[Ocular lesions in HIV-infected patients of the ophthalmic hospitals]. / Porazhenie organa zreniia u VICh-infitsirovannykh patsientov oftal'mologicheskikh statsionarov.

PURPOSE: To analyze ophthalmic pathologies in HIV-infected patients of the ophthalmic hospitals of Perm city. MATERIAL AND METHODS: Medical records of 75 HIV patients registered in Perm Regional Centre for Prevention and Control of AIDS and Infectious Diseases who had received treatment in ophthalmic in-patient clinics of Perm in 2005-2015 years were analyzed retrospectively. Patient examination included traditional ophthalmological methods, as well as immunological (determination of CD4 cells and viral load), serological (detection of antibodies to herpes simplex virus, cytomegalovirus, chlamydia, toxoplasma), general clinical methods, and consultations by allied specialists. RESULTS: Ophthalmopathology requiring in-patient care was detected in 75 people (84 eyes). Men comprised 76%, women - 24%; average age was 32.82±8.68 years. The stage of HIV infection was known in 78.66% of patients: stage II - in 5% of cases, stage III - in 32%, stage IV - in 63%. Co-infection (hepatitis B and C, syphilis, tuberculosis) was detected in 81% of patients. HIV-related diseases (cytomegalovirus and herpes infection, candidiasis, toxoplasmosis) were observed in 48% of individuals. The time of emergence of ocular pathology from the time of HIV detection ranged from 1 day to 14 years. Inflammatory ocular diseases occurred in 55% of cases, dystrophic disorders - in 18%, eye traumas - in 24%, and strabismus - in 3% of patients. Reduced level of CD4 (less than 500 cells/mm3) was more prevalent (72.13%) among the study patients. As of the time of the study, only 36.4% of patients were receiving antiretroviral therapy. CONCLUSION: Among the studied individuals, eye lesion was mostly severe, inflammatory in nature; it occurred more frequently in stage IV HIV patients with reduced number of CD4 lymphocytes who was not receiving antiretroviral therapy.

Regulation of the peritoneal macrophage functional activity by the MP-5 and MP-6 myelopeptides under stress.

In mice, two-hour immobilization stress inhibited zymosan-induced production by macrophages of the oxygen radicals and cytokine IL-1ß. After myelopeptides MP-5 and MP-6 were administered into mice, the stress-induced inhibition of the reactive oxygen species (ROS) and IL-1ß was abrogated. MP-5 peptide stimulated spontaneous ROS production by macrophages and reduced IL-10 production under stress. Thus, under in vivo conditions and under stress, the effect of MP-5 and MP-6 myelopeptides modulates the peritoneal macrophage activity.

[The concentration of cerebral natriuretic peptide in tear and blood serum.]

The natriuretic peptides regulate pressure and water metabolism in human organism. The concentration of N-thermal cerebral natriuretic propeptide (NT-proBNP) in tear and blood serum was analyzed. The sampling included 49 patients divided on three groups. The group I (main) included 14 patients with primary open-angle sub-compensated glaucoma degree I-III. The group II (comparative group) included 15 patients with age-related immature cataract. The group III (control) included volunteers without any visual organ diseases. The concentration of NT-proBNP in tear and blood serum was detected by solid-phase enzyme-linked immunosorbent assay technique using test system NT-proBNP - IFA - BEST¼ («Vektor-Best¼, Russia). The median of content of NT-proBNP in tear turned out to be less than in blood serum in all groups (р1 = 0,00763; р2 = 0,00452; р3 = 0,00029) and made up to in patients with glaucoma 41,3 pg/ml, with cataract - 37.5 pg/ml, in healthy people 0 25.0 pg/ml (inter-quartile range made up to 20-60 pg/ml, 20-65 pg/ml, 10-49,5 pg/ml correspondingly). No differences in concentration of NT-proBNP were established in all groups both for blood serum (p = 0,494) and tear (p = 0,388). The concentration of NT-proBNP in tear correlated with its concentration in blood serum (r = 0,694). However, no dependencies were established from value of intra-ocular pressure (r = 0,168). Therefore, the concentration of NTproBNP in tear is not a perspective laboratory arker for diagnostic of increased intra-ocular pressure and stratification of development of glaucoma.

[THE ANALYSTS OF PROCALCITONIN IN LACRIMAL FLUID AND BLOOD SERUM UNDER UVEITIS].

The analysis was applied to concentration of procalcitonin in lacrimal fluid and blood serum in 15 healthy persons (control group), 16 patients with uveitis (main group) and 14 patients with non-inflammatory pathology of organ of vision (comparison group). The concentration of procalcitonin was detected by immunoenzyme method using commercial test-system "Procalcitonin-IFA-BEST" ("Vector-Best", Russia). The content of procalcitonin in blood serum was low (75% quartile--0.031 ng/ml) and had no significant difference between groups (H-criterion of Kruskal-Wallis test, p = 0.0872). The level of procalcitonin in lacrimal fluid 8-11 times exceeded its concentration in blood serum of all groups (Wilcoxon criterion, p < 0.005). The least content of procalcitonin is detected in lacrimal fluid of patients of comparison group (0.072 ± 0.064 ng/ml). In main and control groups its level was reliably higher (H-criterion of Kruskal-Wallis test, p = 0.0002) and amounted to 0.257 ± 0.146 and 0.198 ± 0.151 ng/ml correspondingly. The correlation analysis established no dependencies between concentration of procalcitonin in tear and blood serum (Spearman correlation coefficient had no exceeding |0.1| in all groups). The development of uveitis is not accompanied by alteration of concentration of procalcitonin in both blood serum and lacrimal fluid. The absence of correlation and higher concentration of procalcitonin in tear as compared with blood serum testify availability of additional source of this protein in lacrimal fluid.

Proliferative response of lymphocyte to pokeweed mitogen depends on the concentration of endogenous cortisol in the early post-traumatic period in patients with penetrating eye injury.

The intensity of lymphocyte proliferation in response to pokeweed mitogen depends on cortisol level in the peripheral blood in the early post-traumatic period of penetrating eye injury. Lymphocyte proliferation in 72- and 96-h cultures from patients with high levels of endogenous hormone was suppressed. In 120-h cultures, the intensity of proliferation remains unchanged. Lymphocyte blast transformation was increased in 120-h cultures from patients with normal cortisol concentration and remained unchanged in case of low cortisol level.

Expression of TLR9 and BD-2 protein genes in corneal cells of mice of different strains with herpetic keratitis.

The dynamics of gene expression of two proteins, TLR9 (one of the key receptors recognizing CpG repeats of herpes virus DNA) and ß-defensin 2 (antibacterial peptide), was studied on the model of herpetic keratitis in C57Bl/6 and BALB/c mice. New data on differences in TLR9 gene expression in mice of the two strains infected with the virus were obtained. Reduced TLR9 gene expression in the cornea of C57Bl/6 mice was associated with their high sensitivity to infection caused by herpes simplex 1 virus.

[Effect of myelopeptides on reactive oxygen species generation and IL-1beta and TNF-alpha production by peripheral blood cells].

Myelopeptides MP-3, MP-5, and MP-6 were found to suppress zymosan-induced production of reactive oxygen species by leukocytes both under one-way introduction and under pretreatment. All of myelopeptides under examination in case of one-way introduction in cultures with zymosan demonstrated a decrease in zymosan-stimulated (1500 mkg/ml) production of IL-1beta, and activation of spontaneous production of this cytokine by whole blood cells. TNF-alpha production under myelopeptide effect was lowered in cultures with 150 mkg/ml zymosan. Under pretreatment myelopeptides did not render effect on IL-1beta and TNF-alpha production, with the exception of single stimulating effect of MP-5 on IL-1beta level in spontaneous cultures. Using comparative analysis the difference in direction and expressivity of effects of various myelopeptides was not revealed that suggests the existence of common mechanism of action in this group of peptide bioregulators.

[Changes in expression level of TLR9 and BD-2 genes in cornea of mice with viral keratitis].

AIM: Study of innate immunity genes expression (TLR9 and BD-2) in epithelial cells of mice cornea on the model of viral keratitis during the application of immunotherapy. MATERIALS AND METHODS: RNA isolation from cornea cells was performed by using RNeasy Mini Kit (Qiagen) and "RIBO-sorb" (ILS, Russia) reagent kits. "Kit for performing PCR-RT in the presence of intercalating dye SYBR Green I" (Syntol, Russia) was used for performing PCR. Data on gene expression is presented in decimal logarithms (relatively to 1 million of beta-actin gene copies). RESULTS: In C57B1/6 mice line at day 1 post infection the virus was detected in 33% of animals, at 3 days - in 90%, at 7 days the number of infected mice started to decline. In cornea of the mice infected with HSV-1 a significant decrease of TLR9 gene expression was observed. Administration of a complex of natural cytokines and antimicrobial peptides (Superlimf preparation) caused statistically significant increase of expression levels of the studied TLR9 and BD-2 genes. CONCLUSION: The model of viral keratitis and expression analysis of recognizing receptor TLR9 and defensin (BD-2) genes can be used to study mechanism of action of various preparations during infection in eye tissues and evaluation of possibility of their use in complex therapy of ophthalmic herpes.

[TLR9 and HBD-2 gene expression by conjunctival and corneal cells in children with dendriform herpetic keratitis].

The current views of the role of innate immunity in protecting the organ of vision from viral infection offer scope for a search for new approaches to diagnosing and treating viral keratitis. TLR9 and HBD-2 gene expression in the conjunctiva and cornea was studied in children with dendriform herpetic keratitis. The latter showed a significant (more than 20-fold) increase in TLR9 gene expression as compared in healthy children. Conventional antiviral therapy for herpetic keratitis resulted in a significant decrease in the level of TLR9 gene expression (by more than two orders of magnitude). The use of the immunomodulator Superlymph in complex treatment normalized the level of TLR9 gene expression in the conjunctiva and cornea; moreover, there was a rapid relief of inflammation signs, an increase in visual acuity, and a reduction in the duration of treatment in patients with herpetic keratitis.

Effects of myelopeptides on immune response and morphometrical manifestations of inflammation in experimental penetrating wound of the eye.

Myelopid and MP-1 myelopeptide increase the count of antibody-producing cells, reduced under the effect of injury and standard therapy, and do not modify the suppression of delayed hypersensitivity. Injections of myelopid and MP-3 together with standard drugs optimized the traumatic inflammation processes.

Mechanism underlying the effect of myelopeptides on lymphocyte proliferation in vitro.

We studied the role of monocytes in the effect of myelopid and myelopeptides MP-1, MP-3, MP-5, and MP-6 on functional activity of peripheral blood lymphocytes in the reaction of blast transformation. Myelopeptides MP-1, MP-3, and MP-6 suppressed blast transformation of lymphocytes. The effect depended on the presence of monocytes in a cell culture.

[Regularities of changes in parameters of humoral immunity in patients with eye diseases]. / Zakonomernosti izmenenii pokazatelei gumoral'nogo immuniteta u bol'nykh s zabolevaniiami organa zreniia.

Changes in the levels of natural antibodies in the lacrimal fluid, saliva, and blood immunoglobulin concentrations were studied in patients with ocular injuries, inflammatory (keratitis, scleritis, uveitis) and noninflammatory diseases of the eyes. These values can be used as additional parameters for the diagnosis and prediction of the treatment efficiency. The immune system functions as a universal system with different compartments, and therefore mathematical relationships between these parameters were studied. Positive linear regression and correlation relationships were detected between -log2 antibody titers in the serum and lacrimal fluid, antibody titers in the saliva and serum, and negative relationships between antibody titers in the lacrimal fluid and IgA concentration and between lacrimal antibody titers and IgA level.