Differences in the Differentiation Potential and Relative Levels of Gene Expression in the Bone Marrow-Derived Fibroblast Colony-Forming Units in Patients during the Onset of Aplastic Anemia Depending on the Disease Severity.

The differentiation potential of individual clones of fibroblast CFU (CFU-F) was studied and the relative expression level of genes was analyzed in the culture of CFU-F from the bone marrow in patients with non-severe and severe forms of aplastic anemia at the onset of the disease. The differentiation potential of CFU-F clones was determined by the relative expression of marker genes using quantitative PCR. In aplastic anemia, the ratio of CFU-F clones with different differentiation potential changes, but the molecular mechanisms of this phenomenon are different in non-severe and severe aplastic anemia. In the culture of CFU-F in non-severe and severe aplastic anemia, the relative expression level of genes associated with the maintenance of the hematopoietic stem cell in the bone marrow niche changes, but the decrease in the expression of immunoregulatory genes occurs in severe form only, which may reflect differences in the pathogenesis of non-severe and severe aplastic anemia.

Bone Marrow Multipotent Mesenchymal Stromal Cells from Patients with Aplastic Anemia Retain Their Ability to Support Hematopoietic Precursors despite Pronounced Changes in Gene Expression.

The properties of bone marrow-derived multipotent mesenchymal stromal cells (MSC) of patients with aplastic anemia at the onset of the disease are studied insufficiently. The aim of this work was to test the ability of MSC from patients with aplastic anemia to maintain hematopoietic precursors and to analyze the expression of genes associated with hematopoiesis and immune response. The ability of MSC to maintain hematopoietic precursors was determined by counting cobblestone area-forming cells; gene expression was analyzed by quantitative PCR. It was shown that MSC of patients with aplastic anemia preserve their ability to maintain hematopoietic precursors. Pronounced changes in the expression of the VEGFA and ANGPT1 genes were found. MSC from aplastic anemia patients with PNH clone significantly differ from those from aplastic anemia patients without PNH clone in terms of the expression of the SDF1, IL1R, and VEGFA genes. Changes in gene expression can be associated with the pathogenesis of the disease.

Immunological Criteria for Predicting Severe and Complicated Forms of Chickenpox.

The aim of the study was to evaluate the levels of mediators of the immune response, cellular immunity, and phagocytic activity in patients with chickenpox with various values of the clinical and laboratory parameters and propose criteria for predicting the severity and complications of the disease. MATERIALS AND METHODS: The blood levels of pro-inflammatory mediators were evaluated by ELISA using monoclonal antibodies (Protein Contour, Russia). RESULTS: The inflammatory mediators and neutrophil chemiluminescence were studied in patients with either presence or absence of Varicella zoster DNA. We found that in patients with positive viral DNA, the levels of IFN-α and IFN-γ were significantly lower compared to patients with negative DNA results. Thus, complications of chickenpox, in particular secondary viral-bacterial pneumonia, can be predicted based on low (less than double-normal) levels of IL-6 and IFN-γ, induced chemiluminescence, CD16, and CD20. This type of immune response indicates the state of immune deficiency with prevailing suppression of the T-effector and phagocytic mechanisms in these patients. CONCLUSION: Prognosis of the development of severe and complicated forms of chickenpox can be based on the insufficiently increased (less than two normal values) levels of IL-6 and IFN-γ, induced chemiluminescence, CD16, and CD20. These relatively low levels are indicative of reduced immune response to the infection, which may require additional immune correcting therapy.

Comparison of polymerase chain reaction and flow cytometry for measuring telomere length of human leukocytes.

Telomere length can be measured by polymerase chain reaction (PCR), allowing to obtain the absolute length of telomeres (ALT) in base pair, and by flow cytometry, which can only estimate the relative telomere length. The aim of the study was to compare the results of the two methods and to develop an accurate and reliable way of converting the relative telomere length to absolute. The peripheral blood from 21 donors was analyzed. Measurement of leukocyte telomere length by flow cytometry was carried out using a commercial Telomere PNA Kit / FITC (Dako, Denmark) with two CytoFLEX flow cytometers (Beckman Coulter, China) and BD FACSCanto II (Becton Dickinson, USA), obtaining the molecular equivalent of fluorescence (MEF). To measure telomere length by real-time PCR, calibrators with a known number of telomeric repeats were prepared. Two quantitative PCRs were carried out: one for telomeric repeats, the other for determining the number of genome-equivalents of DNA, three times for each sample, which made it possible to calculate ALT. A strong direct relationship was found between the MEF obtained with BD FACSCanto II and CytoFLEX (r = 0.97). Analysis of PCR and flow cytometry results showed a significant correlation between ALT and MEF. We calculated the regression equations of ALT and MEF for CytoFLEX - y = 0.0043x (r = 0.84) and for BD FACSCanto II - y = 0.0051x (r = 0.82). Correlation analysis showed a high comparability of telomere lengths measured by two methods. The obtained regression equations allow converting the results of flow cytometry into absolute values, allowing the comparison of the results of different research groups and the use of this method in clinical trials.

Local immunity and microflora of the reproductive tract of gynecological patients before and after immunomodulatory therapy.

Immunomodulatory drugs are widely used as drugs for the treatment of inflammatory diseases of microbial etiology in clinical medicine. The authors evaluated the effectiveness of the therapeutic effect of the Cycloferon immunomodulator in the treatment of chronic purulent-inflammatory diseases of the reproductive tract using clinical, microbiological and immunological parameters. It was shown that under the influence of immunomodulatory therapy, the restoration of the qualitative and quantitative composition of the normoflora was observed with the elimination of etiologically significant microorganisms. The immunocorrective effect of therapy on the indices of local immunity was established: the concentration of lactoferrin increased in the cervical secretion, the level of cytokines IL-1ß and γ-IFN normalized, the amount of secretory IgA increased significantly, which contributed to the enhancement of local protective reactions, as well as the clinical efficacy of therapy, which was manifested in the reduction and/or disappearance of pain syndrome and the absence of relapse for 2 or more years. Conducted researches allow us to speak of immunotropic drugs as a promising direction in the treatment of inflammatory processes of the reproductive tract, a significant advantage of which is rational immunomodulation directly in the focus of inflammation, the availability of treatment, the absence of side effects with adequate therapeutic regimens, which makes it relevant to further study this direction of immunocorrection and its implementation into wide clinical practice.

[A case report of familial dyskeratosis congenital. Case report].

Dyskeratosis congenita (DC) is a hereditary syndrome of bone marrow failure, which develops because of telomeres defects and combines with cancer predisposition. Its classical clinical features are skin pigmentation, nail dystrophy, oral leukoplakia (skin-mucosa triad). The goal is to describe the algorithm of diagnosis, clinical specificities of DC and specific treatment for cases of DC in one family. The present report includes descriptions of diagnosis and treatment of family members diagnosed for the first time as having a DC. The report shows an importance of all diagnostic stages: from a medical history and clinical picture to an application of modern high-tech diagnostic methods (flow-FISH, NGS). The report underlines an importance of diagnosis of all family members for excluding an asymptomatic form after a case of DC has been already detected in that family. A high frequency of a toxicity and secondary neoplasia makes it necessary to realize an individual approach at treatment of each patient with DC (the earliest start of androgen treatment, prompt decision of implementation of allogenic hematopoietic stem cell transplantation). The knowledge of pathogenesis, clinical features and principles of diagnosis and therapy of this disease is relevant to pediatricians and hematologists.

[The capacity of Escherichia coli to form biofilms in presence of nano-particles of aluminum oxide.]

The effect of various concentrations of suspensions of nanoparticles of aluminum oxide of size of 30 and 70 nm on capacity of Escherichia сoli to form biofilm in vitro was examined. The study used highly virulent strain of Escherichia сoli isolated from a patient with chronic pyelonephritis. The strain was characterized by high capacity of forming biofilm. It is established that nanoparticles of aluminum oxide with size of 30 nm inhibited capacity of forming biofilm. At that, a reliable decreasing of analyzed indication in two times occurred and analyzed clones of Escherichia сoli passed to category of microorganisms with average capacity of forming biofilm. It is established that intensity of factor suppression depended both on size and concentration of nanoparticles in medium. The most effective in suppression of capacity of forming biofilm was concentration of nanoparticles of aluminum oxide 0,015 mkg/ml that decreased intensity of factor in two times.

[The dynamics of paroxysmal nocturnal hemoglobinuria clone in patients with aplastic anemia in process of immune suppressive therapy].

The implementation of principles of highly sensitive flow cytometry into diagnostic of paroxysmal nocturnal hemoglobinuria increased rate of detection of paroxysmal nocturnal hemoglobinuria clone in patients with aplastic anemia already at early stages of diagnosis establishment (up to 79%). However, detection of paroxysmal nocturnal hemoglobinuria clone attracts interest not only from point of view of progression in % of patients with aplastic anemia). The occurrence of paroxysmal nocturnal hemoglobinuria clone in patients with aplastic anemia can be accompanied by hidden disorders of haemopoesis with increasing risk in conditions of proliferative stress. Hence, it is necessary to monitor the given clone during all period of observation. The study is a prospective investigation analyzing dynamics of paroxysmal nocturnal hemoglobinuria clone in process of immune suppressive therapy applied to 44 patients with aplastic anemia. The mentioned clone was initially detected in 59.6% of patients. The median of observation amounted to 27 (9-48) months. Depending on size of granulocytic paroxysmal nocturnal hemoglobinuria clone patients were allocated in four conditional groups: group I - from 0.01% to 0.99% (n=11); group II - from 1% to 9.99% (n=8); group III - from10% to 49.9% (n=4); group IV - from 50% and more (n=5). In the course of study the differently directed dynamics of paroxysmal nocturnal hemoglobinuria clone was revealed. In 3 out of 11 patients from group I median of paroxysmal nocturnal hemoglobinuria clone increased from minor values (less than 1%) to 3.55%; at that in one patient occurred total elimination of paroxysmal nocturnal hemoglobinuria clone to 12th month of observation. The noticeable unidirectional dynamics was established in patients of group III: already to 3d month of observation, simultaneously with becoming of remission, median of size of paroxysmal nocturnal hemoglobinuria clone in group diminished from 22.9% (18.39%-24.77%) to 5.6% (1.5%-6.7%). Among patients of groups II and IV paroxysmal nocturnal hemoglobinuria clone remained stable. The development of hemolytic form of paroxysmal nocturnal hemoglobinuria was observed in all patients of group IV i.e. in 18% of patients with aplastic anemia with primarily detected paroxysmal nocturnal hemoglobinuria clone. In the process of observation, in 37% of patients with aplastic anemia without primarily detected paroxysmal nocturnal hemoglobinuria clone its occurrence and persistence (median - 0.34% (0.1%-6.2%)) was noticed. According to the results of study, alteration of sizes of paroxysmal nocturnal hemoglobinuria clone or its occurrence develop in case of response to ISP and, most probably, depend on advantage of growth in the process of repair of normal (GPI positive) or clonal (GPI negative) hemopoiesis. To acquire more reliable conclusions will be possible through development of techniques of molecular diagnostic simultaneously with dynamic observation of course of disease in the given patients.

[The sanitation of staphylococcus bacteria carrying: new possibilities and perspectives.]

The study was carried out to investigate possibility of applying biologically active substances - interferon inductors - for increasing effectiveness of prevention and treatment of torpid forms of infectious process, staphylococcus bacteria carrying in particular, considering effectiveness of their effect on biological characteristics of Staphylococcus aureus. The data are obtained concerning character of effect of pharmaceuticals on persistent characteristics of staphylococcus. These results can be applied in selection of new perspective preparations for sanitation of staphylococcus bacteria carrying.

Spatial and temporal analysis of land cover changes and water quality in the Lake Issaqueena watershed, South Carolina.

Monitoring changes in land cover and the subsequent environmental responses are essential for water quality assessment, natural resource planning, management, and policies. Over the last 75 years, the Lake Issaqueena watershed has experienced a drastic shift in land use. This study was conducted to examine the changes in land cover and the implied changes in land use that have occurred and their environmental, water quality impacts. Aerial photography of the watershed (1951, 1956, 1968, 1977, 1989, 1999, 2005, 2006, and 2009) was analyzed and classified using the geographic information system (GIS) software. Seven land cover classes were defined: evergreen, deciduous, bare ground, pasture/grassland, cultivated, and residential/other development. Water quality data, including sampling depth, water temperature, dissolved oxygen content, fecal coliform levels, inorganic nitrogen concentrations, and turbidity, were obtained from the South Carolina (SC) Department of Health and Environmental Control (SCDHEC) for two stations and analyzed for trends as they relate to land cover change. From 1951 to 2009, the watershed experienced an increase of tree cover and bare ground (+17.4 % evergreen, +62.3 % deciduous, +9.8 % bare ground) and a decrease of pasture/grassland and cultivated land (-42.6 % pasture/grassland and -57.1 % cultivated). From 2005 to 2009, there was an increase of 21.5 % in residential/other development. Sampling depth ranged from 0.1 to 0.3 m. Water temperature fluctuated corresponding to changing air temperatures, and dissolved oxygen content fluctuated as a factor of water temperature. Inorganic nitrogen content was higher from December to April possibly due to application of fertilizers prior to the growing season. Turbidity and fecal coliform bacteria levels remained relatively the same from 1962 to 2005, but a slight decline in pH can be observed at both stations. Prior to 1938, the area consisted of single-crop cotton farms; after 1938, the farms were abandoned, leaving large bare areas with highly eroded soil. Starting in 1938, Clemson reforested almost 30 % of the watershed. Currently, three fourths of the watershed is forestland, with a limited coverage of small farms and residential developments. Monitoring water quality is essential in maintaining adequate freshwater supply. Water quality monitoring focuses mainly on the collection of field data, but current water quality conditions depend on the cumulative impacts of land cover change over time.

[The comparative characteristic of immune status of males with bacterial inflammatory urogenital pathology of different etiology in the city of Orenburg].

The article discusses presence of typical characteristics of parameters of system immunity under gonococcal and nonspecific uretroprostatitis and diagnostic value of these indicators. The reliable differences of immunologic indicators in patients with gonorrhea are established as compared to patients with nonspecific bacterial uretroprostatitis. The study established in peripheral blood the reliable decrease of level of leukocytes, relative amount of monocytes, phagocyte index, functional reserve of leukocytes at the expense of spontaneous and stimulated NBT test, IgA, sIgA. On the contrary, the study detected increasing of level of IgM and lactoferrin in patients with gonorrhea as compared to corresponding indicators in patients nonspecific infections. Under gonorrhea, the largest deviation of indicators from standard values was established for lactoferrin. The detected differences of immunologic parameters can be used as differentiating markers of nonspecific and gonococcal uretroprostatitis and criteria of effectiveness of immune correction.

[Cryptococcosis in hematology practice].

AIM: To study the clinical manifestations of cryptococcosis, its diagnostic features, and treatment results in patients with hemoblastoses. SUBJECTS AND METHODS: The study included adult patients with cryptococcosis treated at the Hematology Research Center (HRC) in 2005 to 2011. The diagnosis of cryptococcosis was established on the basis of isolation of Cryptococcus neoformans from a blood culture or determination of positive cryptococcal antigen in the cerebrospinal fluid (CSF) of patients with infection symptoms. RESULTS: During 7 years, 19 patients aged 19 to 68 years (median 47 years) were diagnosed as having cryptococcosis. In the pattern of cryptococcosis, there was a preponderance of patients with lymphoma (31%) and those with acute lymphoblastic leukemia (26%) at the stages of hemoblastosis remission induction (32%) and consolidation (26%). The diagnosis was made in 9 (47%) patients at the Intensive Care Department, HRC. The major risk factors of cryptococcosis were previous cytostatic drug exposure (68%), use of immunosuppressive and glucocorticoid drugs (63%), and granulocytopenia (42%). Seventeen (78%) patients were diagnosed with cryptococcal meningitis or meningoencephalitis; 1 patient had cryptococcal sepsis and 1 patient had possible cryptococcal pneumonia. All the patients were given antifungal agents. Amphotericin B, fluconazole, and a combination of antimycotics were used as first-line drugs in 16 (84%), 1 (5.5%), and 2 (10.5%), respectively. When their health became better, the patients were treated with voriconazole or fluconazole. Within 30 days after the diagnosis of cryptococcosis, 5 (26%) patients died; of them 2 had tumor progression concurrent with infection. CONCLUSION: In cryptococcosis, the central nervous system is predominantly involved in the infectious process. The determination of cryptococcal antigen in CSF is a necessary diagnostic component in meningitis and meningoencephalitis in patients with blood system tumors, lymphatic ones in particular. When cryptococcosis is timely diagnosed and treated, its mortality, when the tumor is controlled, is lower than that in other invasive mycoses.

[The course of parvovirus B19 infection during pregnancy after kidney transplantation].

Having a tropism for erythroid progenitor cells, parvovirus B19 may cause partial red cell aplasia and thrombocytopenia. Early diagnosis of parvovirus B19 infection in immunocompromised patients is needed for timely antiviral therapy. A high-risk group for parvovirus B19 infection includes patients with blood diseases who receive multiple transfusions of blood components; those who have undergone donor organ transplantation and are long taking immunosuppressive drugs; and pregnant women. These patients require careful virological monitoring for major blood-borne viral infections. This paper describes a clinical case of parvovirus B19 infection in a pregnant woman who has undergone kidney transplantation and is continuously taking immunosuppressive medications. Identification of the cause of severe anemia and timely adequate therapy could lead to the recovery of effective erythropoiesis in the patient.

[Risk factors for violations of the fertility in men living in urban areas].

The immunocytokine status of the reproductive tract was evaluated in men with different forms of gonorrhea, who lived in the urbanized areas of the Orenburg Region. The typical form of gonorrhea, which is a risk for infertility, is accompanied by the reduced levels of lysozyme and IL-10 and the increase content of lactoferrin and proinflammatory cytokines, such as IL-1, Ibeta-6, and IL-8 in the ejaculate.

[Incidence and mortality of dental diseases in patients with hematological disorders].

The study was conducted in 102 patients with hematological disorders admitted to the Research Center of Hematology. It was concluded that more than 50% of them needed oral surgery procedures requiring multidisciplinary approach.

[Long-term results of HBV and HCV infection in patients with blood system diseases].

AIM: To specify trends in clinical and laboratory manifestations of virus hepatitis B and C (HBV and HCV) in patients with blood diseases from the moment of the first positive specific tests for HBV and HCV markers; to assess effects of HBV and HCV infection on efficacy of treatment of blood disease treatment, i.e. lifespan of patients with hematological diseases. MATERIAL AND METHODS: The study enrolled 257 patients: 205 with acute leukemia - AL, 40 with lymphoproliferative diseases, 4 - with CML and 8 - others; 8 healthy bone marrow donors. The patients were admitted to Russian Hematological Research Center in 2004-2006 Follow-up median was 253 days. A total of 7800 biological samples were studied, among them about 4000 tests for HBV DNA and HCV RNA. RESULTS: Positive tests for specific markers of HBV and HCV were absent only in 78 (29.4%) patients. Positive markers of coinfection were detected in 57 (32.8%) of 174 patients with HBV infection and in 81.4% of 70 patients with HCV infection. Probability of detection of HCV markers after positive tests for HBV markers and vice versa is about 3 times higher than probability of their isolated detection. Among patients infected with HBVsymptoms of hepatitis B are likely to appear in 56% patients to day 500 of follow-up from the date of the first positive specific test. Median of the interval between the first positive test for HBV markers and probable clinical signs of hepatitis was 30 days. Among patients with HCV infection, 85% develop hepatitis to follow-up day 300 since the date of the first specific positive test. Almost 100% patients infected with two viruses develop hepatitis to follow-up day 600. Median of the interval between the first positive test for HBV and HCV markers and probable hepatitis picture was 47 days. Overall 3-year survival of AL patients was 40%, of patients with lymphoproliferative diseases - 58%. Overall 7-month survival was 75% in AA patients. HBV infection in patients with blood disease is associated with high risk of death, especially in AA and AL. Association between HCV infection and survival is not proved. CONCLUSION: A high rate of clinical realization of viral hepatitis B and C, especially in coinfection, calls for virological and clinical monitoring of patients with any positive test for HBV and HCV markers.

[Prolonged bone marrow aplasia in patients with acute leukemia after chemotherapy].

AIM: To analyze the causes of prolonged hematopoietic tissue aplasias in patients with acute leukemias (AL) after chemotherapy courses. MATERIALS AND METHODS: Data on 7 patients with acute myeloid leukemia, followed up at the Hematology Departments, Hematology Research Center, Russian Academy of Medical Sciences, over the period 2003 to 2007, who had developed deep bone marrow aplasia (BMA) inadequate to cytostatic drug exposure during chemotherapy, were analyzed. The authors compared in all the patients the values of peripheral blood and bone marrow (BM) puncture specimens and the results of blood tests using the polymerase chain reaction at different AL development stages with the results of an immunohistochemical study using the markers of viruses of hepatitis C and B, a herpes group (EBV, CMV, HSV-1, HSV-2) and parvovirus B19. RESULTS: The marker of hepatitis C was detected in 6 of the 7 patients with prolonged BMA; 3 of these 6 patients showed a simultaneous infection with hepatitis B. Six of the 7 patients were found to have concomitant BM lesion with various herpes group viruses. Two patients had a resistant form of AL. CONCLUSION: Hepatitis C virus infection in patients and the resistant form of the disease were the principal causes of the development of BMA inadequate to cytostatic drug exposure. Affliction of abundant bone marrow cells with herpes group viruses was not a direct cause, but might substantially aggravate BMA.

[Efficiency of cyclosporin A therapy in patients with myelodysplastic syndrome].

AIM: To evaluate the efficacy of cyclosporin A (CsA) in patients with myelodysplastic syndromes (MDS) and to identify determinants of a response to this therapy. SUBJECTS AND METHODS: The efficacy of CsA was evaluated in 52 patients (30 men and 22 women aged 16 to 74 years) with MDS. Thirty-two patients were given CsA as first-line therapy; 20 patients took the agent after prior therapy. CsA was used in a daily oral dose of 5 mg/kg. Its efficacy was evaluated following 3, 6, and 12 months. Actuarial survival was determined by the Kaplan-Meier method. RESULTS: The efficacy of CsA used as first- and second-line therapy was 56 and 55%, respectively; complete remissions were achieved in 19 and 20% of cases. Baseline refractory anemia (RA) transformed to RA with excess blasts (RAEB) in 31% of cases; baseline RAEB did to acute myeloid leukemia in 34%. Overall survival was significantly associated with bone marrow (BM) blast cell percentage (< 5% or > 5%; p = 0.0009), BM cellularity (hypoplasia and focal hypoplasia of hematopoiesis or BM hyperplasia; p = 0.03), focal polyclonal lymphoid infiltration in the BM (p = 0.01) and karyotype anomalies (low, moderate, and high risks; p = 0.001). CONCLUSION: CsA is the drug of choice in treating patients with MDS, including RA, RA with ringed sideroblasts, refractory cytopenia with multilineage dysplasia, with hypoplasia of hematopoiesis, with nodular polyclonal lymphoid infiltration in the BM, a normal karyotype or changes corresponding to a low or moderate IPSS risk.

[Characteristic of biological properties of Neisseria gonorrhoeae and parameters of local immunity during urogenital gonococcal infection].

AIM: To study the prevalence and expression of antilactoferrin, IgA- and slgA-protease activity of gonococci and state of local immunity during various forms of urogenital gonorrhea. MATERIALS AND METHODS: Ability to inactivate lactoferrin (ALfA), IgA and secretory IgA (slgA) was studied in 28 Neisseria gonorrhoeae strains isolated from patients with localized gonorrhea and 26 strains isolated from patients with systemic signs of gonorrhea. State of the local immunity was assessed on the lactoferrin, IgA and slgA levels, which were measured by immunofluorescence assay in ejaculate of 54 patients with gonorrhea and 18 healthy males. RESULTS: Penetrance of ALfA, IgA- and slgA-protease activity of gonococci did not depend from form of infection. Expression of studied characteristics of gonococci as well as combination of ALfA and slgA-protease activities were more prominent in patients with systemic signs of gonorrhea. The same patients had higher level of lactoferrin in semen and, in contrast, lower levels of IgA and slgA compared with patients with localized gonorrhea. CONCLUSION: Strains of gonococci inactivating lactoferrin, IgA and slgA depress mucosal barrier of urogenital biotope and create conditions for the development of disseminated forms of gonococcal infection.

[The study of parameters of mesenchymal stromal cells differentiation in donors and patients with aplastic anemia].

AIM: To examine ability of mesenchymal stromal cells (MSC) of the bone marrow (BM) for differentiation in adipogenic and osteogenic differentiation in donors and patients with aplastic anemia (AA). MATERIAL AND METHODS: We obtained MSC cultures from BM cells of donors and AA patients and induced differentiation of mesenchymal cells with use of relevant reagents. Morphological changes in MSC were studied with light microscopy. A relative level of expression of differentiation marker genes in MSC cultures before and after induction of differentiation was analysed with reverse transcription-polymerase chain reaction. RESULTS: By morphological characteristics, MSC cultures in AA patients before and after differentiation induction do not differ from donor cultures, but relative expression of the genes of differentiation markers demonstrated that expression was different in male and female donors; MSC before and after induction of differentiation differ in donors and AA patients. CONCLUSION: Further studies are needed for detection of functional changes in precursors of stromal microenvironment and understanding of the disease pathogenesis.