— Hunting is an important component of the anthropogenic factor affecting populations of wild animals. In this work, a genetic analysis of mtDNA control region I of the wolf from the Republic of Karelia and the experimental territory (Kondopozhskii district) was carried out on a time interval of ten years. Based on the analysis of data on the population dynamics of the predator and its preys, hunting bag statistic, changes in the spatial and temporal structure, and the genetic characteristics, the degree of human influence on the wolf population in Eastern Fennoscandia was demonstrated. The modern haplotype diversity of the wolf in the region is poor (against the background of the general one typical for the species). In total, two haplotypes widespread in Eurasia were identified here, which is obviously explained by the species history (stages of a strong decline in the number).

Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder caused by WAS gene mutations resulting in haematopoietic/immune cell defects. Recent studies report accelerated death of WAS platelets and lymphocytes. Data on megakaryocyte (MK) maturation, viability and their possible role in thrombocytopenia development in WAS are limited. In this study we evaluate the MK viability and morphology in untreated, romiplostim-treated WAS patients compared with normal controls. The study included 32 WAS patients and 17 healthy donors. MKs were captured from bone marrow aspirates by surface-immobilized anti-GPIIb-IIIa antibody. Viability (by phosphatidylserine [PS] externalization), distribution by maturation stages and size of MK were determined by light microscopy. MK distribution by maturation stages in patients differed from controls. 40 ± 22% of WAS MKs versus 23 ± 11% of normal MKs were at maturation stage 3 (p = 0.02), whereas 24 ± 20% in WAS and 39 ± 14% in controls had megakaryoblast morphology (p = 0.05). Romiplostim treatment changed the MK maturation stages distribution close to normal. PS-positive (PS+) MK in WAS was significantly higher (21 ± 21%) than in healthy controls (2 ± 4%, p < 0.01). WAS patients with more damaging truncating mutations and higher disease score had higher PS+ MK fraction (Spearman r = 0.6, p < 0.003). We conclude that WAS MKs have increased cell death tendency and changes in maturation pattern. Both could contribute to thrombocytopenia in WAS patients.

One of the most ancient aspects of human impact on the wild animals is hunting. The degree of human impact on one of the Eastern Fennoscandian wolf (Canis lupus L.) population based on the analysis of data on the population dynamics of the predator and its preys, “hunting bag” data, changes in the spatial-temporal structure and genetic characteristics, is shown. In the presented work, a genetic analysis of the control region mtDNA of the wolf of the Republic of Karelia (Russia) and the experimental territory (Kondopozhsky district of Karelia) was performed on a time slice of 10 years. The modern mtDNA haplotypic diversity of Russian Karelian and Finnish wolf population, in comparison with the general diversity characteristic for the species, is poor. In total, only two haplotypes common in Eurasia have been identified here, which is obviously explained by the history of the species – the stages of severe population decline.

Background: At present, the diagnosis of lymphoproliferative disorders is based on the combination of blood or bone marrow smear morphology and immunophenotyping by flow cytometry. Immunophenotypic testing by flow cytometry technique is available only in big medical centers, which is not always convenient for a patient. Therefore, development of an available method for preliminary diagnosis of lymphoproliferative diseases not requiring special equipment seems relevant.Materials and methods: Peripheral blood mononuclear cells from 17 patients admitted to the hospital with suspicion of a lymphoproliferative disorder, and 17 healthy donors were studied on a cell biochip for determination of proportions of cells positive for various surface CD antigens. The diagnosis was verified by flow cytometry.Results: Compared to healthy controls and patients with T-cell lymphoproliferative disorders (TCLPD), the patients with B-cell lymphoproliferative disorders (BCLPD) had significantly lower proportion of CD7+ cells (medians, 7% and 73% respectively, p=2×10-6 for comparison with healthy controls; median 7% and 93% for comparison with TCLPD, p=0.032). In addition, the patients with BCLPD had higher proportion of peripheral СD19+ mononuclear cells, compared to that in the patients with TCLPD and healthy donors (medians 84% and 13% for comparison between BCLPD and healthy control, p=2×10-5; 84% and 3% for comparison of BCLPD and TCLPD, p=0.033). The patients with B-cell chronic lymphocytic leukemia had significantly higher CD5+ cells in the cell biochip compared to the patients with other BCLPD (medians 72% and 9%, p=0.024). The patients with TCLPD had significantly lower proportion of CD19+ cells than the healthy controls (medians, 3% and 13%, respectively, р=0.042).Conclusion: The study has demonstrated the potential to use the previously developed cell biochip for diagnosis of lymphoproliferative diseases. The biochip makes it possible to sort out white blood cells according to their surface differentiation antigen for their further morphological analysis. The cell biochip allows for the differential diagnosis between BCLPD and TCLPD and determination the lymphocyte clones based on the expression of immunoglobulin light chains.

Atypical mononuclear cells (AM) appear in significant numbers in peripheral blood of patients with Epstein-Barr virus (EBV)-associated infectious mononucleosis (IM). We investigated the number and lineage-specific clusters of differentiation (CD) expression of atypical mononuclear cells in 110 children with IM using the anti-CD antibody microarray for panning leukocytes by their surface markers prior to morphology examination. The AM population consisted primarily of CD8+ T cells with a small fraction (0%-2% of all lymphocytes) of CD19+ B lymphocytes. AM amount in children with mononucleosis caused by primary EBV infection was significantly higher than for IM caused by EBV reactivation or other viruses and constituted 1%-53% of all peripheral blood mononuclear cells compared to 0%-11% and 0%-8%, respectively. Children failing to recover from classic IM associated with primary EBV infection within 6 months had significantly lower percentage of CD8+ AM compared to patients with normal recovery rate.

Both the ratio of different leukocyte subgroup content and the leukocyte morphology in peripheral blood of newborns are important in diagnosis of several diseases including combined immunodeficiency and neonatal septicemia. There is a need for development of screening methods for parallel study of the leukocyte morphology and population structure in the newborn peripheral blood. We aimed to determine the relative abundance of different leukocyte subsets and to study their morphology in full-term and premature newborn babies and healthy adult volunteers using the cell-binding microarray – a transparent support with immobilized antibodies against leukocyte cluster-of-differentiation antigens. The work was supported by the Scientific council and approved by the ethical committee of the Centre. We have studied the peripheral blood of 12 full-term newborns (38–40 weeks gestation), 9 premature newborns (22–32 weeks gestation) and 18 healthy adults. The relative abundance of the leukocyte and their morphology were determined using the cell-binding microarray including antibodies against CD2, СD3, СD4, CD5, СD7, CD8, CD10, СD11b, CD11c, CD13, CD14, CD15, CD16, CD19, CD20, CD22, CD25, CD33, CD38, CD41a, CD45, CD45RA, CD45RO, CD61, CD64, CD117, CD123, HLA-DR. The percentage of leukocytes positive for every of the studied surface CD antigens among the peripheral blood mononuclear cells of full-term and preterm newborn babies and healthy adults determined on the cell–binding microarray are in good agreement with published flow cytometry data. CD11b+ leukocytes both in premature and full-term newborns included up to 21% myelocytes and 27% metamyelocytes. The reported data can be used as reference values in cell-binding microarray application in diagnosis of combined immunodeficiency or neonatal septicemia.

Hairy cell leukemia (HCL) is a chronic lymphoproliferative disorder constituting about 2% from all leukemia cases and characterized by typical “hairy” morphology of tumor lymphocytes. We describe an HCL case with atypical nuclear shapes (lymphocytes with clover-leaf-like, horse-shoe-like, ring-shaped nuclei and binuclear cells were present). Morphology and immunophenotype of circulating leukemic cells were studied using a cell-binding microarray - a transparent plastic slide with immobilized monoclonal antibodies against surface antigens of lymphocytes. The cell-binding microarray with immobilized anti-CD11c, anti-CD103 and anti-CD123 permits to study a lymphocyte population enriched with hairy cells. Hairy cells with atypical nuclei constituted 3% of all lymphocytes and 15% of all hairy cells. This unusual hairy cell morphology is the first described in Russia and was found in one out of 85 HCL cases in our practice.

Key Clinical MessageThe morphologic diagnosis of hairy cell leukemia coexisting with another lymphoproliferative disorder is hindered by the small size of hairy cell population. It can be simplified by presorting peripheral blood mononuclear cell using an anti‐CD antibody microarray on transparent support (including anti‐CD11c, CD25, CD103, and CD123) before their morphology analysis.

Acute leukemia diagnosis steadily shifts towards automated analysis and precise molecular methods banishing the methods resistant to complete automatization (such as cytochemistry and bone marrow morphology). The reasons for this lie above all in the desire to eliminate the human factor as well as to find a way to cope with increasing work load. The better understanding of the biology of the disease achieved with the advanced molecular technologies led to the inclusion of new recurrent cytogenetic abnormalities, mutations and gene fusions into the revised WHO classification of tumors of the hematopoietic and lymphoid tissues. Wherever possible the acute leukemias are classified by mutations rather than by underlying subtypes according to French-American-British (FAB) classification.1 FAB classification is retained only in the “not otherwise specified” category. These changes give a probably involuntary message that as mutations prevail over morphologic and immunophenotypic characteristics of leukemic cells the latter at a certain point will not be needed for the diagnosis.

We present data on the species composition of helminths in brown bears (Ursus arctos) from the Murmansk Region, Russia. The absence of any information about helminths of brown bear in the region necessitated the conduct of these studies. Samples were collected in 2014 and 2015 in the southern part of the Kola Peninsula from the White Sea coastal habitats. Annually, in the study area, 1–3 bears are legally hunted and biological samples for examination are very difficult to obtain. Therefore, we used fecal samples. We studied 93 feces and identified parasite eggs identified in 43 of them by morphometric criteria. The surveys revealed eggs of the following helminths: Dicrocoelium sp., Diphyllobothrium sp., Anoplocephalidae, Capillariidae, Baylisascaris sp., Strongylida 1, and Strongylida 2. These results represent the first reconnaissance stage, which allowed characterizing the taxonomic diversity and prevalence of parasites of brown bears of the Kola Peninsula.

We have studied the morphology and immunophenotype of circulating tumor cells isolated from peripheral blood of 22 patients with splenic marginal zone lymphoma and show that both of them are highly heterogeneous. Using a cell-binding microarray we have demonstrated that the circulating lymphoma cells are positive for CD19 (100 %), CD20 (100 %), CD22 (100 %), surface IgM (73 %), CD38 (23 %), CD5 (9 %), CD11c (36 %), CD103 (5 %), CD25 (32 %), CD23 (23 %) and these immunophenotypes are confirmed in all cases by flow cytometry.Higher surface density of lymphocyte binding onto anti-CD antibody microarray spots compared to blood smears permits to find circulating lymphoma cells even in leukopenic patients.

Here we describe and qualitatively characterize the distribution of mononuclear cells isolated from bone marrow aspirate of healthy donors and sorted by their surface CD antigens on a cell microarray by morphology (for 11 donors) and by absence or presence of α-naphthyl butyrate esterase and naphthol AS-D chloroacetate esterase (for 9 donors). These data can be used as a reference for the diagnosis of acute myeloid leukemia using the cell microarray.

We describe a method for leukocyte sorting by a microarray of anti-cluster-of-differentiation (anti-CD) antibodies and for preparation of the bound cells for morphological or cytochemical examination. The procedure results in a “sorted” smear with cells positive for certain surface antigens localised in predefined areas. The morphology and cytochemistry of the microarray-captured normal and neoplastic peripheral blood mononuclear cells are identical to the same characteristics in a smear. The microarray permits to determine the proportions of cells positive for the CD antigens on the microarray panel with high correlation with flow cytometry. Using the anti-CD microarray we show that normal granular lymphocytes and lymphocytes with radial segmentation of the nuclei are positive for CD3, CD8, CD16 or CD56 but not for CD4 or CD19. We also show that the described technique permits to obtain a pure leukemic cell population or to separate two leukemic cell populations on different antibody spots and to study their morphology or cytochemistry directly on the microarray. In cases of leukemias/lymphomas when circulating neoplastic cells are morphologically distinct, preliminary diagnosis can be suggested from full analysis of cell morphology, cytochemistry and their binding pattern on the microarray.

We describe an application of a cell-binding microarray – to parallel study of morphology, tartrate-resistant acid phosphatase activity and detection of surface markers on peripheral blood lymphocytes of 90 atients with suspected hairy cell leukemia (HCL). We have formulated the microarray-based diagnostic criteria for hairy cell leukemia, hairy cell leukemia variant (HCLv) and splenic marginal zone lymphoma (SMZL). According to these criteria we have suggested the presence of HCL for 55 patients, HCLv – for 7 patients and SMZL – for 10 patients from the studied cohort. These diagnoses were confirmed by standard diagnostic methods in all cases. These results show that the cellbinding microarray can be used in differential diagnosis of HCL, while the high sensitivity of the microarray permits to detect the leukemic cells in spite of leukopenia and low hairy cell content.