Method of salivary histatins determination.

Histatins are the most significant antimicrobial peptides (AMP) of saliva and there are 12 types of such AMP. Histatin molecules contain relatively high percent of histidine and tyrosine residues. This property allows to use well known from organic chemistry Pauly reaction for detection of protein bounded histidine and tyrosine residues (BHT), which are in fact characterize the summary content of all histatins in saliva. Aim of the present study was comparison of BHT with antimicrobial activity of salivary AMP fraction in patients with inflammatory diseases of upper airways (IDUA). Group of examined persons include 28 patients with different diagnoses: chronic pharyngitis (n=11), chronic tonsillitis (n=7), nasopharyngitis (n=5), pollinosis (n=5). Degree of intensity of inflammatory symptoms was estimated in balls. The algorithm of BHT analysis include following steps: freezing - thawing of saliva; removal of microparticles by centrifugation; separation of fraction lower than100 kDa; dialysis for free amino acids removal; Pauly reaction carrying out. Antimicrobial activities of saliva and its low molecular fractions were estimated towards Candida albicans cells by the spectrophotometric method with bromocresol purpur. Analysis of saliva sediments for coccoid microbiota was carried out by PCR method. Pauly reaction for histatins estimation in saliva of IDUA patients use here for the first time. The histatins levels (BHT) were significantly correlated with the intensity of inflammatory symptoms (r=0,975) and activity of low molecular salivary fraction (AMP activity) (r=0,824). The AMP activity/ BHT ratio, i.e. antimicrobial activity of histatin unit, decreased together with growth of inflammatory symptoms intensity (r=-0,944). Any considerable differences in coccoid microbiota frequency of finding at different diagnoses were not detected. The S. aureus frequency of occurrence was connected neither with inflammatory symptoms intensity (r=0,118), nor with BHT concentration (r=0,318). However S. pyogenes and S. pneumoniae frequencies of occurrence demonstrated the invert correlation towards these indexes: (r=-0,627/-0,614) and (r=-0,827/-0,864). Probably at the exacerbation forms of IDUA the S. pyogenes and S. pneumoniae growth controlled by high levels of histatins.

[Clinical course of acute respiratory infection and the state of microbiocenosis of upper respiratory tract in pregnant women].

AIM: Evaluate clinical features of the course of acute respiratory infection (ARI) and the state of microbiocenosis of nasopharynx of pregnant women at gestational term of 14 - 26 weeks. MATERIALS AND METHODS: At the stage of outpatient therapy of ARI 49 pregnant women at the gestational term of 14 - 26 weeks were examined: group 1--27 individuals consulting at day 1 and receiving Viferon (gel) intranasally for 10 days, group 2--22 individuals consulting at day 2 - 3 and receiving Viferon (gel and suppositorium). Respiratory viral infection was diagnosed by multiplex PCR. Bacteriologic study of discharge from the middle section of the pharynx was performed by the standard technique. RESULTS: ARI in group 1 of pregnant women were regarded as mild severity and persisted for an average of 3 days. In group 2 in 13 (59%) of patients ARI of moderate severity was noted and the recovery occurred at day 6. Pyo-inflammatory diseases of ENT organs appeared in 10 (45.4%) of patients. Respiratory infections were determined in 16 (32.7%) of them: coronaviruses--in 5 (10.2%), rhinoviruses--in 3 (6.2%), respiratory syncytial viruses--in 6 (12.3%), parainfluenza (type 2) and influenza A viruses--1 (2%) both, without significant differences between the observed groups. Streptococcus agalactiae, Staphylococcus aureus, Streptococcus mitis, Moraxella catarrhalis, Corynebacterium spp., Streptococcus pyogenes with the predominance of the latter in 36.7% and 60% of cases, respectively, in pregnant women of the group 2 (p < 0.01), were detected in the microflora of the mucous membrane of the nasopharynx. CONCLUSION: Viferon preparation therapy prescribed at day 1 of outpatient visit of pregnant women may promote mild course o fARI and prophylaxis of complications of upper respiratory tract.

[Specific antibodies in children with recurrent respiratory infections immunized with pneumo 23 and Act-HIB vaccines].

AIM: Assessment of specific IgG levels in children with recurrent respiratory infections after immunization with Pneumo 23 and Act-HIB vaccines. MATERIALS AND METHODS: Concentrations of IgG to polysaccharide capsular antigens and cell wall antigens of Streptococcus pneumoniae and Haemophilus influenzae were measured by solid-phase ELISA in sera of 256 immunized children. RESULTS: Increase of levels of IgG to cell wall antigens and certain polysaccharide antigens was observed in patients with low or intermediate antibody levels at baseline. CONCLUSION: Obtained results allow to conclude that vaccination of children with recurrent respiratory and middle ear infections results in induction of IgG production that proves immunological effect of performed vaccination.

[Use of multiplex PCR method with real-time detection for differential diagnosis of respiratory viral infections].

Multiplex real-time polymerase chain reaction test-system with fluorescent detection (RT-PCR) for simultaneous identification of main agents of acute respiratory viral infections: influenza A (IAV) and B viruses (IBV), parainfluenza viruses types 1, 2, 3, 4 (PIV 1 - 4), adenoviruses (ADV), respiratory syncitial virus (RSV), rhinoviruses (RV) and enteroviruses (EV), in presence internal positive control (IPC) represented by vaccine strain of rubella virus RA 27/3. Using multiplex RT-PCR method, respiratory viruses were detected in 116 out of 226 clinical samples (nasal swabs) obtained from patients with symptoms of acute respiratory infection: in 68 (58.6%) samples--IBV; in 21 (18.1%)--IAV; in 12 (10.3%) --RV; in 6 (5.2%)--PIV 2; in 4--(3.4%) ADV; in 3 (2.6%)--RSV; in 2 (1.7%)--EV; in 2 (1.7%)--PIV 4; in 1 (0.9%)--PIV 3; in 1 (0.9%)--PIV 1. Mixed infection was observed in 4 (3.4%) patients. PCR assay allowed to reveal various respiratory viruses in 51.3% of samples. At the same time samples were tested for the presence of 12 respiratory viruses--IAV, IBV, PIV 1 - 4, RSV, RV, metapneumoviruses, and coronaviruses NL63, 229E and OC43--in the presence of IPC represented by equine arteritis virus using analogous PCR test-system provided by medical center of Leiden university. Results of tests for detection of IAV, IBV, RSV, PIV 1 - 4, and RV, analyzed by both systems, agreed in 94%. Multiplex format of RT-PCR performing significantly reduces time and cost of the test, which make it suitable and effective instrument of epidemiological studies.

[Features of overall and local immunity in patients which chronic rhinitis of different etiology]. / Osobennosti obshchego i mestnogo immuniteta u bol'nykh s khronicheskimi rinitami razlichnoi étiologii.

The systemic and mucosal immune status in 50 patients with all-year-round allergic rhinitis (AYRAR), chronic infectious rhinitis (CIR) and non-atopic eosinophilic rhinitis (NER) was evaluated. Patients with concomitant pathology (bronchicial asthma, chronic bronchitis, sinusitis, etc.) were found to have changes in the systemic immunity. Patients with AYRAR and CIR had a high level of SIgA in their nasal secretions, and in the latter--also an elevated level of local IgG NER patients exhibited a considerable decrease in local production of immunoglobulins. In the nasal secretions of AYRAR and CIR patients high phagocytic activity of neutrophilis was established in the nitro blue tetrazolum test. The results of the study showed the expediency of determination of immune disturbances in these patients with a view to administration of immunocorrective therapy.