Large Lymphocytes Research Articles

B-cell lymphoma-2 downregulation is a useful feature supporting a neoplastic phenotype in mature T-cell lymphomas

Normal T cells express high levels of B-cell lymphoma-2 (BCL2) protein, and data regarding BCL2 expression status and its diagnostic utility in T-cell lymphoma are scarce. We evaluated BCL2 expression in a series of mature T-cell lymphoproliferations (TCLs) including indolent and more recently recognized entities (follicular helper T-cell [TFH] lymphomas). Sixty-six neoplastic biopsies (60 patients) representing mature nodal, extranodal, and leukemia T-cell neoplasms were collected from three institutes (2 US and 1 Japan) and were compared with reactive T cells in 8 benign tissues/blood and 9T cell-rich B-cell proliferations. BCL2 immunostaining was performed and scored based on intensity-weighted H-score (0-300). Next-generation sequencing (NGS; 5 cases), BCL2 gene sequencing, and real-time polymerase chain reaction (PCR; 3 cases) were conducted. Association of H-score with overall survival (using proportional hazards modeling) was assessed in nonleukemic TCLs. Most TCLs showed significantly downregulated median BCL2 H-score (125, range: 18-300) with the exception of T-cell prolymphocytic leukemia and hepatosplenic T-cell lymphoma, both of which showed uniform strong retention of BCL2 as did the 8 reactive tissues (median H-score: 280; p=0.000). Notably all TFH lymphoma CD4 neoplastic T cells, subcutaneous panniculitis-like T-cell lymphoma, CD8 adipocyte-rimming T cells, and T-cell large lymphocyte leukemia with pathogenic STAT5B and TP53 mutation showed BCL2 downregulation. No BCL2 mutations were observed by NGS or sequencing with decreased BCL2 mRNA transcripts by real-time PCR. BCL2 downregulation is pervasive among many TCLs and unrelated to any mutations. There is utility for BCL2 immunostaining in some challenging situations as discussed in this article.

CELLULAR COMPOSITION OF THE LYMPHOID TISSUE OF THE CECAL IMMUNE FORMATIONS IN DUCKS

Observing the quantitative and qualitative composition of immunocompetent cell populations of the lymphoid tissue of the immunogenic organs allows to determine the immune status of the organism in a certain age period. The object of research is determing the cellular composition of the lymphoid tissue of the cecal Peyer’s patches and cecal (apical) diverticula in ducks in age-concerned aspect. Material simples for research were selected from broiler ducks of the Blagovarsky cross. Cytological testss were performed on imprint specimens. Immunoblasts, lymphocytes, proplasmocytes, and plasmocytes, monocytes, and macrophages are distinguished among the cells of the lymphoid tissue of the cecal Peyer’s patches and the cecal diverticula. The content of these cells is not the same. Population of lymphocytes in imprint specimens is the biggest. It consists of subpopulations of small, medium, and large lymphocytes, the ratio is uneven. The largest is a subpopulation of small lymphocytes, and the smallest is a subpopulation of large ones. The total content of lymphocyte in cecal Peyer’s patches and cecal diverticula decreases with age of the subject ducks. The content of small and medium-sized lymphocytes in the cecal diverticula and small lymphocytes in the cecal Peyer’s patches as well decreases. Simultaneously, the content of large lymphocytes in the cecal diverticula, large and medium lymphocytes in the cecal Peyer’s patches increases. The immunoblasts content in the lymphoid tissue of the studied immune formations decreases with age of ducks, while the quantity of macrophages and monocytes conversely increases. Proplasmocytes and plasmocytes are detected in the lymphoid tissue of cecal Peyer’s patches and cecal diverticula from the age of 10 days in ducks. Their content increases significantly with the poultry age. Reticular cells observation if complicated due to their location under a dense layer of lymphoid cells. Fibroblasts, M-cells, erythrocytes, and granulocytes in imprint specimens are detected in trace amounts. The established changes in the cellular composition of the lymphoid tissue of the cecal Peyer’s patches and the cecal diverticula in ducks in the age-related aspect confirm the occurring immune reactions within them. Consideration of these changes will improve the effectiveness of anti-epizootic measures

Exclusive diagnosis of aplastic anemia

As an independent disease, aplastic anemia (AA) has been recognized for more than a century. When AA is diagnosed, other non-AA bone marrow failures should be excluded. It is termed as exclusive diagnosis of AA. The exclusive diagnosis of AA is helplessly based on that there is no parameter by which AA can be sensitively and specifically diagnosed now. So further searching for the meaningful diagnostic parameters of AA should be carried on to establish a direct diagnostic protocol of this disease and make it possible to differentiate it clearly from other bone marrow failure disease such as congenenital bone marrow failure, hypoplastic myelodysplastic syndromes, AA-paroxysmal nocturnal hemoglobinuria syndromes, large granules lymphocyte leukemia, clonal cytopenia of undetermined significance, immunorelated pancytopenia, acute hemopoietic arresting and idiopathic cytopenia of undetermined significance. The new markers and technologies being helpful for distinguishing AA from other bone marrow failures should be used in diagnosing AA. Correct understanding and application of exclusive diagnosis is not only related to the correctness of diagnosis and treatment of excluded diseases, but also to the quality of AA diagnosis, treatment and research.

CHANGES IN MEGAKARYOCYTES IN VARIOUS HEMATOLOGICAL CONDITIONS : A STUDY OF BONE MARROW ASPIRATION IN A TERTIARY CARE CENTRE

INTRODUCTION Dysmegakaryopoiesis is characterized by various Megakaryocytic alterations in bone marrow and includes both dysplastic and non dysplastic features.Dysplastic changes are mostly seen in myelodysplastic syndrome however megakaryocyte alteration have also been noted in some bone marrow aspiration in non myelodysplastic conditions. AIMS OBJECTIVE To evaluate changes in megakaryocytes in various hematological conditions in bone marrow aspiration examination. MATERIAL AND METHODS Study was done at Central Pathology lab in the Department of Pathology GAJRA RAJA MEDICAL COLLEGE GWALIOR over a period of 1.5 year FROM NOV 2019 TO MAY 2021.Total 84 bone marrow were received out of 14 were dilute so excluded from study and results were prepared from 70 cases.Age range was 9 months to 75 years RESULTS AND OBSERVATIONS In this study out of 70 patients, 43 were male and 27 were female.Patients were categorised into neoplastic and non neoplastic category.Out of 70 cases 46 were non neoplastic forming majority while 24 were neoplastic. Out of non neoplastic, megaloblastic anemia was most common while acute leukemia was most frequent in neoplastic category.Bone marrow was evaluated for cellularity. 47 were hypercellular marrow forming majority with 15 cases showing normal cellularity and 08 as hypocellular. Acute leukemia and megaloblastic anemia were two conditions which had hypercellular marrow. The number of the megakaryocytes was considered normal when one megakaryocyte per 3 low power fields was encountered,increased if more than two megakaryocytes per low power field were seen and decreased when one megakaryocyte per five to ten low power fields was documented and absent when no megakaryocytes were seen per ten low power field. Our study showed 27 cases of megakaryocytic hypocellularity, 22 cases normal number of megakaryocytes and 18 as increased number of megakaryocytes. Normally megakaryocytes have four to sixteen nuclear lobes. Immature megakaryocytes were defined as young forms of megakaryocytes with scant bluish cytoplasm and lacking lobulation of the nucleus. Dysplastic megakaryocytes are those with single, multiple/separated nuclei. Micro megakaryocytes were defined as megakaryocytes whose size was that of large lymphocyte or monocyte and which had a single / bi-lobed nucleus. Our study – showed normal megakaryocyte morphology in 32/70 (45%) cases and 35/70 (50%) were with altered morphology. CONCLUSION Dysplastic Morphologic changes in megakaryocyte were not only seen in MDS but also in various non MDS conditions which should be considered during diagnosis.Understanding and detailed knowledge of changes in megakaryocytes,including both cellularity and morphology,can improve the diagnostic accuracy for a wide range of hematological disorders

Debris collected in-situ from spontaneously ruptured atherosclerotic plaque invariably contains large cholesterol crystals and evidence of activation of innate inflammation: Insights from non-obstructive general angioscopy

Background and aimsDevelopment and expansion of cholesterol crystals (CCs) within a lipid rich atherosclerotic core are believed to predispose to plaque rupture. We have used non-obstructive general angioscopy to described a range of appearances of spontaneously ruptured atherosclerotic plaques (SRAPs) in the aorta in-situ, and have confirmed that debris extruding from some SRAPs (puff-chandelier lesions) are rich in cholesterol crystals and leukocytes. The purpose of this study was to characterized the nature of the inflammatory infiltrate of this debris. MethodsDebris was collected from puff-chandelier lesions at the time of angioscopy in patients with known coronary disease. Prepared specimens were examined by light microscopy, and immunostaining was used to detect markers of activation of the innate inflammatory pathway including CD68, NLRP3, caspase-1, IL-1β, IL-18, and IL-6. ResultsWe analysed debris sampled from 20 puff-chandelier lesions. Microscopy confirmed the presence of large CCs, macrophages, fibrin, calcified gruel, lymphocytes, and neutrophils in 100%, 100%, 95%, 25%, 20%, and 15% of the specimens respectively. Immunostaining confirmed the presence of CD68, NLRP3, IL-1β, and IL-6 within the debris in 100%, 90%, 80%, and 80%, of the specimens respectively. CCs, NLRP3, caspase-1, IL-1β, IL-18, were also identified in the cytoplasm of macrophages. ConclusionsDebris from SRAPs with a puff-chandelier appearance invariably contained large CCs associated with a range of activated leukocytes involved in innate inflammation. This observation supports the thesis that the development and enlargement of CCs in the core of lipid rich plaques may precipitate traumatic and inflammatory injury that may lead to plaque rupture.

Clinical and histopathological correlation in patients with chronic tonsillitis

<p class="abstract"><strong>Background:</strong> Chronic tonsillitis is diagnosed when an individual has 7 episodes of tonsillitis in 1 year, 5 episodes in 2 consecutive years, or 3 episodes each year for 3 years consecutively. There is correlation of clinical and histopathological findings in patients with chronic tonsillitis.</p><p class="abstract"><strong>Methods:</strong> It is prospective study which is aimed at correlation of clinical and histopathological findings in chronic tonsillitis patients underwent tonsillectomy. This was carried out in tertiary care hospital, Pune, Maharashtra, India. In our study, 25 patients of chronic tonsillitis underwent tonsillectomy were included. In this we have examined the histopathology of tonsillar tissue section in correlation with the clinical manifestations of tonsillar diseases that were due to infection. </p><p class="abstract"><strong>Results:</strong> In pathological examination of palatine tonsil, underlying subepithelial tissue showed many lymphoid follicles with prominent germinal centres comprising of centrocytes, centroblasts, immunoblasts, small and large lymphocytes in 19 patients (76%). 13 patients (52%) showed prominent germinal centres with bites of salivary gland and muscle tissue. 6 patients (24%) showed prominent germinal centre with mild lymphoplasmocytic inflammatory cell infiltrate. In 4 patients (16%), tissue lined by stratified squamous epithelium showed focal ulceration. 3 patients (12%) showed stratified squamous epithelium with granulation tissue. Crush artifact were noted in 3 patients (12%). 2 patients (8%) had small granules with actinomycotic species colonies within the palatine tonsilar crypts. Only 1 patient (4%) showing keratin impacted in tonsilar tissue.</p><p class="abstract"><strong>Conclusions:</strong> There is strong correlation between histopathological examination suggestive of lymphoid hypertrophy and grading of palatine tonsil hypertrophy.</p>

Hematological and serum biochemical reference intervals of rainbow trout, Oncorhynchus mykiss cultured in Himalayan aquaculture: Morphology, morphometrics and quantification of peripheral blood cells

De novo reference intervals (RIs) for a total of thirty two hematological and serum biochemical attributes were established for rainbow trout (Oncorhynchus mykiss) cultured in Himalayan aquaculture system. For this purpose, long term assessment of hemato-biochemical parameters was carried over a period of one year from March 2019 to February 2020 and a total of 444 blood samples were analysed. Blood examination results were recorded systematically and reference intervals were established, notably for erythrocyte parameters: hematocrit (Hct) 29–40%, hemoglobin 8.32–12.28 g/dL, red blood cell (RBC) count 1.01–2.04 (×106/mm3); leukocyte parameters (x 103/mm3): total leukocytes 31.32–90.60, neutrophils 4.21–18.85, total lymphocytes 20.55–63.63, small lymphocytes 14.86–46.50, large lymphocytes 6.35–22.34 and monocytes 1.22–7.56; thrombocyte count 23.00–68.00 (×106/mm3). RIs were also established for red blood cell indices, vital serum constituents involved in carbohydrate, protein, lipid and nitrogen metabolism including the less known, diagnostically important, serum enzymes and electrolyte concentrations. Principal component analysis revealed that certain serum components were more efficient at distinguishing between the life stages (juvenile, adult) of fish by explaining about 92.7% of variation in the whole dataset compared to the principal hematological components which explained only about 80% of the variation. Significant (P < 0.05) differences were noted for RBC count, total leukocyte count (TLC), total protein, total cholesterol and uric acid with respect to the sex of fish. Moreover, clearly differentiable morphometric and morphological attributes were also noticed among erythrocytes, leukocytes (lymphocytes, neutrophils and monocytes) and thrombocytes. To our knowledge, the present study is the first of its kind that elucidates blood chemistry of cultured rainbow trout, O. mykiss in accordance to the guidelines framed by the American society of veterinary clinical pathologists (ASVCP). RIs reported here can help monitor the fish health status by improving the use of non-lethal diagnostics in piscine medicine.

Sézary syndrome preceded by mycosis fungoides and complicated by tumor lysis syndrome and macrophage activation syndrome

Sir, Mycosis fungoides (MF) and Sézary syndrome (SS) are the most common malignancies of cutaneous T-cell lymphoma [1]. Herein, we report a case of SS complicated by tumor lysis syndrome and macrophage activation syndrome. A 54-year-old patient, followed since October 2017 for mycosis fungoides and undergoing various treatments (PUVA therapy, methotrexate, chlorambucil + prednisone), presented with an aggravation of lesions toward extensive and intensely pruritic. A clinical examination revealed dry erythroderma, scratch marks, wart plaques, an accentuation of frontal wrinkles and nasolabial folds (leonine facies), palmoplantar fissuring keratoderma, xanthopachyonychia of all nails, and a carapace-like appearance of the scalp (Figs. 1 – 3). Generalized lymphadenopathy, hepatomegaly, and a state of anasarca-type edema caused by hypoalbuminemia were also found. A skin biopsy revealed lymphoproliferation of CD4+ T-cells and an aberrant loss of pan-T antigens. The CD4-to-CD8 ratio was at 48.5% and Sézary cells were 6960 (absolute value). A lymph node biopsy showed a dense infiltration of Sézary cells. A PET scan revealed hypermetabolism in the entire skin and at the lymph node level. Tumor lysis syndrome was evident, with high levels of blood uric acid (at 182 mg/L), elevated LDH (at 924 U/L), and functional kidney failure. Macrophage activation syndrome was also present, with fever, anemia and thrombocytopenia, liver cytolysis, hypertriglyceridemia, and hyperferritinemia. The patient received an albumin infusion, oral corticosteroid therapy to treat the syndrome, and rasburicase for hyperuremia. Despite this, the patient died before multiagent chemotherapy could have been started. On rare occasions, SS may be preceded by a prior history of classic MF. The International Society for Cutaneous Lymphomas (ISCL) recommends that such cases be designated “SS preceded by MF” [2]. Traditionally, SS is defined as a leukemic form of CTCL associated with erythroderma. Sézary cells are a population of large lymphocytes in the peripheral blood, with grooved and lobulated nuclei, in the case of SS, numbering 1000 cells/mL or more [2]. The histopathologic findings in the skin often resemble those observed in MF, with less prominent epidermotropism. As in MF, immunohistochemical studies showing a CD4 predominance and loss of pan-T-cell markers may be helpful. Lymph node involvement is characterized by the complete effacement of the nodal architecture by the infiltrating Sézary cells (2). The poor prognostic factors in Sézary syndrome include an advanced stage of the disease, an older age at onset, and large cell transformation [3]. While high response rates may be achieved with systemic chemotherapy, they are frequently short-lived and associated with significant toxicities [2]. The management of SS is complicated and requires multidisciplinary collaboration between dermatologists, hematologists, biologists, and reanimators.

Primary breast CD20-positive extranodal NK/T cell lymphoma with stomach involvement: a case report and literature review

BackgroundWe present a unique case of primary breast CD20-positive extranodal NK/T cell lymphoma with stomach involvement in a young Chinese female patient.Case presentationThe patient presented with a mass in her right breast that rapidly increased in size over approximately 2 months. Upper gastrointestinal endoscopy showed a giant serpentine ulcer in the stomach. Biopsy was performed, and microscopic inspection revealed that the fibrous tissue was diffusely involved by medium to large abnormal lymphocytes. The cytoplasm was low to moderate. The tumor cells had irregular nuclei and inconspicuous nucleoli. The lymphoid cells were strongly immunoreactive to CD20, CD3, CD4, CD56, TIA-1, EBER, and Ki-67 (90%). Epstein-Barr virus genomes were also found in tumor cells by in situ hybridization. A whole-body positron emission tomography (PET)-CT scan revealed intense FDG uptake in the right breast and greater curvature of the stomach. Monoclonal rearrangements of the T cell receptor (TCR-γ) and immunoglobulin heavy chain (IgH) were identified by genetic analysis. Whole-genome next-generation sequencing was performed, and up to 12 gene mutations, including a frameshift mutation in exon 4 of the BCOR (G97Rfs*87; 44.3%) gene and a base substitution mutation (Q61H 17.6%) in exon 3 of the KRAS gene, were detected. Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed using the database for annotation, visualization, and integrated discovery, which showed that rare primary breast CD20-positive extranodal NK/T cell lymphoma had a unique genetic background compared with diffuse large B cell lymphoma and extranodal NK/T cell lymphoma without CD20 expression. The patient received four cycles of the modified SMILE regimen. The second whole-body PET-CT scan revealed that the right breast mass was significantly smaller than before; additionally, FDG uptake in the stomach wall disappeared.ConclusionsSystemic examination, extensive immunohistochemistry, and molecular profiling are essential for an accurate diagnosis. More similar cases are required to clarify the biological pathways and even the potential molecular mechanisms of rare lymphomas, which may help direct further treatment.

SPURIOUS HYPOXEMIA IN A CRITICALLY ILL PATIENT WITH CHRONIC LYMPHOCYTIC LEUKEMIA

TOPIC: Critical Care TYPE: Fellow Case Reports INTRODUCTION: Critically ill patients who require intubation and mechanical ventilation are routinely monitored with arterial blood gas (ABG) analysis to measure arterial oxygen tension (PaO2) as an indicator of adequate gas exchange. The partial pressure of oxygen in an ABG sample can be impacted by many factors including metabolically active cells such as white blood cells or platelets leading to a false measurement of hypoxemia. We report a case of spurious hypoxemia in a critically ill patient due to chronic lymphocytic leukemia. CASE PRESENTATION: 67 year-old male, with no reported past medical history, presented with complaints of acute onset fever, diarrhea and dyspnea. His physical exam was notable for cachexia, tachycardia and tachypnea. On arrival he was hemodynamically unstable and in acute hypoxic respiratory failure, requiring mechanical ventilation. Lab results showed anemia, thrombocytopenia and a leukocyte count of 104,000/mm3. Peripheral blood smear showed large lymphocytes, smudge cells and presence of prolymphocytes. Chest imaging showed left lower lobe pneumonia with emphysematous changes. He was started on broad spectrum antibiotics. Post intubation ABG was notable for PaO2 of 59 but patient's SpO2 was 95%. Repeat ABG showed same results. His vent settings were adjusted including increasing PEEP. His pulse oximetry remained >92% however ABG showed persistent hypoxemia. Patient's PaO2 improved after leukopharesis. However patient succumbed to his clinical condition after family opted for comfort care. DISCUSSION: Spurious hypoxemia occurs when patients with leukocytosis or thrombocytosis have low PaO2 on ABG analysis with a concurrent normal oxygen saturation. It was first described in 1979 as "leukocyte larceny" (1). The pathophysiology is believed to be related to the presence of a high number of metabolically active white blood cells which increase the consumption of the dissolved oxygen in ABG samples (2, 3). The ability to differentiate between true hypoxemia and spurious hypoxemia is important especially when managing critically ill patients with leukocytosis or thrombocytosis. High clinical suspicion for spurious hypoxemia should be considered in patients with white blood cell counts >50,0000/mm3, severe thrombocytosis, high oxygen saturations on pulse oximetry and very low PaO2 on ABG analysis. Some of the methods that have been used to prevent spurious hypoxemia include immediate cooling of ABG samples and rapid analysis of samples, where both methods theoretically slow leukocyte metabolic rate and reduce oxygen consumption. CONCLUSIONS: Early recognition of spurious hypoxemia in patients with leukemia could prevent unnecessary testing and true hypoxemia can be ruled out in the appropriate clinical setting. REFERENCE #1: Hess CE, Nichols AB, Hunt WB, Suratt PM. Pseudohypoxemia secondary to leukemia and thrombocytosis. N Engl J Med. 1979;301(7):361–363. REFERENCE #2: Fox MJ, Brody JS, Weintraub LR. Leukocyte larceny: a cause of spurious hypoxemia. Am J Med 1979;67(5):742–746. REFERENCE #3: Lele A, Mirski M, Stevens R. Spurious hypoxemia. Crit Care Med. 2005;33(8):1854–1856. DISCLOSURES: No relevant relationships by Louis Gerolemou, source=Web Response No relevant relationships by Wael Kalaji, source=Web Response No relevant relationships by Aneeta Kumari, source=Web Response No relevant relationships by Nabil Mesiha, source=Web Response No relevant relationships by Kunal Nangrani, source=Web Response No relevant relationships by Viswanath Vasudevan, source=Web Response

Gamma/Delta (γδ) T Cells: The Role of the T-Cell Receptor in Diagnosis and Prognosis of Hematologic Malignancies.

There are 2 types of T cells: αβ and γδ T cells, named based on the composition of the T-cell receptor. γδ T cells are rare, making up 0.5%-10% of T cells. Although most leukemias, lymphomas, and immune-mediated conditions derive from αβ T cells, a handful of rare but important diseases are generally derived from γδ T cells, particularly primary cutaneous γδ T-cell lymphoma, hepatosplenic T-cell lymphoma, and monomorphic epitheliotropic intestinal T-cell lymphoma. There are also malignancies that may evince a γδ TCR phenotype, including large granulocytic lymphocyte leukemia, T-cell acute lymphobplastic leukemia (T-ALL), and mycosis fungoides, although such cases are rare. In this article, we will review the genesis of the T-cell receptor, the role of γδ T cells, and the importance of TCR type and methods of detection and outline the evidence for prognostic significance (or lack thereof) in lymphomas of γδ T cells. We will also highlight conditions that rarely may present with a γδ TCR phenotype and assess the utility of testing for TCR type in these diseases.

Cell composition of blood for Salvelinus malma (Salmonidae) from the Raduga River (Kamchatka)

Hematopoiesis is a mechanism of multifunctional adaptation in vertebrates, including fish. Adaptive capabilities of populations are studied for a species of genus Salvelinus with high ecological plasticity. Char Salvelinus malma is represented in Kamchatka by the anadromous, river, brook, and several lake forms. Peripheral blood cells are investigated for this species juveniles and adults from the Raduga River in the lower Kamchatka River basin. The fish sampled in August 2013 were divided into two groups: sexually mature individuals (10 specimens) and juveniles (10 specimens). The body length (AC) of mature chars (mean ± standard deviation, range in brackets) was 338.90 ± 12.64 (304.0–380.0) mm, the body weight — 437.40 ± 57.74 (235.0–735.0) g, their sex ratio was close to 1 : 1. The length of juvenile chars was 150.50 ± 9.14 (104.0–195.0) mm, the weight — 33.90 ± 6.19 (10.1–68.8) g. Blood was sampled from the caudal vessel of alive fish immediately after catching, the blood smears were dried and fixed with May-Grunwald dye and then stained with azure-eosin. Composition of leukocytes and erythrocytes was determined on the stained blood smears. The blood cells were classified according to Ivanova (1983) and Zhiteneva et al. (1989). Statistical significance of the blood cells composition differences between fish groups was assessed using the Mann-Whitney U-test, after checking for normal distribution of characteristics (Kolmogorov-Smirnov test). The erythroid row of S. malma was represented by proliferating (erythroblasts, basophilic and polychromatophilic normoblasts), maturing (oxyphilic normoblasts) and mature erythrocytes, the leukocyte row — by blast cells, agranulocytes and granulocytes. The juveniles were characterized by intense proliferative processes (great number of immature erythrocytes, large and medium lymphocytes, presence of blast cells) and appearance of poikilocytes; the sexually mature chars were distinguished by greater number of granulocytes, mainly due to stab and segmented neutrophils (p &lt; 0.01). Parasitological analysis showed different infestation of the juvenile and adult chars by pathogenic helminths (Anisakis simplex, Eubothrium salvelini, Neoechinorhynchus salmonis) that influenced on their blood. Broad adaptive capabilities of S. malma hematopoiesis provided by wide morphological and functional diversity of blood cells are concluded that maintains homeostasis of the species. Blood indices for char from the Raduga River can be used to form reference intervals for monitoring physiological state of this species.

Monitoring of large B-cell lymphoma and T-zone lymphoma in a dog via flow cytometry.

A 12-y-old, castrated male Pomeranian dog was presented because of mandibular lymph node (LN) enlargement. Physical examination and a complete blood count revealed generalized lymphadenopathy and moderate lymphocytosis. Fine-needle aspirate cytology revealed expansion of medium lymphocytes in the right mandibular LN and expansion of large lymphocytes in the left popliteal LN. Flow cytometry identified 2 aberrant lymphocyte populations in both LNs, namely a CD5+CD45- T-cell population, and a large CD21+ B-cell population. Flow cytometry of the peripheral blood revealed an identical population of aberrant CD45- T cells. The patient was diagnosed with concurrent T-zone lymphoma and leukemia, and B-cell lymphoma. Multi-agent chemotherapy was instituted, and serial clinical and flow cytometric analysis revealed complete remission of the neoplastic B cells, but persistence of the neoplastic T cells and persistent lymphadenopathy. This case affirms the diagnostic value of flow cytometry and reveals a unique limitation of the RECIST criteria.

Clinicopathological findings of systemic Epstein-Barr virus-positive T-lymphoproliferative diseases in younger and older adults

BackgroundSystemic Epstein-Barr virus+ T-cell lymphoma (sEBV+ TCL) occurs in childhood and young adults, and is exceptionally rare in older adults.MethodsWe investigated clinicopathological features in 16 patients of various ages with systemic EBV+ CD8+ T-lymphoproliferative diseases.ResultsEight younger patients and four of eight older adults had sEBV+ CD8+ TCL, with invasion by medium-sized to/or large atypical lymphocytes primarily in bone marrow and lymph nodes, hemophagocytic lymphohistiocytosis (HLH), and progressive clinicopathological course. A further two patients demonstrated EBV+ node-based CD8+ large TCL without HLH, while the remaining two had the systemic form of chronic active EBV infection (sCAEBV) with CD8+ small lymphocytes. Past history of sCAEBV-like lesions was observed in one sEBV+ TCL patient (8.3%). Immunohistologically, in 12 sEBV+ TCL patients, atypical lymphocytes were positive for phosphate signal transducer and activator of transcription 3 (66.7%), CMYC (83.3%), and p53 (75%). Strong reactions of programmed cell death-ligand (PD-L)1+ tumor or non-neoplastic cells were detected in nine sEBV+ TCL patients (75%). Clonal peaks of the T-cell receptor (TCR) γ gene were detected in eight sEBV+ TCL patients by polymerase chain reaction. Four younger patients in sEBV+ TCL (33.3%) are in remission with chemotherapies including etoposide, and three of the four underwent allogeneic stem cell transplantation (SCT).ConclusionsEBV+ CD8+ TCL was observed in younger and older adults with less history of sCAEBV. HLH, tumor cell atypia, immunohistological findings, and progressive clinical course were characteristic of sEBV+ CD8+ TCL. Prompt chemotherapy and SCT induced tumor regression in sEBV+ CD8+ TCL patients.

Влияние сукцинатсодержащего препарата на лейкоцитарный состав крови крыс в покое и при плавании с грузом в тесте «до отказа»

This paper studied the dynamics of the blood leukocyte composition in Wistar male rats at rest and when swimming with a weight before and after being administered a succinate-containing drug (Suc, meso-2,3-dimercaptosuccinic acid). Two control groups of animals kept in standard vivarium conditions were selected: those that received Suc 12 hours before the examination (VivC+Suc) and those that did not receive it (VivC). Similarly, two groups of animals were formed that were swimming with a load (4 % of their body weight) to exhaustion: those that received the drug 12 hours before the test (Swim4%+Suc) and those that did not receive it (Swim4%). Unidirectional shifts characterizing the morphofunctional state of white blood cells under experimental conditions were detected. In VivC+Suc animals, compared to VivC, we found an increase in the number of leukocytes due to the growing number of eosinophils and monocytes, large lymphocytes and microlymphocytes with a relative decrease in the number of small lymphocytes and an unchanged level of granulocytes with a tendency towards a decrease in the number of stab neutrophils. In the Swim4%+Suc group, compared to the Swim4%, changes in the number of white blood cells and their subpopulation composition manifested themselves in a similar way to the redistribution of immunocytes identified in the control groups. The increase in the neutrophilto-lymphocyte ratio in the Swim4%+Suc group as an indicator of stress tolerance corresponded to the increase in the swimming time of rats by the factor of 2.8. The differences in the blood cell composition between the VivC+Suc and VivC groups are viewed as the influence of Suc on the body’s preparation to the fulfilment of the protective function under physical load, since the distribution pattern of the blood leukocyte composition in rats from the VivC+Suc group is very similar to that of the Swim4%+Suc group. The practical significance of this study is associated with the search for new biologically active substances that optimally influence the immune system of animals under increased load. New data on the mechanism of blood redistribution in animals under the action of Suc before the swimming test can be used to study the manifestation patterns of acute adaptation effect. For citation: Rubtsova L.Yu., Mongalev N.P., Vakhnina N.A., Shadrina V.D., Chupakhin O.N., Boyko E.R. Effect of a Succinate-Containing Drug on the Blood Leukocyte Composition in Rats at Rest and During a Weight-Loaded Forced Swimming Test. Journal of Medical and Biological Research, 2021, vol. 9, no. 2, pp. 182–191. DOI: 10.37482/2687-1491-Z056

Hematological and cytochemical characteristics of peripheral blood cells in the argus snakehead (Ophiocephalus argus Cantor).

BackgroundThe argus snakehead (Ophiocephalus argus Cantor) is a highly nutritious, freshwater, cultured bony fish with a high economic value. The health of the fish is closely related to its blood cells, which are critical for oxygen transport, natural defense, and immunity. We investigated the morphometry, microstructure, and cytochemical characteristics of the peripheral blood cells of O. argus. Our results may provide the basic reference values needed to monitor the health of this fish for large-scale cultivation.MethodsThe number of blood cells in O. argus were counted on a hemocytometer and their size was measured using a micrometer under light microscope. The morphology and classification of the blood cells were studied using Wright’s staining and the cytochemical characteristics were studied using seven chemical stains including peroxidase (POX), Sudan black B (SBB), periodic acid-Schiff (PAS), acid phosphatase (ACP), alkaline phosphatase (ALP), chloroacetic acid AS-D naphthol esterase (AS-D), and α-naphthol acetate esterase (α-NAE).ResultsThe peripheral blood cells in O. argus can be classified as erythrocytes, leukocytes, and thrombocytes; of which, females had 2.9597 million/mm3, 88,400/mm3, and 43,600/mm3, respectively, and males had 3.0105 million/mm3, 105,500/mm3, and 34,000/mm3, respectively. Leukocytes consisted of neutrophils, monocytes, large lymphocytes, and small lymphocytes. Eosinophils and basophils were not found. Monocytes were the most numerous leukocytes identified, followed by neutrophils and small lymphocytes, while large lymphocytes were the least frequently identified. Cytochemical staining showed that erythrocytes were only positive for PAS staining. Neutrophils were strongly positive for POX, SBB, and ACP, and positive for all the other cytochemical stains. Monocytes were positive for PAS and α-NAE and were weakly positive for ACP and AS-D staining. Large lymphocytes were positive for PAS and were weakly positive for ALP, AS-D, and α-NAE staining. Small lymphocytes were positive for PAS and weakly positive for AS-D and α-NAE staining. Thrombocytes were positive for PAS and were weakly positive for ACP and AS-D, but negative for the remaining cytochemical stains. The morphology of peripheral blood cells in O. argus was generally similar to that of other fish species, while the cytochemical staining patterns showed clear species specificity.

Composite lymphoma of T-cell rich, histiocyte-rich diffuse large B-cell lymphoma and nodular lymphocyte predominant Hodgkin lymphoma: a case report

BackgroundComposite lymphoma is a rare entity where two or more distinct subtypes of lymphoma coexist within a single organ or tissue.Case presentationWe report a new case of a 67-year-old Caucasian male patient, who presented with fatigue, weakness, weight loss, and polyuria. He also had epigastric and left lumbar pain, enlarged spleen, and enlarged left axillary lymph node on examination, with no relevant medical or familial history.A biopsy from the node showed an appearance of T-cell rich, histiocyte-rich diffuse large B-cell lymphoma and nodular lymphocyte predominant Hodgkin lymphoma.The patient was initially treated with adriamycin (doxorubicin), bleomycin, vinblastine, dacarbazine chemotherapy regimen, then switched to rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone regimen.During the therapy, some regression was noticed, especially in the size of the splenic enlargement; however, the patient died 2 months after completing the regimen.ConclusionComposite lymphomas should continue to be studied. Also, treatment is still debatable in type, efficacy, and outcomes.

Canine Multicentric Large B Cell Lymphoma with Increased Mott Cells Diagnosed by Flow Cytometry

A 12-year-old dog was referred due to multiple superficial lymphadenopathy. On cytology, each lymph node showed different cell populations where some of them consisted of intermediate to large lymphocytes with frequent Mott cells. Presence of Mott cells along with immature lymphocytes made the cytological diagnosis challenging, and therefore, supplementary diagnostic tests including PCR for Antigen Receptor Rearrangement (PARR) assay and flow cytometry were performed. This case report illustrates the value of flow cytometry in the diagnosis of lymphadenopathy with ambiguous cytologic findings.

Second Branchial Cleft Cyst: A Case Report

Objective: Branchial anomalies are congenital pathologies that are seen in the lateral region of the neck and are generally benign. The branchial clefts develop in the 2nd-7th weeks of fetal life as embryonal development. The branchial anomalies are caused by non-disappearance, abnormal development and, incomplete emerger of the branchial clefts and pockets during embryonal development. The branchial anomalies are generally seen as the cyst. The most common cyst was the second branchial cleft cyst with 95%. Their diameter is usually a few centimeters.&#x0D; Case: A 37-year old male patient was admitted to the hospital because of a swelling on the left side of his neck for four years. Physical examination revealed a mobile cystic mass in level 2 at the upper left jugular region of the neck. The cystic mass and the surrounding lymph nodes were excised and sent to the pathology laboratory. A cystic mass, approximately 5x4x3.5 cm in size, containing cystic areas was observed macroscopically. Microscopically, the cyst was lined with squamous epithelium and contained large lymphocyte groups in the subepithelial area. The case was reported as the branchial cleft cyst.&#x0D; Conclusion: Branchial cleft cysts should be excised before reaching large sizes, as they may cause pain and pressure on the surrounding tissue. In addition, it should be kept in mind that malignancy may develop from branchial cleft cysts, although rare.

Use of immune repertoire sequencing to resolve discordant microscopic and immunochemical findings in a case of T cell-rich large B cell lymphoma in a young dog

BackgroundLymphocytic neoplasms with frequent reactive lymphocytes are uncommonly reported in dogs, and can pose a diagnostic challenge. Different diagnostic modalities such as cytology, flow cytometry, histopathology, immunohistochemistry, and clonality testing, are sometimes required for a diagnosis. This report illustrates the value of using a multi-modal diagnostic approach to decipher a complex lymphocytic tumor, and introduces immune repertoire sequencing as a diagnostic adjunct.Case presentationA 10-month-old Great Dane was referred for marked ascites. Cytologic analysis of abdominal fluid and hepatic aspirates revealed a mixed lymphocyte population including numerous large lymphocytes, yielding a diagnosis of lymphoma. Flow cytometrically, abdominal fluid lymphocytes were highly positive for CD4, CD5, CD18, CD45, and MHC II, consistent with T cell lymphoma. Due to a rapidly deteriorating clinical condition, the dog was euthanized. Post mortem histologic evaluation showed effacement of the liver by aggregates of B cells surrounded by T cells, suggestive of hepatic T cell-rich large B cell lymphoma. Immune repertoire sequencing confirmed the presence of clonal B cells in the liver but not the abdominal fluid, whereas reactive T cells with shared, polyclonal immune repertoires were found in both locations.ConclusionsT cell-rich large B cell lymphoma is a rare neoplasm in dogs that may be challenging to diagnose and classify due to mixed lymphocyte populations. In this case, the results of histopathology, immunohistochemistry and immune repertoire sequencing were most consistent with a hepatic B cell neoplasm and reactive T cells exfoliating into the abdominal fluid. Immune repertoire sequencing was helpful in delineating neoplastic from reactive lymphocytes and characterizing repertoire overlap in both compartments. The potential pitfalls of equating atypical cytomorphology and monotypic marker expression in neoplasia are highlighted.