Equine Sarcoid

The equine sarcoid is the most common skin neoplasia in the horse. It has a worldwide distribution and can also affect other equids such as donkeys, zebras, and mules. All breeds can develop the disease at any age, with no sex predilection, although geldings seem to be overrepresented. This fibroblastic neoplasm has several clinical presentations and microscopic features and has a nonmetastatic behavior but can be severely locally invasive. In many cases, multiple sarcoids may develop simultaneously or sequentially during their life and spontaneous remission is rarely reported. The etiology is multifactorial and involves bovine papillomaviruses, genetic, and environmental factors. Treatment options include different modalities depending on multiple factors: lesion type, location and extent, individual patient, facilities, owner, and financial issues

Characterization of the immune response against Testudinid herpesvirus 3: new insight

Testudinid herpesvirus 3 (TeHV3) is one of the most lethal viral agents in tortoises worldwide. Although TeHV3 have been extensively studied, only little information is available about host-pathogen interaction. TeHV3 infections in different species of the genus Testudo correlate with various lesions profiles, disease severity and clinical outcome, suggesting the existence of a complex host-pathogen interaction. This might reflect a possible viral-host coevolution (Origgi, 2012).To study the host-pathogen interaction, we previously screened 5.000 clones from a bacteriophage library obtained from the TeHV3 genomic DNA using Testudo graeca seropositive sera. Of the six detected positive clones, only one was confirmed by F.A.C.S. Selected clone was determined to be a concatamer of different TeHV3 genomic fragments including the partial sequence of TE17, UL15, Major capsid protein (MCP), and Glycoprotein B (gB) genes. After complete sequencing of the selected clone, the MCP and the gB were antisenses compared to the phagemid promoter.In order to assess which of the gene fragments among TE17 and UL15 was encoding for the antigenic determinant that was recognized by the anti-TeHV3 tortoise sera, distinct approaches were followed.TE-17 and UL15 fragments were knock out from the original phagemid using the following approaches: a) directed-site mutagenesis, b) molecular cloning, and c) restriction enzymes cloning. All the modified constructs were cloned in two different E. coli cloning vectors (D5α and XL 1-Blue).Transformation of competent cells with the constructs described above did not yield any viable bacteria.Among the different aspects might have influenced transformation success rate, construct size was probably the most relevant (about 9Kb). Furthermore, we could not entirely exclude that genomic DNA editing might have induced mutations in the construct sequence causing toxic effects on the host bacterial cell. Cloning of TE-17 and UL15 gene fragments into different prokaryotic expression vectors is currently under way

Characterization of the immune response against Testudinid herpesvirus 3.

Numerous infectious diseases have been documented in reptiles, however minimal information isavailable concerning their immunological response. One of the most diffuse and lethal reptile pathogenis Testudinid herpesvirus 3 (TeHV3), a Alphaherpesvirinae. All species of tortoises (Testudinide) areconsidered susceptible to TeHV3, however the virus is over represented in the genus Testudo, whichincludes, among others, T. graeca, T. hermanni, T. marginata, and T. horsfieldii, that are popular pets inEurope. Incidence of TeHV3-associated disease is highest right after hibernation (Origgi, 2012).The aim of this work is to partially characterize the immunological response of T. graeca against TeHV3.A bacteriophage library composed of about 5.000 clones containing genomic DNA fragments of TeHV3was produced. Bacteriophages were amplified in a specific strain of E. coli and were screened withTeHV3-seropositive sera from T. graeca. Phagemids were excised from the positive bacteriophages,sequenced, and compare with the TeHV3 genome to identify the encoding genes. Six differentstructural and non-structural proteins have identified as immune relevant. Vero cells where transfectedwith phagemids of the positive clones, to confirm previous results. TeHV3’s proteins expression wasassessed by F.A.C.S using T. graeca seropositive sera. Of all the six selected clones, only that expressingthe partial sequence of the glycoprotein B (gB) showed a positive signal in the F.A.C.S. analysis. Thisresult is consistent with the well-known immunogenicity of gB of other herpesviruses including thoseinfecting humans and with the highly conserved role that gB plays in host-pathogen interaction acrossspecies and evolution (Beals et al., 2016)

TeHV3 outbreak characterization in captive Testudo spp.

Italian Tortoises species are considered either endangered or near threatened according to International Union for Conservation of Nature. When pet tortoises are abandoned or found injured or seized following illegal detention, they are sent to wildlife rehabilitation centers. From 2008, the Testudo spp. population housed in the WWF Vanzago’s oasis exhibited clinical signs compatible with Testudinid herpesviurs 3 (TeHV3) infection.  By the end of 2012 all Testudo had died. The presence of TeHV3 was investigated by molecular biology and pathology. All the tortoises housed in Vanzago resulted ELISA positive for the presence of anti-TeHV3 antibodies except one T. hermanni. Of these, 12 animals died and were all necropsied. Lesion frequency distribution was evaluate by histology. PCR was positive in 8/12 tortoises. To better complement the epidemiological evaluation of the virus in northern Italy, 20 retrospective cases were selected from the archive of the University of Milan. Of these, 5 were TeHV3 PCR positive. Lesions closely resembled those of the Vanzago’s population. These results are consistent with a high prevalence of TeHV3 in northern Italy. The finding of intranuclear inclusion bodies demonstrated to be specific but not sensitive. TeHV3 diagnostic pathological lesions have been reported to vary according with host immune response and by the viral replicative status. Molecular techniques were often necessary to confirm the infection. According to the literature and to our findings, T. hermanni spp. seems the species with higher mortality and lower antibody concentrations when infected with TeHV3.Italian Tortoises species are considered either endangered or near threatened according to International Union for Conservation of Nature. When pet tortoises are abandoned or found injured or seized following illegal detention, they are sent to wildlife rehabilitation centers. From 2008, the Testudo spp. population housed in the WWF Vanzago’s oasis exhibited clinical signs compatible with Testudinid herpesviurs 3 (TeHV3) infection.  By the end of 2012 all Testudo had died. The presence of TeHV3 was investigated by molecular biology and pathology. All the tortoises housed in Vanzago resulted ELISA positive for the presence of anti-TeHV3 antibodies except one T. hermanni. Of these, 12 animals died and were all necropsied. Lesion frequency distribution was evaluate by histology. PCR was positive in 8/12 tortoises. To better complement the epidemiological evaluation of the virus in northern Italy, 20 retrospective cases were selected from the archive of the University of Milan. Of these, 5 were TeHV3 PCR positive. Lesions closely resembled those of the Vanzago’s population. These results are consistent with a high prevalence of TeHV3 in northern Italy. The finding of intranuclear inclusion bodies demonstrated to be specific but not sensitive. TeHV3 diagnostic pathological lesions have been reported to vary according with host immune response and by the viral replicative status. Molecular techniques were often necessary to confirm the infection. According to the literature and to our findings, T. hermanni spp. seems the species with higher mortality and lower antibody concentrations when infected with TeHV3

Evaluation of cytological diagnostic accuracy for canine splenic neoplasms : an investigation in 78 cases using STARD guidelines

Cytology represents a useful diagnostic tool in the preliminary clinical approach to canine splenic lesions, and may prevent unnecessary splenectomy. However, few studies have evaluated diagnostic accuracy of cytology in the diagnosis of canine splenic neoplasms. The aim of this study was to determine overall accuracy, sensitivity, specificity, positive and negative predictive values (i.e. diagnostic accuracy indexes) of cytology for canine splenic neoplasms following Standards for the Reporting of Diagnostic Accuracy Studies (STARD) guidelines. A consecutive series of canine splenic cytological samples was retrospectively retrieved from the database of the Diagnostic Pathology Service of the Department of Veterinary Medicine (DIMEVET\u2014University of Milan). Histopathology was set as the diagnostic reference standard. Cytological cases were enrolled when slides were available for review and when the same lesion was submitted for histopathology. Seventy-eight (78) lesions were included in the study. By histopathology, 56 were neoplastic and 22 were non-neoplastic. Cytology had an overall accuracy of 73.08% (95% C.I. 61.84%-82.50%), sensitivity of 64.29% (95% C.I. 50.36%-76.64%), specificity of 95.45% (95% C.I. 77.16%-99.88%), and positive and negative predictive values of 97.3% (95% C.I. 84.01%-99.60%) and 51.22% (95% C.I. 42.21%-60.15%), respectively. Low sensitivity and negative predictive value were balanced by very high specificity and positive predictive value. When positive for neoplasia, cytology represents a useful diagnostic tool to rule in splenic neoplasia, prompting surgery independently from other diagnostic tests. Conversely, a negative cytological result requires additional investigations to confirm the dog to be disease free

Chromogenic in situ hybridization for the detection of lambda and kappa immunoglobulin light chains as a potential auxiliary diagnostic technique in canine plasmacytomas

The heterogeneous morphologic features of canine plasmacytomas (PCTs) can make their differentiation from other round cell tumors challenging. Immunohistochemistry (IHC) for lambda (\u3bb) and kappa (\u43a) immunoglobulin (Ig) light chains is often equivocal because of high background staining. The chromogenic in situ hybridization (CISH) technique for light chains has shown higher sensitivity compared to IHC in human plasma cell tumors. Therefore, we aimed to validate automated CISH for light chains in canine tissues and to evaluate its diagnostic potential in canine PCTs, in conjunction with routinely used IHC markers. CISH for light chains demonstrated a clear signal in plasma cell populations of canine control tissues (lymph nodes, lymphoplasmacytic inflammation) showing a polyclonal pattern with a prevalence of \u3bb-producing cells. CISH detected monotypic light chain expression in 33 of 53 (62%) PCTs, 31 expressing \u3bb and 2 expressing \u43a. CISH was more sensitive than IHC for \u3bb light chain (58% vs. 47%, respectively) and more easily interpretable given the absence of confounding background staining. The absence of CISH staining for both \u3bb and \u43a in a considerable subset of tumors may be the result of lower light chain production by neoplastic cells. Multiple myeloma oncogene 1 (MUM1) was expressed by all but 2 PCTs (96%), which showed \u3bb expression by CISH and IHC. The identification of poorly differentiated canine PCTs requires the assessment of a panel of IHC markers, with the potential support of CISH for Ig light chains

Effects of age-associated regional changes in aortic stiffness on human hemodynamics revealed by computational modeling

Although considered by many as the gold standard clinical measure of arterial stiffness, carotid-to-femoral pulse wave velocity (cf-PWV) averages material and geometric properties over a large portion of the central arterial tree. Given that such properties may evolve differentially as a function of region in cases of hypertension and aging, among other conditions, there is a need to evaluate the potential utility of cf-PWV as an early diagnostic of progressive vascular stiffening. In this paper, we introduce a data-driven fluid-solid-interaction computational model of the human aorta to simulate effects of aging-related changes in regional wall properties (e.g., biaxial material stiffness and wall thickness) and conduit geometry (e.g., vessel caliber, length, and tortuosity) on several metrics of arterial stiffness, including distensibility, augmented pulse pressure, and cyclic changes in stored elastic energy. Using the best available biomechanical data, our results for PWV compare well to findings reported for large population studies while rendering a higher resolution description of evolving local and global metrics of aortic stiffening. Our results reveal similar spatio-temporal trends between stiffness and its surrogate metrics, except PWV, thus indicating a complex dependency of the latter on geometry. Lastly, our analysis highlights the importance of the tethering exerted by external tissues, which was iteratively estimated until hemodynamic simulations recovered typical values of tissue properties, pulse pressure, and PWV for each age group

Effects of age-associated regional changes in aortic stiffness on human hemodynamics revealed by computational modeling

Although considered by many as the gold standard clinical measure of arterial stiffness, carotid-to-femoral pulse wave velocity (cf-PWV) averages material and geometric properties over a large portion of the central arterial tree. Given that such properties may evolve differentially as a function of region in cases of hypertension and aging, among other conditions, there is a need to evaluate the potential utility of cf-PWV as an early diagnostic of progressive vascular stiffening. In this paper, we introduce a data-driven fluid-solid-interaction computational model of the human aorta to simulate effects of aging-related changes in regional wall properties (e.g., biaxial material stiffness and wall thickness) and conduit geometry (e.g., vessel caliber, length, and tortuosity) on several metrics of arterial stiffness, including distensibility, augmented pulse pressure, and cyclic changes in stored elastic energy. Using the best available biomechanical data, our results for PWV compare well to findings reported for large population studies while rendering a higher resolution description of evolving local and global metrics of aortic stiffening. Our results reveal similar spatio-temporal trends between stiffness and its surrogate metrics, except PWV, thus indicating a complex dependency of the latter on geometry. Lastly, our analysis highlights the importance of the tethering exerted by external tissues, which was iteratively estimated until hemodynamic simulations recovered typical values of tissue properties, pulse pressure, and PWV for each age group.</p

Neoadjuvant and adjuvant doxorubicin chemotherapy in a case of feline soft tissue sarcoma

A 7-year old male neutered domestic shorthair cat was presented with a 2 month history of a slow-growing mass on the right zygomatic area. A CT scan revealed a soft tissue mass in the right zygomatic region with no alterations of the underlying bone and features of local invasiveness. Cytology was suggestive of a mesenchymal tumour and histopathology from an incisional biopsy was consistent with a soft tissue sarcoma (STS). The cat was treated with neoadjuvant intravenous doxorubicin chemotherapy at a dose of 25 mg/m2, every two weeks. The patient experienced a partial response and underwent surgical excision of the tumour. Doxorubicin was continued as an adjuvant treatment for three further chemotherapy sessions, at a dose of 25 mg/m2 every 21 days. Local tumour recurrence was detected on clinical examination and cytologically confirmed 259 days following surgery

Canine smooth muscle tumors: A clinicopathological study

Canine smooth muscle tumors (SMTs) commonly develop in the alimentary and female genital tracts and less frequently in soft tissue. The definition of histological criteria of malignancy is less detailed for SMTs in dogs than in humans. This study evaluated the clinicopathologic features of canine SMTs and compared the veterinary and human medical criteria of malignancy. A total of 105 canine SMTs were evaluated histologically and classified according to both veterinary and human criteria. The Ki67 labeling index was assessed in all SMTs. Estrogen receptor (ER) and progesterone receptor (PR) expression was evaluated for soft tissue SMTs. Follow-up data were available in 25 cases. SMTs were diagnosed in the female genital tract (42%), alimentary tract (22%), and soft tissue (20%). Soft tissue SMTs frequently arose in the perigenital area, pelvic cavity, and retroperitoneum. A subset of soft tissue SMTs expressed ER and/or PR, resembling the gynecologic type of soft tissue SMT in humans. SMTs were less frequently malignant when assessed with human criteria than with veterinary criteria, better reflecting their benign behavior, especially in the genital tract where human criteria tolerate a higher mitotic count for leiomyoma. Decreased differentiation was correlated with increased proliferation, necrosis, and reduced desmin expression. Mitotic count, Ki67 labeling index, and necrosis were correlated with metastases and tumor-related death. Further prognostic studies are warranted to confirm the better performance of the human criteria when assessing SMT malignancy, especially genital cases, to confirm their usefulness in ER/PR-expressing soft tissue SMTs, and to better define the most useful prognostic parameters for canine SMTs