Assessment of Y chromosome copy number alterations in non-neoplastic and neoplastic leukocytes of male dogs.

The loss of the Y chromosome (ChrY), also known as LOY, is a common genetic alteration observed in men. It occurs in non-neoplastic cells as an age-related change as well as in neoplastic cells of various cancer types. While well-documented in humans, LOY has not been extensively studied in non-human mammals. In this study, we developed simple digital PCR-based assays to assess the copy number of ChrY relative to the X chromosome (ChrX) and chromosome 8 (Chr8) to evaluate ChrY numerical alterations in male canine DNA specimens. Using these assays, we analyzed non-neoplastic leukocytes from 162 male dogs without hematopoietic neoplasia to investigate the occurrence of age-related LOY in non-neoplastic leukocytes. Additionally, we examined 101 tumor DNA specimens obtained from male dogs diagnosed with various types of lymphoma and leukemia to determine whether copy number alterations of the ChrY occur in canine hematopoietic cancers. Analysis of the 162 non-neoplastic leukocyte DNA specimens from male dogs of varying ages revealed a consistent ∼1:1 ChrY:ChrX ratio. This suggests that age-related LOY in non-neoplastic leukocytes is rare or absent in dogs. Conversely, a decreased or increased ChrY:ChrX ratio was detected in canine neoplastic leukocytes at varying frequencies across different canine hematopoietic malignancies (P = 0.01, Fisher's exact test). Notably, a higher incidence of LOY was observed in more aggressive cancer types. To determine if this relative LOY to ChrX was caused by changes in ChrY or ChrX, we further analyzed their relative copy numbers using Chr8 as a reference. Loss of ChrX relative to Chr8 was found in 21% (9/41) of B-cell lymphomas and 6% (1/18) of non-T-zone/high-grade T-cell lymphomas. In contrast, a subset (29%, 4/14) of T-cell chronic lymphocytic leukemia showed gain of ChrX relative to Chr8. Notably, no relative LOY to Chr8 was detected indolent hematopoietic cancers such as T-zone lymphoma (0/9) and chronic lymphocytic leukemia of B-cell (0/11) and T-cell origins (0/14). However, relative LOY to Chr8 was present in more aggressive canine hematopoietic cancers, with incidences of 24% (10/41) in B-cell lymphoma, 44% (8/18) in non-T-zone/high-grade T-cell lymphoma, and 75% (6/8) in acute leukemia. This study highlights both similarities and differences in LOY between human and canine non-neoplastic and neoplastic leukocytes. It underscores the need for further research into the role of ChrY in canine health and disease, as well as the significance of LOY across various species.

Progressive and regressive infection with feline leukemia virus (FeLV) in cats in southern Brazil: Prevalence, risk factors associated, clinical and hematologic alterations.

Enzyme-linked immunosorbent assay (ELISA) for viral antigen is commonly used for the diagnosis of progressive feline leukemia virus (FeLV) infection but is not able to determine the true prevalence of infection when used as the sole test. Additional testing to detect proviral DNA will identify regressive (antigen negative) FeLV infections as well as progressive infections. Therefore, this study aimed to determine the prevalence of progressive and regressive FeLV infection, outcome-associated factors, and hematologic changes. A cross-sectional study was performed on 384 cats selected from routine hospital care. Blood samples were subjected to complete blood count, ELISA for FeLV antigen and FIV antibody, and nested PCR amplifying the U3- LTR region and gag gene, which are conserved in most exogenous FeLV. The prevalence of FeLV infection was 45.6% (CI95% 40.6-50.6%). The prevalence of progressive infection (FeLV+P) was 34.4% (CI95% 29.6-39.1%), that of regressive infection (FeLV+R) was 10.4% (CI95% 7.4-13.4%), for discordant but positive results 0.8% (CI95% 0.75-0.84%), for FeLV+P coinfected with FIV 2.6% (CI95% 1.2-4.0%), and FeLV+R coinfected with FIV 1.5% (CI95% 0.3-2.7%). Male cats were three times more likely to be in the FeLV+P group. Cats coinfected with FIV were 4.8 times more likely to belong to the FeLV+R group. In the FeLV+P group, the main clinical changes were lymphoma (38.5%), anemia (24.4%), leukemia (17.9%), concomitant infections (15.4%), and feline chronic gingivostomatitis - FCGS (3.8%). In the FeLV+R group, the main clinical signs were anemia (45.4%), leukemia (18.2%), concomitant infections (18.2%), lymphoma (9.1%), and FCGS (9.1%). Cats in the FeLV+P and FeLV+R groups showed mainly thrombocytopenia (56.6% and 38.2%), non-regenerative anemia (32.8% and 23.5%), and lymphopenia (33.6% and 20.6%). Hemoglobin concentration, packed cell volume (PCV), platelet count, lymphocytes, and eosinophils in the FeLV+P and FeLV+R groups had lower medians than the control group (FeLV/FIV-uninfected, healthy). Erythrocyte and eosinophil counts were statistically different among the three groups, with the medians of the FeLV+P and FeLV+R groups being lower than those of the control group. In addition, the median PCV and band neutrophil counts were higher in FeLV+P than in FeLV+R. Our results show a high prevalence of FeLV, different factors associated with the course of infection, and more frequent and severe hematologic changes in progressive infections compared with regressive infections.

Flow Cytometry in Veterinary Practice.

This article summarizes the current applications of flow cytometry in clinical veterinary medicine, which is largely restricted to the diagnosis of hematopoietic neoplasms (lymphomas and leukemias) of domestic dogs, cats, and horses. A brief background on the technique of flow cytometry and fundamentals of data interpretation are included. Major emphasis is placed on clinical indications for flow cytometry, principles of sample collection and submission, and awareness of diagnostic and prognostic utility. Expectations regarding both the benefits and limitations of flow cytometry in a clinical setting, and its complementary nature with other types of testing, are also reviewed.

Cytokine response as a biomarker for early diagnosis and outcome prediction of stem cell transplant recipients and acute leukemia patients with invasive aspergillosis.

We aimed to determine the role of serum cytokine expression in invasive aspergillosis (IA) diagnosis and outcome prediction in hematologic patients. In this multicenter study, serum cytokines (IL6, IL10, INF-gamma, IL12, IL4, TNF-alpha, IL17, and IL23) were prospectively recruited from all consecutive patients with hematologic malignances at IA diagnosis and compared to control patients matched by center, age, baseline disease, and therapeutic regimen. We included 36 patients with IA and 36 controls. Serum levels of IL6 and IL10 cytokines on day 0 were significantly increased in patients with IA when compared to controls (P = 0.001 and P = 0.025, respectively), even in those who were neutropenic. No differences were observed for the other cytokines. IL6 and IL10 predicted IA with an area under the ROC curve of 0.74 (95% CI 0.62-0.86) and 0.64 (95% CI 0.51-0.77), respectively. The best cutoff point in predicting IA was 20.85 pg/ml for IL6 (sensitivity 72.2%; specificity 77.8%; PPV 76.5% and NPV 73.7%), and 0.045 pg/ml for IL10 (sensitivity 62.9%; specificity 63.9%; PPV 62.9% and NPV 63.9%). IL6 levels were associated with increased mortality, with the best cutoff value being 65.59 pg/ml in mortality prediction. In conclusion, in addition to current tests in place, IL6 and IL10 levels-as measured in plasma-may help clinicians diagnose IA. High levels of IL6 at IA diagnosis are related with worse outcomes. LAY SUMMARY: We evaluated the role of serum cytokine expression in invasive aspergillosis (IA) diagnosis and outcome. Serum levels of IL6 and IL10 are increased in patients with IA compared to controls, and IL6 levels are associated with mortality.

A case of leukaemia cutis in a dog with T-cell chronic lymphocytic leukaemia.

Leukaemia cutis (LC) is the infiltration of neoplastic leukocytes into the skin, characterised by haemorrhagic papules, nodules, and plaques. LC has been reported in human leukaemia patients, but it is extremely rare in dogs. A 13-year-old spayed female Golden Retriever that was previously diagnosed with chronic lymphocytic leukaemia was managed with chlorambucil (20 mg/m2 orally, every 2 weeks) and prednisolone (2 mg/kg orally, every other day) for 8 months; however, immunosuppression was temporarily discontinued because of a bacterial urinary tract infection. Cutaneous signs, including multifocal ecchymosis and white plaques, appeared 1 month after cessation of chemotherapy. Histopathological examination revealed small- to intermediate-sized lymphocytes with mild atypia in a perivascular to interstitial pattern within the superficial dermis. The bands of atypical cells within the superficial dermis were strongly and extensively positive for CD3 on immunohistochemistry. Polymerase chain reaction analysis of the biopsied skin revealed clonal rearrangement of the T-cell receptor gamma locus gene. Given the evidence of clinical signs, peripheral immunophenotyping, histopathology, immunohistochemistry, and clonal gene arrangement, LC was diagnosed. The lesions disappeared when chemotherapy was restarted but were occasionally observed when chemotherapy was stopped. To the authors' best knowledge, this is the first case report of LC in a dog.

Benzyl Isothiocyanate, a Vegetable-Derived Compound, Induces Apoptosis via ROS Accumulation and DNA Damage in Canine Lymphoma and Leukemia Cells.

Treatment of neoplastic diseases in companion animals is one of the most important problems of modern veterinary medicine. Given the growing interest in substances of natural origin as potential anti-cancer drugs, our goal was to examine the effectiveness of benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, against canine lymphoma and leukemia. These are the one of the most common canine cancer types, and chemotherapy is the only treatment option. The study involved established cell lines originating from various hematopoietic malignancies: CLBL-1, GL-1, CLB70 and CNK-89, immortalized noncancerous cell lines: MDCK and NIH-3T3 and canine peripheral blood mononuclear cells (PBMCs). The cytotoxic activity of BITC, apoptosis induction, caspase activity and ROS generation were evaluated by flow cytometry. H2AX phosphorylation was assessed by western blot. The study showed that the compound was especially active against B lymphocyte-derived malignant cells. Their death resulted from caspase-dependent apoptosis. BITC induced ROS accumulation, and glutathione precursor N-acetyl-l-cysteine reversed the effect of the compound, thus proving the role of oxidative stress in BITC activity. In addition, exposure to the compound induced DNA damage in the tested cells. This is the first study that provides information on the activity of BITC in canine hematopoietic malignancies and suggests that the compound may be particularly useful in B-cell neoplasms treatment.

Malic enzyme 2 connects the Krebs cycle intermediate fumarate to mitochondrial biogenesis.

Mitochondria have an independent genome (mtDNA) and protein synthesis machinery that coordinately activate for mitochondrial generation. Here, we report that the Krebs cycle intermediate fumarate links metabolism to mitobiogenesis through binding to malic enzyme 2 (ME2). Mechanistically, fumarate binds ME2 with two complementary consequences. First, promoting the formation of ME2 dimers, which activate deoxyuridine 5'-triphosphate nucleotidohydrolase (DUT). DUT fosters thymidine generation and an increase of mtDNA. Second, fumarate-induced ME2 dimers abrogate ME2 monomer binding to mitochondrial ribosome protein L45, freeing it for mitoribosome assembly and mtDNA-encoded protein production. Methylation of the ME2-fumarate binding site by protein arginine methyltransferase-1 inhibits fumarate signaling to constrain mitobiogenesis. Notably, acute myeloid leukemia is highly dependent on mitochondrial function and is sensitive to targeting of the fumarate-ME2 axis. Therefore, mitobiogenesis can be manipulated in normal and malignant cells through ME2, an unanticipated governor of mitochondrial biomass production that senses nutrient availability through fumarate.

Primary bone marrow T-cell lymphoma in a Golden Retriever.

A 6.2-year-old 28-kg (61.7 lb) intact female Golden Retriever was referred due to persistent and multiple cytopenias noted on a routine CBC prior to a mature ovariohysterectomy procedure. The patient's physical examination was unremarkable, and staging of the thorax and abdomen identified no abnormalities. At the referral hospital, moderate hypercalcemia, borderline anemia, and neutropenia were noted. Assessment of bone marrow samples by cytology, histology, immunohistochemistry, and flow cytometry indicated a T-cell neoplasm. The patient was treated with a multi-agent chemotherapy protocol for 6 months, which induced remission. Nine months after diagnosis, she relapsed with recurrence of hypercalcemia, cytopenias, and clinical illness. Single-agent anthracycline (mitoxantrone) in combination with prednisone therapy was initiated for 3 months. Two months after completion, the patient relapsed again, and palliative therapy with prednisone was elected. The patient was euthanized 16 months after diagnosis due to progressive disease. Post-mortem histopathologic evaluation showed extensive replacement of bone marrow by neoplastic cells, and infiltrates in multiple organs. The neoplasm was diagnosed as lymphoma rather than leukemia due to the lack of abnormal circulating cells throughout the course of disease. The neoplasm was detected only in marrow at the time of initial diagnosis, and the marrow was the most extensively effaced organ at the time of death. Therefore, leukemia or stage V lymphoma was considered unlikely. In patients with a cytopenia and lack of neoplastic leukocytosis or solid tissue masses, primary bone marrow lymphoma should be considered among the differential diagnoses.

Development of Mast Cell and Eosinophil Hyperplasia and HLH/MAS-Like Disease in NSG-SGM3 Mice Receiving Human CD34+ Hematopoietic Stem Cells or Patient-Derived Leukemia Xenografts.

Immunocompromised mouse strains expressing human transgenes are being increasingly used in biomedical research. The genetic modifications in these mice cause various cellular responses, resulting in histologic features unique to each strain. The NSG-SGM3 mouse strain is similar to the commonly used NSG (NOD scid gamma) strain but expresses human transgenes encoding stem cell factor (also known as KIT ligand), granulocyte-macrophage colony-stimulating factor, and interleukin 3. This report describes 3 histopathologic features seen in these mice when they are unmanipulated or after transplantation with human CD34+ hematopoietic stem cells (HSCs), virally transduced hCD34+ HSCs, or a leukemia patient-derived xenograft. The first feature is mast cell hyperplasia: unmanipulated, naïve mice develop periductular pancreatic aggregates of murine mast cells, whereas mice given the aforementioned human cells develop a proliferative infiltrative interstitial pancreatic mast cell hyperplasia but with human mast cells. The second feature is the predisposition of NSG-SGM3 mice given these human cells to develop eosinophil hyperplasia. The third feature, secondary hemophagocytic lymphohistiocytosis/macrophage activation syndrome (HLH/MAS)-like disease, is the most pronounced in both its clinical and histopathologic presentations. As part of this disease, a small number of mice also have histiocytic infiltration of the brain and spinal cord with subsequent neurologic or vestibular signs. The presence of any of these features can confound accurate histopathologic interpretation; therefore, it is important to recognize them as strain characteristics and to differentiate them from what may be experimentally induced in the model being studied.

Leukemic histiocytic sarcoma in a captive common squirrel monkey (Saimiri sciureus) with Saimiriine Gammaherpesvirus 2 (Rhadinovirus), Saimiri sciureus lymphocryptovirus 2 (Lymphocryptovirus) and Squirrel monkey retrovirus (ß-Retrovirus) coinfection.

Hematopoietic neoplasia other than lymphoma and leukemia is uncommon among non-human primates. Herein, we provide the first evidence of occurrence of leukemic histiocytic sarcoma in a captive common squirrel monkey with Saimiriine Gammaherpesvirus 2 (Rhadinovirus), Saimiri sciureus lymphocryptovirus 2 (Lymphocryptovirus), and Squirrel monkey retrovirus (ß-Retrovirus) coinfection.

Bone Marrow Examination: Why, How, and What to Expect from the Pathologist.

This article describes the indications for sampling of bone marrow, the technical aspects of obtaining marrow core biopsies and aspirates, and the preparation of marrow smears. All aspects are illustrated with clinical cases. The information that can be expected from the pathologist's report of marrow samples is outlined, and the clinical features and prognosis of different types of leukemia are detailed.

Morphologic and Immunohistochemical Characterization of Spontaneous Lymphoma/Leukemia in NSG Mice.

The NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ strain (NOD scid gamma, NSG) is a severely immunodeficient inbred laboratory mouse used for preclinical studies because it is amenable to engraftment with human cells. Combining scid and Il2rgnull mutations results in severe immunodeficiency by impairing the maturation, survival, and functionality of interleukin 2-dependent immune cells, including T, B, and natural killer lymphocytes. While NSG mice are reportedly resistant to developing spontaneous lymphomas/leukemias, there are reports of hematopoietic cancers developing. In this study, we characterized the immunophenotype of spontaneous lymphoma/leukemia in 12 NSG mice (20 to 38 weeks old). The mice had a combination of grossly enlarged thymus, spleen, or lymph nodes and variable histologic involvement of the bone marrow and other tissues. All 12 lymphomas were diffusely CD3, TDT, and CD4 positive, and 11 of 12 were also positive for CD8, which together was consistent with precursor T-cell lymphoblastic lymphoma/leukemia (pre-T-LBL). A subset of NSG tissues from all mice and neoplastic lymphocytes from 8 of 12 cases had strong immunoreactivity for retroviral p30 core protein, suggesting an association with a viral infection. These data highlight that NSG mice may develop T-cell lymphoma at low frequency, necessitating the recognition of this spontaneously arising disease when interpreting studies.

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia (ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona) mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/µL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shorly after diagnosis.

Novel Noninvasive Diagnostics.

Molecular diagnostics have revolutionized human oncology to allow early detection, targeted therapy, monitoring throughout treatment, and evidence of recurrence. By identifying genetic signatures associated with cancers, liquid biopsy techniques have been developed to diagnose and monitor cancer in noninvasive or minimally invasive ways. These techniques offer new opportunities for improving cancer screening, diagnosis, and monitoring the impact of therapy on the patients over time. Liquid biopsy also drives drug development programs. Similar diagnostics hold promise for comparable results in the veterinary field. Several noninvasive/minimally invasive techniques have been described in veterinary medicine that could be referred to as liquid biopsy.

Oral lesions and retroviruses in shelter cats / Afecções orais e retroviroses em gatos de abrigo

Oral lesions are common problems in feline medicine worldwide, and may be associated with different causes, such as infectious agents. There are only a few studies reporting the chief oral diseases and the results for retrovirus tests in shelter cats in Brazil, especially in the South region. This study aimed to identify the main inflammatory oral lesions in shelter cats and verify the test results for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) infections. Forty-three felines from private shelters in the central region of Rio Grande do Sul state (RS) that presented clinically evident oral lesions, regardless of age, breed, sex, and neuter status, were used in this survey. Serological tests for FIV and FeLV were performed in all cats, and data regarding the rearing system were collected. Sixteen cats (37.2%) were reared in a free system, whereas 27 (62.8%) were kept under a restrict system. Of the 43 cats with oral lesions, 29 (67.44%) presented only one type of lesion, characterized as periodontitis (n=22, 51.16%), followed by gingivitis (n=6, 13.95%), and stomatitis (n=1, 2.32%). Concomitant stomatitis and periodontitis were found in the 14 remaining cats (100%). With respect to the test results for retrovirus infections, nine (20.93%) of the 43 felines were positive for FIV alone. Co-infection with both viruses was observed in seven cats (16.28%). No cat was seropositive for FeLV valone. None of the six cats that presented gingivitis was positive for FIV and FeLV; one cat with stomatitis was positive for FIV and FeLV; of the 22 cats with periodontitis, six (27.27%) were FIV positive and two (9.09%) were FIV/FeLV positive; and of the 14 cats that presented stomatitis and periodontitis, three (21.43%) were FIV positive and four (28.57%) were FIV/FeLV positive. As for diagnosis, 28 cats (65.1%) presented solely periodontal disease (PD), one cat (2.32%) had feline chronic gingivostomatitis (FCG) alone, and 14 (32.5%) had both PD and FCG. The results obtained show that the main oral lesions found in shelter cats in the central region of RS were gingivitis, stomatitis, and periodontitis. Periodontitis, in association or not with stomatitis, was the most frequently observed oral cavity lesion in FIV- and/or FeLV-positive cats. Other factors may contribute to installation of inflammatory oral diseases in shelter cats because most cats with oral cavity lesions tested negative for retrovirus infections.(AU)

As afecções orais são problemas comuns em medicina felina em diferentes locais do mundo e podem estar relacionadas a diferentes causas, como agentes infecciosos. Poucos estudos foram encontrados no Brasil sobre o levantamento das principais doenças orais e dos resultados de testes para retrovírus em gatos de abrigos, principalmente na região Sul. Diante disso, o objetivo deste artigo foi identificar as principais afecções orais inflamatórias em gatos de abrigos e verificar os resultados dos testes para o vírus da imunodeficiência felina (FIV) e o vírus da leucemia felina (FeLV). Foram incluídos 43 felinos provenientes de abrigos privados localizados na região central do Rio Grande do Sul (RS) que apresentavam lesões orais clinicamente evidentes, independente de idade, raça, gênero e estado reprodutivo. Em todos os gatos foram realizados testes sorológicos para FIV e FeLV e obtidas informações referentes ao sistema de criação. Em 16 gatos (37,2%), o sistema de criação era livre, enquanto em 27 (62,8%) era restrito. Dos 43 gatos com lesões orais, em 29 (67,44%) foi verificado somente um tipo de lesão, caracterizado como periodontite (n=22, 51,16%), seguido de gengivite (n=6, 13,95%) e estomatite (n=1, 2,32%). Lesões concomitantes de estomatite e periodontite foram encontradas nos 14 gatos (100%) restantes. Quanto aos resultados dos testes para retrovírus, nove (20,93%) dos 43 felinos testados, foram positivos somente para FIV. Em sete gatos (16,28%) foi observada coinfecção pelos dois vírus. Em nenhum gato foi observado soropositividade somente para FeLV. Dos seis gatos com gengivite, nenhum foi positivo para FIV e FeLV; um gato com estomatite foi positivo para FIV e FeLV; dos 22 gatos com periodontite, seis (27,27%) foram FIV positivos e dois (9,09%) FIV/FeLV positivos; e dos 14 com estomatite e periodontite, três (21,43%) foram FIV positivos e quatro (28,57%) FIV/FeLV positivos. Quanto ao diagnóstico, em 28 gatos (65,1%) foi observada somente doença periodontal (DP), em um (2,32%) somente gengivoestomatite crônica felina (GECF) e em 14 gatos (32,5%) DP e GECF. Diante dos resultados obtidos, pode-se concluir que as principais lesões orais encontradas em gatos de abrigos da região central do RS foram gengivite, estomatite e periodontite; a periodontite associada ou não a estomatite foi a lesão oral mais frequente nos gatos positivos para FIV e/ou FeLV. Acredita-se que outros fatores possam contribuir na instalação de doenças orais em gatos de abrigos, já que houve predomínio de gatos com resultados negativos nos testes para os retrovírus.(AU)

Implementation of total body photon irradiation as part of an institutional bone marrow transplant program for the treatment of canine lymphoma and leukemias.

A total body irradiation (TBI) protocol was developed to support a bone marrow transplant (BMT) program for the treatment of canine hematologic malignancies. The purpose of this prospective study is to describe implementation of the protocol and resultant dosimetry. Nongraphic manual treatment planning using 6 MV photons, isocentric delivery, 40 × 40 cm field size, wall-mounted lasers to verify positioning, a lucite beam spoiler (without use of bolus material), a dose rate of 8.75 cGy/min at patient isocenter, and a source-to-axis distance of 338 cm were used for TBI. A monitor unit calculation formula was derived using ion chamber measurements and a solid water phantom. Five thermoluminescent dosimeters (TLDs) were used at various anatomic locations in each of four cadaver dogs, to verify fidelity of the monitor unit formula prior to clinical implementation. In vivo dosimetric data were then collected with five TLDs at various anatomic locations in six patients treated with TBI. A total dose of 10 Gy divided into two 5 Gy fractions was delivered approximately 16 h apart, immediately followed by autologous stem cell transplant. The mean difference between prescribed and delivered doses ranged from 99% to 109% for various sites in cadavers, and from 83% to 121% in clinical patients. The mean total body dose in cadavers and clinical patients when whole body dose was estimated by averaging doses measured by variably placed TLDs ranged from 98% to 108% and 93% to 102% of the prescribed dose, respectively, which was considered acceptable. This protocol could be used for institutional implementation of TBI.

Plasma ammonia concentration after L-asparaginase therapy in 27 dogs with high-grade lymphoma or leukemia.

OBJECTIVES: To establish the occurrence of increased plasma ammonia concentration after L-asparaginase (L-asp) administration in dogs with high-grade lymphoma or leukemia; to identify risk factors for the development of hyperammonemia after L-asp administration; and to determine occurrence of adverse events related to hyperammonemia. DESIGN: Prospective case controlled study of sequentially enrolled dogs between May 2011 and March 2012. SETTING: A university veterinary teaching hospital. ANIMALS: Twenty-seven dogs with high-grade lymphoma or leukemia. INTERVENTIONS: All dogs received L-asp intramuscularly at a median dose of 400 IU/kg. MEASUREMENTS AND MAIN RESULTS: Plasma ammonia concentrations were measured at baseline, 16 hours, and 48 hours after L-asp therapy. Clinicopathological abnormalities were assessed to determine risk factors for the development of hyperammonemia. Adverse events following L-asp were recorded. Median plasma ammonia concentrations at baseline, 16 hours, and 48 hours were 26 µmol/L (44 µg/dL), 98 µmol/L (166.9 µg/dL), and 67 µmol/L (114 µg/dL), respectively. Median plasma ammonia concentrations at 16 and 48 hours after administration were significantly increased compared to baseline. Six dogs had adverse events following L-asp administration. No significant clinical signs were noted that could clearly be attributed to hyperammonemia. No risk factors for developing hyperammonemia were identified; however, there was a positive correlation between the development of hyperammonemia at 16- and 48-hour time points. CONCLUSIONS: Subclinical hyperammonemia in dogs with lymphoma or leukemia after L-asp administration appears to be common. No risk factors were identified for the development of hyperammonemia after L-asp treatment, and severe adverse events were rare.

Effects of pre-analytical variables on flow cytometric diagnosis of canine lymphoma: A retrospective study (2009-2015).

Flow cytometry (FC) is increasingly being used for immunophenotyping and staging of canine lymphoma. The aim of this retrospective study was to assess pre-analytical variables that might influence the diagnostic utility of FC of lymph node (LN) fine needle aspirate (FNA) specimens from dogs with lymphoproliferative diseases. The study included 987 cases with LN FNA specimens sent for immunophenotyping that were submitted to a diagnostic laboratory in Italy from 2009 to 2015. Cases were grouped into 'diagnostic' and 'non-diagnostic'. Pre-analytical factors analysed by univariate and multivariate analyses were animal-related factors (breed, age, sex, size), operator-related factors (year, season, shipping method, submitting veterinarian) and sample-related factors (type of sample material, cellular concentration, cytological smears, artefacts). The submitting veterinarian, sample material, sample cellularity and artefacts affected the likelihood of having a diagnostic sample. The availability of specimens from different sites and of cytological smears increased the odds of obtaining a diagnostic result. Major artefacts affecting diagnostic utility included poor cellularity and the presence of dead cells. Flow cytometry on LN FNA samples yielded conclusive results in more than 90% of cases with adequate sample quality and sampling conditions.

Carcinocythemia: First report in a cat and literature review.

A 6-year-old female neutered European Shorthair cat was presented with a 2-day history of lethargy and hyporexia. On physical examination, the cat was slightly depressed and had a 2.5 cm nodule in the left 3rd mammary gland. The hemogram revealed mild leukocytosis with mature neutrophilia and moderate thrombocytopenia. On blood smear evaluation, rare pleomorphic cells, possibly of epithelial origin, were observed mainly at the feathered edge. The animal died about 12 hours after presentation, and a necropsy was performed. On histopathology, the mammary nodule was diagnosed as a tubulopapillary adenocarcinoma with vascular invasion and widespread metastases. Immunocytochemical tests for cytokeratins (AE1/AE3) confirmed the epithelial phenotype of the neoplastic cells observed on the blood smear. The present report describes a feline mammary carcinoma with widespread metastases and the presence of malignant epithelial cells in the peripheral blood referred to as carcinocythemia. This condition has been previously described in people and dogs. To the author's knowledge, this is the first reported case of feline carcinocythemia. As in other species, the phenomenon was associated with a terminal phase of systemic malignancy.