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Introduction. Optimal therapy for elderly patients with chronic lymphocytic leukemia (CLL) is the subject of intensive research.Aim – to study the safety of obinutuzumab, as well as the selection of the optimal scheme of its use in patients with CLL, complicated by diabetes mellitus, renal insuffi ciency, cardiac comorbidity.Materials and methods. The study included primary patients with CLL having indications requiring therapy. The inclusion criteria were: Cumulative Illness Rating Scale, CIRS) (CIRS) > 6 and or glomerular fi ltration rate (GFR) < 70 mL/min, the lower limit of GFR was not restricted. The study focused on patients with diabetes mellitus, renal failure and signifi cant cardiac pathology. Patients with Richter’s syndrome, CNS involvement, HBs-antigen, and 17p deletion were not included. In the fi rst cycle obinutuzumab was administered at a dose of 100 or 25 mg on the fi rst day and 900 or 975 mg on the second day, then at a dose of 1000 mg on days 8 and 15. For all subsequent cycles, obinutuzumab was given at a dose of 1000 mg on day 1. The dosage of chlorambucil was 10 mg/m2 from days 1 to 7. Treatment cycles in totals of 6 were repeated every 28 days.Results. The study included 90 patients. Median age was 73.5 years, range – 60–89 years, there were 49 men (54 %) and 41 women (46 %). Twenty-four patients (27 %) had stage C, IGHV unmutated status was detected in 76 % of patients. The median creatinine clearance was 48.6 mL/min (25–110). The median CIRS score was 3 (range – 1–14). Thirty-one patients (34 %) had signifi cant cardiovascular comorbidity (previous myocardial infarction, coronary artery stenting or bypass, HF ≥ II NYHA, peripheral artery disease) as well as hemodynamically signifi cant valvular disease. Fifteen patients (17 %) had diabetes mellitus and 71 patients (79 %) had creatinine clearance < 70 ml/min. Infusion reactions to obinutuzumab grade ≥ II were reported in 29 patients (32 %). Hospitalization on the day of administration or the next day after the fi rst administration was required in 5 cases (5.5 %). Twenty-seven (30 %) patients could not complete 6 cycles. The largest number of patients (14 people, 15.5 %) stopped treatment after 1 course. The causes were the development of persistent cytopenia (n = 4), grade IV reaction to obinutuzumab (n = 3), patient’s refusal (n = 2), infectious complications (n = 2), severe tumor lysis syndrome (n = 1), acute pancreatitis (n = 1) and toxicodermia (n = 1). The leading cause of premature discontinuation on subsequent cycles was persistent neutropenia. Progression during treatment occurred in 3 patients only. Overall survival was signifi cantly predicted by CIRS (maximum discriminatory value of 3, p = 0.013) as well as GFR < 50 mL/min (p = 0.03). No other associations were identifi ed. At least 1 episode of grade III–IV neutropenia occurred in 41 % of patients. Grade IV neutropenia was associated with creatinine clearance < 60 mL/min (p = 0.05), baseline neutrophil level < 2 × 109/L (p = 0.0001), baseline monocyte level < 0.3 × 109/L (p = 0.007) and age > 70 years (p = 0.01). The effectiveness of treatment was evaluated in patients who completed at least 3 cycles of therapy. Complete remission was achieved in 26 patients (35 %), partial remission – in 41 (54 %), stabilization – in 4 (5 %), progression was noted in 3 (4 %). Sixteen patients (18 %) were not available to respond to the assessment. Minimal residual disease < 0.01 % in the bone marrow after completion of treatment was found in 17 patients (19 %), within 0.01–0.9 % – in 25 (28 %) patients. The median follow-up from the date of therapy initiation was 39.7 months (range – 0.6–72 months). The median relapse-free survival was not reached, and 2- and 3-year survival rates were 81 and 62 %, respectively. Poor relapse-free survival signifi cantly correlated with unmutated IGHV genes (HR = 2.4, 95 % CI: 1.12–5.0, p = 0.02) and partial response as opposed to complete response (HR = 3.35; 95 % CI: 1.45–7.7, p = 0.03).Conclusion. The results of our study have practical implications as obinutuzumab is actively integrated into modern treatment regimens. Infusion reactions pose a high risk of complications in elderly patients. The dose of obinutuzumab on day 1 of administration in elderly patients, should not exceed 25 mg. The G-Clb regimen may not be optimal in patients over 75 years of age due to the unpredictable risk of complications. In patients at high risk of neutropenia, it may be appropriate to consider primary prophylaxis. ClbG is an effective regimen that resulted in high rate of MRD-negative responses and prolonged relapse-free survival.

Objective. To determine clinical course and state of cellular immunity in young children with rotavirus infection.Children characteristics and research methods. The scientists examined children without infectious pathology and with rotavirus infection (20 patients in each group) using general clinical methods. Rotavirus infection was diagnosed by polymerase chain reaction and immunochromatography. Cellular immunity parameters were determined by flow cytometry.Results. All the children under observation had a moderate form of the disease with symptoms of exsicosis of the II degree. Changes in the immune status were mainly of a regulatory, adaptive nature, which contributed to the favorable course of rotavirus infection in children, however, the dynamics of the number of cells expressing Toll-like receptors indicates the immunosuppressive properties of rotavirus.Conclusion. Currently, rotavirus infection in young children is typical with watery diarrhea as the most pronounced and long-lasting clinical symptom. Shifts in immunogram indices in general indicate a deficiency of the cellular link of immunity and a violation of its regulation with simultaneous activation of the immune system in an effort to achieve the eradication of the rotavirus with immunosuppressive properties.

Purpose: to improve the complex therapy of rotavirus infection in infants by including interferon and to evaluate its clinical and immunological effectiveness.Materials and methods. Infants without infectious pathology and with rotavirus infection, who were on standard therapy and treatment, including standard therapy and interferon (20 in each group), were examined using general clinical methods. The diagnosis of rotavirus infection was carried out by molecular-genetic method and immunochromatography. Indicators of cellular immunity were determined by flow cytometry. Statistical processing of the results was carried out using the STATISTICA 8.0 program for related and independent samples using the nonparametric Mann-Whitney test and χ2.Summary. In all infants with rotavirus infection, the disease occurred in a moderate form with symptoms of grade II exicosis. In the group of patients who received an additional interferon drug as part of complex therapy, the duration of intoxication symptoms was significantly less, and in the hemogram at discharge from the hospital, the relative content of neutrophils was higher and monocytes lower compared to infants who were only on standard treatment, which corresponded to the norm. In the acute period of rotavirus infection, the relative level of lymphocytes decreased, and NK cells and T-NK cells increased compared to healthy infants. The relative number of cells expressing TLR-3 at the beginning of the disease was lower than in the control group. The inclusion of interferon in the complex therapy contributes to normalization of the relative number of NK cells, while maintaining an increased content of T-NK cells in both relative and absolute terms compared to healthy infants, and a reduced content of cells expressing TLR-3.Conclusion. Currently, rotavirus infection in infants is typical, the most pronounced and long-lasting clinical symptom of the disease is watery diarrhea. The use of interferon in complex therapy helps to reduce the duration of intoxication and normalize the hemogram. Changes in the immune status in the acute period of rotavirus infection indicate increased antigenic stimulation of the cell link and inhibition of the synthesis of pro-inflammatory cytokines, including interferons, under the influence of rotavirus, which is the justification for the use of interferon therapy.

Recent studies have revealed how the freshwater biota of Lake Baikal responds to climate change and anthropogenic impacts. We studied phyto- and zooplankton, as well as phyto- and zoobenthos, in the open coastal waters of the southern basin of the lake and of Listvennichny Bay. A total of 180 aquatic organism taxa were recorded. The response of the Baikal ecosystem to climate change can be traced by changes in the species composition of planktonic communities of the lake’s open coasts in summer. The key species were thermophilic the Anabaena lemmermannii P. Richt. (Fij = +0.7) blue-green algae, the Asplanchna priodonta Gosse (Fij = +0.6) rotifers in 2016, the Rhodomonas pusilla (Bachm.) Javorn. (Fij = +0.5) cold-loving algae, and the Cyclops kolensis Lilljeborg (Fij = +0.9) copepods in the past century. The proportion of Chlorophyta decreased from 63% to 17%, the Cyanophyta increased from 3% to 11% in the total biomass of phytoplankton, and the proportion of Cladocera and Rotifera increased to 26% and 11% in the biomass of zooplankton, respectively. Human activity makes an additional contribution to the eutrophication of coastal waters. The Dinobryon species, the cosmopolitan A. formosa and F. radians, dominated phytoplankton, and filamentous algae, Spirogyra, dominated at the bottom in the area with anthropogenic impact. The trophic level was higher than at the unaffected background site: the saprobity index varied from 1.45 to 2.17, the ratio of eutrophic species to oligotrophic species ranged from 1:2 to 3:1, and the ratio of mesosaprobiont biomass to endemics biomass ranged from 2:1 to 7:1. Currently, the boundaries of eutrophication zones of shallow waters in Lake Baikal are expanding, and its coastal zone has acquired features typical of freshwater bodies of the eutrophic type.

We have reconstructed major demographic and phylogenetic events for two closely related copepod species: Epischura baikalensis Sars, 1900 (Copepoda, Calanoida), a key endemic species of the plankton community from Lake Baikal, and E. chankensis Rylov, 1928, a key zooplankton species from Lake Khanka. The reconstructions were based on the nucleotide sequences of mitochondrial DNA (mtDNA) cytochrome oxidase subunit 1 (CO1) using the BEAST software package and recently identified calibrations for the Baikal amphipods Gmelinoides fasciatus. The time span to a common ancestor for E. baikalensis and E. chankensis is 13.7 million years (95% confidence interval is 8.3–21.4 million years. Both species studied have experienced abrupt changes in the effective population size in the past. These changes are likely associated with climate and geological shifts in their habitat areas. For E. baikalensis, these changes can be associated with alternating glacier and interglacial periods as well as an increase in depth of Lake Baikal; for E. chankensis with formation and increase in the area of Lake Khanka, which was also due to climate and geological shifts in the Khanka basin.

Profound immunological dysfunction is the key factor determining the development of infectious complications in chronic lymphocytic leukemia (CLL). The aim of this work is to assess the features of the subpopulation composition of T-lymphocytes (T-helpers (Th), cytotoxic T-lymphocytes (Tcyt), T regulatory cells (Treg), T-NK cells, naive Th, Th-memory, activated T-lymphocytes, TCRγδ cells) and NK cells in peripheral blood of patients with newly diagnosed chronic lymphocytic leukemia (CLL) and receiving ibrutinib therapy. Hematological and immunophenotypic studies have been performed in 30 patients with previously untreated CLL, 122 patients on ibrutinib therapy and 20 healthy donors. The subpopulation composition of T-lymphocytes (Th, Tcyt, Treg, T-NK, naive T-helpers, memory T-helpers, TCRγδ cells, activated T-lymphocytes) and NK cells has been assessed on flow cytometer (FACSCanto II (BD)) using the following panel of monoclonal antibodies: CD45, CD19, CD3, CD4, CD5, CD8, TCRγδ, CD127, CD16, CD56, CD57 CD45RA, CD45R0, HLA-DR, CD25. Compared to controls all CLL samples were found to have higher the absolute number of T-lymphocytes, NK cells and their subpopulations, T-helpers (especially of memory T-cells), cytotoxic T-cells, regulatory T-cells, TCRγδ T-cells, activated T-lymphocytes, increased cytotoxic potential of NK cells in previously untreated CLL patients. Patients who received ibrutinib therapy have registered a positive trend towards recovery of the subpopulation composition of T-lymphocytes and NK-cells. CLL patients have been found to have quantitative and functional changes in the subpopulations of T-lymphocytes and NK cells, indicating dysregulation of the immune response, and a high risk of developing infections. Monitoring of immunological parameters for ibrutinib therapy make possible to estimate impact of ibrutinib on the adaptive anti-CLL immune response.

We studied the population structure of the Cyclops kolensis.Lill, 1901 copepod in Lake Baikal. For analysis, we used the fragment sequences of the cytochrome oxidase subunit 1 (CO1) of mitochondrial DNA. Based on these sequences, we constructed a haplotype tree for Baikal individuals as well as a phylogenetic tree for the sequences of Baikal and non-Baikal members of C. kolensis using the molecular clocks. In general, the distribution of haplotypes across the lake basins indicates the absence of geographical subdivision, although there are traces of the past local isolations as well as possible recent introductions from the water bodies near Lake Baikal. We have determined the time of the divergence between the Baikal representatives and the European ones (from the lakes of Moscow and Kiev), which dates back to 1.13 million years (95% CI: 0.67-1.63 million years). The divergence of the Baikal group of haplotypes is 0.31 million years (CI: 0.16 - 0.48 million years), which may be the time when the species entered the lake.

Aim. To develop reference intervals for white blood cell subpopulations in peripheral blood using Cytomics FC500 flow cytometer and CytoDiff™ reagent (Beckman Coulter). Materials & Methods. The trial included the analysis of blood samples of healthy donors (n = 315) using Cytomics FC500 flow cytometer and CytoDiff™ reagent cocktail composed of 6 antibodies in 5 colors (CD36-FITC, CD2-PE, CD294-PE, CD19-ECD, CD16-Cy5, CD45-Cy7) and enabling to count 17 cell populations. Results. The data obtained by means of multicolor flow cytofluorometry included the reference values of white blood cell populations in peripheral blood. In 1 first-time donor a lymphoproliferative disease was detected. It was subsequently confirmed by immunophenotyping; B-cell chronic lymphocytic leukemia was diagnosed. Conclusion. Multicolor flow cytofluorometry using CytoDif-f™ is considered to be a new step toward an improved WBC differential evaluation aimed mainly at reducing the volume of blood smear analysis using light microscopy at large laboratories, enhancing objectivity, precision and reproducibility of results. WBC differential extended with the count of lymphocyte and monocyte subpopulations can be regarded as modern donor blood screening to detect changes in the pattern of lymphocyte subpopulations as grounds for further examination of donors.

Background & Aims. This paper presents the results of the observational study of ibrutinib in patients with chronic lymphocytic leukemia (CLL), conducted in SP Botkin Municipal Clinical Hospital. The main objective was the analysis of complications of ibrutinib and identification of factors, influencing the dosage regimen; the secondary objective was the estimation of the total response to treatment, event-free and overall survival. Materials & Methods. The study included 96 patients with CLL with indications for ibrutinib therapy. The median age was 64,9 years (range 32-91 years), the study population consisted of 69 (72 %) men and 27 (28 %) women. The condition of 25 (26 %) patients according to the ECOG scale was of > 3 points. The disease of stage C were diagnosed in 36 (37 %) patients. Deletion of 17p/TP53 mutations were detected in 29 (33 %) of 87 patients. Seventy patients had refractory CLL. The median of the number of the lines of the previous therapy was 3 (range 1-9). Adverse events were assessed in accordance with the CTCAE criteria, version 4.0; the bleeding severity was evaluated using ITP-specific bleeding score; hematological complications were classified according to the recommendations of IWCLL-2008. Results. Ibrutinib was administered at a dosage of 420 mg per day daily until progression or intolerable toxicity. The median duration of ibrutinib therapy was 10.3 months. brutinib was shown to have moderate toxicity, mostly of grade I or II. The bleeding was the most frequent complication. Of the hematological complications, thrombocytopenia was the most common (35 %); neutropenia < 1 x 109/L was observed in 4 patients. GIT complications were identified in 51 (53 %) patients. Atrial fibrillation was registered in 5 patients, who initially had sinus rhythm. The total of 144 infections were diagnosed in 64 (66 %) patients. Severe infections (> grade III) developed in 26 % of patients. The treatment response was assessed in 92 patients. The overall response to treatment was 89 %. Complete remission, partial remission and partial remission with lymphocytosis were achieved in 4 (4 %), 57 (62 %), and 21 (23 %) patients, respectively. The event-free survival and overall survival by the month 10 was 90 % and 91 %, respectively. For this observation period, ECOG status and the number of the lines of therapy prior to ibrutinib had the prognostic value. Conclusion. Ibrutinib was shown to have high efficiency in relapsed/refractory forms of CLL. The nature of the ibrutinib toxicity is fundamentally different from that of the conventional chemotherapy. The frequency of ibrutinib therapy complications and patients' non-compliance depends on the intensity of the previous treatment of CLL. Despite a short observation period, it can be concluded that ibrutinib had the greatest impact on the patient's quality of life when administered for the first relapse. The low toxicity of ibrutinib is likely to allow the combination with other antitumor agents.

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 review is devoted to the experimental studies of mitogenetic radiation (weak UV-chemiluminescence of biological objects) conducted between 1923 and 1948 years. In this period UV-radiation of various biological objects (so called inductors) and its influence on mitotic rate of other biological objects (detectors) were investigated very actively. Very promising results were obtained including the finding of the peptide tumor marker in blood, which was called cancer quencher due to its ability to quench UV-chemiluminescence of blood. Later these researches were interrupted and almost abandoned. The relevance of the scientific problems addressed in these works to this day is stated, the key experiments and the most valuable results obtained between 1923 and 1948 are described in details and subsequent researches are briefly traced up to the present. The prospects of future development are discussed.

It has been proposed that circulating HSCs play a role in graft survival after liver transplantation. The aim was to analyze the relationship between the number of HSCs before and after LDLT and liver function, immune biomarkers, and clinical outcomes in pediatric patients. We studied 15 pairs of adult healthy liver donors and pediatric recipients with ESLD. The CD34/CD45+ cell number was measured in the blood via flow cytometry, and plasma levels of immune biomarkers - via ELISA. CD34/CD45+ cell number in the recipients decreased within the first week after LDLT. The cell number before LDLT was negatively correlated with the plasma levels of CRP and the development of graft dysfunction in the early post-transplant period. After LDLT, the CD34/CD45+ cell number was positively correlated with the pretransplant plasma level of sCD40L, a T-cell activation marker. In adult liver donors, the cell number did not change within the first week after liver resection and was lower than in pediatric recipients. The results suggest that in pediatric recipients, the HSC number may be associated with graft function and could be regarded as a potential predictor of the clinical outcome after LDLT.

A metamaterial absorber is realized for microwave and terahertz ranges using two-dimensional bianisotropic omega-elements by thorough analysis of their distribution on substrate as well as properly choosing omega's geometrical and material parameters. Simulations and experimental studies of the metamaterial confirm its low reflectance in a wide microwave range and high absorbance at the resonance.

The systems comprising arrays of microhelical resonators, GaAs slabs and air spacers were formed with use of the rolling-up and 3D printing technologies and studied. The systems demonstrate the interplay of the half-wave resonance of helices, waveguide and Fabry-Perot resonances, which result in ultrasharp high regular peaks in polarization spectra. The background mechanisms of the peaks arising in different systems are discussed.

Electromagnetic metamaterials opened the way to extraordinary manipulation of radiation. Terahertz (THz) and optical metamaterials are usually fabricated by traditional planar-patterning approaches, while the majority of practical applications require metamaterials with 3D resonators. Making arrays of precise 3D micro- and nanoresonators is still a challenging problem. Here we present a versatile set of approaches to fabrication of metamaterials with 3D resonators rolled-up from strained films, demonstrate novel THz metamaterials/systems, and show giant polarization rotation by several chiral metamaterials/systems. The polarization spectra of chiral metamaterials on semiconductor substrates exhibit ultrasharp quasiperiodic peaks. Application of 3D printing allowed assembling more complex systems, including the bianisotropic system with optimal microhelices, which showed an extreme polarization azimuth rotation of 85° with drop by 150° at a frequency shift of 0.4%. We refer the quasiperiodic peaks in the polarization spectra of metamaterial systems to the interplay of different resonances, including peculiar chiral waveguide resonance. Formed metamaterials cannot be made by any other presently available technology. All steps of presented fabrication approaches are parallel, IC-compatible and allow mass fabrication with scaling of rolled-up resonators up to visible frequencies. We anticipate that the rolled-up meta-atoms will be ideal building blocks for future generations of commercial metamaterials, devices and systems on their basis.

The properties of an artificial anisotropic structure formed by microhelices have been numerically simulated by the example of a specially designed sample. The sample contains paired helices with right- and left-handed twisting directions, due to which the metamaterial chirality is compensated for. Helices are characterized by precalculated optimal parameters; as a result, the permittivity and permeability of metamaterial are approximately equal. Therefore, the wave impedance of the structure is close to the impedance of free space in the frequency range near 2 THz and the reflection of normally incident electromagnetic waves is weak. A boundary problem is solved and an analytical expression is obtained for the reflection and transmission coefficients of an electromagnetic wave passing through the structure. The simulated properties of the structure are compared with experimental results.

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal stem cell disorder characterized by partial or absolute deficiency of glycophosphatidyl-inositol (GPI) anchor-linked surface proteins on blood cells. A lack of precise diagnostic standards for flow cytometry has hampered useful comparisons of data between laboratories. We report data from the first study evaluating the reproducibility of high-sensitivity flow cytometry for PNH in Russia.PNH clone sizes were determined at diagnosis in PNH patients at a central laboratory and compared with follow-up measurements in six laboratories across the country. Analyses in each laboratory were performed according to recommendations from the International Clinical Cytometry Society (ICCS) and the more recent 'practical guidelines'. Follow-up measurements were compared with each other and with the values determined at diagnosis.PNH clone size measurements were determined in seven diagnosed PNH patients (five females, two males: mean age 37 years); five had a history of aplastic anemia and three (one with and two without aplastic anemia) had severe hemolytic PNH and elevated plasma lactate dehydrogenase. PNH clone sizes at diagnosis were low in patients with less severe clinical symptoms (0.41-9.7% of granulocytes) and high in patients with severe symptoms (58-99%). There were only minimal differences in the follow-up clone size measurement for each patient between the six laboratories, particularly in those with high values at diagnosis.The ICCS-recommended high-sensitivity flow cytometry protocol was effective for detecting major and minor PNH clones in Russian PNH patients, and showed high reproducibility between laboratories.

Fluorescence and phosphorescence quenching processes of polar and non-polar luminescent probes associated with human serum albumin (HSA) in phosphate buffer at pH 7.4 were studied. Stern–Volmer quenching constants of anthracene and pyrene fluorescence and eosin phosphorescence and rate constants for quenching of eosin triplet states were determined. The polarity index of pyrene bound to HSA was obtained as a function of thallium nitrate concentration. The influences of structural changes in the proteins that were stimulated by heavy-metal salts and of screening of protein charges by salt ions on quenching processes of singlet and triplet states of the probes were found.

In the present paper, a theoretical and experimental study of a highly absorptive, weakly reflective coating designed and fabricated on the basis of 3D THz resonant elements is reported. Transmission and reflection of electromagnetic waves from the metamaterial-substrate structure involving a highly absorptive, weakly reflective array of artificial bi-anisotropic elements were analyzed. The samples contained paired right-handed and left-handed helices, due to the fact that the chirality was compensated. The parameters of helices were optimized to achieve roughly identical values of dielectric permittivity and magnetic permeability. As a result, the metamaterial exhibited weak reflectivity in the vicinity of resonance frequency. On the other hand, effective resonance properties of the helices were tuned to ensure substantial absorption of THz radiation. Analytical expressions for the coefficients of radiation reflection and transmission in the samples were derived by solving a boundary-value problem for the propagation of electromagnetic waves in the metamaterial-substrate system. Simulated properties of fabricated structures were compared with experimental data.

The article presents diagnostic of night paroxysmal hemoglobinuria. The night paroxysmal hemoglobinuria is an orphan disease characterized by absence of GPI-anchor on blood cells as a result of mutation of PIG-A gene on the short arm of X-chromosome. The particular proteins bounded with GPI-anchor implement function of defense from activation of components of complement and development of membrane-attacking complex. The erythrocytes exposed to destruction in bloodstream are among the most impacted. Therefore, one of the main signs of night paroxysmal hemoglobinuria is complement-depending intravascular hemolysis which indicators for a long time played a key role in diagnostic of night paroxysmal hemoglobinuria. The actual technique of diagnostic of night paroxysmal hemoglobinuria is flow cytometry. The analysis of night paroxysmal hemoglobinuria clone is recommended to patients with hemolysis of unclear genesis, thrombosis of cerebral and abdominal veins, thrombocytopenia and macrocytosis and also patients with AA, myelodysplastic syndrome, myelofibrosis. The international protocol recommended by the International Society of Clinical Cytometry (2010) is implemented to diagnose night paroxysmal hemoglobinuria. The original technique of evaluation of reticulocytes was developed with purpose to detect night paroxysmal hemoglobinuria clone. The high correlation was substantiated between size of night paroxysmal hemoglobinuria clone measured among reticulocytes according to proposed mode and night paroxysmal hemoglobinuria clone measured among granulocytes and monocytes detected according international standardized approach.

The chiral properties of an artificial anisotropic structure composed of microhelices are numerically simulated using the example of a sample developed by a team of authors from the Institute of Semiconductor Physics of the Russian Academy of Sciences. It is shown that this artificial structure can exhibit strong chiral properties in the THz range. Analytical expressions for the dielectric, magnetic, and chiral susceptibilities of the structure are derived on the condition of strong gyrotropy. The calculated values of the angle of the electromagnetic-wave polarization plane rotation and the circular dichroism are compared with the experimental results.

Chiral metamaterials of the terahertz range are formed by means of rolling of strained nanofilms. Structural elements of these metamaterials are metal-semiconductor microhelices. Resonant optical activity of new metamaterials in the terahertz range is demonstrated. Arrays of model wire helices are formed and tested in the microwave range.

The achievement of molecular remission is associated with increasing of survival of patients with chronic lymphatic leukemia. The important direction of research is seeking of parameters applicable to forecast of response to therapy. The purpose of the study was evaluating prognostic significance of indicator of minimal residual disease detected by technique of flow cytofluorometry of peripheral blood of patients with chronic lymphatic leukemia during therapy application. The sampling included 112 patients with chronic lymphatic leukemia aged from 43 to 82 years. All patients were given treatment consisted of 6 courses of immune chemotherapy combined with fludarabine with cyclophosphan and rituximab. The samples of peripheral blood were analyzed after 3 courses during therapy and after 6 courses after completion of treatment. The cells were analyzed using 5 and 6 color flow cytometry for the purpose of detection of immune phenotype associated with chronic lymphatic leukemia. The evaluation of minimal residual disease was implemented according international standardized protocol (Rawstron A.C. et al. 2007; 21 (5): 956-64). The minimal residual disease negative status was reached in 87 (78%) patients during evaluation of response after 6th course of treatment. The implementation of indicators of residual disease after 3 courses with fludarabine, cyclophosphan and rituximab permitted to sort out two groups of patients with chronic lymphatic leukemia i.e 67 patients with low (0.12%) level of minimal residual disease and 45 patients with high (0.12%) level of tumor cells. The rate of molecular remission after completion of treatment. in the given groups consisted 100% and 44% correspondingly. The study demonstrates possibilities of early immune phenotype evaluation of minimal residual disease to forecast differences in response to treatment in patients with chronic lymphatic leukemia that makes it possible to avoid undesirable toxicity of therapy or to choose method of consolidation.

The techniques of flow cytofluorometry and immune cytochemistry combined with routine cytological analysis were applied to 81 patients with non-Hodgkin's lymphomas to diagnose the malignant lymphomas and to determine their immune phenotypes. The accuracy of proposed techniques combination to diagnose the malignant lymphoma consists 98% and to determine the immune phenotype - 90%. The presented combination of routine cytological analysis combined with techniques of flow cytofluorometry and immune cytochemistry is the express-technique to diagnose lymphoma and its immune phenotype the day of patient visit to the doctor. This approach significantly increases the role of cytological method in diagnostics of lymphomas. The technique makes it possible to analyze simultaneously huge quantity number of lymph nodes, including deep-seated and not only the superficial lymph nodes.

The article considers the main positions concerning the clinical laboratory diagnostics as an independent clinical specialty and the principles of professional training and improvement of specialists. The basic issues complicating the training and improvement of personnel to be kept in line with actual needs of laboratory service of public health system are discussed. Among them are the availability of laboratory academic sub disciplines demanding a profound special theoretical education and technical skills; the need to account in the process of professional training the variety of forms, sizes and types of laboratory structures in different medical institutions; the need of special training programs for numerous specialists with non-medical basic education. The combination of the present system of postgraduate training of specialists on chairs of state educational organizations with initiative involvement of specialists in various public forms of permanent professional improvement (professional scientific societies meetings, research conferences, internet seminars, etc.) is supported Along with a positive appraisal of the existing system of training in the state educational institutions and corresponding regulation documents, a critique is expressed regarding certain actual documents which improperly limit the administrative functions of physicians of clinical laboratory diagnostics and complicate training of bacteriologists for clinical laboratories.

The terahertz optical activity of 2D arrays of metal-semiconductor microhelices was investigated by Fourier spectroscopy and free electron laser methods. We found strong resonance at first harmonic with wavelength twice helix length and weaker resonances at the second and third harmonics. The resonances are present in the transmittance, reflectance, and absorption spectra. For linear polarized radiation, a chirality of the media leads to their ellipticity at resonance frequency and rotation of the polarization plane in opposite directions (up to 16.5 degrees) on both sides of the resonance. It has been shown by direct interferometric method on free electron laser that the refraction index of the investigated layer has negative values for definite circular wave at certain frequencies. Its minimal value is −0.11.

High power THz applications on the Novosibirsk terahertz free electron laser are described.

In the past decade, a notable advance has been made in the understanding of the pathogenesis of NK/T-cell lymphomas; however, their diagnosis remains difficult because of their rarity and clinical and morphological variabilities. The paper generalizes the ten-year experience of the Hematology Research Center, Ministry of Health of Russia, in diagnosing and treating hepatosplenic T-cell lymphoma (HSTL), considers the problems of differential diagnosis with other hematological diseases occurring with similar clinical and laboratory symptoms, and lays down current approaches to the diagnosis and treatment of this condition. A clinician's view of the problem of diagnosis and treatment of this disease is given. HSTL is shown to be a heterogeneous group of diseases differing in a T-cell receptor chain gene rearrangement, the clinical course of the disease, and overall survival (OS). According to our data, 3-year OS was 12%; the median survival was 26 months. Two-year OS for γδ and αβ HSTL was equal to 25 and 70%, respectively. The difference in OS for the variants of HSTL failed to reach statistical significance (because the sample might be insufficient).За последнее десятилетие достигнуты значительные успехи в понимании патогенеза NK/Т-клеточных лимфом, однако их диагностика остается затруднительной вследствие их редкости и клинико-морфологического разнообразия. В статье обобщен десятилетний опыт диагностики и лечения гепатолиенальной Т-клеточной лимфомы (ГЛТЛ) в ФГБУ 'Гематологический научный центр' Минздрава России, рассмотрены вопросы дифференциальной диагностики с другими гематологическими заболеваниями, протекающими со сходными клинико-лабораторными симптомами, сформулированы современные подходы к диагностике и лечению. Представлен взгляд клинициста на проблему диагностики и лечения данного заболевания. Показано, что ГЛТЛ - гетерогенная группа заболеваний, различающихся по реаранжировкам генов цепей Т-клеточного рецептора, клиническому течению заболевания, общей выживаемости (ОВ). По нашим данным, 3-летняя ОВ составила 12%, медиана продолжительности жизни - 26 мес. Для вариантов γδ и αβ ГЛТЛ 2-летняя ОВ равна 25 и 70% соответственно. Различие ОВ для вариантов ГЛТЛ не достигало статистической значимости (возможно, в силу недостаточного объема выборки).

Novel approach to controlling polarization of THz radiation is introduced. Polarization plane rotation of transmitted wave by a monolayer of precise microhelices was demonstrated by experiments on the Novosibirsk free electron laser. It had strongly resonant character. Modulation of polarization opens up new possibilities in communication, biomedical, chemistry, security, imaging and spectroscopy applications.

A comparison between phase characteristics of the laser multimode beam and random light fields is carried out theoretically. It is shown that probability density of phase distribution has considerable distinctions. The peculiarities of the wavefront dislocation structure in investigating fields are considered.

Introduction. Paroxysmal nocturnal hemoglobinuria (PNH) is a rare hematopoietic stem cell disorder resulting from the somatic mutation of the X-linked phosphatidylinositolglycan complementation Class A (PIG-A) gene. PIGA mutations in PNH patients lead to a glycosylphosphatidylinositol (GPI)-linked membrane proteins expression deficiency.In PNH, there is a partial or absolute inability to make GPI-anchored proteins including complement-defense structures such as CD55 and CD59 on RBCs and WBCs. Clinical features of PNH include intravascular hemolysis, bone marrow failure, and thrombosis, all major causes of morbidity and mortality. Flow cytometry (FCM) plays a key role in the laboratory investigation of PNH, and rapid diagnosis of this condition is highly desirable. A definitive diagnosis of PNH can be established by demonstrating the absence of cell membraneGPI-anchored proteins from granulocytes or red blood cells (RBC) according to ICCS Guidelines for the diagnosis and monitoring of PNH by flow cytometry. It has been also described that the expression of CD16 can be decreased on PNH-affected granulocytes. Recently new method for extended flow WBC differential was introduced by Beckman Coulter. This method uses flow cytometric analysis with CytoDiff™** reagents which is a 5-color/6-marker reagent that provides a 10-part cytometric differential from whole blood specimens and comprises CD36-FITC, (CD2+CD294)-PE, CD19-ECD, CD16-PC5, and CD45-PC7 (Beckman Coulter). The protocol allows detection of mature neutrophils, total lymphocytes, total monocytes, eosinophils, basophils, immature granulocytes, B lymphocytes, CD16-negative T/NK lymphocytes, CD16-positive T/NK lymphocytes, CD16 positive and CD16 negative monocytes, and blasts cells with lineage orientation. This method also allows the detection of the abnormal antigen expression on WBC, for example low CD16 expression on neutrophils. In case of abnormal low CD16 expression on segmented neutrophils they will be classified as Immature Granulocytes (Imm Gran). The aim of the study was to evaluate the efficacy of CytoDiff** analysis of peripheral blood for PNH screening detecting low CD16 expression on neutrophils. Methods: EDTA-anticoagulated blood samples from 53 patients with PHN suspicion were prospectively included in the study. Analysis of the PNH clones was conducted in according with international protocol, using CD235a for RBC gating, CD15-PE/CD45-PC7 for granulocyte gating, CD64-PC5/CD45-PC7 for monocyte gating and GPI-anchored proteins CD59-PE, FLAER-FITC/CD24-PC5 and FLAER-FITC/CD14-PE for RBCs, granulocytes and monocytes accordingly. For extended flow WBC differential analysis the blood samples were stained with the CytoDiff** panel, lyzed with Versalyse (Beckman Coulter) and 20 000 leucocytes were analyzed on a FC500 Flow Cytometer (Beckman Coulter) using CytoDiff** CXP software. Results:Totally 53 patients with PHN suspicion were prospectively included in the study. All these patients were characterized by anemia, thrombocytopenia and/or leucopenia. PNH diagnosis was confirmed in 6 patients and in other 7 patients the final diagnosis was aplastic anemia with PNH clone. For all 13 patients (4 males, 9 females, with median age of 41.5 years) with confirmed presence of PNH clone CytoDiff** reported increased number of Imm Gran (range 3-45%). Microscopy analysis did not detect the presence of Imm Grans in the slide, so we concluded that falsely reported increased Imm Gran count was due to the decreased expression of CD16 on Neutrophils. Good correlation (r=0.9257) was observed between Imm Gran count and the size of granulocytic PNH clone. Conclusion: Our data demonstrate that CytoDiff** analysis, which is able to detect a wide spectrum of normal and pathological cells in peripheral blood allows determination of CD16-low expression on neutrophils and thus provide efficient screening for suspicious of PNH in patients with anemic syndrome. ** Not available in the United States and other geographies. Disclosures Lugovskaya: Beckman Coulter: Research Funding.

The problems of the atom beam formation and its application to the micro- and nanoelectronic manufacturing are considered. The method introducing a useful information into the structured atom beam and focusing this beam is described. This method named the atom projection is relevant to atom optics. Negative detuned laser radiation field controls the beam. Features of the cooling, structuring, information input, focusing processes are discussed. The scheme of the plant using the atom beam with adjustable density distribution. The atom beam cross-section and the processed area are shaped according to the desired profile. The scheme of the atom projector and the functions of its units are described. The spatial resolution of processing and the resolution of surface analysis are evaluated to be about 10 nm and 1 nm correspondingly. Presented process is maskless, in-situ, with high output rating. The atom projector can be applied for the manufacturing, analysis and in-line reconstruction of the IC as well as for basic researchers of matter. Authors consider the nanoelectronic chip manufacturing as the most perspective application for the atom projector.

The problems of neutral atom beam formation and its application to the microelectronic manufacturing are considered. The plant using the focused neutral atom beam is presented. This high-vacuum plant is designed for complex CIP's processing. The scheme of this plant is given. It is divided into the following sections: atom generation, beam preparing, its structuring, compression, and surface treatment. The preparing section is described in detail. The beam is controlled by laser radiation resonant to basic transition in the atom absorption spectrum. The plant forms precisely structured low-temperature atom beams and controls atom density distribution in flow by rather simple projective methods. It allows discussing the possibility of the new, effective and high-automated manufacturing and analysis of micro-object surfaces. Resolution of the suggested method of treatment is proved to be limited only by characteristics of the equipment used. Calculation shows that at modern level of laser technique the spatial resolution of such a process can reach about 10 nm. The resolution of surface analysis is evaluated to be about 1 nm. Probable applications of the presented new technological process for integrated manufacturing, analysis and in-line reconstruction of the IC of super high integration by methods of liquid-free in-situ processing are considered.

One proposes the method of neutral atom beam formation due to effect of neutral atoms long-time localization in the minimums of potentials of intense standing wave of resonant light with simultaneous influence of cooling radiation. For fields of complicated configurati on the resonant light pressure force and the momentum diffusion tensor of two-level atom ensemble are presented. Mathematical modeling is based on the solving of equations for their spatial terms. One shows that with cooling time about 2 - 3 ms and with manipulation of intensity of co-propagated light wave, the arbitrary given cross-section atom density distribution with the contrast up to 1000 can be produced. One demonstrates that size of localization region can be essentially decreased by sweeping of cooling light carrier frequency in the range about 2 - 3 widths of moving atoms spectral transition lines. Low temperature cooling of atoms in the beam and 'soft' output from interaction region allow us to produce the well-manipulated practically aberration-off process. In this case transverse compression of image in cross-section of atomic beam can reach to 106 with given level of contrast. Calculations shows that at modern level of laser technique the spatial resolution of such process can reach to 30 - 50 A. The possibility of using of given method for high-resolution development and the analyses of micro-object surfaces is discussed.© (1997) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Introduction Paroxysmal nocturnal hemoglobinuria (PNH) is a rare and life-threatening hematopoietic stem cell disease caused by a partial or absolute deficiency of proteins linked to the cell surface membrane via a glycosylphosphatidyl-inositol anchor, which leads to complement-induced intravascular hemolysis mediated via the membrane attack complex. Multiparameter high-sensitivity flow cytometric measurement of PNH clones is the method of choice for the diagnosis of PNH, as recommended by the International Clinical Cytometry Society (ICCS). After publication of the ICCS guidelines, screening of patients considered at high risk of PNH was commenced in Russia. Data are presented on PNH clone size distribution across patients with relevant ICD-10 diagnostic codes (based on patients′ initial assumed diagnoses). Methods Patients were tested for the presence and size of PNH clones using high-sensitivity flow cytometry across nine laboratories. PNH clone evaluations were performed as described in the ICCS guidelines: CD59/CD235a monoclonal antibodies for RBC; CD45/CD15/CD24/FLAER for granulocytes and; CD45/CD64/CD14/FLAER or CD45/CD33/CD14/FLAER for monocytes. The sensitivity for PNH clone detection was 0.01%. Changes in PNH clone size were evaluated among patients who had follow-up studies after initial measurements. Results 1889 patients were assessed between October 2011 and June 2013 (Table 1). Suspected PNH and bone-marrow disorders (AA, MDS, cytopenia) were the most common reasons for PNH testing. The greatest proportions of patients with PNH clones were among those with of an initial assumed diagnosis of AA or PNH. Notably, around 40% of patients with an initial assumed diagnosis of PNH actually had no detectable PNH clones. Most patients with small clone sizes (< 1%) were in the AA, MDS and hemolytic anemia groups. Overall, mean clone sizes were slightly higher in monocytes (31.5%) than in granulocytes (30.1%) across the diagnostic categories. While there was generally a good correlation between clone size measurements in granulocytes and monocytes (linear regression r2 = 0.9851), 10% of PNH-positive patients had detectable clones only in one of these leucocyte populations (i.e. either in monocytes or in granulocytes, but not both). PNH clones in RBCs were generally lower than in granulocytes. Repeat clone size measurements were performed in 316 patients over a mean follow-up period of 7.8 months. In patients with initial clone sizes 50%, clones tended to increase. PNH clones were not changed at all in 98 patients at follow-up, among whom 48% were patients with AA. Conclusion These screening data confirm the utility of high-sensitivity flow cytometry testing in high-risk patient groups to ensure early and accurate diagnosis and to aid in the effective clinical management of patients. Disclosures: Babenko: Alexion: Research Funding. Sipol:Alexion: Research Funding. Borisov:Alexion: Employment. Naumova:Alexion: Research Funding. Boyakova:Alexion: Research Funding. Glazanova:Alexion: Research Funding. Chubukina:Alexion: Research Funding. Pronkina:Alexion: Research Funding. Popov:Alexion: Research Funding. Mustafin:Alexion: Research Funding. Fidarova:Alexion: Honoraria. Lisukov:Alexion: Honoraria. Kulagin:Alexion: Honoraria.

We show that THz chiral metamaterials consisting of large regular arrays of metal–semiconductor microhelices can be manufactured from strained heterofilms by directional rolling. We also report the results of an experimental study of the electromagnetic properties of such arrays.