozygous deficiency in the general population, does not result in a substantial increase in susceptibility to meningococcal disease.

Complement C8↗

Molecular bases of combined subtotal deficiencies of C6 and C7: their effects in combination with other C6 and C7 deficiencies.

Combined subtotal deficiency of C6 and C7, in which both proteins are expressed at very low levels, has been observed in homozygous form in two families. A defect at the 5' splice donor site of intron 15 of the C6 gene explains the low molecular weight of the C6 protein and is probably responsible for its low expressed concentration. The C7 defect is more enigmatic: the protein is of normal molecular weight, low circulating concentration, and altered isoelectric point. An Arg > Ser codon substitution in exon 11 is the only molecular alteration within the mature C7 protein. These defects are associated with a characteristic set of polymorphic DNA markers in the C6/C7 region, forming a distinct haplotype. The haplotype has been found in combination with a number of other haplotypes containing defective genes that lead either to C6 or C7 deficiency, but with different consequences. Where it is combined with a C6-deficient gene, the serum C7 levels can be surprisingly high, possibly because there is no C6 generating C56 to consume the C7. In contrast, where the C7 genes are both defective (but still partially functional), there may be a profound deficit of circulating C7 because there is ample C6 to produce C56 and consume the already small amount of C7. Each molecular defect has also been found in isolation and has the expected effect.

Amino Acid Sequence↗

Outbreak of West Nile virus infection, Volgograd Region, Russia, 1999.

From July 25 to October 1, 1999, 826 patients were admitted to Volgograd Region, Russia, hospitals with acute aseptic meningoencephalitis, meningitis, or fever consistent with arboviral infection. Of 84 cases of meningoencephalitis, 40 were fatal. Fourteen brain specimens were positive in reverse transcriptase-polymerase chain reaction assays, confirming the presence of West Nile/Kunjin virus.

Animals↗

[The prospects for the diagnosis of bacterial meningitis].

The samples of spinal fluid arriving to the Clinical Infectious Hospital in 1994-1996 with the clinical diagnosis "generalized form of meningococcal infection" or "purulent meningitis of unclear etiology" were studied. The etiological agent was bacteriologically identified in 35% of 487 patients (in 25% of cases Neisseria meningitidis, in 7% of cases Streptococcus pneumoniae and in 2% of cases Haemophilus influenzae, type b, were detected). The method of latex agglutination, used in this study, was highly specific (100%) and moderately sensitive (67%); this method made it possible to diagnose 25% of cases additionally (N. meningitidis in 15% of cases, S. pneumoniae in 5% of cases and H. influenzae in 3% of cases). Diagnostics with the use of PCR was characterized by high specificity (> 97%) and sensitivity (> 85%) relatively to the "golden standard" of microbiological diagnostics. There were few false positive results (3 samples), caused probably by contamination at the moment of taking the samples. For this reason the results obtained by PCR could be used for diagnostic purposes even in cases of negative results given by other methods. Tests with the use of PCR made it possible to diagnose 29% more cases additionally (in 26% of cases N. meningitidis DNA and in 3% of cases S. pneumoniae DNA were detected. Thus the complex of methods used in this study permitted the detection of the etiological agent altogether in 87% of cases.

Antigens, Bacterial↗

[Isolation of the West Nile Fever virus from human patients during an epidemic outbreak in the Volgograd and Astrakhan regions].

Two strains of West Nile virus LEIV 27889 Vig and Ast 986 were isolated from the brain of a dead subject and from the blood of a patient, respectively, during an outbreak of serous meningitis and meningoencephalitis in July-September, 1999, in the Volgograd region, Krasnodar territory, and Astrakhan region. These strains reacted with convalescent sera in hemagglutination inhibition test, which proves their etiological role in this outbreak.

Antigens, Viral↗

[Pilot experience of studying calicivirus infection in Moscow children].

An RT-PCR-based assay for detecting calicivirus RNA was developed. Samples of feces of 124 children patients with acute gastroenteritis were tested for the presence of NLV-1, NLV-2, and SLV. Calicivirus infection was detected in 6.4% of cases of children with acute gastroenteritis. All isolates were non-homogeneous Norwalk like Viruses 2 genotype with maximum nucleotide homology to the strains, isolated in UK and Japan. The use of the RT-PCR assay developed in our laboratory allowed the etiology of 22.2% of cases of acute gastroenteritis of uncertain etiology to be identified.

Acute Disease↗

[Evaluation of the diagnostic efficacy of a test system for detecting hepatitis virus C RNA by polymerase chain reaction "AmpliSens(TM) HCV-240/BKO-440)"].

The test system developed at the Central Research Institute of Epidemiology, Ministry of Health of the Russian Federation for identification of hepatitis C virus RNA was studied. The sensitivity of the test system which the rate of similar results was 100% with its 5-fold reproduction was evaluated. That was 5 x 103 genomic equivalents (or international units) per ml of a sample. A scheme for evaluation of the reproductibility of test systems based on the polymerase chain reaction (PCR) by using model samples is proposed. Whether it can be used for intra- and extra-laboratory assessment of the quality of PCR analyses is discussed.

Hepacivirus↗

[Stability of hepatitis C virus RNA in human blood plasma in a panel of control samples].

The stability of hepatitis C virus (HPC) RNA concentration in 5 human plasma samples after storage at +22 degrees C for two months, at -20 degrees C, and +4 degrees C for six months after 10 freezing-unfreezing cycles was evaluated. In this study, the concentration of HCV RNA in the samples was stable after six months of storage at -20 degrees C. The concentration of HCV RNA decreased on the average of 92% after 2-month storage at +22 degrees C. After six months of storage at +4 degrees C and after 10 freezing-unfreezing cycles, that decreased by 28 and 42%, respectively. Based on their own findings, the authors developed a HCV-RNA panel containing 5 positive human plasma samples with RNA levels of 103-105 IU/ml. The panel may be recommended both for the standardization of PCR kits and for the intra- and interlaboratory quality control of PCR laboratories.

Cryopreservation↗