Assessment of alendronate and dietary treatment in the management of feline idiopathic ionised hypercalcaemia and ionised hypercalcaemia associated with chronic kidney disease: 29 cases (2016-2022).

OBJECTIVES: This retrospective cohort multicentre study aimed to characterise the signalment, clinicopathologic data, complications and the association between treatment and outcome (the reduction in ionised calcium concentration) in cats with idiopathic or chronic kidney disease-associated ionised hypercalcaemia managed with alendronate and standard treatment or standard treatment alone, and to compare the outcome between the two groups. MATERIALS AND METHODS: Medical records for cats diagnosed with idiopathic or chronic kidney disease-associated ionised hypercalcaemia were retrospectively reviewed. Cats treated with alendronate and standard treatment were assigned to the alendronate group and cats treated with standard treatment alone were assigned to the control group. The standard treatment was defined as dietary modification and/or monitoring of ionised calcium concentrations and management of complications secondary to hypercalcaemia. The follow-up period was selected as 6 months. RESULTS: Twenty-nine cats were enrolled in the study. Nine cats were included in the control group and 20 cats in the alendronate group. A significant reduction in serum ionised calcium was observed in both groups between the baseline and the follow-up visit; however, this reduction did not differ significantly between both groups (the mean change in the ionised calcium concentration in alendronate and control group was -0.18 and -0.17, respectively). Suspected bisphosphate-related osteonecrosis of the jaw was reported in one cat receiving alendronate. CLINICAL SIGNIFICANCE: In this study, similar reduction in serum ionised calcium concentration was observed in cats with ionised hypercalcaemia treated with diet alone and in cats treated with diet and alendronate. These results should be interpreted with caution, as the study was underpowered for meaningful statistical comparison. Cats receiving alendronate should be monitored for the development of adverse reactions, including osteonecrosis of the jaw.

Weight loss outcomes are generally worse for dogs and cats with class II obesity, defined as > 40% overweight.

In pet dogs and cats, adiposity is most-often estimated clinically using a 9-category body condition score (BCS), with BCS 9 equating to ~ 40% overweight. Animals that are more overweight (> 40%) are seen in clinical practice but are not appropriately depicted by descriptions in the existing categories. To determine whether being > 40% overweight has clinical relevance, this study aimed to compare the outcomes of weight management in animals that were > 40% overweight with those < 40% overweight. Records of dogs and cats attending a specialist obesity care clinic, where adiposity is determined using dual-energy X-ray absorptiometry (DXA), were reviewed. Animals were assigned to two classes (class I ≤ 40% overweight: 118/398 [40%] dogs and 68/116 [59%] cats; class II, > 40% overweight: 180/398 [60%] dogs and 48/116 [41%] cats) based on DXA results, and weight loss outcomes were compared. Fewer class II dogs obesity completed weight management than class I dogs (P < 0.001), rate of weight loss was also slower (P = 0.012) and lean tissue loss greater (P < 0.001). Compared with class I, cats with class II obesity lost more weight (P = 0.048) albeit over a longer period (P = 0.043) leading to greater lean tissue loss (P = 0.004). Approximately half the pets presenting to a specialist clinic were have class II obesity (> 40% overweight), and some weight loss outcomes are worse for these animals.

An in vivo genome-wide CRISPR screen identifies the RNA-binding protein Staufen2 as a key regulator of myeloid leukemia.

Aggressive myeloid leukemias such as blast crisis chronic myeloid leukemia and acute myeloid leukemia remain highly lethal. Here we report a genome-wide in vivo CRISPR screen to identify new dependencies in this disease. Among these, RNA-binding proteins (RBPs) in general, and the double-stranded RBP Staufen2 (Stau2) in particular, emerged as critical regulators of myeloid leukemia. In a newly developed knockout mouse, loss of Stau2 led to a profound decrease in leukemia growth and improved survival in mouse models of the disease. Further, Stau2 was required for growth of primary human blast crisis chronic myeloid leukemia and acute myeloid leukemia. Finally, integrated analysis of CRISPR, eCLIP and RNA-sequencing identified Stau2 as a regulator of chromatin-binding factors, driving global alterations in histone methylation. Collectively, these data show that in vivo CRISPR screening is an effective tool for defining new regulators of myeloid leukemia progression and identify the double-stranded RBP Stau2 as a critical dependency of myeloid malignancies.

Comparison of Different Small Clinical Hematology Laboratory Configurations With Focus on Remote Smear Imaging.

CONTEXT.­: Stand-alone clinical sites (eg, infusion centers) are becoming increasingly common. These sites require timely hematology analysis. Here we compare performance and costs of currently available analysis configurations with special focus on a proposed alternative using a minimal hematology analyzer plus a digital imaging device, allowing for remote oversight and interpretation. OBJECTIVES.­: To determine whether low-volume laboratories might realize savings while gaining function by substituting commonly used configurations with a proposed alternative. DESIGN.­: To evaluate the performance of the proposed alternative configuration, blood counts with automated differentials produced by a Sysmex XE5000 (complete blood count reference method) were compared with cell counts from the Sysmex pocH-100i, CellaVision DM96 preclassified differentials, and DM96 reclassified differentials (differential reference method) by using standard regression analyses, 95% CIs, and truth tables. Financial cost modeling used staffing practices, test volumes, and smear production rates observed at remote clinics performing on-site hematology analysis within the University of California at San Diego Health system. RESULTS.­: Differential blood count parameters showed excellent correlation between the XE5000 and preclassification DM96 with R2 > 0.95. For blasts/abnormal cells, immature granulocytes, and nucleated red blood cells, the DM96 showed higher sensitivity and similar specificity to the XE5000. Cost modeling revealed that decreased personnel costs through remote monitoring of results facilitated by the DM96 would lead to lower operational costs relative to more conventional analysis configurations. CONCLUSIONS.­: A digital imaging instrument with an inexpensive hematology analyzer provides similar information to a complex hematology analyzer and allows remote review of the blood smear findings by experts, leading to significant cost savings.

Impact of Integrating Rumke Statistics to Assist with Choosing Between Automated Hematology Analyzer Differentials vs Manual Differentials.

BACKGROUND: To optimize precision of nucleated blood cell counting, the clinical laboratory scientist should post the automated differential rather than the manual differential if the former is within the 95% CI of the latter, as determined by the "Rumke statistic." The objective of this study was to determine the potential impact of real-time, computer-assisted use of Rumke statistics for more judicious use of the automated vs digitally imaged, manual differential. METHODS: Complete blood counts with automated differentials produced by a XE5000™ hematology analyzer (Sysmex) were compared with both the DM96 (CellaVision™ AB) preclassification differentials and the posted reclassified manual differentials, using the Rumke 95% CIs as calculated using the Clopper-Pearson method. RESULTS: A total of 44.7% of manual differentials had no statistical or clinical justification over the automated differential. In addition, 31.1% of manual differentials had statistical discrepancies between the instrument absolute neutrophil count (IANC) and manual absolute neutrophil count (ANC). Nineteen of these IANC/manual ANC discrepant samples had ANCs below 1500/µL, a decision level for cancer treatment. Holding the IANC when it is less than 2000/µL until after manual smear review would have prevented the posting of any IANC vs manual ANC discrepant results at the 1500/µL ANC decision threshold. CONCLUSIONS: A real-time operator alert concerning the statistical identity of imaging device differentials vs automated differentials could have reduced manual differentials by nearly 45%. Not posting unnecessary manual differentials for the cases with IANC/manual ANC discrepancies would have likely reduced clinical error/confusion.

A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study.

In chronic lymphocytic leukemia (CLL) the level of minimal residual disease (MRD) after therapy is an independent predictor of outcome. Given the increasing number of new agents being explored for CLL therapy, using MRD as a surrogate could greatly reduce the time necessary to assess their efficacy. In this European Research Initiative on CLL (ERIC) project we have identified and validated a flow-cytometric approach to reliably quantitate CLL cells to the level of 0.0010% (10(-5)). The assay comprises a core panel of six markers (i.e. CD19, CD20, CD5, CD43, CD79b and CD81) with a component specification independent of instrument and reagents, which can be locally re-validated using normal peripheral blood. This method is directly comparable to previous ERIC-designed assays and also provides a backbone for investigation of new markers. A parallel analysis of high-throughput sequencing using the ClonoSEQ assay showed good concordance with flow cytometry results at the 0.010% (10(-4)) level, the MRD threshold defined in the 2008 International Workshop on CLL guidelines, but it also provides good linearity to a detection limit of 1 in a million (10(-6)). The combination of both technologies would permit a highly sensitive approach to MRD detection while providing a reproducible and broadly accessible method to quantify residual disease and optimize treatment in CLL.

Successful treatment of both double minute of C-MYC and BCL-2 rearrangement containing large B-cell lymphoma with subsequent unfortunate development of therapy-related acute myeloid leukemia with t(3;3)(q26.2;q21).

Double minute chromosomes (DMs), although relatively frequently encountered in solid tumors, are rare in hematologic neoplasms such as acute myeloid leukemia (AML), and even rarer in lymphoid neoplasms. t(3;3)(q26.2;q21) is a very rare genetic alteration observed in myeloid neoplasm. Herein we report an interesting and unique case of concomitant C-MYC DMs and t(14;18)-containing large B-cell lymphoma, which was successfully treated with R-hyper-CVAD; unfortunately, the patient has developed a therapy-related AML (t-AML) 2 years since the start of his lymphoma treatment. His t-AML contains both t(3;3)(q26.2;q21) and monosomy 7, and the patient died of AML 10 months after the initial diagnosis of t-AML despite clinical remission. To the best of our knowledge, this is the first reported case of C-MYC DM-containing de novo large B-cell lymphoma, which was successfully treated with complete remission, but unfortunately died of t-AML harboring t(3;3)(q21;q26).

Lis1 regulates asymmetric division in hematopoietic stem cells and in leukemia.

Cell fate can be controlled through asymmetric division and segregation of protein determinants, but the regulation of this process in the hematopoietic system is poorly understood. Here we show that the dynein-binding protein Lis1 is critically required for hematopoietic stem cell function and leukemogenesis. Conditional deletion of Lis1 (also known as Pafah1b1) in the hematopoietic system led to a severe bloodless phenotype, depletion of the stem cell pool and embryonic lethality. Further, real-time imaging revealed that loss of Lis1 caused defects in spindle positioning and inheritance of cell fate determinants, triggering accelerated differentiation. Finally, deletion of Lis1 blocked the propagation of myeloid leukemia and led to a marked improvement in survival, suggesting that Lis1 is also required for oncogenic growth. These data identify a key role for Lis1 in hematopoietic stem cells and mark its directed control of asymmetric division as a critical regulator of normal and malignant hematopoietic development.

Myeloid neoplasm with t(3;8)(q26;q24): report of six cases and review of the literature.

Balanced translocation between chromosomes 3q26 and 8q24 is a very rare event. Here we report six patients with t(3;8)(q26;q24) either as a sole or as a part of genetic abnormalities. Five of the six patients were men with ages ranging from 41 to 84 years old. One patient had a long history of granulocyte colony stimulating factor (G-CSF) treatment. Three of the patients were initially diagnosed with acute myeloid leukemia, two with myelodysplastic syndrome and one with chronic myelogenous leukemia with blast crisis. The peripheral blood in all patients showed severe to moderate anemia; one had absolute neutropenia, one with neutrophilia; four had thrombocytopenia, two with thrombocytosis. The bone marrows from all patients showed dysmegakaryopoiesis with additional erythroid (three patients) and granulocytic (two patients) dysplasia. Cytogenetics revealed t(3;8)(q26;q24) as the sole abnormality in three patients. The majority of patients (4/6) had a poor clinical course, with an average survival of 10 months.

A rare and unique case of aggressive IgE-λ plasma cell myeloma in a 28-year-old woman presented initially as an orbital mass.

A 28-year-old African-American woman presented with new onset of left exophthalmos and diplopia. Computed tomography of the head showed a solitary mass in the left orbit. Excisional biopsy revealed a diffuse infiltrate composed of exclusively λ-restricted monotypic plasma cells based on morphology and immunohistochemistry, consistent with a plasma cell neoplasia. A subsequent staging bone marrow biopsy showed involvement of the bone marrow by λ-restricted monotypic plasma cells, consistent with a plasma cell myeloma. Serum protein electrophoresis and immunofixation studies on the peripheral blood showed a monoclonal band of IgE-λ; thus, an IgE-λ plasma cell myeloma was established. Additional clinical and radiologic workups showed multiple lytic bone lesions, diffuse lymphadenopathy, a pelvic mass, multiple mesenteric soft tissue nodules, and multiple pulmonary nodules, although none of the aforementioned sites was biopsied. The patient was treated with a combination of multiple chemotherapeutic agents and localized radiation due to the aggressive nature of the disease. To the best of our knowledge, this is the first case of an IgE plasma cell myeloma in a patient under the age of 30 years old presenting as a mass in an extramedullary site.

ROR1 is expressed on hematogones (non-neoplastic human B-lymphocyte precursors) and a minority of precursor-B acute lymphoblastic leukemia.

ROR1 is a receptor tyrosine kinase expressed during embryogenesis, on chronic lymphocytic leukemia (CLL) and in other malignancies. Hematogones (non-neoplastic B-lymphocyte precursors) express surface ROR1 at an intermediate stage of maturation that lacks CD34 or TdT. The neoplastic counterpart to hematogones is precursor-B acute lymphoblastic leukemia (B-ALL), but less than 10% of B-ALL express surface ROR1, and these ROR1+ B-ALL cases have an unusually high frequency of lacking CD34 and/or having t(1;19), a chromosomal translocation that defines a specific subtype of B-ALL.

t(4;22)(q12;q11.2) involving presumptive platelet-derived growth factor receptor A and break cluster region in a patient with mixed phenotype acute leukemia.

The patient is a 45-year-old woman with a history of breast cancer who had been treated 1 year ago with radiation and chemotherapy. Flow cytometric analysis of bone marrow aspirate revealed 81% blasts positive for CD4, CD11c (partial), CD13, CD19 (partial), cytoplasmic CD22, CD34, CD36, CD45, cytoplasmic CD79a, CD117 (partial), HLA-DR, and terminal deoxynucleotide transferase, consistent with a mixed phenotype acute leukemia (B/myeloid lineage). Conventional karyotypic analysis revealed a t(4;22)(q12;q11.2) in 12 of 13 cells analyzed. Fluorescence in situ hybridization analysis using a dual-color, dual-fusion break cluster region/ABL probe set showed no break cluster region/ABL translocation but an extra break cluster region signal in 85% (170/200) of cells, consistent with a translocation involving the break cluster region gene at 22q11.2. A FIP1L1/CHIC2/platelet-derived growth factor receptor α deletion/fusion probe showed signal separation in 96.5% (193/200) of interphase nuclei. Reverse transcriptase-polymerase chain reaction using sense break cluster region primers and an antisense platelet-derived growth factor receptor α primer resulted in a product of approximately 590 base pairs, consistent with the presence of a break cluster region/platelet-derived growth factor receptor α fusion gene. Because of the presumptive platelet-derived growth factor receptor α translocation and its sensitivity to tyrosine-kinase inhibitor, the patient was treated with imatinib mesylate, cytarabine, and idarubicin as induction and maintenance therapy; and she has remained free of disease for 5 months since the initial diagnosis.

Monoclonal antibodies in the treatment of hematologic malignancy.

Methods to generate monoclonal antibodies to antigens of neoplastic cells have revolutionized our understanding of cancer cell growth and differentiation, diagnosis, and treatment. Monoclonal antibodies derived by immunizing animals (mostly mice) with mammalian cells or molecules have been critical reagents for the discovery and characterization of many key molecules involved in the behavior of neoplastic cells. Now, over 30 years later, monoclonal antibodies are widely used in the differential diagnosis of cancer and are key elements in the treatment of many forms of cancer. This review will focus on the roles that monoclonal antibodies play in the treatment of hematological malignancies. In particular, we will focus on acute myeloid leukemia and mature B-cell neoplasms.

Systemic mastocytosis in association with chronic lymphocytic leukemia and plasma cell myeloma.

Systemic mastocytosis with associated clonal haematological non-mast cell lineage disease (SM-AHNMD) is a heterogeneous group of mast cell disorders with different clinical, pathologic and underlying molecular characteristics. While myelomonocytic/myeloid neoplasia overwhelmingly predominates the AHNMD component, lymphoproliferative disorders rarely occur as an AHNMD component of SM-AHNMD. Here we report two cases of SM-AHNMD, in which the AHNMD component is chronic lymphocytic leukemia in one case, and concurrent chronic lymphocytic leukemia as well as plasma cell myeloma in another case. To the best of our knowledge, this is the first case report of SM-AHNMD with chronic lymphocytic leukemia and plasma cell dyscrasia simultaneously.

Characteristics of lymphocyte subsets in a normal Afro-Caribbean population and the implications in HIV management.

The Caribbean relies on normal lymphocyte subsets ranges established in other geographical locations and from different racial ethnic groups for the basis of the clinical management of HIV and AIDS with Highly Active Anti Retroviral Therapy (HAART). Normal ranges of these parameters have not been previously established in an Afro-Caribbean population. So we set out to determine how the normal lymphocyte subset ranges compare to those reported in other, races and geographical locations. A prospective study was done on 112 healthy Afro-Caribbean clients who attended the Blood Collection Unit, Queen Elizabeth Hospital, Barbados from July 15th to November 12th 2004. Analysis for lymphocyte subsets was done by flow cytometry, which allows simultaneous identification and enumeration of total T Helper, cytotoxic T, natural killer and B lymphocyte cells. HIV-1, Hepatitis B and C, HTLV-1 and full blood count test were done as part of the normal screening routine of all blood donors. Absolute white blood cell counts and percentage lymphocytes for males and females were not significantly different, and the absolute and percentage of the T-Helper CD4 positive lymphocyte cells were not significantly higher in females than in males. Absolute Cytotoxic T cell CD8 positive lymphocyte cells were higher in males than in females. CD56 Natural Killer cells absolute and percentage counts were higher in males than females however CD19 B Lympocyte absolute and percentage counts were not different between the two sexes in this sample population. Compared to published normal ranges published by the WHO and CDC, there were no significantly differences observed in any of the lymphocyte subsets. These finding are very similar to what has been reported in previous studies. We conclude that WHO/CDC recommendations established for the treatment and monitoring of HIV/AIDS patients based on CD4 levels can be safely utilized in our population.

Quality control of the total lymphocyte count parameter obtained from routine haematology analyzers, and its relevance in HIV management.

Lylmphocyte subsets/CD4 T Helper cell enumeration in HIV care and treatment in resource constrained settings can be difficult to ascertain as a result of the lack of the necessary instrumentation, capacity and infrastructure. However. it is imperative to gain such information for patient monitoring in HIV. The Total Lymphocyte Count (TLC) is useful as a surrogate marker for CD4 count as recommended by the World Health Organisation (WHO) and to calculate CD4% for pacdiatric use. This study therefore sets out to determine and compare the accuracy of the total lymphocyte counts obtained from three haematology analysers designated A. B and C. that are in regular use for routine haemnatological parameters at the main referral hospital in Barbados. West Indies. The TLC of 263 HIV treatment naive individuals attending the HIV Reference Unit in Barbados were enumnerated on the three haematology analysers. The lymphosumn (Sum of lymphocyte subsets: T-helper cell. T-cytotoxic cells. B lymphocytes and Natural killer cells) should be equal to the TLC. and is derived by immunophenotypic analysis on a 4-colour flowcytometer. Machine C had the highest positive correlation between the TLC and the lymphosumn with and R' of 0.9031 compared to machine A with an R values of 0.7119 and Machine B with R(2) values of 0.637. These results show that there can be dramatic inaccuracies when using routine haematology analysers for both routine use. as a surrogate marker of CD4 or for derivation of CD4% in HIV management. It further demonstrates that all haematology analyzers require some form of Quality control. The possible lack of accuracy of the TLC by haematology analysers should be taken into consideration when following the recommendations of the WHO in resource poor settings or using it as a denominator for calculating CD4%.

FLAG chemotherapy followed by allogeneic stem cell transplant using nonmyeloablative conditioning induces regression of myelofibrosis with myeloid metaplasia.

A 38-year-old woman with agnogenic myeloid metaplasia complicated by the poor prognostic factors of severe osteosclerosis, prominent hepatosplenomegaly, and profound anemia was treated with FLAG chemotherapy to decrease her organomegaly before undergoing a nonmyeloablative allogeneic stem cell transplant from a matched-sibling donor. The patient's pre- and post transplant course were complicated by an autoimmune disorder and her post transplant course was complicated by severe hepatic and gastrointestinal GVHD. A technetium-99m sulfur colloid scan 4 months post transplant and bone marrow studies 8 months post transplant demonstrated intramedullary hematopoiesis, complete resolution of marrow fibrosis, and partial resolution of osteosclerosis.