[Correlation between the receptor specificity of pandemic influenza A (H1N1)pdm09 virus strains isolated in 2009-2011 and the structure of the receptor-binding site and the probability of fatal primary viral pneumonia].

The receptor specificity (RS) of pandemic influenza A(H1N1) pdm09 virus strains deposited into the State Collection of Viruses of the Russian Federation, D. I. Ivanovsky Research Institute of Virology, Ministry of Health and Social Development of Russia, in the 2009-2010 and 2010-2011 epidemic seasons to a panel of 9 sialoglycopolymers (SGP). The strains were divided into 3 groups according to the W(3/6) index proposed by the authors, which was equal to the amount of reactivities to unbranched alpha2-3-SGP to that of reactivities to unbranched alphal-6-SGP: W(3/6) < or = 1.0; 1.0 < W(3/6) < or = 1.5. The W(3/6) < or = 1.5 group showed a predominance of a2-3-RS, attended by the high incidence of fatal primary viral pneumonias (FPVP) (60.0%) and amino acid replacements in the HA1 receptor-binding site (RBS) (80.0%): D222{G, N} and Q223R. The 1.0 < W(3/6) < or = 1.5 group was characterized by mixed alpha2-3/alpha2-6-RS with the incidence of FPVP (29.7%) and amino acid replacements in the HA1 RBS (40.5%) (D222{G, N, V} and Q223), respectively. In the W(3/6) < or = 1.0 group, alpha2-6-RS was prevalent, FPVPs were absent and amino acid replacements in HA1 RBS (D222{G, E}) were seen only in 6.0% of cases. The number of strains with increased specificity to alpha2-3-sialosides increased in the 2010-2011 epidemic season as compared to the previous season. With their further spread among the population, there may be a rise in cases of severe primary viral pneumonias with possible fatal outcomes, which can be, however, accompanied by a decrease in the capacity of mutants to air-dropwise transmission.

[Pandemic influenza in Russia: specific features of clinical course and the absence of early etiotropic therapy as a risk factor of severe forms of the disease].

AIM: To assess efficacy and safety of ingavirin in the treatment of the flu caused by pandemic virus of flu A (H1N1) sw1 in hospitalized patients compared with oseltamivir. MATERIAL AND METHODS: A population-based comparative multicenter trial included 194 patients with verified diagnosis of the flu aged 18-60 years with marked clinical symptoms, body temperature over 38 degrees C and duration of the disease 48 hours maximum. The patients were randomized into 2 groups: group 1 (n=152) received ingavirin (90 mg once a day), group 2 received oseltamivir (n=42) in a dose 150 mg twice a day. Duration of the course was 5 days. RESULTS: Ingavirin and oseltamivir normalized body temperature within treatment hours 24-36 if therapy was initiated in the first disease hours 27.0 +/- 10.0 and 31.9 +/- 10.4. Mean duration of the fever for ingavirin was 35.1 +/- 14.5 hours, for oseltamivir--26.3 +/- 13.0 hours (p < 0.817). The antiviral medicines significantly reduced duration of intoxication (head ache, weakness), catarrhal symptoms (cough, tracheitis, rhinitis), rate of complication vs. patients untreated with antivirus drugs (n=30). CONCLUSION: The results of the treatment show safety and efficacy of ingavirin in uncomplicated flu caused by pandemic virus of flu A (H1N1) sw1 in inpatients. Early etiotropic therapy is a basic treatment policy able to reduce the number of severe complications and lethality.

[Pathological anatomy in A/H1N1 influenza].

Forty autopsy cases died from A/H1N1 influenza in Moscow, the Moscow Region, and Chita were analyzed. Major changes were found to develop in the trachea and lung. The lung showed the pattern of exudative and proliferative stages of diffuse alveolar lesion. In addition, the signs of viremia-associated infectious-toxic shock, as suggested by pronounced changes in the brain, kidneys, liver, spleen, adrenals, and lymph nodes were revealed. Bacterial bronchopneumonia was detected only in 20% of cases.

[Pathomorphology of the lung in severe influenza A/H1N1].

Thirty-five autopsy cases of influenza A/HIN1 in Moscow were analyzed. Primary pathological changes were found in the trachea and lung. The lung showed a pattern of the exudative and proliferative stages of diffuse alveolar damage. In addition, there were signs of infectious-toxic shock associated with viremia, as suggested by pronounced changes in the brain, kidneys, liver, spleen, adrenals, and lymph nodes. Bacterial bronchopneumonia was detected in only 20% of cases.

[Outcomes of severe pandemic influenza A/H1N1/2009].

AIM: To study the outcomes of severe pandemic influenza A/H1N1/2009. SUBJECTS AND METHODS: The study enrolled 24 patients, including 8 males and 16 females (10 of whom were pregnant), aged 17 to 58 years, with a laboratorily verified diagnosis of pandemic influenza A/H1N1/2009, treated at the intensive care unit for the significant symptoms of acute respiratory failure (RF). Real-time RT-PCR was used to verify the diagnosis. Organs and tissues from deceased patients were histologically studied; chest computed tomography, body plethysmography, fibrobronchoscopy, breath test, and 6-minute walk test were performed in the late period. RESULTS: Within the first 30 days, a fatal outcome caused by therapy-resistant progressive RF was observed in 33% of the patients with pandemic influenza treated at the intensive care unit. Diffuse alveolar damage caused by influenza virus, which gives rise to hyaline membranes, underlies RF. Lung tissue fibrosis formed in recovered patients. CONCLUSION: The severity of pandemic influenza A/H1N1/2009 was determined by massive bilateral pneumonia, interstitial (alveolar) pulmonary edema, formation of diffuse bilateral lung fibrosis at the outcome of severe virus pneumonia (acute respiratory distress syndrome) with a decrease in vital and diffusing capacities, thereby generating a need to follow up this patient category and, possibly, to elaborate special rehabilitation programs.

[Detection of the circulation of Crimean-Congo hemorrhagic fever virus in the piedmont steppes of the North Caucasus].

The laboratory verified cases of Crimean-Congo hemorrhagic fever (CCHF) in the piedmont steppes of the North Caucasus (Malgobeksky District, Republic of Ingushetia) are first described. The source of the first infection was Ixodidae ticks; three subsequent sources were contacts with the bloody discharges from patients. CCHF virus genome was detected in the blood of the cattle from an epidemic focus and in the pools of the Ixodes ticks Haemaphysalis parva Neum., 1897 and Boophilus annulatus Say, 1821, taken from cattle. The problem of including the piedmont steppes of the North Caucasus into the CCHF nosological area is discussed.

[Hepatitis C virus proteins in the liver cells in acute hepatitis C: their association with liver damage and with the outcome of the disease].

The accumulation of individual hepatitis C virus (HCV) proteins in the liver cells of patients with acute hepatitis C (AHC) and their association with the course and outcome of the disease were studied. AHC protein expression in the cryostat liver sections from 20 patients with AHC was estimated by immunohistochemical assay using original monoclonal antibodies to 5 HCV proteins (core, NS3, NS4A, NS4B, and NS5A). The results of HCV detection in the patients were compared with their biochemical, clinical, and morphological findings. HCV proteins were totally revealed in the livers of all the patients, individual proteins were identified with a frequency of 89-95%, which is significantly more than those in patients with chronic hepatitis C. The hepatic expression of core protein was shown to inversely correlate with the duration of an acute period. There was a direct relationship between the accumulation of core, NS3, and NS5A proteins and the liver tissue damage caused by stepwise necrosis rather than intralobular necrosis. The presumed convalescence was ascertained to be associated with the larger count of hepatocytes containing the proteins NS4A and NS3 early after AHC manifestation.

[Diagnostic significance of type IV collagen and hyaluronic acid in the serum of patients with chronic hepatitis C for staging hepatic fibrosis].

AIM: To evaluate diagnostic value of serologic fibrosis markers (hyaluronic acid--HA and type IV collagen C-IV) in patients with chronic hepatitis C (CHC) and hepatic cirrhosis (HC). MATERIAL AND METHODS: HA and C-IV were measured in 88 CHC patients with fibrosis stage 1 (n = 63) and 3 (n = 25), 13 patients with acute hepatitis C (AHC), 28 patients with hepatic cirrhosis (HC), 19 patients with pulmonary fibrosis (PF). The control group consisted of 32 healthy subjects. RESULTS: HA concentrations in the serum of CHC patients with mild to severe inflammation and fibrosis (F1 and F3) were normal (100 ng/ml). For HC diagnosis, HA test proved highly sensitive and specific (in HA 100 ng/ml sensitivity was 100%, specificity 84.6%), but this method cannot stage hepatic fibrosis. HA test was inferior to C-IV test. A mean C-IV concentration in the serum of CHC patients at the stage of marked fibrosis (F3) is significantly higher than in F1, in HC (A) patients higher than in patients with CHC F3. CONCLUSION: It is shown than concentration of C-IV above 196 ng/ml can differentiate fibrosis stage 1 from stage 3 with specificity 58.7 and sensitivity 88%.

[Changes of humoral and cellular immunity in chronic hepatitis C patients of different staging]. / Izmenenie pokazatelei gumoral'nogo i kletochnogo immuniteta u patsientov s khronicheskim gepatitom C pazlichnoi tiazhesti.

The purpose of the present study was to investigate the influence produced by viral proteins in the hepatic cells and RNA of hepatitis C virus (HCV) on the indices of T- and B-cell response in 52 patients with chronic hepatitis C (CHC). A relative count of peripheral-blood lymphocytes (PBL), expressing antigens CD3+, CD4+, CD8+, CD16+, CD20+ and CD95+ was estimated. The repertoire of antibodies to HCV proteins was specified. The thus obtained data were compared with an activity and a disease stage by using the histological diagnosis and alanine-amino-transferase (ALT) level as well as with the presence of HCV RNA in the serum and viral protein of the liver. Such comparison of data and the use of the correlation analysis made it possible to establish that the antibodies to NS5 protein were detected reliably more often in patients with a more pronounced hepatic fibrosis, with a higher ALT activity and with expression of HCV proteins in the liver. At the same time, the presence of proteins in the liver and of RNA in the serum were accompanied by a more active humoral response to the non-structure proteins of NS4 and NS5 as well as by more profound discrepancies of the immunity T-cell chain (a lowered ratio of CD4+/CD8+ and a smaller content of CD95+). There were no differences between PBL of the studied populations in patients with various activities and an HCV stage. A relatively bigger quantity of CD95(+)--positive PBL was found to be reliably higher in patients with viremia but lower in those cases, in which HCV proteins were detected in the liver. This confirms the inhibiting ability of HCV proteins to the Fas-mediated apoptose of PBL in CHC patient.

[Analysis of hepatitis C virus proteins in hepatic cells of patients with chronic hepatitis C]. / Analiz belkov gepatita C v kletkakh pecheni bol'nykh khronicheskim gepatitom C.

A correlation between the detection of proteins and an activity of the pathological process was analyzed in a study of the content of the C virus hepatitis (CVH) proteins in hepatic cells of patients with chronic C hepatitis (CCH). The expression of CVH proteins in frozen sections of biopsy samples of 69 CCH patients was evaluated by using the immune-histological method involving original monoclonal antibodies (MCA) to 5 CVH proteins. The results of the detection of proteins in patients were compared with an activity and stage of CCH (by using histological tests and a level of alanine aminotransferase--AAT). A set of the CVH proteins were found in the liver of 74% of patients, i.e. core proteins, NS3, NS4A, NS4B and NS5A--in 28, 43, 43, 55 and 58%, respectively. All studied proteins were detected in the cytoplasm of hepatocytes. Proteins were found in the liver more often as compared with the detection rate of CVH RNA in the blood serum (61%). This demonstrates a high sensitivity of the discussed test at detecting the CVH infection. The accumulation of the core protein was shown to correlate with the presence of the replicative form of CVH RNA in the liver and with a higher level of AAT. The quantity of NS5 A-expressing cells correlated directly with a CCH stage. The quantity of NSB- and NS3-positive hepatocytes correlated negatively with an activity of the inflammatory-and-necrotic processes in the liver. Hyper-fermentation was found more often among the antigen-positive patients. The CCH histological activity was proven to be reliably higher at a simultaneous detection of CCH proteins in the liver and of CVH RNA--in the serum.

[Prospects for interferon therapy in chronic hepatitis C with normal transaminase levels]. / Vozmozhnosti interferonoterapii pri khonicheskom gepatite C s normal'nym urovnem transaminaz.

AIM: To study effectiveness of recombinant interferon-alpha (Ifna) in patients with chronic hepatitis C with normal transaminases. MATERIAL AND METHODS: 26 patients with positive tests for abHCV and RNA HCV with a normal level of alaninaminotransferase (AlAT) entered the study. 16 patients of the test group received recombinant Ifna three times a week in a dose 3 mln IU. Mean duration of interferon therapy was 7.44 +/- 4.27 months. Control group consisted of 10 patients with natural course of the disease. Biochemical, virusological and morphological examinations were conducted in all the patients. The patients were followed up for 1.5-2 years, on the average. RESULTS: Treatment with recombinant Ifna was well tolerated. A persistent positive virusological response after the treatment was achieved in 25% of the test patients. They exhibited improvement of histological structure of the liver. In control group the dynamics was opposite. CONCLUSION: In patients with chronic hepatitis C with normal transaminases level interferon therapy improves structure of hepatic tissue. Puncture biopsy of the liver is the main method which determines the validity of the treatment and its efficacy.

[The role of hepatitis C virus RNA detection in various substrates of patients with chronic hepatitis C]. / Znachenie viiavleniia RNK virusa gepatita C v razlichnykh substratakh u bol'nykh khronicheskim gepatitom C.

Puncture biopsy of the liver and blood count were made in 72 patients with chronic hepatitis C (CHC). Morphological alterations in the liver were assessed by Knodell index. The blood serum, lymphocytes and hepatic tissue were examined for a genome form of hepatitis C virus (HCV) RNA, blood lymphocytes and hepatic tissue--for a relevant replication form. HCV RNA was detected using "nested" RT-PCR. Only 26% patients had symptoms of asthenovegetative and dyspeptic syndromes. Normal alaninaminotransferase (ALT) level was observed in 24% patients, the rest had it high. HCV RNA was encounted more frequently in hepatic tissue than lymphocytes or serum (83, 68 and 46%, respectively). A replication form of HCV RNA was present in hepatic tissue of 31% patients and was absent in the lymphocytes. The incidence of the RNA detection was not related either to the disease symptoms or morphological alterations in hepatic tissue. The occurrence of the genome and replication forms in hepatic tissue does not correlate to ALT level. HCV RNA occurs more often in the serum, blood lymphocytes and in three substrates simultaneously in patients with hyperalatemia.

[The course and outcome of chronic hepatitis B in children]. / Techenie i iskhody khronicheskogo gepatita B u detei.

Overall 98 children aged 1 to 14 years suffering from chronic hepatitis B (CHB) were followed up clinically for 1 to 6 years. CHB was diagnosed on the basis of the clinical and laboratory data. In the majority of the children, the diagnosis was verified by the results of a histological study of liver biopsy specimens. Chronic active hepatitis (CAH), was revealed in 27 children, chronic persistent hepatitis (CPH) in 31. CHB was marked by the presence of HBe-antigenemia in 89 patients (90.8%). The studies have demonstrated that CHB associated with HBe-antigenemia runs its course with insignificant clinical manifestations and enzymic exacerbations without jaundice. In the presence of persistent HBs-antigenemia, the natural course of CHB (in CAH and CPH) is characterized by seroconversion (from HBeAg to anti-HBe) with a simultaneous decrease and normalization of aminotransferase activity and a reduction of the pathological process activity in the liver (transformation of CAH to CPH). Seroconversion and clinico-biochemical amelioration supervene at different observation periods (after 1-6 years) and do not depend on the initial activity of hepatitis. As the observation period increases, the rate of anti-HBe appearance in the blood rises, amounting to 90% with the observation period exceeding 5 years. The conclusion is made that CHB patients do not need active drug therapy but require long and permanent observation.

[A comparative experimental study of pathogenic properties of new strains of California encephalitis virus serogroups]. / Sravnitel'noe izuchenie patogennykh svoistv novykh shtammov virusa serogruppy Kaliforniiskogo éntsefalita véksperimente.

Experiments in noninbred mice, Syrian hamsters and grey monkeys were made to characterize the pathogenic properties of new strain of the California encephalitis serogroup isolated for the first time on the island of Taimir, in the Murmansk, Leningrad, Tver regions and in Karelia. All the strains displayed marked tropism for the CNS. The strains isolated in the northern regions of this country turned out more pathogenic for the animals. The strain Leiv-12812 Kl (isolated in the Tver region) differed from the remaining ones in more pronounced pathogenicity for the monkeys, as well as in the site and intensity of brain lesions. The persistence of the virus in hamsters organs may play a role of a reservoir of infection in nature.

[Chronic hepatitis delta--the course and approaches to therapy]. / Techenie khronicheskogo gepatita del'ta i podkhody k lecheniiu.

Variants of chronic hepatitis delta (CHD) course were studied in 94 patients. Slow progress (for decades) occurred most frequently, while rapid progress to hepatic cirrhosis (for 1-2 years) or benign course presenting as greater than 3-year remission were rare findings. In addition to routine therapeutic modalities, except corticosteroids, 43 CHD patients received recombinant alpha 2-interferon (reaferon). Follow-up results evaluated separately for hepatic cirrhosis and free of it CHD patients support clinical promise of Soviet recombinant alpha 2-interferon against CHD.

[Use of personal protective equipment by pathologists and legal physicians working with particularly dangerous infectious diseases]. / Primenenie sredstv individual'noi zashchity patologoanatomami i sudebnymi ékspertami pri rabote s osobo opasnymi infektsiami.

The possibility of pneumosuit use constructed in the Institute to protect the pathology department staff at the hospitals for infectious diseases from HIV virus was studied. The suit is a hose-type suit for protection of respiratory organs and skin using the ventilated underhood space and autonomic air supply. Both the detailed description of this suit and its disinfection scheme developed and tested by authors are presented. The comparative physio-hygienic assessment of pneumosuit and standard antiplaque suit demonstrated a number of advantages of the former during 2 hours work at pathology department. The authors recommend the pneumosuit as an equipment for the protection of skin and respiratory organs of pathologists and legal physicians working with particularly dangerous infections.