[Enzyme immunoassay system for serological diagnosis of hemorrhagic fever with renal syndrome based on inactivated purified Puumala virus (Hantaviridae: Orthohantavirus)].

INTRODUCTION: Hemorrhagic fever with renal syndrome (HFRS) is the most common zoonotic human viral disease in the Russian Federation. More than 98% of the HFRS cases are caused by Puumala orthohantavirus (PUU). Effective serological tests are required for laboratory diagnosis of HFRS. OBJECTIVE: Construction of an enzyme immunoassay (ELISA) test system for detection of specific antibodies using standard antigen in the form of highly purified inactivated PUU virus as immunosorbent. MATERIALS AND METHODS: Preparation of PUU virus antigen, designing the ELISA for detection of specific antibodies, developing parameters of the ELISA system, parallel titration of HFRS patients sera by fluorescent antibody technique (FAT) and the new ELISA. RESULTS AND DISCUSSION: For the first time, ELISA based on purified inactivated PUU virus as standard antigen directly absorbed onto immunoplate was developed. Parallel titration of 50 samples from HFRS patients blood sera using FAT and the developed ELISA showed high sensitivity and specificity of this ELISA, with 100% concordance of testing results and significant level of correlation between the titers of specific antibodies in the two assays. CONCLUSION: The ELISA based on purified inactivated PUU virus as an immunosorbent can be effectively used for HFRS serological diagnosis and for mass seroepidemiological studies.

[Experience of application of IgY-technology for laboratory diagnostics of viral infections.]

INTRODUCTION: The well-known advantages of class Y antibodies (IgY) from egg yolks of immunized hens in comparison with class G antibodies (IgG) of laboratory animals traditionally used in laboratory diagnosis of infectious diseases determine the stable interest of researchers in using IgY for these purposes (IgY technology). Over the past 20 years, the obvious benefits of IgY technology have been demonstrated for a number of viral and bacterial infections. Goals and objectives. Construction of ELISA systems based on specific IgY for laboratory diagnosis of infections caused by tick-borne encephalitis virus, yellow fever virus, poliovirus. MATERIAL AND METHODS: Obtaining yolk preparations of immunized chickens, obtaining highly purified IgY preparations (salting out, affinity chromatography), constructing ELISA systems for determining virus-specific antigens, testing the parameters of ELISA systems. RESULTS AND DISCUSSION: For the first time in laboratory practice, ELISA systems based on the use of specific polyclonal IgY were designed for laboratory diagnosis of topical human viral infections caused by flaviviruses and enteroviruses: determination of antigens of tick-borne encephalitis virus, yellow fever virus, 3 types of poliovirus. It was experimentally shown that these ELISA systems have high sensitivity and specificity, which allows them to be used for the semiquantitative and quantitative determination of antigens of these viruses in various materials (infected cell cultures, vaccines, etc.). CONCLUSION: The ELISA systems developed on the basis of specific IgY for determination of viral antigens can be effectively used for laboratory diagnosis of a number of viral infections, for the validation and control of vaccine preparations.

[Development of inactivated cultural yellow fever vaccine].

INTRODUCTION: The only currently available live vaccine against yellow fever (YF) based on chicken embryos infected with an attenuated 17D strain of the YF virus is one of the most effective vaccine preparations. However, the live vaccine is associated with "viscerotropic syndrome" (approximately 0.4 cases per 100 000 vaccinated). Therefore, the development and introduction of highly purified inactivated vaccine against YF is intended to ensure the maximum safety of vaccination against one of the most common human viral diseases.Goals and objectives. Development and evaluation of immunogenicity of the cultural inactivated vaccine against YF at the laboratory model level. MATERIAL AND METHODS: Adaptation of 17D strain of YF virus to Vero cell culture, cultivation, removal of cellular DNA, inactivation with ß-propiolactone, concentration, chromatographic purification, determination of protein and antigen of YF virus, assessment of immunogenicity in mice in parallel with commercial live vaccine. RESULTS AND DISCUSSION: Immunogenicity: the determination of specific antibodies of class G (IgG) and virus neutralizing antibodies in the sera of immunized mice showed high level of antibodies exceeding that of immunized with commercial live vaccine. The optimal dose of antigen in the vaccine (total protein) was 50 µg/ml (5 µg/0.1 ml -dose and volume per 1 vaccination of mice). Thus, the laboratory version of cultural inactivated vaccine against YF is as effective (and even superior) as the commercial live vaccine. CONCLUSION: Laboratory version of cultural inactivated vaccine against YF, which is not inferior in immunogenicity (in animal model) to commercial live vaccine, has been developed.

[Production of the polyclonal enterovirus antibodies of chicken (IgY) and its evaluation as alternative to the rabbit enterovirus neutralizing sera].

Experimental data show the usefulness of the Leghorn chicken as a producer of the enterovirus neutralizing antibodies (IgY). The resulting serum is not inferior to the specific activity of the commercial rabbit enterovirus diagnostic sera (EDS) in the neutralization reaction. The IgY have lower backgrounds than mammalian IgG and do not cause toxic effect to cell culture. Compared with the conventional manufacturing method EDS IgY, preparation process is much more effective: the number of serum producers is significantly lower, whereas the yield of the product is higher. Reduction of the volume of the immunogens, immunization cycle, and number of injections is also an advantage of this manufacturing method.

[STRUCTURAL, FUNCTIONAL, AND VEGETATIVE CONDITIONS OF PATIENTS WITH STABLE ARTERIAL HYPERTENSION, ITS MASKED FORM, AND WHITE COAT HYPERTENSION].

The character of changing of functional and vegetative characteristics in different clinical variants of arterial hypertension thus far remains obscure. The aim of this work was to study patients with stable arterial hypertension (SAH), its masked form (MH), and white coat hypertension (WCH). We examined 125 patients who were divided into these 3 groups based on results of 24 hr AP monitoring by cardiac rhythm measurement and echocardiography. The study revealed different risk factors of each of the three forms of AH. The main risk factors of WCH are age and disturbed rhythm variability, those of MH are overweigh, obesity, and myocardial dysfunction, while SAH is associated with the advanced age, overweight, obesity, the history of concomitant cardiovascular diseases, and disordered daily profile of AP. It is concluded that different clinical variants of AH result from the changes in the functioning of cardiovascular and vegetative nervous systems associated with different risk factors.

[State of the vegetative nervous system and the risk of repeated hospitalization after elimination of hypertonic crisis].

Observation of 58 outpatients with arterial hypertension (AH) revealed a group of 25 subjects who needed repeated hospitalization within 1 year after hypertonic crisis due to unstable conditions (group 1). The two groups were comparable in terms of office AP measurements, but 24 hr AP, systolic AP and its variations were significantly higher while daily index was lower in group 1. Moreover, patients of this group had decreased variability of the cardiac rhythm with increased activity of the suprasegmental level and reduced parasympathetic activity. A rise in BMI in these patients correlated with the state of sympathetic component of the vegetative nervous system. It is concluded that normalization of office AP can not be regarded as evidence of benign course of the disease; evaluation of daily AP characteristics, BMI, and balance of vegetative nervous system is needed to stratify risks for the outpatients with AH followed up after the treatment of hypertonic crisis.

The 2010 outbreak of poliomyelitis in Tajikistan: epidemiology and lessons learnt.

A large outbreak of poliomyelitis, with 463 laboratory-confirmed and 47 polio-compatible cases, took place in 2010 in Tajikistan. Phylogenetic analysis of the viral VP1 gene suggested a single importation of wild poliovirus type 1 from India in late 2009, its further circulation in Tajikistan and expansion into neighbouring countries, namely Kazakhstan, Russia, Turkmenistan and Uzbekistan. Whole-genome sequencing of 14 isolates revealed recombination events with enterovirus C with cross-overs within the P2 region. Viruses with one class of recombinant genomes co-circulated with the parental virus, and representatives of both caused paralytic poliomyelitis. Serological analysis of 327 sera from acute flaccid paralysis cases as well as from patients with other diagnoses and from healthy people demonstrated inadequate immunity against polio in the years preceding the outbreak. Evidence was obtained suggesting that vaccination against poliomyelitis, in rare cases, may not prevent the disease. Factors contributing to the peculiarities of this outbreak are discussed. The outbreak emphasises the necessity of continued vaccination against polio and the need, at least in risk areas, of quality control of this vaccination through well planned serological surveillance.

[An ELISA system based on the specific class Y (IgY) antibodies from egg yolks for the quantitative determination of D-antigen in inactivated poliovirus vaccines].

The results of the construction of the first Russian ELISA system for the quantitative determination of D-antigen of 1-3 poliovirus types in the preparations of inactivated poliovirus vaccines are presented. For the first time, this kind of system is based on the use of specific antibodies of class Y (IgY) from egg yolks of immunized hens. It was shown that this ELISA system is specific, sufficiently sensitive, and can be used for quantitative determination of D-antigen of 1-3 poliovirus types in inactivated poliovirus vaccines.

[Effect of cardiovascular functional disorders on cerebral oxygenation in children].

The article deals with data of cerebral oximeter INVOS Somanetics (US) used for comparison the degree of brain oxygenation reduction, ejection fraction and stroke volume relative to age norms in children with dilated cardiomyopathy.

[Obstructive sleep apnea syndrome in patients with acute myocardial infarction: timing of onset of the disease, clinico-functional and some other characteristics].

With the aim of elucidating special features of the course of acute myocardial infarction (MI) in patients with obstructive sleep apnea syndrome (OSA) we retrospectively studied clinico-functional parameters of 476 patients including 64 (13.3%) who had episodes of apnea during sleep. Frequency of OSA was 1.3 times greater in overweight patients, and 2.3 times greater in those with history of diabetes mellitus. Development of MIs in patients with OSA more often occurred during night or early morning (relative risk 6.00-7.41) and depended on the severity of OSA. Patients with MI and OSA had lowering of blood levels of high density lipoproteins, elevation of levels of triglycerides and high sensitive C-reactive protein. We noted the role of excessive neurohumoral activity in these processes. A conclusion was made about necessity of timely diagnosis of OSA during MI and timely correction of the revealed pathophysiological traits.

[Esophagoplasty in combined scarry lesions of the esophagus, laryngopharynx and the trachea in children].

The experience of plasty of the pharynx and esophagus with graft of the free small intestine segment was presented in 12 children after burn stricture by the potassium permanganate. The technical aspects of operation using methods of the reconstructive vascular surgery were described. Good short-term result was obtained in all the patients. The long-term results were investigated during 15 years. It was observed, that the graft diverticulum developed by 4-7 years in 3 patients, whom repeated operation should be performed. According to the authors, the regional plasty of the esophagus by free revasculizated small intestine graft was really effective surgical supply and could be considered as the method of choice in limited scarry lesions of the pharynx and cervical esophagus, but the method should be improved.

[Repeated plasty of the esophagus in children with the application of transplant from the ileum].

The authors give topographic-anatomical substantiation of a possibility to employ the ileum for esophageal replacement and describe a technique of forming such transplant. Their clinical experience includes 9 operated children. Earlier they had endured from 1 to 4 attempts of esophageal replacements by different traditional methods. In all children operated upon good results were obtained. It allows the ileum to be included in the arsenal of possible methods of esophageal replacement and considering it as an alternate variant in nonstandard situations.

[Effect of acute operational stress on the exchange of biogenic amines in prostate gland and steroidogenesis (experimental study)].

Patterns of metabolic imbalance of biogenic amines in the tissue of the prostate, leading to violations of hemodynamics and trophism, were revealed in experimental study on laboratory animals. At the same time the stress was acute inducer of functional testosterone deficiency, which persisted throughout the observation period. Based on experimental data obtained, pathogenetic role of stress-induced metabolic imbalance of biogenic amines and steroidogenesis as possible early pathophysiological mechanisms of prostate diseases in humans is proved.

[Total risk for coronary events in cardiac patients due to snoring, daytime drowsiness, and obstructive sleep apnea syndrome].

AIM: To assess the risk of cardiovascular events due to obstructive sleep apnea syndrome (OSAS) in persons without organic heart diseases. SUBJECTS AND METHODS: Two hundred and forty men complaining about their sleep snoring and/or daytime drowsiness and 209 men without this symptomatology were examined. The presence of OSAS was verified by cardiorespiratory monitoring. Blood lipid composition, blood pressure, heart rate, and body mass index were determined in all the examinees. The risk for cardiovascular diseases (CVD) was assessed using the SCORE and PROCAM scales. RESULTS: When only the subjective manifestations of OSAS were present, there was a preponderance of hypercholesterolemia and an increased risk for CVD as possible markers of preclinical atherosclerosis. Hypertension and smoking, as well as a combination of 4 traditional CVD risk factors are recognized the worst. At the same time, the PROCAM scale allows identification of a high-risk group, the size of which is 1.6 times more than that of the group rated by the SCORE scal CONCLUSION: OSAS is an important CVD risk factor and the PROCAM scale is more preferable to identify a group at high risk for CVD among subjects with OSAS.

[Management of arrhythmia in patients with arterial hypertension].

We estimated the risk of arrhythmic complications after the use of cardiovascular medications in a prospective study including 123 patients with arterial hypertension and recurrent atrial fibrillation. Dispersion characteristics of P wave and QT interval were studied in patients with left ventricular concentric remodeling, concentric hypertrophy, and eccentric hypertrophy. The role of therapy with beta-blockers. ACE inhibitors, and preductal in the reduction of the risk of atrial fibrillation and ventricular arrhythmia was evaluated.

[Endoscopic treatment of children with esophagus stenoses of different etiology].

Nowadays not only diagnostic but also therapeutic interventions are widely used in pediatric endoscopy. The authors analyze 25 cases of esophageal stenosis of different genesis (caustic, peptic, anastomotic and congenital esophageal obstructions) in children aged from 1.6 to 15 years. The endoscopic treatment was successfully performed by mechanical dilators with a guidewire and balloon dilator in all patients. The age less than 2 years, previous complications, subtotal stenosis can not be considered absolute contraindication for endoscopic treatment.

[Disturbances of heart rhythm in patients after myocardial infarction with obstructive sleep apnea during outpatient rehabilitation].

UNLABELLED: We studied specific features of heart rhythm disturbances observed during outpatient rehabilitation of 432 patients after first myocardial infarction. In 94 of these patients (21.8%) we diagnosed obstructive sleep apnea syndrome (OSAS). We noted substantial preponderance of pathologic ectopy related to OSAS with most significant detection of frequent ventricular extrasystoles. Their paired variant and polymorphic forms in patients with OSAS were 3.4 and 7.4 times, respectively, more frequent than in patients without OSAS. Frequency of ventricular ectopy rose with increase of severity of OSAS and was maximal during nocturnal hours. Analysis of correlations revealed association between OSAS and frequencies of ventricular and supraventricular extrasystoles. CONCLUSION: After myocardial infarction patients should be thoroughly examined for timely detection of OSAS and corresponding correction of their rehabilitation program.

[Obstructive sleep apnea syndrome, arterial hypertension, and obesity: clinical and functional aspects].

A total of 196 patients with increased body mass index (BMI) were included in the study designed to elucidate specific features of arterial hypertension (AH) and obstructive sleep apnea syndrome (OSAS). 65 of them did not experience OSAS episodes. All the patients were examined using 24-hour ECG and respiration monitoring combined with veloergometry (VEM). It was shown that OSAS alone in the absence of AH was associated with a 1.17-fold decrease of the physical load threshold compared with the respective control values. In the patients suffering AH and having high BMI, this parameter further decreased (by 1.22 and 1.35 times, respectively). In addition, they experienced changes in the period of recovery from veloergometry when their heart rate decreased but insignificantly by the end of the first minute after the termination of exercises. It is concluded that OSAS and AH influence tolerability, of physical exercises especially in the patients with elevated BMI.

[Temperature regime of biological tissue under photodynamic therapy].

An analytical model is proposed to calculate heating of human skin cover under laser light action of photodynamic therapy. A photosensitizer of "Fotolon" is taken as an example. Temperatures of skin surface and of deep dermis regions are studied as a function of time under pulsed and stationary irradiation of skin surface at the wavelength of 665 nm corresponding to the maximum of the photosensitizer absorption band. It is shown that, under the action of a short light pulse, the photosensitizer can lead to an essential temperature rise of dermis due to a considerable increase in its absorption coefficient. However, this rise does not destruct tissue cells because of the short action. Under stationary irradiation, the photosensitizer concentration has a low effect on the temperature regime of tissue. This is related with the specific features in heating of the medium by red light, where the main thermal process in skin is heat transfer over tissue volume from epidermis having a substantially larger absorption coefficient than that of dermis in the said spectral range. The role of blood perfusion in dermis and its effect on the temperature regime of tissue are evaluated.