Publications by authors named "Barbara Brunetti"

36 Publications

Fibroblast-associated protein-α expression and BPV nucleic acid distribution in equine sarcoids.

Vet Pathol 2021 Jun 15:3009858211022696. Epub 2021 Jun 15.

Department of Veterinary Medical Sciences, 9296University of Bologna, Italy.

Sarcoids are the most common cutaneous tumor of equids and are caused by bovine papillomavirus (BPV). Different clinical subtypes of sarcoids are well characterized clinically but not histologically, and it is not known whether viral activity influences the clinical or histological appearance of the tumors. The aim of this study was to verify whether the development of different clinical types of sarcoids or the presence of certain histological features were associated with BPV distribution within the tumor. The presence of BPV was assessed by polymerase chain reaction (PCR) and visualized in histological sections by chromogenic in situ hybridization (CISH) in 74 equine sarcoids. Furthermore, to better characterize the molecular features of neoplastic cells, immunohistochemistry for S100, smooth muscle actin-α (αSMA), and fibroblast-associated protein-α (FAPα) was performed. The presence of BPV was confirmed in all tissues examined by either or both PCR and CISH (72/74, 97% each). Of 70/74 CISH-positive cases, signal distribution appeared as either diffuse (61/70, 87%) or subepithelial (9/70, 13%); the latter was more frequently observed in the verrucous subtype. However, no statistically significant association was found between clinical subtypes and specific histological features or hybridization pattern. Moreover, CISH signal for BPV was not detected in the epidermis overlying sarcoids nor in the tissue surrounding the neoplasms. By immunohistochemistry, αSMA confirmed the myofibroblastic differentiation of neoplastic cells in 28/74 (38%) sarcoids. Using tissue microarrays, FAPα labelling was observed in neoplastic fibroblasts of all sarcoids, suggesting this marker as a potential candidate for the immunohistochemical diagnosis of sarcoids.
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http://dx.doi.org/10.1177/03009858211022696DOI Listing
June 2021

Epidemiologic case investigation on the zoonotic transmission of Staphylococcus aureus infection from goat to veterinarians.

Zoonoses Public Health 2021 May 5. Epub 2021 May 5.

Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.

Staphylococcus aureus infection led to a case of goat abortion, and four veterinarians contracted S. aureus infection from the goat during and after the abortion. Three veterinarians assisted a doe during the dystocic delivery of a dead foetus. Seventy-two hours after the dystocia, which ended with the goat's death, the veterinarians who assisted during the kidding and the veterinarian who performed the necropsy showed the presence of multiple, isolated, painful pustules 1-5 mm in diameter located along their forearms and knees. S. aureus was isolated from the pustules of the veterinarians, the placenta and uterus of the goat, the organs (brain, thymus gland, abomasum, liver and spleen) of the foetus, the scrotum and eye swabs of the buck, and mammary pustules of another goat from the same herd. Histological analysis revealed purulent metritis and inflammation of the placental cotyledons. Additional investigations eliminated the chances of other infections. S. aureus isolates recovered from the veterinarians, goats, foetus and buck were sensitive to the tested anti-microbials and did not encode staphylococcal enterotoxin genes (sea, ser, sep, see, seg and sei). The isolates were closely related, as indicated by the results of Fourier-transform infrared spectroscopy and comparative whole-genome sequencing analysis. The results of this study clearly support the hypothesis that an episode of professional zoonosis was caused by S. aureus infection during the abortion and also highlight the need for bacterial subtyping in epidemiological surveys.
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http://dx.doi.org/10.1111/zph.12836DOI Listing
May 2021

Immunohistochemical Screening of HER2 in Canine Carcinomas: A Preliminary Study.

Animals (Basel) 2021 Apr 3;11(4). Epub 2021 Apr 3.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, 40064 Bologna, Italy.

HER2 overexpression has been reported in various human and canine tumours. The aim of this study is to investigate the expression of HER2 protein in different histotypes of canine carcinomas in order to identify potential tumours that could benefit from the HER2-targeted therapy. Eighty-two (82) canine carcinomas (squamous cell, gastro-intestinal, rectal, pulmonary, prostatic, urothelial, and ovarian) from paraffin-embedded samp les were immunohistochemically evaluated. The degree of HER2 expression was scored based on the ASCO/CAP 2018 guidelines. Intestinal carcinomas were those with greater HER2 overexpression (3+) with 81% of positive cases, followed by 42% of rectal carcinomas and 28% of squamous cell carcinomas. These observations suggest that HER2 overexpression could be a driver in the oncogenesis of several types of canine carcinomas and lay the foundations for the identification of different types of canine carcinomas that could benefit from HER2-targeted therapy.
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http://dx.doi.org/10.3390/ani11041006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8065471PMC
April 2021

The Site of Origin of Canine Abdominal Masses Correlates with the Risk of Malignancy: Retrospective Study of 123 Cases.

Animals (Basel) 2021 Mar 30;11(4). Epub 2021 Mar 30.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, 40064 Bologna, Italy.

The detection of an abdominal mass represents a common finding in clinical practice. The aim of this study was to retrospectively describe the tissue distribution and diagnosis of abdominal masses amenable to surgical removal in a canine population. Dogs with abdominal masses with a minimum diameter of 3 cm were selected. Cases were classified, based on the anatomical location, as splenic, gastrointestinal, hepatobiliary, genital, and masses not associated with any organ. Masses were surgically removed and formalin-fixed for the histological examination. Collected data were statistically analyzed. A total of 123 masses were collected from 122 dogs. Sixty-nine masses were classified as malignant neoplasia, 15 as benign, and 39 as non-neoplastic. The abdominal masses were 5.8-fold more likely to be malignant if located in the gastrointestinal tract ( = 0.01). A significant association between the size and the site of the masses was identified, the masses not associated with any organ being larger than the genital and splenic lesions ( = 0.008). This case series describes the most frequent location in association with the histopathological diagnosis of canine abdominal masses and suggests that the gastrointestinal location was related to a higher risk of representing a malignant neoplasm.
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http://dx.doi.org/10.3390/ani11040962DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8065481PMC
March 2021

High Intrinsic Expression of P-glycoprotein and Breast Cancer Resistance Protein in Canine Mammary Carcinomas Regardless of Immunophenotype and Outcome.

Animals (Basel) 2021 Mar 2;11(3). Epub 2021 Mar 2.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, 40064 Bologna, Italy.

P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are major actors in multidrug resistance (MDR) phenomenon in both human and canine mammary carcinomas (CMCs). The aim of this study was to investigate an association between the intrinsic expression of P-gp and BCRP compared to the immunophenotypes and outcome in CMCs. Fifty CMCs were evaluated at immunohistochemistry (IHC) for P-gp, BCRP, Estrogen receptor alpha (ER), Progesterone receptors (PR), Human Epidermal Growth Factor Receptor type 2 (HER2), basal cytokeratins 5/6 (CK5/6), Epidermal Growth Factor Receptor 1 (EGFR), and Ki67 proliferation index. P-gp and BCRP positive cases were, respectively, 52% and 74.5%, with a significantly higher expression of BCRP than P-gp. Five immunophenotypes were defined in 37 out of 50 CMCs: 9 (24.3%) Luminal A, 5 (13.5%) Luminal B, 9 (24.3%) HER2 overexpressing, 9 (24.3%) Triple-negative basal-like, and 5 (13.5%) Triple-negative non-basal-like. In all CMCs at least one marker was expressed. Follow-up data were available for 25 animals. The average cancer-specific survival was 739 ± 444 days. A number of CMCs bear a high expression of P-gp and BCRP but no significant association was found between their expression and the immunophenotypes, Ki67 index, the histological grade, and tumor-related death.
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http://dx.doi.org/10.3390/ani11030658DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8001331PMC
March 2021

Unusual Myoid Differentiation in a Canine Benign Mixed Mammary Tumour.

Case Rep Vet Med 2021 23;2021:6615256. Epub 2021 Jan 23.

Department of Veterinary Medical Sciences, University of Bologna, Italy.

This report describes an unusual mesenchymal differentiation in a canine benign mixed mammary tumour. A 13-year-old crossbreed female dog was submitted to surgery to remove an inguinal mammary nodule. The tumour was composed of mammary epithelium and mesenchymal populations, not only of cartilage and bone but also of myoid cells. PTAH demonstrated cross striation of striated muscle, and immunohistochemistry highlighted striated muscle expressing desmin and calponin, and smooth muscle expressing desmin, SMA, and calponin. The tumour was diagnosed as a benign mixed tumour with leio- and rhabdomyoid differentiation. There was no tumour recurrence after one year of clinical follow-up. In conclusion, the well-differentiated features of leiomyocytes and rhabdomyocytes and the growth pattern define the benign origin of the mesenchymal component, which has been confirmed by a benign outcome; therefore, the knowledge of this kind of differentiation is helpful to avoid misdiagnoses.
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http://dx.doi.org/10.1155/2021/6615256DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7850841PMC
January 2021

Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines.

BMC Vet Res 2021 Jan 18;17(1):30. Epub 2021 Jan 18.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, Italy.

Background: Doxorubicin (DOX) is widely used in both human and veterinary oncology although the onset of multidrug resistance (MDR) in neoplastic cells often leads to chemotherapy failure. Better understanding of the cellular mechanisms that circumvent chemotherapy efficacy is paramount. The aim of this study was to investigate the response of two canine mammary tumour cell lines, CIPp from a primary tumour and CIPm, from its lymph node metastasis, to exposure to EC50 DOX at 12, 24 and 48 h of treatment. We assessed the uptake and subcellular distribution of DOX, the expression and function of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP), two important MDR mediators. To better understand this phenomenon the effects of DOX on the cell cycle and Ki67 cell proliferation index and the expression of p53 and telomerase reverse transcriptase (TERT) were also evaluated by immunocytochemistry (ICC).

Results: Both cell lines were able to uptake DOX within the nucleus at 3 h treatment while at 48 h DOX was absent from the intracellular compartment (assessed by fluorescence microscope) in all the surviving cells. CIPm, originated from the metastatic tumour, were more efficient in extruding P-gp substrates. By ICC and qRT-PCR an overall increase in both P-gp and BCRP were observed at 48 h of EC50 DOX treatment in both cell lines and were associated with a striking increase in the percentage of p53 and TERT expressing cells by ICC. The cell proliferation fraction was decreased at 48 h in both cell lines and cell cycle analysis showed a DOX-induced arrest in the S phase for CIPp, while CIPm had an increase in cellular death without arrest. Both cells lines were therefore composed by a fraction of cells sensible to DOX that underwent apoptosis/necrosis.

Conclusions: DOX administration results in interlinked modifications in the cellular population including a substantial effect on the cell cycle, in particular arrest in the S phase for CIPp and the selection of a subpopulation of neoplastic cells bearing MDR phenotype characterized by P-gp and BCRP expression, TERT activation, p53 accumulation and decrease in the proliferating fraction. Important information is given for understanding the dynamic and mechanisms of the onset of drug resistance in a neoplastic cell population.
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http://dx.doi.org/10.1186/s12917-020-02709-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7814552PMC
January 2021

HER2 Overexpression and Amplification in Feline Pulmonary Carcinoma.

Vet Pathol 2021 May 19;58(3):527-530. Epub 2021 Jan 19.

9296University of Bologna, Bologna, Italy.

HER2 is overexpressed, amplified, and mutated in a subset of human lung cancer. The aim of this study was to investigate HER2 protein overexpression and gene amplification in feline pulmonary carcinomas. Thirteen pulmonary carcinomas were selected and TTF-1 and HER2 expression was evaluated by immunohistochemistry. Fluorescence in situ hybridization (FISH) was performed with a probe and a BAC probe for the feline chromosome E1p1.12-p1.11 region. Twelve adenocarcinomas and 1 squamous cell carcinoma were diagnosed. TTF-1 was positive in 7 carcinomas (58%). HER2 was overexpressed in 2 (15%), equivocal in 5 (38%), and negative in 6 cases (46%). FISH analysis of was indeterminate in 2 cases. Three pulmonary carcinomas (27%) had amplification and 8 cases were not amplified (73%). The significant correlation between HER2 protein overexpression and gene amplification are promising preliminary data, but study of additional cases is needed to confirm HER2 as a target for possible innovative treatments.
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http://dx.doi.org/10.1177/0300985820988147DOI Listing
May 2021

p53, ER, and Ki67 Expression in Canine Mammary Carcinomas and Correlation With Pathological Variables and Prognosis.

Vet Pathol 2021 Mar 19;58(2):325-331. Epub 2020 Nov 19.

9296University of Bologna, Bologna, Italy.

Using immunohistochemistry, 170 canine mammary carcinomas were evaluated for p53, ER (estrogen receptor), and Ki67. Of the 170 tumors, 89 were grade I (52.3%), 36 were grade II (21.2%), and 45 were grade III (26.4%). Eight cases (0.5%) were positive for p53 and 69/170 cases (40.5%) were positive for ER. Ki67 values were 24 ± 18% (mean ± SD). Using a cutoff value of 33.3% Ki67-positive neoplastic nuclei, 38/159 (23.8%) were classified as high proliferative and 121/159 (76.2%) as low proliferative. p53-positive cases had significantly higher Ki67 expression and higher histological grade. ER expression was not correlated with p53 expression but was significantly related to low Ki67 values and low histological grade. Moreover, ER-positive cases had significantly longer survival compared to ER-negative tumors, and ER expression had better correlation with tumor-related survival than histological grade. In summary, p53 accumulated in a small subset of canine mammary tumors and was associated with higher proliferative activity and higher histological grade. ER expression was confirmed as a differentiation marker associated with more favorable prognosis and biological behavior. The combined use of these 3 markers could be used in addition to histological grade to predict the biological behavior of canine mammary carcinomas.
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http://dx.doi.org/10.1177/0300985820973462DOI Listing
March 2021

Canine Mammary Carcinoma With Vacuolated Cytoplasm: Glycogen-Rich Carcinoma, a Histological Type Distinct From Lipid-Rich Carcinoma.

Vet Pathol 2021 01 18;58(1):63-70. Epub 2020 Nov 18.

9296University of Bologna, Bologna, Italy.

Lipid-rich carcinoma is a rare histotype of canine mammary tumors with cytoplasmic vacuolation. In humans, glycogen-rich carcinoma, secretory carcinoma, and myoepithelial neoplasms are included in the differential diagnosis for lipid-rich carcinoma. The aim of the study was to investigate the existence of histotypes other than lipid-rich in canine mammary carcinomas with vacuolated cytoplasm using a diagnostic algorithm based on histopathology, histochemistry, immunohistochemistry, and ultrastructure and to evaluate the molecular phenotype of these neoplasms. Ten mammary carcinomas were collected, histologically reviewed, and subjected to histochemistry (PAS, PAS with diastase, Alcian blue, Sudan III [1 case], and Congo red [1 case]); immunohistochemistry for CK19, CK5/6, CK14, p63, calponin, vimentin, ER, PR, and HER2; and transmission electron microscopy (TEM). Cytokeratin immunolabeling demonstrated the epithelial origin of all tumors. Sudan III and TEM confirmed the diagnosis of lipid-rich carcinoma in 8 tumors (one amyloid-producing). One tumor was reclassified as a glycogen-rich carcinoma based on PAS reactivity that was diastase-labile, and a second tumor was reclassified as a carcinoma-and-malignant myoepithelioma based on the differentiation markers. Lipid-rich carcinomas were basal-like (5/8), null-type (2/8), and luminal A phenotype (1/8). The glycogen-rich carcinoma was basal-like, while the carcinoma-and-malignant myoepithelioma was luminal A. Vacuolated morphology of neoplastic cells in canine mammary carcinoma can indicate either a neoplasm of luminal epithelial origin with cytoplasmic lipid or glycogen, or vacuolated neoplastic suprabasal myoepithelial cells. Glycogen-rich carcinoma is a novel histological type that should be considered in the differential diagnosis for canine mammary carcinomas with vacuolated cytoplasm.
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http://dx.doi.org/10.1177/0300985820969962DOI Listing
January 2021

X-Linked Duchenne-Type Muscular Dystrophy in Jack Russell Terrier Associated with a Partial Deletion of the Canine Gene.

Genes (Basel) 2020 10 8;11(10). Epub 2020 Oct 8.

Institute of Genetics, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.

A 9-month old male Jack Russell Terrier started showing paraparesis of the hindlimbs after a walk. Hospitalized, the dog went into cardiac arrest, and later died. Necroscopic examination revealed a severe thickness of the diaphragm, esophagus, and base of the tongue, leading to the diagnosis of muscular dystrophy. The histology confirmed the marked size variation, regeneration, and fibrosis replacement of the skeletal muscle fibers. Immunohistochemistry demonstrated the absence of dystrophin confirming the diagnosis. Transmission electron microscopy showed disarrangement of skeletal muscle fibers. Finally, whole-genome sequencing identified a ~368kb deletion spanning 19 exons of the canine dystrophin () gene. This pathogenic loss-of-function variant most likely explains the observed disease phenotype. The X-chromosomal variant was absent in seven controls of the same breed. Most likely, this partial deletion of the gene was either transmitted on the maternal path within the family of the affected dog or arose de novo. This study revealed a spontaneous partial deletion in gene in a Jack Russell Terrier showing a Duchenne-type muscular dystrophy due to non-functional dystrophin.
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http://dx.doi.org/10.3390/genes11101175DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7600251PMC
October 2020

P-Glycoprotein and Breast Cancer Resistance Protein in Canine Inflammatory and Noninflammatory Grade III Mammary Carcinomas.

Vet Pathol 2019 11 16;56(6):840-847. Epub 2019 Sep 16.

Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.

P-glycoprotein (P-gp/ABCB1) and breast cancer resistance protein (BCRP/ABCG2) expression are frequently related to multidrug resistance (MDR) in neoplastic cells. Canine inflammatory and grade III noninflammatory mammary carcinomas (IMC and non-IMC) are aggressive tumors that could benefit from chemotherapy. This study describes the immunohistochemical detection of P-gp and BCRP in 20 IMCs and 18 non-IMCs from dogs that had not received chemotherapy. Our aim was to determine if P-gp and BCRP expression was related to the "inflammatory" phenotype, to establish a basis for future studies analyzing the response to chemotherapy in dogs with highly malignant mammary cancer. Immunolabeling was primarily membranous for P-gp with a more intense labeling in emboli, and immunolabeling was membranous and cytoplasmic for BCRP. P-gp was expressed in 17 of 20 (85%) IMCs compared to 7 of 18 (39%) non-IMCs ( = 0.006). BCRP was expressed within emboli in 15 of 19 (79%) emboli in IMC, 12 of 15 (80%) primary IMCs, and 12 of 18 (67%) non-IMCs, without statistically significant differences ( > .05). All IMCs and 67% of non-IMCs expressed at least 1 of the 2 transporters, and 63% (12/19) of IMCs and 39% (7/18) of non-IMCs expressed both P-gp and BCRP. P-gp and BCRP evaluation might help select patients for chemotherapy. P-gp, expressed in a significantly higher percentage of IMCs vs non-IMCs, might play a specific role in the chemoresistance of IMC.
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http://dx.doi.org/10.1177/0300985819868647DOI Listing
November 2019

HER2 Amplification Status in Feline Mammary Carcinoma: A Tissue Microarray-Fluorescence In Situ Hydridization-Based Study.

Vet Pathol 2019 Mar 1;56(2):230-238. Epub 2018 Nov 1.

1 Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.

Human epidermal growth factor receptor 2 (HER2) is a tyrosine kinase receptor overexpressed in a subset of breast cancer due to HER2 gene amplification. HER2 protein is expressed in feline mammary carcinomas, but little is known about its cytogenetic alterations. The aim of this study was to evaluate HER2 gene amplification status and its correlation with HER2 protein expression in feline mammary carcinomas. Feline mammary carcinomas were retrospectively selected and immunohistochemically (IHC) evaluated for HER2 protein expression. All the HER2 IHC-positive (3+) and equivocal (2+) cases and a subset of negative cases (0/1+) were selected for fluorescence in situ hybridization (FISH). Dual-core tissue microarrays were prepared for FISH. IHC and FISH were evaluated according to the 2013 American Society of Clinical Oncology/College of American Pathologists guidelines. The study included 107 feline mammary carcinomas from 88 queens. HER2 protein expression was positive (3+) in 7 cases (6.5%), equivocal (2+) in 48 cases (45%), and negative (0/1+) in 52 cases (48.5%). HER2 status was indeterminate in 8 feline mammary carcinomas (12%), amplified in 3 (4%), equivocal in 4 (6%), and nonamplified in 53 (78%). HER2 gene amplification and protein expression were significantly positively correlated ( R = 0.283; P < .0001). HER2 gene is amplified in a subset of feline mammary carcinomas despite the HER2 positive or equivocal protein expression, but it remains to be determined if the HER2 amplification is a gene alteration that drives mammary tumor carcinogenesis or only a bystander passenger mutation.
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http://dx.doi.org/10.1177/0300985818808531DOI Listing
March 2019

Glomeruloid Microvascular Proliferation, Desmoplasia, and High Proliferative Index as Potential Indicators of High Grade Canine Choroid Plexus Tumors.

Vet Pathol 2018 05 5;55(3):391-401. Epub 2018 Feb 5.

2 Division of Neurological Sciences, Department of Clinical Research and Veterinary Public Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland.

Choroid plexus tumors (CPT) are intraventricular neoplasms accounting for 10% of all primary central nervous system tumors in dogs. They are frequently classified according to the human WHO classification into choroid plexus papilloma (CPP, grade I), atypical CPP (aCPP, grade II), and choroid plexus carcinoma (CPC, grade III). Histological features observed in canine CPT such as increased vascular density (IVD) and glomeruloid microvascular proliferation (GMVP) are not part of the WHO classification. This multi-centric study aimed to investigate tumor-associated vascular hyperplasia in dogs by determining the prevalence of GMVP and IVD in 52 canine CPT and their association with tumor grade. In addition, the expression of angiogenic factors was assessed by immunohistochemistry in 25 tumors to investigate the pathogenesis of tumor-associated vascular hyperplasia. Based on the classical histological hallmarks, this study of 52 CPT identified 22 (42%) CPP (grade I) and 30 of (58%) CPC (grade III). GMVP was more prevalent in CPC (13/30; 43%) than CPP (1/22; 4%), whereas IVD occurred to a similar extent in CPP and CPC. Desmoplasia was more common in CPC (19/30; 63%) than CPP (2/22; 9%), and similarly, the proliferative index (PI) of neoplastic epithelium was significantly higher in CPC (5.14%) than CPP (0.94%). The majority of CPT expressed platelet-derived growth factor (PDGF), PDGFRα, PDGFRβ, and vascular endothelial growth factor (VEGF) irrespective of tumor grade or tumor-associated vascular hyperplasia. These results suggest that tumor-associated GMVP, desmoplasia, and PI may serve as histological indicators of malignancy in CPT.
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http://dx.doi.org/10.1177/0300985817754124DOI Listing
May 2018

Wearable Wireless Tyrosinase Bandage and Microneedle Sensors: Toward Melanoma Screening.

Adv Healthc Mater 2018 04 18;7(7):e1701264. Epub 2018 Jan 18.

Department of Nanoengineering, University of California, San Diego La Jolla, CA, 92093, USA.

Wearable bendable bandage-based sensor and a minimally invasive microneedle biosensor are described toward rapid screening of skin melanoma. These wearable electrochemical sensors are capable of detecting the presence of the tyrosinase (TYR) enzyme cancer biomarker in the presence of its catechol substrate, immobilized on the transducer surface. In the presence of the surface TYR biomarker, the immobilized catechol is rapidly converted to benzoquinone that is detected amperometrically, with a current signal proportional to the TYR level. The flexible epidermal bandage sensor relies on printing stress-enduring inks which display good resiliency against mechanical deformations, whereas the hollow microneedle device is filled with catechol-coated carbon paste for assessing tissue TYR levels. The bandage sensor can thus be used directly on the skin whereas microneedle device can reach melanoma tissues under the skin. Both wearable sensors are interfaced to an ultralight flexible electronic board, which transmits data wirelessly to a mobile device. The analytical performance of the resulting bandage and microneedle sensing systems are evaluated using TYR-containing agarose phantom gel and porcine skin. The new integrated conformal portable sensing platforms hold considerable promise for decentralized melanoma screening, and can be extended to the screening of other key biomarkers in skin moles.
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http://dx.doi.org/10.1002/adhm.201701264DOI Listing
April 2018

Is Serum Total LDH Evaluation Able to Differentiate between Alimentary Lymphoma and Inflammatory Bowel Disease in a Real World Clinical Setting?

PLoS One 2016 17;11(3):e0151641. Epub 2016 Mar 17.

Medical Imaging, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.

Context: An increase in enzyme lactate dehydrogenase (LDH) in serum is a negative prognostic factor for survival in cats affected by lymphoma. Measuring LDH at the time of diagnosis has been studied for differentiating neoplastic disease from non-neoplastic disease in dogs. Inflammatory bowel disease (IBD) and alimentary lymphoma are common diseases in cats.

Objective: The aim of this study was to determine whether elevation of total LDH occurred in cats with alimentary lymphoma and non-neoplastic gastrointestinal disease, such as IBD, and to evaluate whether this enzyme is useful in supporting the differential diagnosis of these specific diseases.

Materials And Methods: A prospective non-randomized controlled study was carried-out in a real world setting of three Italian private veterinary clinics. Seventy-one client-owned cats with a history of chronic gastrointestinal symptoms were enrolled; 33 cats were histologically diagnosed as having alimentary lymphoma and 38 cats as having IBD. Serum samples of total LDH analysis were measured.

Results: Gender (P = 0.016) and age (P = 0.046) were found to be significant factors influencing the differentiation of serum total LDH between cats with alimentary lymphoma and those with IBD. Despite low diagnostic accuracy in the overall population (63%), a cut-off value of serum total LDH ranging from 0.85- to 1.04-times the upper reference limit showed good capability (accuracy >80%) of differentiating these two conditions in neutered males and cats younger than 8 years of age (AUC: 0.805, 0.833; sensitivities: 76.9%, 83.3%; specificities: 80.0%, 76.5%; PPV: 76.9%, 55.6%; NPV: 80.0%, 92.9%; respectively).

Conclusions: Although our study showed that gender and age are significant factors in differentiating serum total LDH between cats with alimentary lymphoma and those with IBD, this test had poor diagnostic accuracy in differentiating between these two conditions in the overall population.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0151641PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4795755PMC
August 2016

Electrochemical signatures of multivitamin mixtures.

Analyst 2015 Nov;140(22):7522-6

Department of Nanoengineering, University of California San Diego, 9500 Gilman Drive #0448, La Jolla, CA 92093, USA.

The ability of cyclic square wave voltammetry to identify distinct fingerprints of multiple vitamins, in a single voltammetric run, is demonstrated. This method represents an efficient alternative to more common techniques for fast screening of complex vitamin mixtures or commercial tablets due to its low cost, high speed and sensitivity.
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http://dx.doi.org/10.1039/c5an01964hDOI Listing
November 2015

Gastric inflammatory pseudotumour secondary to Actinomyces hordeovulneris infection in a cat.

Ir Vet J 2015 26;69(1):12. Epub 2016 Aug 26.

Department of Veterinary Medical Sciences, School of Agriculture and Veterinary Medicine, Alma Mater Studiorum - University of Bologna, Ozzano dell'Emilia (BO), 40064 Italy.

Background: The authors report the first case of feline gastric actinomycosis associated with infection by Actinomyces hordeovulneris.

Case Presentation: A 4-year-old, neutered male, semi-feral European cat, with a 1 year history of chronic vomiting, was referred to the clinic. Abdominal ultrasound examination identified a hypoechoic focal transmural thickening with loss of normal wall layering and hyperechoic speckles at the gastric body. Initial gastroscopic examination showed a tumour-like gastric mass with an ulcerated depression at the level of the greater curvature. Histologic examination of endoscopic biopsy specimens was consistent with a severe lymphoplasmacytic gastritis. After 2 months, due to persistence of abdominal discomfort, surgical exploration and intraoperative sampling of gross abnormalities was recommended. Full thickness gastric wall biopsies, and fine needle aspiration of the gastric thickening and gastric lymph node, were performed. Histopathological examination identified a transmural pyogranulomatous gastritis. Aspirate samples of the gastric wall cultured positive, with colony morphology, biochemical testing and PCR of the 16 s rRNA gene compatible with Actinomyces hordeovulneris. After 4 months of treatment with cefovecin (8 mg/kg subcutaneously every 14 days), the vomiting completely resolved, as well as the ultrasonographic gastric alteration.

Conclusion: This case report of feline gastric actinomycosis, caused by Actinomyces hordeovulneris, suggests that gastric bacterial infection should be considered in cases of focal gastric wall thickening associated with chronic vomiting in the cat, which may otherwise closely resemble neoplastic disease. Once a diagnosis of actinomycosis was obtained, a correct treatment with antibiotic therapy can resolve it.
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http://dx.doi.org/10.1186/s13620-016-0071-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5002114PMC
August 2016

Deletion in the EVC2 gene causes chondrodysplastic dwarfism in Tyrolean Grey cattle.

PLoS One 2014 14;9(4):e94861. Epub 2014 Apr 14.

Institute of Genetics, Vetsuisse Faculty, University of Bern, Bern, Switzerland.

During the summer of 2013 seven Italian Tyrolean Grey calves were born with abnormally short limbs. Detailed clinical and pathological examination revealed similarities to chondrodysplastic dwarfism. Pedigree analysis showed a common founder, assuming autosomal monogenic recessive transmission of the defective allele. A positional cloning approach combining genome wide association and homozygosity mapping identified a single 1.6 Mb genomic region on BTA 6 that was associated with the disease. Whole genome re-sequencing of an affected calf revealed a single candidate causal mutation in the Ellis van Creveld syndrome 2 (EVC2) gene. This gene is known to be associated with chondrodysplastic dwarfism in Japanese Brown cattle, and dwarfism, abnormal nails and teeth, and dysostosis in humans with Ellis-van Creveld syndrome. Sanger sequencing confirmed the presence of a 2 bp deletion in exon 19 (c.2993_2994ACdel) that led to a premature stop codon in the coding sequence of bovine EVC2, and was concordant with the recessive pattern of inheritance in affected and carrier animals. This loss of function mutation confirms the important role of EVC2 in bone development. Genetic testing can now be used to eliminate this form of chondrodysplastic dwarfism from Tyrolean Grey cattle.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0094861PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3986253PMC
May 2015

Pathology in practice. Productive osteoblastic osteosarcoma with metastases in a guinea pig.

J Am Vet Med Assoc 2013 Sep;243(6):801-3

Department of Veterinary Medical Sciences, Division of Veterinary Pathology, University of Bologna, Italy.

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http://dx.doi.org/10.2460/javma.243.6.801DOI Listing
September 2013

Molecular portrait-based correlation between primary canine mammary tumor and its lymph node metastasis: possible prognostic-predictive models and/or stronghold for specific treatments?

BMC Vet Res 2012 Nov 12;8:219. Epub 2012 Nov 12.

Department of Veterinary Medical Sciences, University of Bologna, Via Tolaradi Sopra 50, Ozzano Emilia, Bologna 40064, Italy.

Background: This study aimed to evaluate the relationship between the molecular phenotype of the primary mammary tumor and its related lymph node metastasis in the dog to develop prognostic-predictive models and targeted therapeutic options.

Results: Twenty mammary tumor samples and their lymph node metastases were selected and stained by immunohistochemistry with anti-estrogen receptor (ER), -progesterone receptor (PR), -human epidermal growth factor receptor 2 (c-erbB-2), -cytokeratin 5/6 (CK 5/6), -cytokeratin 14 (CK14), -cytokeratin 19 (CK 19) and -protein 63 (p63) antibodies. Four phenotypes (luminal A, luminal B, c-erbB2 overexpressing and basal-like) were diagnosed in primary tumors and five (luminal A, luminal B, c-erbB-2 overexpressing, basal-like and normal-like) in the lymph node metastases. Phenotypic concordance was found in 13 of the 20 cases (65%), and seven cases (35%) showed discordance with different lymph node phenotypic profile from the primary tumor.

Conclusions: The phenotype of the primary tumor assumes a predictive-therapeutic role only in concordant cases, meaning that both the primary tumor and its lymph node metastasis should be evaluated at the same time. A treatment plan based only on the primary tumor phenotype could lead to therapeutic failures if the phenotype of the lymph node metastasis differs from that of the primary tumor.
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http://dx.doi.org/10.1186/1746-6148-8-219DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3538651PMC
November 2012

Morphology of the myoepithelial cell: immunohistochemical characterization from resting to motile phase.

ScientificWorldJournal 2012 5;2012:252034. Epub 2012 Aug 5.

Department of Veterinary Medical Science, University of Bologna, 40064 Ozzano Emilia, Italy.

Myoepithelium is present in canine mammary tumors as resting and proliferative suprabasal and spindle and stellate interstitial cells. The aim of this paper was to evaluate a panel of markers for the identification of four different myoepithelial cell morphological types in the normal and neoplastic mammary gland and to investigate immunohistochemical changes from an epithelial to a mesenchymal phenotype. Cytokeratin 19 (CK19), cytokeratin 5/6 (CK5/6), cytokeratin 14 (CK14), estrogen receptor (ER), p63 protein, vimentin (VIM), and α-smooth muscle actin (Alpha-SMA) antibodies were used on 29 neoplasms (3 benign and 3 malignant myoepithelial tumors, 7 carcinomas in benign-mixed tumors and 16 complex carcinomas) and on normal tissue of mammary glands. All these antibodies were also tested on 3 mammary tissues from animals with no mammary pathology. The myoepithelial markers were well expressed in the suprabasal cells and gradually lost in the motile types, with the stellate cells maintaining only VIM expression typical of mesenchyma. ER labeled some resting and motile myoepithelial cells. On the basis of our results, we propose a transition from myoepithelial immotile cells into migratory fibroblast-like cells. This transition and the characterization of an immunohistochemical panel for resting and motile myoepithelial cells shed more light on the biological behavior of myoepithelial cells.
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http://dx.doi.org/10.1100/2012/252034DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3420080PMC
March 2013

Comparative spatiotemporal analysis of the intrathecal immune response in natural listeric rhombencephalitis of cattle and small ruminants.

Comp Immunol Microbiol Infect Dis 2012 Sep 24;35(5):429-41. Epub 2012 Apr 24.

Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Switzerland.

This study examined the spatiotemporal immune response in listeric rhombencephalitis of ruminants in situ. Our data support the view that astrocytes facilitate the containment of infectious lesions. Results on the natural disease recapitulate observations in experimental rodent models and suggest that the mounted adaptive lymphocytic response of ruminants is effective in eliminating Listeria monocytogenes (LM). However, our data indicate earlier participation of the adaptive immune response, a stronger B lymphocyte contribution and a more protracted macrophage infiltration in the natural disease than it has been deduced from experimental models. Therefore, such models should be complemented by studies in natural host systems. Various macrophage and microglia subsets are involved in listeric rhombencephalitis and their differential contribution may account for species differences in clinical course and outcome of infection as might species differences in the B-cell response. Future functional ex vivo and in vitro studies are necessary to further investigate the findings obtained in the present study.
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http://dx.doi.org/10.1016/j.cimid.2012.03.009DOI Listing
September 2012

Uncertainty of measurement and conformity assessment: a review.

Anal Bioanal Chem 2011 Jun 25;400(6):1729-41. Epub 2011 Feb 25.

DiSTAM, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy.

The uncertainty of measurement is the key indicator of the quality of any experimental result. Proper consideration of this uncertainty is imperative when testing a sample against legal/compositional limits. This task can be quite challenging when the entity measured in the investigated sample is so close to the limit that its uncertainty, however estimated, critically affects decision-making. This explains the many literature contributions discussing the problem. Even though some of the most authoritative organisations have issued specific guidelines aimed at assisting the staff involved in such measurements, several aspects of conformity testing are still debated in the literature. In this review, after a short outline of existing information, somewhat more detailed insight is given into the guidelines of ASME, ISO, and Eurachem/CITAC, because they are the most useful tools for operators of testing and calibration laboratories. Some aspects of Council Directive 96/23/EC are also discussed. Insight into the contents of the mentioned documents enables emphasis of analogies and discrepancies.
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http://dx.doi.org/10.1007/s00216-011-4776-yDOI Listing
June 2011

Seminoma in a koi carp Cyprinus carpio: histopathological and immunohistochemical findings.

Dis Aquat Organ 2010 Oct;92(1):83-8

Servizio di Prova di Anatomia Patologica, Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale, Facoltà di Medicina Veterinaria, Alma Mater Studiorum Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.

A spontaneous seminoma in a 3 yr old male koi carp Cyprinus carpio L. is described. The animal, presenting a symmetric abdominal enlargement, showed a celomatic multinodular, white-yellowish and firm mass that infiltrated the liver and the intestine wall. Histologically, the neoplasm was non-encapsulated and poorly demarcated, showed invasive growth and was characterized by a lobular architecture, subdivided by abundant fibro-connective septa. Large necrotic and calcified areas together with small aggregates of residual spermatids were present. We diagnosed a classical seminoma with a diffuse pattern. Neoplastic cells cross-reacted with vimentin, placental alkaline phosphatase, and c-KIT. An immunohistochemical phenotypization of the tumor was performed to exclude other celomatic neoplasms and to compare this seminoma with those reported in mammals and humans.
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http://dx.doi.org/10.3354/dao02273DOI Listing
October 2010

About estimating the limit of detection of heteroscedastic analytical systems.

Anal Chim Acta 2009 Nov 30;655(1-2):30-7. Epub 2009 Sep 30.

Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Università degli Studi di Milano, Via Celoria 2, 20133 Milano (I), Italy.

The limit of detection is a fundamental figure of merit in chemical analysis. Many different approaches are available for its estimation, but only few allow analysing data characterized by a significant change of precision with concentration. Among these, the ISO 11843-2 approach is certainly the most suitable. However, its implementation may not look easy to operators needing to develop the necessary spreadsheets by commercial software packages. This paper is aimed to verifying if alternative approaches are available for getting acceptably approximate estimates of the limit of detection (LOD). Pragmatically, they were set-up by simply adapting some of the approaches already available in the literature, but compatible only with homoscedastic analytical systems. The results obtained by these alternative approaches when analysing a series of calibrations relevant to the electroanalytical determination of hexavalent chromium in some water samples were compared to those furnished by the ISO approach. The results allowed confirming the critical role of the numerosity of the available data on the consistency of any estimate. Even when using the ISO approach, reliable estimates were only obtained by performing a number of calibrations usually uncommon under standard routine conditions. In such a situation, the differences between the results obtained by all the examined approaches appear less important.
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http://dx.doi.org/10.1016/j.aca.2009.09.036DOI Listing
November 2009

Quantitative analysis of telomerase in feline mammary tissues.

J Vet Diagn Invest 2009 May;21(3):369-73

Department of Veterinary Public Health and Animal Pathology, Section of General Pathology and Pathologic Anatomy, Faculty of Veterinary Medicine, University of Bologna, Via Tolara di Sopra, Bologna, Italy.

The purpose of this study was to validate immunohistochemistry (IHC) as an alternative to telomerase repeat amplification protocol (TRAP) analysis to detect telomerase activity. TRAP-enzyme-linked immunosorbent assay (ELISA) reactivity was compared with telomerase reverse transcription (TERT) IHC staining in 22 feline mammary tissues (6 normal mammary glands, 2 dysplastic mammary glands, 1 fibroadenoma, and 13 malignant neoplasms [6 solid mammary carcinomas, 2 squamous-cell carcinomas, 4 tubulopapillary mammary carcinomas, and 1 mammary carcinosarcoma]). TERT IHC staining revealed enzymatic expression in nuclear, nucleolar, cytoplasmic, and combined nuclear and nucleolar staining patterns that were separately quantified by image analysis and expressed as the absolute number (average) of positive cells or percentage of positive cells with respect to overall cellularity. With TERT IHC staining, the absolute number and percentage of cells with positive nuclei and nucleoli within the same cell were the variables with the greatest discrimination between benign and malignant mammary lesions (analysis of variance [ANOVA], average P < 0.0001; percentage P < 0.001). For TRAP-ELISA-positive versus TRAP-ELISA-negative tissues, a positive test result provided greater differentiation between malignant versus benign mammary lesions (ANOVA, average P = 0.00038; percentage P = 0.0022). The same IHC pattern of expression showed a proportional and significant (average P = 0.004; percentage P = 0.002) but low (average R = 0.60; percentage R = 0.63) correlation with TRAP-ELISA by the Pearson test. The correlation coefficients obtained show that IHC and TRAP cannot be considered interchangeable because the 2 methods are more complementary than exclusive.
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http://dx.doi.org/10.1177/104063870902100312DOI Listing
May 2009

Immunohistochemical characterization of mammary squamous cell carcinoma of the dog.

J Vet Diagn Invest 2008 Nov;20(6):766-73

Department of Veterinary Public Health and Animal Pathology, Division of Veterinary Pathology, Via Tolara di Sopra 50, Ozzano dell'Emilia, Bologna, Italy.

Squamous cell carcinoma of the mammary gland is rare in both veterinary and human medicine. Whereas human metaplastic and squamous variants are known, the objectives of the current study were to ascertain the presence of such entities in canine mammary tumors and to distinguish them from other (epidermal, sweat gland) squamous tumors that may develop in the same area. A panel of antibodies (anti-cytokeratin [CK] 19, CK 14, CK 5/6, pancytokeratin, and vimentin) was used on 18 mammary gland malignancies with squamous features and 16 malignant skin tumors (11 squamous cell carcinomas of the skin and 5 sweat glands). Fifteen of the 18 mammary carcinomas were classified as metaplastic carcinomas, and the remaining 3 were classified as squamous cell carcinomas. The 2 most useful markers to establish the histogenesis of mammary tumors were pancytokeratin and CK 19. All other antibodies were equally expressed (CK 14 and 5/6) in all histotypes. The antibody panel discriminated primary epidermal squamous tumors (pancytokeratin positive and CK 19 negative) from gland-derived squamous neoplasms (pancytokeratin positive and CK 19 positive) but failed to distinguish primary mammary tumors from other squamous tumors of glandular origin.
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http://dx.doi.org/10.1177/104063870802000608DOI Listing
November 2008

Amperometric detection of carbohydrates and thiols by using a glassy carbon electrode coated with Co oxide/multi-wall carbon nanotubes catalytic system.

Talanta 2008 Jul 28;76(2):454-7. Epub 2008 Mar 28.

Department of Food Science and Technology, University of Milan, Via Celoria 2, 20133 Milan, Italy.

A glassy carbon electrode coated with cobalt oxide/multi-wall carbon nanotubes (MWCNT) system was used for the detection of carbohydrates and thiols. The modification of the glassy carbon electrode increased the anodic current response of these organic compounds and decreased their overvoltage. The amperometric responses were extremely stable with no loss of sensitivity over many days of storage. Such attractive performance characteristics indicate great promise for using this new catalytic system for monitoring in fast and simple way compounds of great interest for food industry, biotechnology and clinical diagnostics.
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http://dx.doi.org/10.1016/j.talanta.2008.03.031DOI Listing
July 2008