Expression of Pattern Recognition Receptors by Nociceptive Metasympathetic Neurons.

Immunohistochemical assay with double label and confocal laser scanning microscopy showed that innate immunity receptor TLR4 is expressed predominantly in neurons of the intestinal Auerbach (myenteric) plexus, while vanilloid nociceptive receptor TRPV1 is expressed by neurons of Meissner (submucous) plexus. Immunohistochemical analysis with triple labeling revealed coexpression of TLR4 and TRPV1 in enteric neurons of rat colon. The results attest to a possibility of functional interaction between Toll-like and vanilloid receptors in the neuron level.

[Relapsing (recurrent) disease caused by Borrelia miyamotoi].

AIM: To clarify the clinical, laboratory, and epidemiological characteristics of relapsing Ixodes tick-borne borreliosis (ITB) caused by Borrelia miyamotoi. SUBJECTS AND METHODS: Retrospective clinical observation was made in 79 inpatients of the Republican Infectious Diseases Hospital (Udmurt Republic), who had been diagnosed with B. miyamotoi-caused disease verified by real-time polymerase chain reaction. The latter and enzyme immunoassay ruled out possible vector-borne coinfections (ITB caused by B. burgdorferi sensu lato; tick-borne encephalitis; anaplasmosis; and ehrlichiosis). RESULTS: The recurrent course of the disease was observed in 8 (10%) of the 79 patients. The relapsing fever curve was noted in 6 of the 8 patients; 4 patients had 2 episodes of fever and 2 patients had 3 episodes; the wave-like continuous type of fever cannot enable one to estimate the specific number of episodes in 2 more cases. Relapses occurred in all the 8 patients before antibiotic treatment. Febrile syndrome (weakness, headache, chill, fever, sweating, dizziness, nausea, vomiting, myalgia, and arthralgia) was leading in patients with relapses. These patients were less frequently observed to have signs of organ dysfunctions than those with one episode of fever. The values of clinical and biochemical blood tests and urinalyses were normal and near-normal in the majority of patients on hospital admission. CONCLUSION: Relapsing B. miyamotoi infection cases detected in the directed study proved to be unrecognized by practical health authorities during the first and sometimes second episodes of fever. This indicates that the prevalence of this disease is essentially underestimated and there is a need to increase physicians' alertness and awareness and to introduce adequate diagnostic methods.

[Non-invasive techniques in diagnosis of steatosis and fibrosis in nonalcoholic fatty liver disease].

UNLABELLED: Diagnosis and treatment of nonalcoholic fatty liver disease (NAFLD) is one of the most actual problems of modern hepatology. Ultrasound elastometry is alternative method to determine the density of the liver tissue. The efficiency of the method was confirmed by a large amount of data in patients with viral hepatitis. However, the diagnostic value of elastometry was not studied enough in patients with NAFLD. THE AIM OF THE RESEARCH: to study the possibility of non-invasive diagnostic methods of examination in a complex assessment of the liver in patients with NAFLD. MATERIALS AND METHODS: We studied 24 patients with signs of NAFLD and a long persistent elevation of liver enzymes. All patients underwent a needle biopsy of the liver. We conducted a comparative analysis of the results of the morphological study of the liver tissue with ultrasound elastometry, computed tomography of the liver, biochemical blood tests. RESULTS AND DISCUSSION: Among biochemical liver enzymes ALT was a marker of NAFLD which characterized the transformation of steatosis to steatohepatitis. The combined ALT and GGT increase reflected the high activity of inflammation in the liver tissue. Elastometry results were comparable to the histological features in all liver fibrosis stages, but the maximum diagnostic accuracy was observed at the late stages. The area under the ROC-curves showed the highest precision in the F2-F3 stages, the lowest value - in the F1 stage. Imprecision of the method in the early stages of fibrosis may be due to the hepatic steatosis in NAFLD and high biochemical activity with cholestasis signs. Patients with early-stage liver fibrosis diagnosed with elastometry need additional examination using other methods of noninvasive diagnostic tools.

[Mosquito complex (Diptera, Culicidae) in a West Nile fever focus in the Volgograd Region. II. Host-feeding patterns of mosquitoes in different habitats].

Host preference of the mosquitoes collected in the urban and rural habitats of Volgograd and its suburbs was studied by the precipitation reaction test. Human and avian blood was detected in Cx. pipiens, Cx. modestus, Ae. vexans, Ae. behningi, Ae. caspius, Ae. sticticus, and females of the Anopheles maculipennis. The proportion of the mosquitoes fed on birds was similar in the urban and rural biotopes whereas that of the mosquitoes feeding on humans was significantly higher in Volgograd than in its environs. The increase in the number of human blood-fed mosquitoes in the city resulted mainly from the females collected in its multi-storied buildings.

Economic evaluation of Haemophilus influenzae type b vaccination in Moscow, Russian Federation.

OBJECTIVE: To estimate the cost-effectiveness of providing Haemophilus influenzae type b (Hib) vaccine to children in Moscow in routine immunization services. METHODS: The incidence of Hib meningitis among children aged <5 years in Moscow was obtained from a prospective surveillance study undertaken during October 1999-September 2001, with treatment cost data collected for all cases. Sequelae in surviving children were assessed in December 2002. The costs of Hib vaccination in Moscow were estimated assuming a vaccine price of US dollar 5 per dose and the same four-dose schedule and 97% coverage as for diphtheria-tetanus-pertussis vaccine. The most uncertain variables were varied in a sensitivity analysis. RESULTS: The annual incidence of Hib meningitis was 5.7 per 100,000 children <5 years. The average treatment cost for an acute Hib meningitis case was US dollar 1296. For a patient with sequelae, the average additional lifetime discounted treatment cost was US dollar 15,820. The total annual cost of Hib vaccination of infants in Moscow was estimated as US dollar 1.5 million per year. In the base case analysis, the cost-effectiveness ratios amount to US dollar 77,503 per Hib meningitis case averted and US dollar 10,842 per discounted disability adjusted life year averted. The break-even vaccine price, where the annual vaccination costs equal annual treatment costs averted, is only US dollar 0.04 per dose in the base case scenario. If discounted indirect costs are included, the break-even vaccine price is US dollar 0.5 per dose. CONCLUSION: In Moscow, the incidence of Hib meningitis is low and the costs of hospitalization and subsequent medical treatment are relatively inexpensive. Given these factors, Hib vaccine at US dollar 5 per dose would not be a cost-effective option in Moscow at the present time.

[Multilocus sequence-typing for characterization of Moscow strains of Haemophilus influenzae type b]. / Kharakteristika moskovskikh shtammov Haemophilus influenzae tipa b metodom mul'tilokusnogo sekvenirovaniia-tipirovaniia.

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.

[Characteristics of antitoxic and antibacterial immune response in nontoxic forms of oropharyngeal diphtheria]. / Pokazateli antitoksicheskogo i antibakterial'nogo immunnogo otveta pri netoksicheskikh formakh difterii rotoglotki.

The enzyme immunoassay system (EIA) for differentiation of antibodies in therapeutic heterogeneous antitoxic serum and antibodies to Corynebacterium diphtheriae toxigenic strains in patients and carriers was developed. The use of EIA permitted the dynamic evaluation of the characteristics of humoral antitoxic and antibacterial immune response in 50 patients with the localized and disseminated forms of stomatopharyngeal diphtheria and 14 "healthy" carriers of toxigenic C. diphtheriae. As revealed in this study, the symptoms of the disease in patients with disseminated forms of stomatopharyngeal diphtheria developed in the presence of statistically significant low quantitative values of antitoxic and antibacterial antibodies to C. diphtheriae antigens. In the group of patients with the localized forms of the disease the initially low level of antitoxic antibodies was detected with the concentration of antibacterial antibodies remaining unchanged. During the period of convalescence the levels of antitoxic antibodies in both groups reached those of healthy persons. In case of localized forms of the disease the level of antibacterial antibodies decreased as compared with healthy persons, starting from the second week of the disease. The period of convalescence in the disseminated forms was characterized by the low concentration of antibacterial antibodies. Carrier state was formed in the presence of high levels of antitoxic antibodies and significantly low levels of antibacterial ones.

Isolation, crystallization, and investigation of ribosomal protein S8 complexed with specific fragments of rRNA of bacterial or archaeal origin.

The core ribosomal protein S8 binds to the central domain of 16S rRNA independently of other ribosomal proteins and is required for assembling the 30S subunit. It has been shown with E. coli ribosomes that a short rRNA fragment restricted by nucleotides 588-602 and 636-651 is sufficient for strong and specific protein S8 binding. In this work, we studied the complexes formed by ribosomal protein S8 from Thermus thermophilus and Methanococcus jannaschii with short rRNA fragments isolated from the same organisms. The dissociation constants of the complexes of protein S8 with rRNA fragments were determined. Based on the results of binding experiments, rRNA fragments of different length were designed and synthesized in preparative amounts in vitro using T7 RNA-polymerase. Stable S8-RNA complexes were crystallized. Crystals were obtained both for homologous bacterial and archaeal complexes and for hybrid complexes of archaeal protein with bacterial rRNA. Crystals of the complex of protein S8 from M. jannaschii with the 37-nucleotide rRNA fragment from the same organism suitable for X-ray analysis were obtained.

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.

Divergence of the indole alkaloid pattern in two somatic hybrid plant cell subcultures of Rauvolfia serpentina x Rhazya stricta.

The alkaloid pattern in Rauvolfia serpentina x Rhazya stricta somatic hybrid cell subcultures R x R17 K was studied and 11 compounds were identified on the basis of their spectral data. Among them, 1,2-dehydroaspidospermidine, rhazinilam, stemmadenine and tabersonine were reported as typical of Rhazya species while vomilenine and sarpagine are characteristic of Rauvolfia alkaloid metabolism. The alkaloid pattern in R x R17 K subcultures was compared with that in the other hybrid cell subcultures (R x R17 M) which were developed from the same origin hybrid cultures but have been maintained separately for about ten years. The data presented here exhibit pronounced divergence of the alkaloid patterns in R x R17 K and R x R17 M cell subcultures.

[Multilocus sequencing--a new method of genotyping bacteria and first results of its use]. / Mul'tilokusnoe sekvenirovanie--novyi metod genotipirovaniia bakterii i pervye rezul'taty ego primeneniia.

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.

[The characteristics of the humoral antibacterial immunity of young children with respiratory organ diseases]. / Osobennosti gumoral'nogo antibakterial'nogo immuniteta u detei rannego vozrasta s zabolevaniiami organov dykhaniia.

The state of antibacterial humoral immunity in young children with acute bronchitis, acute obstructive bronchitis and bronchial asthma at the period of exacerbation has been shown with the use of the enzyme immunoassay. The concentration of antibodies to endotoxin positively correlates with the severity of clinical manifestations of the endogenic intoxication of the body. As the inflammatory process in the bronchial tree increases, the spectrum bacterial agents to which elevated concentrations of specific antibodies can be detected becomes wider, and this finds its maximum reflection in bronchial asthma.

Hemosorption of restored long-preserved blood.

Incubation of long-preserved donor blood erythrocytes (over 21 days) with rejuvenating solution Erythropifaden resulted in an increased ATP concentration to 2.03 +/- 0.34 mmol/l; increased 2,3-DPG to 3.89 +/- 0.51 mmol/l, and the number of erythrocyte discoid forms was raised to 30%. Further blood perfusion through SKN-D hemosorbent allowed to reduce the following adenine level to zero; riboxine by 90; citrate and lactate by 50%. The plasma K+ was decreased from 29.8 +/- 1.25 to 13.0 +/- 0.87 mmol/l, while blood pH rose from 6.675 +/- 0.018 to 7.310 +/- 0.028. At the end of the perfusion, the ATP level was seen to further increase to 2.53 +/- 0.28 mmol/l, and 2,3-DPG to 4.99 +/- 0.7 mmol/l. The indices of osmotic resistance and deformability of erythrocytes were normalized. The discocyte number reached 40%. The amount of red blood cell irreversible forms declined from 26 to 13% without any increment plasma free hemoglobin. The data obtained confirm the high efficacy of the combination of the rejuvenating and hemosorption procedures when applied to stored long-preserved blood.

[Effect of pH on the regulatory characteristics of energy metabolism in human erythrocytes]. / Vliianie pH na reguliatornye kharakteristiki énergeticheskogo metabolizma éritrotsitov cheloveka.

The glucose consumption rate versus ATP content in human red cells (regulatory patterns of glycolysis) and ATP concentration versus glucose uptake rate in red cell suspension (regulatory patterns of total ATPases), when the rate of glucose uptake is constant and lower than the rate of glucose consumption at physiological conditions, were measured at different pH values. The shape of both types of kinetic curves was found to be dependent on the pH of the incubation medium but the same for the red cells taken from different donors. It is supposed that at alkaline pH, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase reactions become the rate-limiting steps of glycolysis instead of hexokinase and phosphofructokinase under physiological conditions.

[Quantitative model of human erythrocyte glycolysis. II. Effect of arsenate on glycolysis. Experimental study of the relationship between the rate of glycolysis and the ATP concentration]. / Kolichestvennaia model' glikoliza éritrotsita cheloveka. II. Vliianie arsenata na glikoliz. Eksperimental'noe issledovanie zavisimosti skorosti glikoliza ot kontsentratsii ATF.

The effect of arsenate on human erythrocyte glycolysis was studied. The stationary rate of glucose consumption rises and then drops with a gradual increase of arsenate concentration while ATP and glucose-6-phosphate concentrations drop monotonically. A plot of glucose consumption rate against ATP concentration gives a bell-like curve. This curve is the same for different donors if it is plotted in relative units with values at zero arsenate concentration taking for 100%, while the absolute values obtained for separate donors are very different. The normal (physiological) point is situated on the steeply descending part of the curve.