In vitro quality control analysis after processing and during storage of feline packed red blood cells units.

BACKGROUND: During the storage of packed red blood cells (pRBC), packed cell volume (PCV), bacterial contamination and percentage of haemolysis [percentage of free haemoglobin (HGB) in relation to the total HGB] are important quality parameters. Both PCV and haemolysis are indicators of the cellular integrity of stored units. There are no published experimental studies that evaluated these parameters during storage of feline pRBC using SAGM (adenine, dextrose, mannitol and sodium chloride) as the additive solution. The present study aims to (1) evaluate the quality of feline pRBCs stored in SAGM; (2) test for the semi-closed system's suitability for use and risk of bacterial contamination; (3) establish the maximum storage time that may be appropriate to meet the criteria established by the United States Food and Drug Administration (US-FDA) guidelines for human blood banking; and (4) evaluate the need to calculate the percentage of haemolysis prior to the administration of units stored for more than 4 weeks. Four hundred eighty nine feline pRBC units were analyzed. Bacterial culture, PCV and percentage of haemolysis were determined within 6 h after processing (t0). One hundred and eighty units were re-tested for haemolysis and PCV after 29-35 days of storage (t1) and 118 units after 36-42 days (t2). RESULTS: Bacterial contamination was not detected in any pRBC unit. Mean PCV at t0 was 52.25% (SD: ±5.27) and decreased significantly (p < 0.001) during storage to 48.15% (SD: ±3.79) at t1 and to 49.34% (SD: ±4.45) at t2. Mean percentage of haemolysis at t0 was 0.07% (SD: ±0.06) and increased significantly (p < 0.001) to 0.69% (SD: ±0.40) at t1 and to 0.81% (SD: ±0.47) at t2. In addition, 13.88% and 19.49% of pRBC units exceeded 1% haemolysis at t1 and t2, respectively. CONCLUSIONS: According to the US-FDA guidelines for human blood banking that recommend a maximum of 1% haemolysis, the results of this study show that all feline pRBC units with less than 24 h of shelf life have low levels of haemolysis. However, units preserved up to 28 days can only be administered if tested for haemolysis before use, since 13.88% units exceeded the 1% limit. The semi-closed system was considered safe for use as bacterial contamination was not detected in any pRBC unit.

Volume-dependent hemodynamic effects of blood collection in canine donors - evaluation of 13% and 15% of total blood volume depletion.

BACKGROUND: There is no consensus regarding the blood volume that could be safely donated by dogs, ranging from 11 to 25% of its total blood volume (TBV). No previous studies evaluated sedated donors. AIM: To evaluate the hemodynamic effects of blood collection from sedated and non-sedated dogs and to understand if such effects were volume-dependent. MATERIALS AND METHODS: Fifty three donations of 13% of TBV and 20 donations of 15% TBV were performed in dogs sedated with diazepam and ketamine. Additionally, a total of 30 collections of 13% TBV and 20 collections of 15% TBV were performed in non-sedated dogs. Non-invasive arterial blood pressures and pulse rates were registered before and 15 min after donation. RESULTS: Post-donation pulse rates increased significantly in both sedated groups, with higher differences in the 15% TBV collections. Systolic arterial pressures decreased significantly in these groups, while diastolic pressures increased significantly in 13% TBV donations. Non-sedated groups revealed a slight, but significant, SBP decrease. No clinical signs related to donations were registered. CONCLUSION: These results suggest that the collection of 15% TBV in sedated donors induces hemodynamic variations that may compromise the harmlessness of the procedure, while it seems to be a safe procedure in non-sedated dogs.

Aberrant P-cadherin expression is associated to aggressive feline mammary carcinomas.

BACKGROUND: Cadherins are calcium-dependent cell-to-cell adhesion glycoproteins playing a critical role in the formation and maintenance of normal tissue architecture. In normal mammary gland, E-cadherin is expressed by luminal epithelial cells, while P-cadherin is restricted to myoepithelial cells. Changes in the expression of classical E- and P-cadherins have been observed in mammary lesions and related to mammary carcinogenesis. P-cadherin and E-cadherin expressions were studied in a series of feline normal mammary glands, hyperplastic/dysplastic lesions, benign and malignant tumours by immunohistochemistry and double-label immunofluorescence. RESULTS: In normal tissue and in the majority of hyperplastic/dysplastic lesions and benign tumours, P-cadherin was restricted to myoepithelial cells, while 80% of the malignant tumours expressed P-cadherin in luminal epithelial cells. P-cadherin expression was significantly related to high histological grade of carcinomas (p <0.0001), tumour necrosis (p = 0.001), infiltrative growth (p = 0.0051), and presence of neoplastic emboli (p = 0.0401). Moreover, P-cadherin positive carcinomas had an eightfold likelihood of developing neoplastic emboli than negative tumours. Cadherins expression profile in high grade and in infiltrative tumours was similar, the majority expressing P-cadherin, regardless of E-cadherin expression status. The two cadherins were found to be co-expressed in carcinomas with aberrant P-cadherin expression and preserved E-cadherin. CONCLUSIONS: The results demonstrate a relationship between P-cadherin expression and aggressive biological behaviour of feline mammary carcinomas, suggesting that P-cadherin may be considered an indicator of poor prognosis in this animal species. Moreover, it indicates that, in queens, the aberrant expression of P-cadherin is a better marker of mammary carcinomas aggressive behaviour than the reduction of E-cadherin expression. Further investigation with follow-up studies in feline species should be conducted in order to evaluate the prognostic value of P-cadherin expression in E-cadherin positive carcinomas.

Common phenotypic and genotypic antimicrobial resistance patterns found in a case study of multiresistant E. coli from cohabitant pets, humans, and household surfaces.

The objective of the study described in this article was to characterize the antimicrobial resistance profiles among E. coli strains isolated from cohabitant pets and humans, evaluating the concurrent colonization of pets, owners, and home surfaces by bacteria carrying the same antimicrobial-resistant genes. The authors also intended to assess whether household surfaces and objects could contribute to the within-household antimicrobial-resistant gene diffusion between human and animal cohabitants. A total of 124 E. coli strains were isolated displaying 24 different phenotypic patterns with a remarkable percentage of multiresistant ones. The same resistance patterns were isolated from the dog's urine, mouth, the laundry floor, the refrigerator door, and the dog's food bowl. Some other multiresistant phenotypes, as long as resistant genes, were found repeatedly in different inhabitants and surfaces of the house. Direct, close contact between all the cohabitants and the touch of contaminated household surfaces and objects could be an explanation for these observations.

A retrospective histopathological survey on canine lymphomas subtypes of Porto District.

Background: Lymphomas are dogs' most common hematopoietic neoplasms and represent a heterogeneous group, as occurs in humans. Considering the role of dogs as models of human lymphomas and the geographical correlation of the cases of canine and human lymphoma, it is important to continuously assess the epidemiological distribution of lymphoma subtypes in dogs. Aim: This study aimed to provide a survey of canine lymphoma subtypes diagnosed from 2005 to 2016 in the academic veterinary pathology laboratory of the University of Porto. Methods: A total of 75 canine lymphomas diagnosed by histopathology in the Porto district were included. All cases were immunophenotyped by CD3 and PAX5, classified according to the current classification WHO and coded with Vet-ICD-O-canine-1. Results: Mixed breed dogs were most common (28%), followed by Cocker Spaniels (12%), Boxers (9%), and Labrador Retrievers (6%). The mean age was 9.2 years (SD = 3.3) (10.7 years for small, 8.9 years for medium and large, and 5.7 years for giant breed dogs, p < 0.05). Regarding sex, there was no difference in frequencies or mean age. B-cell lymphomas were more common (57.4%) than T-cell lymphomas (37.3%), and 5.3% were classified as non-B/non-T-cell lymphomas. Of the cases, 49% had a multicentric distribution, followed by splenic (22%), cutaneous (12%), alimentary (12%), and extranodal (3%) forms. The most common B-cell subtypes were diffuse large B-cell lymphoma (DLBCL) (16.3%) and large immunoblastic lymphoma (14%), while T-zone lymphoma (21.4%) and intestinal lymphoma (18%) were the most common T-cell lymphoma subtypes. Conclusion: Our study shows that the Porto district follows the international trend of higher prevalence of B-cell lymphomas in dogs, especially of the DLBCL subtype.

Evaluation of bacterial growth, effects on albumin, and coagulation factors in canine fresh frozen plasma administered as continuous rate infusion exposed to room temperature for 12 hours.

OBJECTIVES: To determine the risk of bacterial growth and to analyze the stability of albumin and coagulation factors in canine fresh frozen plasma (FFP) units exposed to room temperature (24°C) administered as a continuous rate infusion (CRI) for 12 hours. DESIGN: Ex vivo study. SETTING: University teaching hospital and pet blood bank. ANIMALS: None. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: An FFP CRI was simulated to replicate the standard routine procedure used in dogs. Plasma samples were collected before starting the CRI (H0), after 4 hours (H4), and after 12 hours (H12). Bacterial culture of FFP was performed and albumin concentration and specific activity levels for factors V, VII, VIII, and IX were measured and compared. All plasma culture results were negative. There were no statistically significant differences at any time point in the factor VIII activity (median 105.5% [range, 75.6%-142.0%] at H0; median 107.8% [range, 75.0%-172.7%] at H4; and median 112.1% [range, 81.7%-171.0%] at H12); factor IX activity (median 119.3% [range, 89.1%-175.9%] at H0; median 123.1% [range, 72.5%-172.7%] at H4; and median 118.3% [range, 86.6%-177.5%] at H12); or albumin concentration (median 21.0 g/L [range, 17.0-23.0 g/L] at H0 and median 20.0 g/L [range, 17.0-24.0 g/L] at H12). A slight but significant increase in factor V activity was observed when comparing H0 (median 107.0% [range, 71.0%-159.0%]) to H4 (median 117.7% [range, 71.0%-176.7%]) (P = 0.002) or H12 (median 116.2% [range, 71.0%-191.6%]) (P = 0.001). A slight but significant increase in factor VII activity was observed when comparing H0 (median 115.4% [range, 70.6%-183.7%]) to H4 (median 118.2% [range, 82.7%-194.6%]) (P = 0.005); H0 to H12 (median 128.7% [range, 86.4%-200.0%]) (P < 0.001); and H4 to H12 (P = 0.002). CONCLUSIONS: FFP CRI at room temperature for 12 hours could be considered safe with regard to risk for bacterial growth and also effective by providing albumin and clotting factors.

Vet-OncoNet: Developing a Network of Veterinary Oncology and Reporting a Pioneering Portuguese Experience.

Vet-OncoNet is a replicable tripartite animal cancer database with the scientific and academic purposes of collecting data and producing evidence-based knowledge for cancer science in general. Inspired by the One Health vision, Vet-OncoNet uses business intelligence tools to optimize the process of capturing, treating, and reporting animal cancer data to a national level in three interfaces: ACR (animal cancer registry, pathology-based), COR (clinical oncology registry, vet practice-based) and RFR (risk factor registry, owner-based). The first results show that skin and mammary gland are by far the most affected systems. Mast cell tumors and complex adenoma of the mammary gland are the most frequent histologic type in dogs, while in cats they are squamous cell carcinomas, tubular adenocarcinoma of the mammary gland and lymphomas. Regarding COR, it provides valuable information on the landscape of veterinary oncology practices, therapeutics options, outcomes and owners' drivers' adherence towards therapies, which range from 30% up to 80% upon vet practices. Furthermore, being aware of the role of animals within the family and as possible sentinels of environmental risks to cancer in humans, the network built an interface (Pet-OncoNet) dedicated to owners and a database (RFR) that receives information regarding pets and owners' daily habits.

Stability of coagulation factors in feline fresh frozen plasma intended for transfusion after 1 year of storage.

OBJECTIVES: The most common use of plasma transfusion is for haemostatic purposes, but coagulation factor activities in stored feline plasma are unknown. The concentration and stability of coagulation factors I (fibrinogen), II, V, VII, VIII, IX, X, XI and XII in feline fresh frozen plasma (fFFP) stored for 1 year were studied. METHODS: Fifty-five units of fFFP were produced from 55 fresh whole-blood donations obtained from indoor healthy blood donor cats. Twenty-one units were stored for <2 weeks (T0) and 34 were stored for 1 year (T1). After the completion of storage, specific coagulation factor activities for factors II, V, VII, VIII, IX, X, XI and XII were tested using modified one-stage activated partial thromboplastin or prothrombin time assays. Fibrinogen was determined using the Clauss method. RESULTS: Significantly decreased activities were observed for factors II (T0: 101.94% ± 19.06%; T1: 73.23% ± 39.06% [P = 0.001]), VII (T0: 102.78% ± 24.69%; T1: 60.08% ± 38.17% [P <0.001]), VIII (T0: 77.52% ± 30.39%; T1: 50.32% ± 23.8% [P = 0.001]), XI (T0: 88.76% ± 22.73%; T1: 66.28% ± 22.2% [P = 0.001]) and XII (T0: 89.50% ± 21.85%; T1: 55.46% ± 23.18% [P <0.001]) when comparing units at time 0 and after 1 year of storage. No significant difference was observed for factors IX (T0: 84.86% ± 29.35%; T1: 71.37% ± 22.23% [P = 0.064]) and X (T0: 96.24% ± 25.1%; T1: 83.91% ± 49.54% [P = 0.236]). Unexpectedly, a significant increase was observed for factor V (T0: 71.94% ± 24.14%; T1: 97.89% ± 62.33%; P = 0.046). Fibrinogen was 2.76 ± 1.09 g/l at T1. Factors VIII, XII and VII had the lowest mean activities after 1 year. CONCLUSIONS AND RELEVANCE: Although a decrease in most coagulation factors activities was noted with storage, 1-year-old fFFP was haemostatically active in vitro. The most suitable factors for quality control assessment of fFFP are factors VII and VIII. Approximately 13-20 ml/kg of fFFP is required to administer a minimum of 10 IU/kg coagulation factor activity.

Feline blood donation adverse reactions: classification and description of acute and delayed reactions in a donor population.

OBJECTIVES: This article aims to analyse the safety of feline blood donation by describing the frequency and nature of any adverse reactions and their causes, as well as propose measures to decrease the incidence of adverse reactions. METHODS: In this prospective study, any blood donor adverse reactions detected by the clinical staff during and immediately after donation were recorded. The owners of the cats were also surveyed by a veterinary practitioner or veterinary nurse 5 days after donation, using a predefined questionnaire to assess for any clinical or behavioural changes. Data were collected between January 2019 and March 2020 from blood donors enrolled in an animal blood bank programme. RESULTS: Of 3690 blood donations from 1792 feline donors assessed, post-donation reactions were reported in 1.14% (n = 42): 0.22% (n = 8) were acute reactions, which included weakness, pallor, tachypnoea and open-mouth breathing; and 0.92% (n = 34) were delayed post-donation reactions, with 0.16% involving cutaneous (haematomas and skin rashes, n = 6), 0.68% involving behavioural (n = 25) and 0.08% involving digestive (emesis and inappetence, n = 3) signs. CONCLUSIONS AND RELEVANCE: The low incidence of post-donation reactions in this study is encouraging, suggesting that a well-established protocol and competent staff can help to ensure a high level of safety in a feline donor programme and, in turn, increase the confidence of cat owners.

Cross Species Analysis and Comparison of Tumors in Dogs and Cats, by Age, Sex, Topography and Main Morphologies. Data from Vet-OncoNet.

The animal cancer burden is essential for the translational value of companion animals in comparative oncology. The present work aims to describe, analyze, and compare frequencies and associations of tumors in dogs and cats based on the Animal Cancer Registry created by Vet-OncoNet. With 9079 registries, regarding 2019 and 2020, 81% (n = 7355) belonged to dogs. In comparison, cats have a general one-year right advance in the mean age of cancer diagnosis compared to dogs. The multivariate topography group analysis shows a distinct pattern between the two species: dogs have higher odds of cancer in the genito-urinary system, spleen, soft tissue tumors and skin, while cats show higher odds for tumors in the eyes, digestive organs, nasal cavity, lymph nodes, bones and mammary glands. Regarding morphologies, dogs are overrepresented in mast cell tumors (MCT), melanomas, and hemangiosarcomas. While cats are overrepresented in fibrosarcomas, lymphomas (T and B-cell), in malignant mammary tumors, and squamous cell carcinoma (SCC). Females have greater odds only in the mammary gland, with males having greater odds in six of twelve topographies. This study is the first outcome of continuous animal cancer registration studies in Portugal.

Quantitative assessment of infusion pump-mediated haemolysis in feline packed red blood cell transfusions.

OBJECTIVES: Haemolysis caused by the use of peristaltic infusion pumps (PIPs) has been described in human and canine packed red blood cells (pRBCs). The aim of this study was to evaluate the effects of two different linear PIPs on the haemolysis of feline pRBC units stored for a long time. METHODS: Feline pRBC units stored with adenine, dextrose, mannitol and sodium chloride (SAGM) were manufactured. After 35-42 days of storage at 2-4°C, a line administration system with a 180 µm filter was attached to every pRBC bag, the system was drained by gravity alone (8 drops/min) and a 1.3 ml sample was collected (G). A NIKI V4 pump was then used at a flow rate of 25 ml/h, the flow was stopped when the infusion system was filled with blood coming from the infusion pump and another 1.3 ml sample was collected (NK). Finally, an Infusomat FmS pump was evaluated, collecting another 1.3 ml sample (IM). Packed cell volume (PCV) was measured in all samples by microhaematocrit centrifugation, total haemoglobin (HGB) was measured using a specific haemoglobin analyser and, after centrifugation, free HGB was determined by spectrophotometry. The percentage of haemolysis was calculated. Friedman's test was used to compare the samples. RESULTS: Fifteen feline pRBC units were evaluated. The average degree of haemolysis for sample G (gravity-assisted) was 1.12%. Comparison of the degree of gravity-assisted haemolysis with haemolysis in PIP NK (1.13%) and IM (1.14%) samples revealed no significant differences, with differences of only 0.01% and 0.02%, respectively. CONCLUSIONS AND RELEVANCE: The results of this study demonstrate that the use of two common PIPs in veterinary hospitals does not produce levels of haemolysis that are significantly different than that caused by gravity alone during transfusion of feline pRBCs at a rate of 25 ml/h.

Coordinated expression of galectin-3 and galectin-3-binding sites in malignant mammary tumors: implications for tumor metastasis.

Galectin-3 is a glycan-binding protein that mediates cell-cell and/or cell-extracellular matrix (ECM) interactions. Although galectin-3 is implicated in the progression of various types of cancers, the mechanisms by which galectin-3 enhances metastasis remain unclear. In order to elucidate the role of galectin-3 in the complex multistage process of cancer metastasis, we examined galectin-3 and galectin-3-binding site expression in a series of 82 spontaneous canine mammary tumors (CMT) and two CMT cell lines. Benign CMT tumors exhibited strong nuclear/cytoplasmic galectin-3 immunostaining, whereas malignant CMT tumors and metastases exhibited dramatically decreased galectin-3 expression with the majority of the immunostaining confined to the cytoplasm. Interestingly, intravascular tumor cells overexpressed galectin-3 regardless of their location. CMT-U27 xenografts displayed the same pattern of galectin-3 expression found in spontaneous malignant CMT. In parallel with the downregulation of galectin-3, malignant CMT displayed an overall loss of galectin-3-binding sites in the ECM and focal expression of galectin-3-binding sites mainly detected in intravascular tumor cells and endothelium. Furthermore, loss of galectin-3-binding sites was correlated with the downregulation of GLT25D1, a ß (1-O) galactosyltransferase that modifies collagen, and upregulation of stromal galectin-1. Finally, GLT25D1 mRNA expression was strikingly downregulated in malignant CMT-U27 compared with the benign cell line, and its expression was further decreased in a galectin-3 knockdown CMT-U27 cell line. We therefore hypothesized that the loss of galectin-3-binding sites in the ECM in conjunction with the overexpression of galectin-3 in specific tumor cell subpopulations are crucial events for the development of mammary tumor metastases.

Determination of urokinase-type plasminogen activator serum levels in healthy and oncologic cats.

The urokinase plasminogen activator system (uPAS) has been poorly investigated in veterinary oncology. The aim of this study was to determine uPA serum concentrations in healthy and oncologic cats to understand the potential value of uPA as a cancer biomarker. Serum samples were collected from 19 healthy cats and 18 cats with spontaneous malignant neoplasms and uPA was measured through a specific enzyme-linked immunosorbent assay kit. The differences between uPA values and their relation with intrinsic factors and clinicopathological parameters were analyzed using an analysis of variance (ANOVA) and independent t-test. The average serum concentration of uPA in cancerous cats (0.54 ± 0.22 ng/mL) differed from that of healthy cats (1.10 ± 1.16 ng/mL) but was not significantly influenced by cats' clinicopathological parameters or by the presence of metastases. This study describes, for the first time, the serum concentrations of uPA in cats and proposes directions for future studies to uncover the relevance of uPAS in feline carcinogenesis.

Le système activateur de plasminogène de type urokinase (uPAS) a été peu étudié en oncologie vétérinaire. L'objectif de la présente étude était de déterminer les concentrations sériques d'uPA chez des chats en santé et oncologiques afin de comprendre la valeur potentielle d'uPA comme marqueur de cancer. Des échantillons de sérum furent prélevés de 19 chats en santé et de 18 chats avec des néoplasmes malins spontanés et l'uPA fut mesuré à l'aide d'une trousse immuno-enzymatique. Les différences entre les valeurs d'uPA et leur relation avec des facteurs intrinsèques et des paramètres clinico-pathologiques furent analysées par analyse de variance (ANOVA) et test de t indépendant. La concentration moyenne d'uPA chez les chats avec cancer (0,54 ± 0,22 ng/mL) différait de celle des chats en santé (1,10 ± 1,16 ng/mL) mais n'était pas influencée de manière significative par les paramètres clinico-pathologiques des chats ou la présence de métastases. Cette étude décrit, pour la première fois, les concentrations sériques d'uPA chez les chats et propose des orientations pour des études ultérieures afin de révéler la pertinence d'uPAS dans la carcinogénèse chez les chats.(Traduit par Docteur Serge Messier).

In vitro hemolysis of stored units of canine packed red blood cells.

BACKGROUND: Hemolysis is an important quality parameter of packed red blood cells (pRBCs) that is used to assess the cellular integrity of stored blood units. According to human standards, hemolysis at the end of storage must not exceed 1%, as otherwise it may be responsible for decreased transfusion effectiveness and acute life-threatening reactions. OBJECTIVES: This prospective study was designed to evaluate the hemolysis of canine pRBCs stored in an additive solution containing adenine, dextrose, mannitol, and sodium chloride, and to assess its associations with storage time, duration of the collection process, collection disturbances, and with the final volume and PCV of the pRBCs units. METHODS: One hundred eighty pRBCs units were collected from canine donors. Hemolysis of the pRBCs units was determined immediately after processing (t = 0). The units were then stored and retested (t = 1) either before administration (during weeks 2, 3, 4, 5, or 6 of storage) or at the end of the storage period (42 d) if not used. RESULTS: Mean hemolysis at t = 0 was 0.09% (SD 0.06) and increased during storage, at a more pronounced rate from the 5th (mean values of 0.52%, SD 0.29) to the 6th week (1.2%, SD 0.72). Almost 51% of the units with 36-42 days of shelf-life showed more than 1% hemolysis. Disturbances in the collection process, the volume of the whole blood units, and the volume of stored pRBCs units or their PCV were not related to pRBCs hemolysis. CONCLUSIONS: According to human blood bank recommendations regarding acceptable hemolysis, canine pRBCs stored for more than 35 days should be tested to ensure <1% hemolysis prior to administration.

E-cadherin, beta-catenin, invasion and lymph node metastases in canine malignant mammary tumours.

Recent studies of canine malignant mammary tumours suggest that reduction of E-cadherin and/or beta-catenin correlates with invasive behaviour and lymph node metastasis. The aims of this study were to examine the interrelationships between the expression of E-cadherin and beta-catenin, and the relationship between the expression of E-cadherin and/or beta-catenin and the mode of growth and metastatic capacity of canine malignant mammary tumours. 90 spontaneous malignant tumours and local and regional lymph nodes were studied. A significant relationship was evidenced between membranous expression of E-cadherin and beta-catenin (p=0.0027), but not between E-cadherin and cytoplasmic beta-catenin. Only E-cadherin as a separate factor was significantly related to tumour invasion (p=0.0072) and lymph node metastasis (p=0.0001). Neither membranous nor cytoplasmic beta-catenin expression was significantly related to either of these phenomena.

Use of the optical disector in canine mammary simple and complex carcinomas.

Grading of canine mammary carcinomas (CMC) is associated to subjective assessments made by the pathologists. Due to its unbiased nature, stereology can be used to objectively quantify morphological parameters associated with grading and malignancy. However, the use of stereology in CMC has not been fully disclosed. The nuclear numerical density [NV (nuclei, tumor)] is a cellularity-associated parameter that can be estimated by the optical disector. Herein, it was estimated in 44 CMC and its association with clinicopathologic factors - such as tumor size, histological subtype and grade, vascular/lymph node invasion, nuclear pleomorphism, and survival - was evaluated. Considering all the cases, the mean NV (nuclei, tumor) was 1.6 × 106 ± 0.5 × 106 nuclei/mm3 . Lower values were attained in complex carcinomas, comparing to simple carcinomas, in tumors smaller than 5 cm, with low mitotic activity and in those with high nuclear pleomorphism. No statistically significant association with grade or vascular/lymph node invasion was observed, but tumors with disease progression had lower nuclear densities. The NV (nuclei, tumor) and the correlated parameters mirror to some extension those in human breast cancer, suggesting an interesting interspecies agreement. This first estimation of the nuclear numerical density in CMC highlights the feasibility of the optical disector and their utility for objective morphological assessments in CMC. The association between nuclear numerical density and disease progression warrants future studies.

Serum levels of urokinase-type plasminogen activator in healthy dogs and oncologic canine patients.

AIM: Urokinase plasminogen activator (uPA) has been scarcely studied in veterinary oncology. The aim of this study was to determine the uPA serum concentrations in healthy and oncologic canine patients and to investigate its potential value as a tumor biomarker. MATERIALS AND METHODS: Serum uPA concentrations of healthy and oncologic canine patients were measured by enzyme-linked immunosorbent assay. Their relationships with the dogs' health status and tumor characteristics were analyzed through ANOVA and independent t-test. RESULTS: There were no significant differences between mean serum values (±standard deviation) of healthy dogs (0.19±0.13 ng/ml) and oncologic canine patients (0.22±0.33 ng/ml), or between dogs with benign or malignant tumors, and with or without metastases, although the latter tended to show higher uPA serum levels. CONCLUSION: This is the first study describing the uPA serum levels in dogs. Although its results do not support uPA as a tumor biomarker, higher uPA levels in dogs with metastatic neoplasms may reflect the role of the enzyme in tumor progression.

Distribution of feline AB blood types: a review of frequencies and its implications in the Iberian Peninsula.

OBJECTIVES: The objective of this study was to document the prevalence of feline blood types in the Iberian Peninsula and to determine the potential risk of incompatibility-related transfusion reactions in unmatched transfusions and the potential risk of neonatal isoerythrolysis (NI) in kittens born to parents of unknown blood type. METHODS: Blood samples were obtained from blood donors of the Animal Blood Bank (BSA-Banco de Sangue Animal). Blood typing was performed using a card method (RapidVet-H Feline Blood Typing; MDS). RESULTS: The studied population comprised 1070 purebred and non-purebred cats from Portugal and Spain aged between 1 and 8 years. Overall, frequencies of blood types A and B were 96.5% and 3.5%, respectively. No AB cats were found. Based on these data, the potential risks of NI and transfusion reactions in unmatched transfusions were calculated to be 6.8% and 2.8%, respectively. CONCLUSIONS AND RELEVANCE: Unlike previous studies, no type AB cats were found in this study. Although the calculated potential risks of transfusion reaction in unmatched transfusions and neonatal isoerythrolysis were low, blood typing prior to blood transfusion and blood typing of cats for breeding purposes are highly recommended.

MIB-1 labelling indices according to clinico-pathological variables in canine mammary tumours: a multivariate study.

The relationship between MIB-1 labelling indices (LI), as detected by immunohistochemical methods, and other clinico-pathological characteristics was studied in a series of 77 malignant mammary tumours surgically removed from 47 female dogs. The immunostaining was assessed on the basis of the estimated percentage of positive cells in the areas of highest labelling. Multivariate logistic regression demonstrated no influence of breed, age, previous pregnancies, previous progestin administration, histological type or location of the tumour on MIB-1 LI. MIB-1 LI was significantly related to the size of the tumour, necrosis, invasive growth and histological grade, but not with ulceration, lymph node metastasis, skin fixation or E-cadherin expression. The significant relationship between MIB-1 LI and other known factors of poor prognosis suggests that a high LI may have prognostic value in canine malignant mammary tumours.

Serum Galectin-3 Levels in Dogs with Metastatic and Non-metastatic Mammary Tumors.

Galectin-3 is implicated in tumor progression and metastasis. High levels of galectin-3 have been reported in intravasated cells in primary and metastatic tumor sites of canine malignant mammary tumors (CMMT). Nevertheless, it is still unknown whether this increase is limited to the site of the lesion or if it is a systemic feature. To better understand the pattern of the expression of galectin-3 and to investigate the possibility of using serum galectin-3 levels as a relevant biomarker in this disease, galectin-3 concentrations were determined in a series of sera from CMMT-bearing female dogs. None of the dogs included in the study had detectable metastases at the time of presentation. Animals were retrospectively divided into two groups dependent on whether or not they developed metastatic lesions during a 25-month follow-up period. Samples were collected from all dogs before surgery, 1 month after resection of the primary tumor and every 3 months during the postoperative period. Galectin-3 levels were significantly higher 1 month after than at the time of surgery (p=0.0058). Higher galectin-3 was found in samples collected 7 (p=0.0007), 10 (p=0.0061) and 13 months (p=0.0052) after surgery from dogs of the metastatic group when compared to those remaining free of development of detectable metastases. In conclusion, increased serum galectin-3 levels seem to be present in both metastatic and non-metastatic cases during the postoperative period, however, while in non-metastatic cases the values tend to return to baseline levels after surgery, in metastatic cases, levels remain persistently elevated.