Topoisomerase IIα immunoexpression in feline mammary carcinomas: A correlation with Ki67 immunoexpression and the mitotic count.

The aim of the study was to compare the immunohistochemical expression of topoisomerase IIα protein (Topo IIα) with Ki67 expression and mitotic count in feline mammary carcinomas (FMCs). Topo IIα is considered as a proliferation indicator as well as a molecular target of anthracycline chemotherapy. The studied material included 70 FMCs from female cats treated with mastectomy. Primary mouse monoclonal antibodies directed against Topo IIα and Ki67 were used in immunohistochemical reactions. The number of mitotic figures was counted at 400× magnification in a field of 2.37 mm2. Immunohistochemical reaction for Topo IIα occurred in cell nuclei. The Topo IIα index ranged from 6.12% to 54.60% and was positively correlated with the values of the Ki67 index (r = 0.7193) and the mitotic count (r = 0. 2858). This indicates the potential possibility of use of the immunohistochemical expression of Topo IIα to assess the rate of proliferation in FMCs. The wide range of expression of Topo IIα in individual tumorus found in the conducted studies allows us to hypothesize that its assessment could be used as a predictive marker in chemotherapy of FMCs with the use of anthracyclines. However, this requires confirmation in clinical trials.

Molecular Confirmation of Taenia crassiceps Cysticercosis in a Captive Ring-Tailed Lemur (Lemur catta) in Poland.

(1) Background: Taenia crassiceps is a cosmopolitan tapeworm endemic to the northern hemisphere with an indirect lifecycle. Its definitive hosts are carnivores, and its intermediate hosts are rodents and rabbits. Nonhuman primates in zoos appear to be highly susceptible to T. crassiceps cysticercosis. The aim of this study was to confirm the presence and the molecular characterization of T. crassiceps cysts isolated from a captive ring-tailed lemur. (2) Methods: Surgery revealed multifocal, transparent saccules containing several thin-walled tapeworm cysticerci. In some of the metacestodes, single or multiple exogenous buds from daughter cysticerci were spotted. A molecular analysis was performed to confirm our morphological examinations, using two protocols to obtain the partial nad1 and cox1 genes of the Taenia sp. (3) Results: On the basis of morphological features and molecular analysis, the cysticerci were identified as T. crassiceps metacestodes, and products taken from the PCRs were sequenced. With respect to interpreting the sequencing results of the obtained amplicons, we compared them with data in the GenBank database, proving that, in this case, the causative agent was indeed T. crassiceps. (4) Conclusions: The received data can be used to supplement descriptions of this species. To the best of our knowledge, this is the first case of cysticercosis caused by T. crassiceps in a nonhuman primate in Poland.

Molecular Confirmation of Massive Taenia pisiformis Cysticercosis in One Rabbit in Poland.

The aim of this study was to provide molecular characterization, together with phylogenetic analysis, of Taenia pisiformis cysts isolated from rabbit. On the basis of morphological features and molecular analysis, the cysticerci were identified as T.pisiformis metacestodes. PCR was performed with three different protocols to obtain partial sequences of 12S ribosomal RNA (12S rRNA), NADH dehydrogenase subunit 1 (nad1), and cytochrome oxidase subunit 1 (cox1) of Taenia spp. The products from the PCRs were sequenced. Interpretation of the sequencing results of the obtained amplicons, by comparing them with the GenBank database, proved that the causative agent, in this case, was T. pisiformis. The phylogenetic analysis of the received sequences identified a new haplotype. The received data can be used to supplement the species description. To our knowledge, this is the first molecular confirmation of T. pisiformis metacestodes infection in the rabbit, in Poland.

Evaluation of the Proliferative Activity of Diffuse Large B-Cell Lymphoma (DLBCL) in Dogs with Respect to Patient Eligibility for Anthracycline-Based Chemotherapy.

Different types of canine lymphoma respond differently to chemotherapy and have different prognoses. Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma in dogs. Topoisomerase II alpha (TOPIIα) protein has been shown to be a proliferation marker associated with prognostic significance. The aim of the study was to determine the relationship between TOPIIα expression, mitotic count (MC), and Ki67 antigen index in DLBCL in dogs, taking into account the applicability of these parameters to select the chemotherapy protocol with emphasis on the use of anthracycline drugs. Samples of formalin-fixed paraffin-embedded lymph nodes from 34 dogs with DLBCL were immunohistochemically labelled with anti-TOPIIα and Ki67. The number of positive cells and the intensity of the reaction were taken into account in order to assess TOPIIα expression. MC was estimated in the hematoxylin and eosin-stained slides in the area of 2.37 mm2. Positive association between TOPIIα and MC, but no association between TOPIIα and Ki67 was found. It can be concluded that the immunohistochemical determination of TOPIIα as a molecular target for drugs from the anthracycline group may be used in association with MC to establish a diagnostic-clinical protocol for selecting dogs with DLBCL for treatment with anthracycline drugs.

Study of DNA topoisomerase IIa expression in canine lymphomas and its potential role as a marker of sensitivity to anthracycline-based chemotherapy in dogs.

INTRODUCTION: Canine lymphoma remains one of the most chemotherapy-responsive neoplasia in dogs. Many factors affect the prognosis in dogs treated for lymphoma, but indications for a specific treatment regimen in individual animals with lymphoma are poorly defined. Topoisomerase IIα (TOPIIα) is a key enzyme in DNA replication and a molecular target for TOPIIα inhibitors, including anthracyclines. The aim of this study was to determine the expression of TOPIIα in canine malignant lymphomas. The relationship between TOPIIα expression in canine lymphomas and potential sensitivity of neoplastic cells to anthracycline-based chemotherapy is discussed. MATERIALS AND METHOD: Samples of formalin-fixed paraffin-embedded lymph nodes from 47 dogs with different subtypes of non-Hodgkin's (34 B-cell and 13 T-cell) lymphoma were immunohistochemically labeled with anti-TOPIIα. The number of positive cells and the intensity of the reaction were taken into account in order to assess TOPIIα expression. RESULTS: TOPIIα expression was evident in all cases, although differences in the number of positive cells and intensity of the reaction were demonstrated between B-cell and T-cell lymphoma groups as well as within individual groups. Based on the established scoring system, in the B-cell lymphoma group statistically higher expression of TOPIIα was found compared to the T-cell lymphoma group (P = 0.006). In B-cell lymphoma group moderate (41.18%) and strong (32.35%) TOPIIα expression predominated, whereas among T-cell lymphoma group the majority were cases with a weak (46.15%) TOPIIα expression. CONCLUSION: These preliminary results indicate that further studies are needed to determine the prognostic value of TOPIIα expression with regard to the sensitivity of canine B-cell lymphomas to anthracycline-based chemotherapy regimen. Nevertheless, this study indicates the possibility of choosing the appropriate treatment of canine lymphoma based on TOPIIa expression.

Seroprevalence of 12 serovars of pathogenic Leptospira in red foxes (Vulpes vulpes) in Poland.

BACKGROUND: Leptospira spp. infect humans and a wide range of domestic and wild animals, but certain species such as small rodents and red foxes (Vulpes vulpes) play a particular role as reservoirs and transmission of leptospirosis as they easily adapt to many habitats including human environments. To investigate the significance of red foxes in the epidemiology of leptospirosis in Poland, a seroprevalence survey was conducted. During the 2014-2015 hunting season, blood samples of 2134 red foxes originating from the central-eastern part of Poland were collected. Serum samples were tested by a microscopic agglutination test for the presence of specific antibodies to Leptospira serovars Icterohaemorrhagiae, Grippotyphosa, Sejroe, Tarassovi, Pomona, Canicola, Hardjo, Ballum, Australis, Bataviae, Saxkoebing and Poi. RESULTS: Antibodies to at least one serovar were detected in 561 sera (26.3%). The highest seroprevalence was found in the Subcarpathia (41.6%) and Warmia-Masuria (40.3%) provinces. Antibodies were mainly directed against serovars Poi (12.4%), Saxkoebing (11.3%), and Sejroe (6.0%). CONCLUSIONS: Exposure of red foxes to certain Leptospira serovars seems to be common in central and eastern Poland. In addition, the high prevalence of antibodies against Leptospira spp. in foxes may indicate a potential risk of infection for humans and other species coming into contact with these animals.