[Flow cytometry evaluation of the rhesus monkey cellular immunity following the Zaire ebolavirus (Filoviridae; Ebolavirus: Zaire ebolavirus) experimental infection].

INTRODUCTION: The outbreaks of the Zaire ebolavirus (ZE) disease (ZED) that have arisen in the last decade determine the need to study the infection pathogenesis, the formation of specific immunity forming as well as the development of effective preventive and therapeutic means. All stages of fight against the ZED spread require the experimental infection in sensitive laboratory animals, which are rhesus monkeys in case of this disease .The aim of the study is to evaluate the rhesus monkey cellular immunity following the ZE experimental infection by the means of flow cytometry (cytofluorimetry). MATERIAL AND METHODS: Male rhesus monkeys were intramuscularly infected by the dose of 15 LD50 (dose of the pathogen that causes 50% mortality of infected animals) of the ZE, the Zaire strain (ZEBOV). Levels of 18 peripheral blood lymphocyte populations of the animals before the ZE experimental infection and at the terminal stage of the disease were assessed using flow cytometry. RESULTS AND DISCUSSION: The certain changes in the levels of the lymphocyte populations were observed following infection, indicating simultaneous activation and suppression of the immune system during ZED. The increase in content was observed for T-lymphocytes, T-helper and cytotoxic T-lymphocytes expressing the corresponding markers of early activation. The decrease was recorded for T-lymphocytes and double-positive T-lymphocytes expressing corresponding markers of late activation, as well as natural killer cells expressing CD8 (p < 0.05). CONCLUSION: For the first time in the Russian Federation, the rhesus monkey cellular immunity before and after the ZE experimental infection was assessed using flow cytometry.

[The choice of individual treatment strategy in geriatric patients with benign prostatic hyperplasia with consideration of indications and contraindications for elective isolated and simultaneous interventions].

AIM: To develop a rationale treatment strategy for elective isolated and simultaneous interventions in geriatric patients with benign prostatic hyperplasia (BPH) aimed at decreasing of complications rate and grade, based on current approach to indications and contraindications. MATERIALS AND METHODS: From 2000 to 2015 yy. a total of 639 elderly and senile patients with BPH were undergone to isolated simple prostatectomy, transurethral prostatic resection and simultaneous surgery. Two timeframe were allocated, from 2000 to 2006 and from 2007 to 2015 yy. During the first period, the conventional indications and contraindications were used for determination of the type and volume of the the surgery. During the second period, a novel authors approach to the structure indications and contraindications for the surgical treatment was introduced. RESULTS: For the first time in the geriatric urology a modern structure of the indications and contraindications for elective interventions in patients with BPH has been developed, which allowed to clarify the treatment strategy on the individual basis. The new criteria for absolute and relative contraindications are included as well as, for the first time, temporary, specific and organizational contraindications for the simple prostatectomy and TURP. There is a need for review some conventional contraindications for the TURP in elderly and senile patients, considering that in specialized geriatric urological hospital for patients with severe concomitant diseases (therapeutic, neurologic, etc.), TURP should be the method of choice regardless prostate volume, often exceeding 60 cc. The use of a new structure of indications and contraindications allowed to establish absolute contraindications for the elective surgery in 19%, relative contraindications in 21% and temporary contraindications in 8% cases, respectively. These patients were undergone to palliative surgeries or drug treatment which helped to prevent possible postoperative complications and mortality. CONCLUSION: In order to determine the treatment strategy in elective isolated and simultaneous interventions in geriatric patients with BPH it is necessary to consider the current general and specific indications as well as absolute, relative, specific, temporary and organizational contraindications. The introduction of the novel structure of the indications and contraindications for the elective isolated and simultaneous surgeries with priority of TURP had a pronounced clinical effect, contributing to the marked reduction of postoperative complications and deaths in geriatric patients with BPH.

[Systematic approach in performing isolated and simultaneous operations in elderly patients with prostate adenoma].

AIM: To develop a scientifically based system for treating elderly patient with prostate adenoma (AP), aimed at reducing the incidence and severity of complications and mortality after isolated and simultaneous operations. MATERIALS AND METHODS: Between 1995 and 2015, 1170 elderly and older patients with prostate adenoma (PA) underwent isolated adenomectomy (AE) or transurethral resection of the prostate (TURP) or simultaneous operations (SO). Simultaneous interventions were performed in 335 (28.6%) patients. RESULTS: Immediate treatment results were significantly better in patients who underwent isolated TURP or TURP as a part of SO than that of those who had isolated AE or AE as a part of SO. SO was not associated with greater incidence of local or general postoperative complications and mortality compared with isolated AE or TURP. CONCLUSION: Continuous advancements in diagnostic and treatment methods contributed to a significant reduction in the incidence of postoperative complications and deaths in geriatric patients with PA. The study findings confirm the superiority of TURP over AE, as previously reported by other authors.

Safety and immunogenicity of GamEvac-Combi, a heterologous VSV- and Ad5-vectored Ebola vaccine: An open phase I/II trial in healthy adults in Russia.

Ebola hemorrhagic fever, also known as Ebola virus disease or EVD, is one of the most dangerous viral diseases in humans and animals. In this open-label, dose-escalation clinical trial, we assessed the safety, side effects, and immunogenicity of a novel, heterologous prime-boost vaccine against Ebola, which was administered in 2 doses to 84 healthy adults of both sexes between 18 and 55 years. The vaccine consists of live-attenuated recombinant vesicular stomatitis virus (VSV) and adenovirus serotype-5 (Ad5) expressing Ebola envelope glycoprotein. The most common adverse event was pain at the injection site, although no serious adverse events were reported. The vaccine did not significantly impact blood, urine, and immune indices. Seroconversion rate was 100 %. Antigen-specific IgG geometric mean titer at day 42 was 3,277 (95 % confidence interval 2,401-4,473) in volunteers immunized at full dose. Neutralizing antibodies were detected in 93.1 % of volunteers immunized at full dose, with geometric mean titer 20. Antigen-specific response in peripheral blood mononuclear cells was also detected in 100 % of participants, as well as in CD4+ and CD8+ T cells in 82.8 % and 58.6 % of participants vaccinated at full dose, respectively. The data indicate that the vaccine is safe and induces strong humoral and cellular immune response in up to 100 % of healthy adult volunteers, and provide a rationale for testing efficacy in Phase III trials. Indeed, the strong immune response to the vaccine may elicit long-term protection. This trial was registered with grls.rosminzdrav.ru (No. 495*), and with zakupki.gov.ru (No. 0373100043215000055).

[Sensitivity and specificity of the ELISA Kit for the detection of antidobies to Junin virus].

The goal of this work was to describe methodological approaches to determination of sensitivity and specificity of the enzyme-linked immunosorbent assay kit (ELISA Kit) for detection of the specific anti-Junin virus (JV) antibody. Comparison of ELISA to plaque reduction neutralization test (PRNT) showed direct relationship between antibody titers in the samples of serum of immunized animals, determined by either PRNT or ELISA methods. The obtained results provided an opportunity to form the panels of positive and negative serum samples to determine the sensitivity and specificity of the ELISA Kit. Sensitivity of the ELISA Kit was at least 98% when studying the samples of serum of immunized guinea pigs and rabbits (determined as positive in PRNT). The sensitivity of the ELISA Kit was at least 68% when studying the samples determined by PNRT as uncertain positive. The specificity was 98%. The specificity of the ELISA Kit was 98%.

[Study of sensitivity of laboratory animals to a causative agent of argentine hemorrhagic fever].

AIM: Study sensitivity of laboratory animals to a causative agent ofArgentine hemorrhagic fever. MATERIALS AND METHODS: Junin virus strain XJ P37 was obtained from the State Collection of Causative Agents of Viral Hemorrhagic Fevers of the Pathogenicity Group I of Scientific Research Center of the 33rd Central Scientific Research Test Institute (SRC of the 33rd CSRTI). Junin virus strain XJ P37 culture with biological activity of 5.2 1g PFU x ml was used in the experiments. Mice (2 - 4 and 7 - 14 days old), guinea pigs (250 - 300 g), 1.8 - 2.5 kg shinshilla breed rabbits, 2.0 - 3.0 kg javanese macaque monkeys were obtained from vivarium of the SRC of the 33rd CSRTI. Vero (B) and GMK-AH-1 (D) cell cultures were obtained from cell culture collection of the SRC of the 33rd CSRTI. Biological activity calculation of Junin virus was carried out by Kerber in I.P. Amsharin modification. RESULTS: Lethality in animals was from 12.5 to 50% after intranasal and intraperitoneal infection of guinea pigs, intramuscular, intraperitoneal and subcutaneous infection of rabbits, intracerebral and intranasal infection of mice at the doses from 0.4 to 1.0 x 10(5) PFU. Death of infected monkeys after intramuscular administration of the virus at 1.0 x 10(4) PFU dose was not observed. In 100% of surviving animals formation of virus-neutralizing antibodies was registered. CONCLUSION: Evaluation of sensitivity of laboratory animals to Junin virus has shown that intracerebrally infected mice may be used to maintain causative agent culture, infected guinea pigs - to prepare virus-containing cultures and modelling infection exacerbation in humans. Intramuscularly infected rabbits may be used to obtain hyper-immune sera.

[Optimization of conditions of quantitative evaluation of argentine hemorrhagic fever causative agent].

AIM: Optimization of conditions of quantitative evaluation of Argentine hemorrhagic fever causative agent. MATERIALS AND METHODS: Junin virus (XJ P37 strain) was obtained from National collection of viral hemorrhagic fever causative agents of the 1st pathogenicity group of the 33rd Central Research Testing Institute. Junin virus (XJ P37 strain) culture with biological activity of 5.2 lg PFUxm(-1) was used in the experiments. Vero B, 6619-1(D) and GMK-AH-1(D) were obtained from collection of cell culture of the Research Scientific Centre of the 33rd Central Research Testing Institute. Calculation of biological activity of Junin virus during titration in cell cultures was carried out by Kerber method with modification by I.P. Ashmarin. RESULTS: During incubation for 4 - 7 days after the infection of cell monolayer the determined biological activity was 4.4 - 6.4 lg PFUxml(-1); the size of the formed negative colonies--(1.5 +/- 0.5) mm. CONCLUSION: The conditions of quantitative evaluation of Argentine hemorrhagic fever were optimized by negative colonies method (using 5 - 7 day Vero B cell culture monolayer with staining of monolayer on day 5 of secondary incubation, recording of results at day 7 after the infection).

[Angiographic evaluation of coronary vessels in patients with diabetes mellitus type 2 and myocardial infarction].

High risk of vascular complications in diabetes foundation gave the American Heart Association ranked diabetes to diseases of the cardiovascular system. More than 60% of patients with type 2 diabetes life expectancy will be reduced by the development of their coronary artery disease, diabetes was an independent factor that increases the risk of myocardial infarction (2-3 times), stroke (more than 2 times) independently of other known risk factors for cardiovascular disease, and acute coronary events are characterized by the extremely serious nature against the background of pronounced coronary atherosclerosis. In the present study, a comparative assessment of the frequency of coronary vessels of different severity in patients with myocardial infarction in the presence of diabetes and without. Status of the coronary vessels were studied by means of selective coronary angiography. The study found that in patients with myocardial infarction in combination with type 2 diabetes is much more frequent multivessel disease and coronary delay the passage of contrast material than patients with MI without diabetes. These features and characteristics of coronary lesions in patients with diabetes and myocardial infarction may explain why more severe myocardial infarction with frequent development of coronary events in these patients. After analyzing the data obtained, it can be assumed relationship progression of vascular lesions with concomitant coronary glycemic decompensation.

[Estimation of the risk and prevention of postoperative esophagogastroduodenal hemorrhage in elderly urological patients].

Open transvesical adenomectomy was made in 853 old males with prostatic adenoma in Samara Hospital for Veterans from 1995 to 2010. The age of the patients ranged from 60 to 89 years, of them 389 (45.6%) were from 60 to 75 years of age, 464 (54.4%) - 75 to 89 years. Elective simultaneous operations were conducted in 104 (12.2%) of the above patients. It is shown that early postoperative esophagogastroduodenal hemorrhage after adenomectomy occurs in 7% old patients. Endoscopic parietal pH-metry is "a gold standard" in selective sensitive diagnosis of acid-dependent conditions in presenile and senile patients with prostatic adenoma. The proposed method of prediction of the risk of early postoperative esophagogastroduodenal hemorrhage provides realization of the principle of a differential approach to effective prevention of the hemorrhage in old patients with prostatic adenoma. Complex preoperative preparation using adequate antisecretory therapy in old patients with prostatic adenoma with the risk of early postoperative esophagogastroduodenal hemorrhage decreases hemorrhage rate from 7 to 0.4%. Prevention of early postoperative esophagogastroduodenal hemorrhage in old patients with prostatic adenoma should become as obligatory as prevention of cardiovascular, thromboembolic, pulmonary and pyoseptic complications.

[Elective simultaneous operations in elderly and senile urological patients].

Surgeons of Samara Regional Veterans' Hospital made elective-simultaneous operations in 229 presenile and senile patients. Surgery was performed in the urological department in 1995-2010. It is shown that over 12% patients of urological geriatric departments need elective situltaneous operations, of them 8.7% are mild, 78.6%--moderate and 12.7%--severe. The results of simultaneous operations in 133 patients and staged operations in 60 control patients (adenomectomy and Vinkelman) differed insignificantly (p > 0.05). Such achievement was realized due to advanced prophessional education of all medical personnel of the department, introduction of new techniques in examination and treatment: accurate definition of absolute and relative contraindications for simultaneous surgery, effective preoperative preparation, low-invasive operations, different variants of non-endotracheal anesthesia. In addition to a positive clinical effect, new technology of simultaneous operations reduces 2.3-fold economic costs vs staged surgery, improves significantly quality of life of the above patients.

[Stabilization of peroxidase conjugates used in enzyme immunoassay systems to detect Ebola and Marburg virus antigens].

The time course of changes in the activity of solutions of horseradish peroxidase conjugates with immunoglobulins against Ebola and Marburg fevers was studied in the presence of different components. The series of the conjugates of ELISA kits for the detection of Ebola and Marburg virus antigens, which were prepared on the basis of the designed stabilizing solution, preserved at less than 90% of its baseline activity during 10 months at a storage temperature of 2 to 8 degrees C.

[Chemotherapy for Bolivian hemorrhagic fever in experimentally infected guinea pigs].

The efficacy of the antiviral agents nucleoside analogues manufactured in Russia and foreign countries against Bolivian hemorrhagic fever in the treatment and rapid prevention regimens was studied in guinea pigs. In rapid prevention and treatment of guinea pigs, the intraperitoneal injection of 1/10 of the equivalent dose of vero-ribavirin for this animal species was ascertained to protect 70% of the animals infected with Machupo virus strain Carvallo in doses 8 and 10 LD50.

[Use of guinea pigs to evaluate the efficacy of a heterological immunoglobulin against Bolivian hemorrhagic fever].

The use of guinea pigs as a laboratory model was proven to be appropriate in investigating the protective properties of a heterological immunoglobulin against Bolivian hemorrhagic fever at the preclinical stage of the study. A highly pathogenic Machupo virus strain that caused guinea pigs' death with respect with an agent's dose was cultivated. Injection of 1.0 ml of the immunoglobulin provided a 100% protective effect for the guinea pigs infected with the highly pathogenic Machupo virus strain in a dose of 10 LD50.

[Use of guinea pigs for assessing the efficacy of vaccines against Lassa fever]. / Ispol'zovanie morskikh svinok dlia otsenki éffektivnosti vaktsin protiv likhoradki Lassa.

The use of guinea pigs as a laboratory model was proven to be appropriate in investigating the vaccines developed against Lassa fever at the preclinical study stage. An adapted variant of Lassa virus was cultivated, which caused death of guinea pigs with respect for an agent's dose. Finally, it was shown to be possible to investigate the immunogenic and protective properties of the inactivated antigen of Lassa virus in experiments with guinea pigs.