Prevalence of Golden retriever in European dogs with lymphoma: preliminary data

Introduction. Canine breeds, being genetic clusters, are good models for studies on genetic predisposition. Golden retriever (GR) has been described with a high incidence of both lymphoma overall (19%) and T zone lymphoma (TZL, 40%) with differences in different geographical areas in US. This breed predisposition is confirmed in Japanese but not in European (EU) case series although specific studies are still lacking.Aim of the present study is to investigate the prevalence of GR in a huge case series of canine lymphomas from different EU countries and to compare prevalence of different subtypes with studies in extra-EU countries, in order to support a possible different genetic predisposition.Materials and methods. Signalment data on 1734 consecutive cases of canine lymphoma collected from 9 different European countries are retrospectively analysed. When subtypes are available, cases are furtherly separated in three subtype groups: 1) B-cell lymphoma, 2) T-cell lymphoma-high grade, 3) TZL. Odds ratio (OR) for different lymphoma subtypes are calculated in comparison with mixed breed population, considered as control.Results. Overall prevalence of GR is 5.19% (range 1.59-7.32%) of lymphoma cases and differs from that reported in American and Japanese caseloads. Prevalence slightly varies among EU countries and no subtypes predilection is found if compared with mixed breed. Concerning Italian cohort, GR is not predisposed to develop a lymphoma when normalized for the breed prevalence (OR=1.49, 95% confidence interval=0.87-2.55, p=0.14).Discussion. Prevalence of lymphoma in EU population of GR is much lower than that of US. No predisposition is identified in EU GR for TZL differently from US and Japan. Being genetic of European GR population quite different from American and Japanese ones this suggest a possible different genetic predisposition. Slight differences in GR lymphoma prevalence among European countries likely reflects different breed distribution rather than different genetic predisposition

Loss of cd45 protein in canine small clear cell/t-zone lymphoma is due to absence of gene transcription.

Canine small clear cell/T-zone lymphoma (TZL) is a peculiar lymphoma subtype characterized by an indolent clinical course and aberrant CD45-negative phenotype, easily recognized by flow cytometry (FC). Recent studies have described clinical presentation and behavior, but to date the mechanisms for CD45-negativity have never been investigated.Aim of this study is to confirm the lack of surface protein using a different technique from FC and to investigate if CD45-absence in TZL is linked to the lack of the corresponding transcript and gene.40 TZL cases and 17 controls (7 T-high grade lymphoma, 10 reactive lymphnodes) were included in the present study. Immunohistochemistry was performed with a different antibody respect with FC to confirm CD45 surface protein absence. Total RNA and genomic DNA were extracted from lymph-nodes aspirates. CD45 transcript amount was investigated by quantitative real-time RT-PCR and the corresponding gene fragment was analyzed by quantitative real-time PCR. ΔΔCt method was used for the relative quantification of transcript amount and DNA load compared to housekeeping genes.All TZL cases were negative for CD45 at immunohistochemistry. CD45 transcript amount was significantly lower in TZL compared to controls (p=0.000). This difference was not significant (p=0.584) for CD45 DNA load, that was similar between TZL and controls.These results highlight that CD45 protein is lacking on cell surface and gene transcription is absent in TZL, whereas the corresponding gene is not deleted. The data here reported support further studies for clarifying possible genomic or epigenomic factors involving CD45 gene transcription and for better clarifying the possible role of CD45 in lymphomagenesis

Flow cytometry for feline lymphoma: a retrospective study about pre-analytical factors possibly affecting the quality of samples

Introduction Flow cytometry (FC) is an increasingly required technique on which veterinary oncologists rely to have an accurate, fast, minimally invasive lymphoma or leukemia diagnosis. FC has been studied and applied with great results in canine oncology, whereas in feline oncology the use of this technique is still to be experienced. This is mainly due to a supposed discomfort in sampling, because of the high prevalence of intra-abdominal lymphomas. The purpose of the present study is to investigate whether any pre-analytical factor might affect the quality of suspected feline lymphoma samples for FC analysis.Methods 97 consecutive samples of suspected feline lymphoma were retrospectively selected from the authors’ institution FC database. The referring veterinarians were recalled and interrogated about several different variables, including signalling, features of the lesion, features of the sampling procedure and the experience of veterinarians performing the sampling. Statistical analyses were performed to assess the possible influence of these variables on the cellularity of the samples and the likelihood of being finally processed for FC.Results None of the investigated variables significantly influenced the quality of the submitted samples, but the needle size, with 21G needles providing the highest cellularity (Table 1). Notably, the samples quality did not vary between peripheral and intra-abdominal lesions. Sample cellularity alone influenced the likelihood of being processed. About a half of the cats required pharmacological restraint. Side effects were reported in one case only (transient swelling after peripheral lymph node sampling).Conclusions FC can be safely applied to cases of suspected feline lymphomas, even for intra-abdominal lesions. 21G needle should be preferred for sampling. This study provides the bases for the spread of this minimally invasive, fast and cost-effective technique in feline medicine

Immunophenotypical and cytomorphological examination of feline peripheral blood in patients with suspect leukemia

To date, knowledge about leukemias in feline patients is poor. Classification of leukemias is mostly based on morphological, immunological and genetic aspects in dogs and humans [Harvey, 2012]. However, in cat poor literature about classification of leukemias via flow cytometric (FC) immunophenotyping is available and just a few publications of case reports are present . We retrospectively selected 44 cases of leukemias from our FC service database from 2009 to 2017 with the aim to describe the major immunological features and define the prevalence of different subtypes. Blood and/or bone marrow submitted for FC analysis with a prevalent hematological presentation were selected. Cases with an evident lymph node enlargement or with clinical aspects suggesting a solid lesion (lymphoma) were excluded. Cases were classified depending on antigen expression and hematologic features in five groups: chronic lymphocytic leukemias (CLL), acute lymphoblastic leukemias (ALL), acute myelogenous leukemias (AML), acute undifferentiated leukemias (AUL) and undetermined ALL vs lymphoma stage V.26 cases (59%) were classified as acute leukemias whereas 11 cases (25%) were classified as chronic leukemias; 7 cases (16%) were classified as undetermined. Among acute leukemias 10 (38.5%) were AML, 10 (38.5%) AUL and 6 (23%) ALL. All ALL were of T cell origin (CD3+ or CD5+). Among chronic leukemias, 10 (91%) were of T cell origin and among these, 80% expressed CD4 (T-helper lymphocytes), while 20% were double negative (CD4- CD8-).These results confirmed that T cell leukemias are more frequent than B ones in the cat with a prevalent T-helper phenotypes as previously described. AMLs were highly represented, but the lack of an adequate panel of specific antibodies for myeloid lineage rendered a high number of AUL in our caseload. Similarly, the lack of an anti-feline CD34 antibody did not permit differential diagnosis of acute leukemia vs lymphoma with blood involvement in a remarkable percentage of cases without an evident nodal enlargement and without an extreme leukocytosis

Evaluation of CD45 protein expression and transcript in canine small clear cell/T zone lymphoma.

Canine small clear cell lymphoma is a peculiar lymphoma entity with T-zone histopathological pattern andindolent clinical course. From an immunophenotypic point of view the main feature is the lack of CD45 staining by flow-cytometry (FC), which accounts for >95% of cases. Underlying mechanisms have never been investigated.Aim of this work was to evaluate CD45 protein and mRNA expression in small clear cell lymphoma.Lymph nodes of 18 cases and 11 controls, with either reactive hyperplasia or CD45-positive high grade T-cell lymphoma, were investigated. FC was performed on lymph node fine needle aspiration and CD45 median fluorescence intensity (MFI) was then evaluated on small clear cells and normal residual T-lymphocytes. CD45 surface expression was also evaluated by immunohistochemical reaction on paraffin wax-embedded lymph node sections.Quantitative real-time RT-PCR was performed on cases and controls. Total RNA was isolated from cell suspension in RNA later. The generated CD45cDNA was amplified and ΔΔCt method was used for the relative mRNA quantification.CD45-MFI in neoplastic cells was <1% compared to normal residual T-lymphocytes in the same sample. Cells were also negative for CD45 stain on histopathological preparations. RT-PCR showed a significantly lower amount of CD45 transcript in neoplastic samples compared to controls, likely due to the residual population.Results showed the lack of CD45 surface antigen and the virtually absence of CD45-mRNA in small clear cell lymphoma. We hypothesize a possible genomic/epigenomic aberration; further studies are in progress to investigate the pathogenesis of this aberrancy and the possible linkage to lymphomagenesis

Breed-associated risks for developing canine lymphoma differ among countries: an European canine lymphoma network study

Canine breeds may be considered good animal models for the study of genetic predisposition to cancer, as they represent genetic clusters. From epidemiologic and case collection studies it emerges that some breeds are more likely to develop lymphoma or specific subtypes of lymphoma but available data are variable and geographically inconsistent. This study was born in the context of the European Canine Lymphoma Network with the aim of investigating the breed prevalence of canine lymphoma in different European countries and of investigating possible breed risk of lymphoma overall and/or different lymphoma subtypes

Gold meets enamine catalysis in the enantioselective α-allylic alkylation of aldehydes with alcohols

Synergetic gold(I)- and organo-catalysis proved to be efficient towards the intramolecular stereoselective a-allylic alkylation of aldehydes with simple primary and secondary alcohols. High diastereo- and enantioselectivity were recorde

Unstabilized DNA breaks in lymphocytes of patients with different subsets of systemic sclerosis

The clastogenic effects on DNA, proven by the presence of micronuclei (MN) and the protective cellular mechanisms normally used to stabilize DNA breaks were investigated in three subsets of patients with systemic sclerosis (SSc). The frequency of MN found in cultures of peripheral lymphocytes in patients with anticentromere and antitopoisomerase I antibodies was significantly higher than that in the control group. The group with anticentromere antibody showed a significantly higher frequency of MN than did the subjects with antitopoisomerase antibody (4.22% versus 2.34%, P < 0.001). Patients with anti-RNA polymerase III, instead, had a low prevalence of typical micronucleated cells (0.98%), not significantly different from that of the healthy controls (0.82%). Moreover, when MN was characterized for the presence or absence of DNA fragments with free 3'-OH ends by digoxigenin-dUTP (DIG-dUTP) using terminal deoxynucleotidil transferase, its frequency was found to be increased in the groups with anticentromere and antitopoisomerase I antibodies with respect to that in the controls. The increase was significantly higher in the lymphocytes of the patients with anticentromere than in those with antitopoisomerase I antibody (35% versus 20.08%, P < 0.001). Nonetheless, the prevalence of unstable DNA fragments in patients with anti-RNA polymerase III antibody was low (2.05%) and not significantly different from that of the control group (1.18%). Our results indicate that there is a clastogenic effect on DNA and an interference in the protective cellular mechanisms normally stabilizing DNA breaks only in some subsets of SSc patients. The clastogenic effects on DNA, proven by the presence of micronuclei (MN), and the protective cellular mechanisms normally used to stabilize DNA breaks were investigated in patients with systemic sclerosis (SSc). The frequency of micronucleated cells found in cultures of peripheral lymphocytes in patients was significantly higher than in the control group. The patient group with anti-centromere antibodies showed a significantly higher frequency of micronucleated cells than that observed in the patients with anti-topoisomerase I antibodies (4.22% versus 2.34%, p < 0.001). Moreover, we attempted to characterize MN for the presence or absence of DNA fragments with free 3'-OH ends by digoxigenin-dUTP (DIG-dUTP) using terminal deoxynucleotidil transferase. It was found that the frequency of MN containing DNA fragments with 3'-OH free ends (unstable fragments) increased in SSc patients compared to that observed in the control group. Moreover, this increase was significantly higher in lymphocytes of the patients with anti-centromere antibodies than in those with anti-topoisomerase I antibodies (35% versus 20.08%, p < 0.001). Our results indicate that in SSc patients there is an interference in the protective cellular mechanisms, normally stabilizing DNA break

Breed-associated risks for developing canine lymphoma differ among countries : an European canine lymphoma network study

Canine breeds may be considered good animal models for the study of genetic predisposition to cancer, as they represent genetic clusters. From epidemiologic and case collection studies it emerges that some breeds are more likely to develop lymphoma or specific subtypes of lymphoma but available data are variable and geographically inconsistent. This study was born in the context of the European Canine Lymphoma Network with the aim of investigating the breed prevalence of canine lymphoma in different European countries and of investigating possible breed risk of lymphoma overall and/or different lymphoma subtypes