[Cell technologies as a basis for the development of regenerative principles for the treatment of lacrimal gland diseases]. / Kletochnye tekhnologii kak osnova razrabotki regeneratornykh printsipov lecheniya zabolevanii sleznoi zhelezy.

The lacrimal gland (LG) is a tubuloacinar exocrine gland composed of acinar, ductal, and myoepithelial cells. Three-dimensional distribution of acinar lobules, ducts, and myoepithelial cells is necessary for the effective functioning of the organ. LG is the main organ of immune surveillance of the ocular surface system. The embryogenesis of the gland is regulated by the interaction of genetic mechanisms, internal epigenetic (enzyme systems, hormones) and exogenous factors. There is no doubt that there is a clear genetic program for the implementation of the complex process of embryonic development. The mechanisms regulating LG organogenesis initiate the work of a huge number of structural oncogenes, transcription and growth factors, etc. Studying the expression and selective activity of regulatory genes during organ development, their participation in the differentiation of different cell types is a current trend at the nexus of clinical genetics, molecular biology, embryology and immunocytochemistry. Due to its relatively simple structure and accessibility, human LG is a suitable object for potential application in regenerative medicine. Development of a universal protocol for obtaining functional differentiated secretory epithelium of LG capable of expressing tissue-specific markers is an urgent task. Determining the nature and origin of stem cells and progenitor cells will allow the isolation and multiplication of these cells in culture. After obtaining a functionally active culture of LG cells, it is possible to create a model of autoimmune diseases.

[The role of miRNA in the pathogenesis of diseases associated with functional dysregulation of the lacrimal gland]. / Rol' mikroRNK v patogeneze zabolevanii, svyazannykh s narusheniem funktsii sleznoi zhelezy.

At this time, the mechanism causing lacrimal gland dysfunction is not understood completely. In diseases associated with lacrimal gland involvement (Sjogren's syndrome, sarcoidosis, IgG4-associated disease, etc.) patients have been observed to experience elevated cellular apoptosis, active production of autoantibodies to glandular tissue, increased level of pro-inflammatory cytokines, functional disruption of signaling molecules leading to changes in tear production. Difficulties in differential diagnosis of lacrimal gland dysfunction in above-listed diseases are associated, on the one hand, with similarity of the clinical picture of ophthalmological manifestations, and on the other hand - with complicated morphological interpretation of changes in the glandular tissues. In this view, miRNA is a promising diagnostic and prognostic marker that would help with differential diagnosis as well as with choosing the treatment tactics. Methods of molecular profiling and identification of "molecular phenotypes" of lacrimal gland and ocular surface damage will allow the use of miRNA as biomarkers and prognostic factors for personalized treatment.

[Evolution of dry eye disease diagnostics]. / Evolyutsiya metodov diagnostiki sindroma sukhogo glaza.

This article reviews modern functional and instrumental examination methods included in the diagnostic algorithm for dry eye disease. The described methods can serve as an objective criterion for the effectiveness of the therapy.

[Predictive significance of genetic analysis of the development of dry eye disease of different origin]. / Prediktivnaya znachimost' geneticheskogo analiza razvitiya sindroma «sukhogo glaza¼ razlichnogo geneza.

One of the etiological causes of dry eye disease (DED) is systemic autoimmune diseases (AID): primary Sjögren's syndrome (PSS), rheumatoid arthritis (RA); their manifestation may begin with ophthalmic symptoms. The relationship of PSS and RA with genetic factors is proven. The contribution of polymorphic markers of the genes THBS1, MUC1, TRIM21, STAT4, PTPN22 in the development of these diseases is established, as well as their connection with the development of DED. A panel of genetic markers for evaluating the risk of developing DED in PSS and RA is developed, and its sensitivity and specificity is determined. PURPOSE: The aim of the study was to determine the prognostic significance of a panel of polymorphic gene markers in the development of dry eye syndrome in patients with primary Sjögren's syndrome and rheumatoid arthritis over a five-year follow-up period. MATERIAL AND METHODS: Patients with a verified diagnosis of PSS and RA without signs of DED were examined (n=35 and n=42, respectively). The control group included 82 volunteers without AID and DED. The observation period was 5 years. Every year all study subjects underwent an ophthalmological clinical and functional examination. RESULTS: Dry eye disease had developed in groups of patients with AID with predisposing genotypes of polymorphic markers of the genes THBS1, MUC1, TRIM21, STAT4, PTPN22. The peak of DED development in these patients was in the third year of the follow-up. As a result of ROC analysis, it was found that the sensitivity and specificity of determining the predisposing genotypes of polymorphic markers of the THBS1, MUC1, TRIM21, STAT4, PTPN22 genes was 68 and 87%, respectively (p<0.0001). CONCLUSION: Genetic research methods are essential for minimally invasive early diagnosis of dry eye disease, and can subsequently become the basis for a personalized approach to its treatment.

[Association of polymorphic markers rs7947461 of the TRIM21 gene and rs33996649 of the PTPN22 gene with the risk of developing exogenous dry eye syndrome]. / Assotsiatsiya polimorfnykh markerov rs7947461 gena TRIM21 i rs33996649 gena PTPN22 s riskom razvitiya sindroma sukhogo glaza ekzogennoi etiologii.

In this age of technological advancement, an increasing number of people is being exposed to external risk factors of damaging their ocular surface (wearing contact lenses, electromagnetic radiation from computers, mobile devices, etc.). However, the presence of external factors does not lead to a 100% risk of developing the dry eye disease (DED). The trigger mechanism in the development of autoimmune lesions of the ocular environment in some systemic diseases is known to be associated with molecular genetic factors. The search for molecular genetic disorders is based on the analysis of polymorphic markers of a number of genes responsible for the state of the eye surface. PURPOSE: To study the relationship of polymorphic markers rs7947461 of the TRIM21 gene and rs33996649 of the PTPN22 gene with the risk of developing dry eye syndrome of exogenous etiology. MATERIAL AND METHODS: The study included 57 people with exogenous risk factors for DED development. The control group included volunteers without a history of ophthalmic pathologies (n=75). Genotyping was done by real-time polymerase chain reaction followed by melting curve analysis. Statistical processing of data was done using the Statistica 6.1 RUS software for statistical analysis. RESULTS: In the course of the study, 31 patients of the main group were diagnosed with DED and separated into the 1st subgroup; DED diagnosis was not confirmed in 26 patients, who were put into the 2nd subgroup. The 1st subgroup showed a significant increase in the frequency of predisposing genotypes of the TRIM21 and PTPN22 genes. The relative risk of developing DED turned out to be 2.5 and 4.86 times higher, respectively. In the 2nd subgroup, no statistically significant data was found on the presence of predisposing genotypes of polymorphic markers of the TRIM21 and PTPN22 genes (p=0.3). CONCLUSION: The revealed association of polymorphic markers rs7947461 of the TRIM21 gene and rs33996649 of the PTPN22 gene with the risk of developing DED of exogenous etiology puts these loci as possible markers for diagnosing this pathology.

The Role of Polymorphic Markers rs1478604, rs2292305, and rs2228262 in THBS1 Gene in the Development of Autoimmune Dry Eye Syndrome.

An association of polymorphic marker rs2228262 in the THBS1 gene with the risk of developing dry eye in Sjögren syndrome was revealed. Confocal microscopy data suggest that this polymorphic marker is responsible for the high probability of corneal nerve fiber lesion in Sjögren syndrome even in the absence of clinical and functional signs of dry eye syndrome. A significant correlation was established between polymorphic markers rs1478604, rs2228262 in THBS1 gene and the coefficients of anisotropy and orientation symmetry of corneal nerve fibers. These results allow considering these polymorphic markers as a genetic factor of predisposition to dry eye syndrome in patients with Sjögren syndrome.

[Association of polymorphisms of the TRIM21 gene with the severity of dry keratoconjunctivitis in rheumatoid arthritis and Sjogren's disease]. / Vzaimosviaz' polimorfizmov gena TRIM21 s tiazhest'iu sukhogo keratokon''iunktivita pri revmatoidnom artrite i bolezni Shegrena.

Ophthalmologic manifestation of Sjogren's disease (SD) and rheumatoid arthritis (RA) is dry keratoconjunctivitis (dry eye disease; DED). PURPOSE: To study the relationship of polymorphic markers rs7947461 (C/T), rs915956 (C/T), rs4144331 (C/A) of the TRIM21 gene with the severity of DED in patients with RA and SD. MATERIAL AND METHODS: The study included 70 patients with RA (n=27) and SD (n=43). The control group consisted of volunteers without a history of RA or SD (n=35). Alleles of the polymorphic marker C660T rs7947461 of the TRIM21 gene were identified using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method; alleles of the polymorphic marker rs915956 (C/T) and rs4144331 (C/A) of the TRIM21 gene were identified by analyzing DNA melting curves. RESULTS: An association was found between the predisposing genotype (TT) of rs7947461 polymorphic marker and the risk of developing severe DED. The AA genotype of rs4144331 polymorphic marker was found only in severe DED (c2=7.74; OR=17.46, CI95%=1.96-318.38, p=0.02). CONCLUSION: An association was established between rs7947461 (rs660) and rs4144331 and the risk of developing severe DED.

[Polymorphic markers of certain genes in the development of dry keratoconjunctivitis in patients with rheumatoid arthritis and Sjogren's syndrome]. / Polimorfnye markery nekotorykh genov v razvitii sukhogo keratokon''iunktivita u patsientov s revmatoidnym artritom i sindromom Shegrena.

The article reviews literature on relationships between polymorphic variants of the genes THBS1, GTF2I, MUC1, TRIM21, STAT4, PTPN22 with clinical features of dry keratoconjunctivitis in rheumatoid arthritis and Sjogren's syndrome. The development and implementation of a method for analyzing polymorphic gene variants used to diagnose dry keratoconjunctivitis in rheumatoid arthritis and Sjogren's syndrome will allow assessment of the possibility of developing dry keratoconjunctivitis and/or its progression in patients with autoimmune diseases or in people at risk. Determination of clinical and morphological regularities of dry keratoconjunctivitis in accordance with the revealed molecular and genetic changes will contribute to better understanding of the etiology and pathogenesis of ophthalmological manifestations of autoimmune diseases, and will also help improve the diagnostics and prognosis of dry keratoconjunctivitis.

[Interpretation of the results of immunohematological tests in hematological patients.]

The correct determination of the blood types of the recipient and the donor is very importante for the choice of blood components for transfusion. As a result of the study, it was established that 18.0% of patients, admitted to the hematology clinic, have difficulties in interpreting of the results of immunohematological tests. Most often, a double population of red blood cells was detected when determining antigens of the Rhesus system (10.9%), auto- (3.9%) and alloantibodies (2.8%). The proposed algorithm for the selection of donor red blood cells in difficult diagnostic cases helps to prevent the development of post-transfusion complications.

[Molecular genetic examination for dry eye syndrome prognosis]. / Molekuliarno-geneticheskie issledovaniia v prognoze techeniia sukhogo keratokon''iunktivita.

The article reviews the results of Russian and foreign studies concerning the search of susceptibility genes for widespread multifactorial diseases: Sjogren's syndrome and rheumatoid arthritis. Studying regularity patterns of genome changes in autoimmune processes with ophthalmic manifestations and their correlation with the severity of dry keratoconjunctivitis will contribute to a better understanding of the etiology and pathogenesis of eye manifestations of the diseases, and will also allow the development of new effective methods of diagnosis and targeted therapy.

[The methods of diagnostic of immune thrombocytopenia of fetus and newborn].

The thrombocytopenia is found in 1%-5% of newborns. Depending on mechanisms of pathogenesis of thrombocytopenia is divided on immune and non-immune one. The reaction of of interaction between antibodies and antigens of superficial structures of cells are in the basis of immune destruction of thrombocytes. During intrauterine period and period of newborness auto-, trans- and alloimmune alternatives of development of thrombocytopenia can be observed. The neonatal alloimmune thrombocytopenia is registered with rate of 1 case per 800-1000 newborns. The study was targeted to developing algorithm of diagnostic of immune thrombocytopenia, detecting main diagnostic criteria, exploring clinical significance of results of laboratory tests. The methods of study included typing of genes of HPA system using polymerase chain reaction with detection of results in real-time mode, detection of compatibility of HPA-genotypes of mother and child using flow cytometry technique. The following criteria of diagnostic of neonatal alloimmune thrombocytopenia: 1. detection of incompatible combination of HPA genes in mother and child (HPA-1bb/HPA-1ab; HPA-5aa/HPA-5ab; HPA-15ab/HPA- 15ab); 2. detection in blood serum of mother antibodies adsorbing more than on 3% of thrombocytes of child; 3. absence of antithrombocyte antibodies in mother and child (coefficient of auto-sensitization is less than 5%). The immune genesis of thrombocytopenia is established in 40% 0f children with low number of thrombocytes at birth. In 50% of cases the cause was determined as anti-thrombocyte alloantibodies with diagnosis neonatal alloimmune thrombocytopenia. Also in 50% of cases decreasing of number of thrombocytes in children occurred as result of impact of autoantibodies of mother with diagnosis "transimmune thrombocytopenia". The considered laboratory methods have high specificity and permit to properly diagnose immune causes of thrombocytopenia in newborns.

Δ6-fatty acid desaturase and fatty acid elongase mRNA expression, phagocytic activity and weight-to-length relationships in channel catfish (Ictalurus punctatus) fed alternative diets with soy oil and a probiotic.

A time-course feeding trial was conducted for 120 days on juvenile channel catfish (Ictalurus punctatus) to study the effects of diets differing in oil source (fish oil or soy oil) and supplementation with a commercial probiotic. Relative levels of Δ6-fatty acid desaturase (Δ6-FAD) and fatty acid elongase (FAE) expression were assessed in brain and liver tissues. Both genes showed similar expression levels in all groups studied. Fish weight-to-length relationships were evaluated using polynomial regression analyses, which identified a burst in weight and length in the channel catfish on day 105 of treatment; this increase was related to an increase in gene expression. Mid-intestinal lactic acid bacterium (LAB) count was determined according to morphological and biochemical criteria using API strips. There was no indication that intestinal LAB count was affected by the modified diets. The Cunningham glass adherence method was applied to evaluate phagocytic cell activity in peripheral blood. Reactive oxygen species (ROS) generation was assessed through the respiratory burst activity of spleen macrophages by the NBT reduction test. Probiotic-supplemented diets provided a good substrate for innate immune system function; the phagocytic index was significantly enhanced in fish fed soy oil and the probiotic, and at the end of the experimental period, ROS production increased in fish fed soy oil. The substitution of fish oil by soy oil is recommended for food formulation and will contribute to promoting sustainable aquaculture. Probiotics are also recommended for channel catfish farming as they may act as immunonutrients.

[The detection of thrombocyte-bound IgG in patients with throbocytopenia].

The article presents the mode of detection of anti-thrombocyte auto-antibodies using flow cytometry technique. The values of IgG conjugated with thrombocytes are established in healthy persons. The rate of frequency of auto-antibodies to thrombocytes is analyzed in patients with primarily diagnosed immune thrombocytopenia, with relapse of disease and after therapy of immune thrombocytopenia. The carried out studies demonstrate that flow cytometry technique has a high sensitivity to detect thrombocyte-related antibodies and can be applied under examination of patients with thrombocytopenia.

[The interpretation of immunophenotyping results during diagnostics of lymphatic proliferative disease using immunophenotyping count].

The article considers immune phenotyping heterogeneity of chronic lymphatic leukemia detected using basic diagnostic markers ofcell. The results of analysis of immune phenotypes of 108 patients with B-cell lymphatic proliferative diseases made it possible to establish that the atypical is related most rarely to indicators of expression of monotypic immunoglobulines and CD5 and most frequently to CD23, FMC7, CD22 and CS79b. During the present observation, the immune phenotyping count made up "3" or "2"points and the atypical alternative was registered among 10% of all examined patients with chronic lymphatic leukemia. It is demonstrated that patients with chronic lymphatic leukemia and with lower immune phenotyping count are characterized by major intensity of tumor substrate.

[Efficacy of photon therapy in combined postoperative treatment of children with congenital obstructive uropathy and secondary pyelonephritis].

A total of 110 children aged from 5 to 15 years with different forms of congenital obstructive uropathy and secondary pyelonephritis received standard combined treatment (n = 71) or standard treatment combination with photon therapy 3 months after discharge from hospital. Postoperative combined treatment including photon therapy promoted uneventful course of the postoperative period, normalized immunological indices, achievement of persistent remission.

[New immunoglobulin for treatment of anthrax].

AIM: To experimentally assess activity and safety of anti-anthrax intravenous immunoglobulin manufactured on standard technology. MATERIALS AND METHODS: Plasma from selected donors vaccinated with combined anthrax vaccine was tested by enzyme immunoassay. Samples of plasma with increased titer of anti-anthrax antibodies were merged in one manufacturing load and fractionated in ethanol at negative temperature according to standard technology. Formulation of intravenous immunoglobulin was manufactured according to standard technology of acid-enzyme hydrolysis. RESULTS: Proved medical technology of donors immune plasma fractionation provided 4 - 8-fold concentration of anti-anthrax antibodies. The finished product contained 5% of protein and was apyrogenic, non-toxic, thermostable, electrophoretically homogenous, had pH 6.65 and meet the requirements for manufacturing batches of human intravenous immunoglobulin. CONCLUSION: Protective effects of experimental human anti-anthrax immunoglobulin were comparable with control biological--equine anti-anthrax immunoglobulin for intramuscular use.

Immunologic parameters evaluations in Nile tilapia (Oreochromis niloticus) exposed to sublethal concentrations of diazinon.

Fish resistance to microorganisms depends basically on the immune response. Although there are several studies on the diazinon mammalian immunotoxicity, in the case of fish there are only few. The aim of present study was to evaluate the effect of diazinon on immunological parameters (relative spleen weight, splenocytes count, lysozyme activity, respiratory burst and IgM concentration) in Nile tilapia. Diazinon at sublethal concentrations (0.39 and 0.78 mg/L) did not alter RSW, splenocytes count or lysozyme activity. However, at the highest concentration tested (1.96 mg/L) diazinon significantly increased respiratory burst and IgM concentration. In summary, diazinon (and perhaps other pesticides) could alter immunological response and induce oxidative stress.

Organosilica composite for preconcentration of phenolic compounds from aqueous solutions.

A new adsorbent is proposed for the solid-phase extraction of phenol and 1-naphthol from polluted water. The adsorbent (TX-SiO(2)) is an organosilica composite made from a bifunctional immobilized layer comprising a major fraction (91%) of hydrophilic diol groups and minor fraction (9%) of the amphiphilic long-chain nonionic surfactant Triton X-100 (polyoxyethylated isooctylphenol) (TX). Under static conditions phenol was quantitatively extracted onto TX-SiO(2) in the form of a 4-nitrophenylazophenolate ion associate with cetyltrimethylammonium bromide. The capacity of TX-SiO(2) for phenol is 2.4 mg g(-1) with distribution coefficients up to 3.4 x 10(4) mL g(-1); corresponding data for 1-naphthol are 1.5 mg g(-1) and 3 x 10(3) mL g(-1). The distribution coefficient does not change significantly for solution volumes of 0.025-0.5 L and adsorbent mass less than 0.03 g; 1-90 microg analyte can be easily eluted by 1-3 mL acetonitrile with an overall recovery of 98.2% and 78.3% for phenol and 1-naphthol, respectively. Linear correlation between acetonitrile solution absorbance (A(540)) and phenol concentration (C) in water was found according to the equation A(540) = (6 +/- 1) x 10(-2) + (0.9 +/- 0.1) C (micromol L(-1)) with a detection range from 1 x 10(-8) mol L(-1) (0.9 microL g(-1)) to 2 x 10(-7) mol L(-1) (19 microL g(-1)), a limit of quantification of 1 microL g(-1) (preconcentration factor 125), correlation coefficient of 0.936, and relative standard deviation of 2.5%. A solid-phase colorimetric method was developed for quantitative determination of 1-naphthol on adsorbent phase using scanner technology and RGB numerical analysis. The detection limit of 1-naphthol with this method is 6 microL g(-1) while the quantification limit is 20 microL g(-1). A test system was developed for naked eye monitoring of 1-naphthol impurities in water. The proposed test kit allows one to observe changes in the adsorbent color when 1-naphthol concentration in water is 0.08-3.2 mL g(-1).

[Immunogenetics of congenital obstructive uropathies in children].

The authors have examined immunogenetically 136 children of east slavonic nationality aged 5-15 years with different forms of congenital obstructive uropathy. Positive and negative association was found of some HLA antigenes, their phenotypical combinations and haplotypical combinations with diseases in the group of patients with congenital obstructive uropathies and in patients with obstructive pyelonephritis. Different immunogenetic picture was seen in patients with congenital hydronephrosis and in children with pyeloectosy. Different distribution of immunogenetic markers can be used for differential diagnosis, definition of the risk of progression of hydronephrotic transformation, complication of obstructive pathology with secondary infectious process and objective validation of therapeutic policy in children with congenital obstructive uropathy.