Publications by authors named "Serenella Papparella"

30 Publications

  • Page 1 of 1

spp.-Infected Dogs Have Circulating Anti-Skeletal Muscle Autoantibodies Recognizing SERCA1.

Pathogens 2021 Apr 12;10(4). Epub 2021 Apr 12.

Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, 80137 Naples, Italy.

spp. infection is associated with an inflammatory myopathy (IM) in dogs. The pathomechanism underlying this disorder is still elusive, however, the pattern of cellular infiltration and MHC I and II upregulation indicate an immune-mediated myositis. This study aimed to investigate the presence of autoantibodies targeting the skeletal muscle in sera of leishmania-infected dogs and individuate the major autoantigen. We tested sera from 35 leishmania-infected dogs and sera from 10 negative controls for the presence of circulating autoantibodies with indirect immunofluorescence. Immunoblot and mass spectrometry were used to identify the main target autoantigen. Immunocolocalization and immunoblot on immunoprecipitated muscle proteins were performed to confirm the individuated major autoantigen. We identified circulating autoantibodies that recognize skeletal muscle antigen(s) in sera of leishmania-infected dogs. The major antigen was identified as the sarcoplasmic/endoplasmic reticulum Ca-ATPase 1 (SERCA1). We also found that canine SERCA1 presents several identical traits to the calcium-translocating P-type ATPase of . In the present study, we defined circulating anti-SERCA1 autoantibodies as part of the pathogenesis of the leishmania-associated IM in dogs. Based on our data, we hypothesize that antigen mimicry is the mechanism underlying the production of these autoantibodies in leishmania-infected dogs.
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http://dx.doi.org/10.3390/pathogens10040463DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8070147PMC
April 2021

Implication of the NLRP3 Inflammasome in Bovine Age-Related Sarcopenia.

Int J Mol Sci 2021 Mar 30;22(7). Epub 2021 Mar 30.

Department of Veterinary Medicine and Animal Production, Unit of Pathology, University of Naples "Federico II", 80137 Naples, Italy.

Sarcopenia is defined as the age-related loss of skeletal muscle mass, quality, and strength. The pathophysiological mechanisms underlying sarcopenia are still not completely understood. The aim of this work was to evaluate, for the first time, the expression of NLRP3 inflammasome in bovine skeletal muscle in order to investigate the hypothesis that inflammasome activation may trigger and sustain a pro-inflammatory environment leading to sarcopenia. Samples of skeletal muscle were collected from 60 cattle belonging to three age-based groups. Morphologic, immunohistochemical and molecular analysis were performed to assess the presence of age-related pathologic changes and chronic inflammation, the expression of NLRP3 inflammasome and to determine the levels of interleukin-1β, interleukin-18 and tumor necrosis factor alpha in muscle tissue. Our results revealed the presence of morphologic sarcopenia hallmark, chronic lymphocytic inflammation and a type II fibers-selective NLRP3 expression associated to a significant decreased number of immunolabeled-fibers in aged animals. Moreover, we found a statistically significant age-related increase of pro-inflammatory cytokines such as interleukin-1β and interleukin-18 suggesting the activation of NLRP3 inflammasome. Taken together, our data suggest that NLRP3 inflammasome components may be normally expressed in skeletal muscle, but its priming and activation during aging may contribute to enhance a pro-inflammatory environment altering normal muscular anabolism and metabolism.
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http://dx.doi.org/10.3390/ijms22073609DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036417PMC
March 2021

Immunopathological Response, Histological Changes, Parasitic Burden, and Egg Output in Sheep Naturally Infected by .

Animals (Basel) 2021 Feb 19;11(2). Epub 2021 Feb 19.

Department of Veterinary Medicine and Animal Production, Unit of Pathology, University of Naples "Federico II", 80137 Naples, Italy.

The aim of this study was to investigate the correlation between infection by (class Trematoda) and the animal host response in terms of macroscopic lesions, the immunopathological response, and histological changes in the livers of naturally infected sheep. Twenty-four sheep were selected on the basis of positive fecal egg counts (FECs). Gross and histological injuries were scored. A positive significant association was observed between the number of adult worms recovered from the liver, FEC, macroscopic lesions, fibrosis, and bile duct hyperplasia. A significant negative association was observed among these variables and the degree of leukocyte infiltration. In addition, immunophenotyping of the inflammatory cells was carried out using primary antibodies against T cell epitopes (CD3+, CD4+, and CD8+), B cell epitopes (CD79α), and the ionized calcium-binding adapter molecule 1 (IBA-1) antigen. Independently of the severity of the infection, the predominant cell population was CD3-positive and associated with lesser numbers of CD79α- and Iba-I-positive cells. An increase in Iba-1-positive cells was observed in the livers of animals with a high worm burden. Our results provide a reference basis to better understand the local immune response in sheep naturally infected by in relation to the FEC and parasitic burden.
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http://dx.doi.org/10.3390/ani11020546DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7923235PMC
February 2021

RNAScope Hybridization as a Novel Technique for the Assessment of c-KIT mRNA Expression in Canine Mast Cell Tumor.

Front Vet Sci 2021 16;8:591961. Epub 2021 Feb 16.

Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Naples, Italy.

RNA is considered as an indicator of the dynamic genetic expression changes in a cell. RNAScope is a commercially available hybridization assay for the detection of RNA in formalin-fixed paraffin-embedded tissue. In this work, we describe the use of RNAScope as a sensitive and specific method for the evaluation of messenger RNA (mRNA) in canine mast cell tumor. We investigated the expression of mRNA with RNAscope in 60 canine mast cell tumors (MCTs), comparing it with the histological grade and KIT immunohistochemical expression patterns. Our results showed an overall good expression of mRNA in neoplastic cells if compared with control probes. We also observed a statistically significant correlation between histological grade and mRNA expression. No correlations were found between KIT protein immunohistochemical distribution pattern and mRNA expression or histological grade. Our results provide a reference basis to better understand mRNA expression in canine MCTs and strongly encourage further studies that may provide useful information about its potential and significant role as a prognostic and predictive biological marker for canine MCTs clinical outcome.
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http://dx.doi.org/10.3389/fvets.2021.591961DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7921150PMC
February 2021

The correlation between cardiac and skeletal muscle pathology in animal models of idiopathic inflammatory myopathies.

Acta Myol 2020 Dec 1;39(4):313-319. Epub 2020 Dec 1.

Department of Veterinary Medicine and animal production, Unit of Pathology. University of Naples "Federico II", Naples, Italy.

Idiopathic inflammatory myopathies (IIMs) represent a heterogeneous group of disorders in which skeletal muscle is inappropriately targeted by the immune system. IIMs are characterized by inflammation of muscle and varying degrees of muscle dysfunction. Extra-muscular manifestations may involve heart, skin, joints, lungs, and gastrointestinal tract. Cardiovascular involvement is a feared event because is one of the leading causes of mortality in IIM patients. As the myocardium shares many features with the skeletal muscle, it is supposed that it can be affected by the same inflammatory processes, which take place during the different forms of IIMs. However, the full extent of this link and the mechanisms behind it are still not fully understood. Animal models have greatly improved our understanding of IIM pathomechanisms and have proven to be a useful tool for discovering therapeutic drug targets. Here we report the evidence of heart muscle involvement in different animal models of spontaneous IIMs, assuming a common autoimmune mechanism and presenting them as study models for human pathology.
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http://dx.doi.org/10.36185/2532-1900-035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7783441PMC
December 2020

Pneumoperitoneum Modifies Serum and Tissue CCL2-CCL5 Expression in Mice.

JSLS 2020 Apr-Jun;24(2)

Department of Woman, Child, and General and Specialized Surgery, University of Campania "Luigi Vanvitelli," Naples, Italy.

Background And Objectives: Laparoscopy is the preferred method when operating in the abdomen. In this study, we evaluated systemic and morphological peritoneal cytokine modifications (RANTES/CCL5 and MCP-1/CCL2) due to CO pneumoperitoneum in rats.

Methods: Twenty-five prepubertal Sprague-Dawley rats were randomized into three groups. Pneumoperitoneum lasting 30 minutes, was induced with a flow of 0.5 L/min, in two groups (S1 and S2, n = 20), at a P/CO of 6 and 10 mm Hg, respectively. In the control group (C, n = 5), only anesthesia was carried out. All animals were sacrificed after 24 hours. The serum of the rats was collected for ELISA, and the levels of the cytokines RANTES and MCP-1 were investigated. An immunohistochemical analysis of RANTES and MCP-1 was performed on samples of the peritoneum, and the morphological evaluation was conducted with a blinded evaluation by two independent, experienced pathologists by using a grading system (0, 1+, 2+, 3+: no, faint, moderate, and strong reactivity, respectively).

Results: RANTES mean levels were significantly different in the S1, S2, and C groups (70.3 ± 2.26, 58.23 ± 4.32, 29.66 ± 4.03, respectively, = .0001). The levels of MCP-1 were 32.1 ± 1.63 in the S1 group, 27.0 ± 9.26 in the S2 group, and 16.4 ± 9.55 in the C group ( = .159). Normal control peritoneum showed little reactivity, whereas a moderate to strong cytoplasmic reaction to anti-CCL5/CCL2 antibodies was observed in mesothelial and inflammatory cells in the S1 and S2 groups.

Conclusion: CO pneumoperitoneum evokes an inflammatory response by modifying plasma RANTES levels and peritoneal CCL5/CCL2 expression.
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http://dx.doi.org/10.4293/JSLS.2020.00017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7242020PMC
November 2020

Autophagy and NLRP3 inflammasome crosstalk in neuroinflammation in aged bovine brains.

J Cell Physiol 2020 06 5;235(6):5394-5403. Epub 2020 Jan 5.

Department of Veterinary Medicine and Animal Production, University of Naples "Federico II", Naples, Italy.

NLRP3 inflammasome is a multiprotein complex that can sense several stimuli such as autophagy dysregulation and increased reactive oxygen species production stimulating inflammation by priming the maturation of proinflammatory cytokines interleukin-1β and interleukin-18 in their active form. In the aging brain, these cytokines can mediate the innate immunity response priming microglial activation. Here, we describe the results of immunohistochemical and molecular analysis carried out on bovine brains. Our results support the hypothesis that the age-related impairment in cellular housekeeping mechanisms and the increased oxidative stress can trigger the inflammatory danger sensor NLRP3. Moreover, according to the recent scientific literature, we demonstrate the presence of an age-related proinflammatory environment in aged brains consisting in an upregulation of interleukin-1β, an increased microglial activation and increased NLRP3 expression. Finally, we suggest that bovine may potentially be a pivotal animal model for brain aging studies.
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http://dx.doi.org/10.1002/jcp.29426DOI Listing
June 2020

Muscular Sarcocystosis in Sheep Associated With Lymphoplasmacytic Myositis and Expression of Major Histocompatibility Complex Class I and II.

Vet Pathol 2020 03 5;57(2):272-280. Epub 2019 Dec 5.

Department of Veterinary Medicine and Animal Production, Unit of Pathology, University of Naples Federico II, Naples, Italy.

Sarcocystosis is a protozoal disease affecting a wide range of animals. The aims of this study were to characterize the following in sheep: (1) the muscle pathology in infection, (2) the inflammatory infiltrate and its relationship to severity of infection, and (3) immune markers expressed by parasitized muscle fibers and parasitic cysts. Skeletal muscle samples from 78 sheep slaughtered in southern Italy were snap frozen and analyzed by histopathology, immunohistochemistry, and immunofluorescence. Polymerase chain reaction (PCR) and sequencing were used for Sarcocystis species identification. All 40 muscle samples tested were PCR-positive for Histologically, cysts were identified in 76/78 cases (97%), associated with an endomysial infiltrate of lymphocytes and plasma cells. The T cells were predominantly CD8+, with fewer CD4+ or CD79α+ cells. Eosinophils were absent. Notably, sarcolemmal immunopositivity for major histocompatibility complex (MHC) I and II was found in 76/78 cases (97%) and 75/78 cases (96%), respectively, both in samples with and in those without evident inflammatory infiltrate. The number of cysts was positively correlated with inflammation. In addition, MHC I was detected in 55/78 cyst walls (72%), and occasionally co-localized with the membrane-associated protein dystrophin. The findings suggest that muscle fibers respond to the presence of cysts by expression of MHC I and II. The possible role of MHC I and II in the inflammatory response and on the cyst wall is also discussed.
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http://dx.doi.org/10.1177/0300985819891257DOI Listing
March 2020

Long term breeding of the Lmna G609G progeric mouse: Characterization of homozygous and heterozygous models.

Exp Gerontol 2020 02 30;130:110784. Epub 2019 Nov 30.

CNR - Institute of Molecular Genetics "Luigi Luca Cavalli-Sforza"- Unit of Bologna, Bologna, Italy; IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy. Electronic address:

The transgenic Lmna progeric mouse represents an outstanding animal model for studying the human Hutchinson-Gilford Progeria Syndrome (HGPS) caused by a mutation in the LMNA gene, coding for the nuclear envelope protein Lamin A/C, and, as an important, more general scope, for studying the complex process governing physiological aging in humans. Here we give a comprehensive description of the peculiarities related to the breeding of Lmna mice over a prolonged period of time, and of many features observed in a large colony for a 2-years period. We describe the breeding and housing conditions underlining the possible interference of the genetic background on the phenotype expression. This information represents a useful tool when planning and interpreting studies on the Lmna mouse model, complementing any specific data already reported in the literature about this model since its production. It is also particularly relevant for the heterozygous mouse, which mirrors the genotype of the human pathology however requires an extended time to manifest symptoms and to be carefully studied.
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http://dx.doi.org/10.1016/j.exger.2019.110784DOI Listing
February 2020

The T197A Knock-in Model of Gene to Study the Effects of p27 Restoration .

Mol Cancer Ther 2019 02 13;18(2):482-493. Epub 2018 Nov 13.

Department of Experimental and Clinical Medicine, University "Magna Graecia," Catanzaro, Italy.

The CDK inhibitor, p27, encoded by the gene can negatively modulate cell proliferation. The control of p27 activity during the cell cycle is regulated at multiple levels, including transcription, translation, and protein stability. The last residue of p27 (threonine 198 in human, threonine 197 in mouse) is involved in the control of protein stability. We have generated a murine knock-in model () in which threonine 197 is replaced by alanine, which renders p27 protein highly unstable due to a high rate of proteasomal degradation. Expectedly, mice present with increased body size and weight, organomegaly, and multiple organ hyperplasia, similar to what is observed in mice. We investigated the effects exerted by the restoration of normal levels of p27 protein in the tissue of mice. We found that proteasome inhibition with bortezomib rescues the hyperplasia induced by the lack of p27 expression in but not in mice. However, BAY 11-7082, a proteasome inhibitor that stabilizes IκB but not p27, fails to rescue hyperplasia in mice. Bortezomib increases p27 half-life and reduces the proliferation in MEFs derived from but not from mice, whereas BAY 11-7082 had no effect on the protein levels of p27 and on the proliferation rate of MEFs.The results presented here demonstrate that mice represent an attractive model to investigate whether the targeting of p27 degradation machinery might prove beneficial in the treatment of a variety of human proliferative disorders caused by increased turnover of p27 protein.
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http://dx.doi.org/10.1158/1535-7163.MCT-18-0134DOI Listing
February 2019

HMGA2 cooperates with either p27 deficiency or Cdk4 mutation in pituitary tumorigenesis.

Cell Cycle 2018 30;17(5):580-588. Epub 2018 Jan 30.

a CNR - Institute of Experimental Endocrinology and Oncology - c/o Department of Molecular Medicine and Medical Biotechnologies , University of Naples "Federico II" , Naples , Italy.

We have previously reported a critical role of HMGA proteins in pituitary tumorigenesis since either the Hmga1 or Hmga2 gene overexpression/activation induces the development of mixed growth hormone/prolactin cell pituitary adenomas by activating the E2F transcription factor 1, and then enhancing the G1/S transition of the cell cycle. Consistently, amplification and overexpression of the HMGA2 gene was found in human pituitary prolactinomas. Since impairment of the cell cycle control represents a feature of experimental and human pituitary adenomas, we have investigated the possible synergism between the alterations of other cell cycle regulators, such as p27 deficiency or Cdk4 mutation, with Hmga2 overexpression in pituitary tumorigenesis. Therefore, we crossed the Hmga2/T mice, overexpressing the truncated/active form of the Hmga2 gene, either with the knockout mice for p27, or with the knockin mice for the Cdk4 mutation, both developing pituitary adenomas. Increased incidence and decreased latency in the development of pituitary lesions appeared in double mutant Hmga2/T;Cdk4 mice, and increased features of invasiveness and atypia were observed in pituitary tumors of both Hmga2/T;p27-ko and Hmga2/T;Cdk4 double mutant mice as compared with single mutant compounds. Interestingly, most of these mice develop pituitary adenomas with high Ki67 index, extrasellar expansion and brain tissue infiltration, representing good mouse models for human aggressive pituitary adenomas. Taken together, the results reported here indicate a cooperation between HMGA2 overexpression and either p27 or CDK4 impairment in promoting pituitary tumor development and progression.
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http://dx.doi.org/10.1080/15384101.2017.1403682DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5969540PMC
September 2019

Inflammatory Myopathy in Horses With Chronic Piroplasmosis.

Vet Pathol 2018 01 18;55(1):133-143. Epub 2017 Jul 18.

2 Unit of Pathology, Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Naples, Italy.

Horses affected by chronic piroplasmosis may develop poor performance and muscle atrophy. Here we investigate the pathological and immunopathological aspects of myopathy occurring in chronic equine piroplasmosis. The study included 16 horses serologically positive for equine piroplasms presenting with clinical signs and supporting serum biochemical evidence of a myopathy. Skeletal muscle was evaluated by histopathology, immunohistochemistry, indirect immunofluorescence, and molecular detection of piroplasms and inflammatory cytokines in skeletal muscle. Histologic lesions included muscle fiber atrophy (100% of cases), degenerative changes (13/16, 81%), and perivascular perimysial and endomysial lymphocytic infiltrates (81% of cases). In 15 cases (94%), muscle fibers had strong immunostaining for major histocompatibility complex classes I and II. T lymphocyte populations were mainly CD3+, CD8+, and CD4+ in equal proportions, with a lower number of CD79α+ cells. The serum from affected horses was tested by indirect immunofluorescence for binding of IgG, IgM, or IgA to sections of normal equine muscle to detect circulating autoantibodies against muscle antigen(s). In all cases, distinct sarcolemmal staining was detected in sections incubated with serum from affected horses, in contrast to sections incubated with phosphate-buffered saline or equine control sera. Reverse transcription polymerase chain reaction (RT-PCR) testing of muscles from affected animals revealed a significant increase of interferon-γ, interleukin-12, and tumor necrosis factor-α gene expression compared to healthy controls. Theileria equi or Babesia caballi was not detected in samples of affected muscle by RT-PCR. Thus, inflammatory myopathy associated with equine piroplasmosis may involve an autoimmune pathogenesis with upregulation of inflammatory cytokines that may cause myofiber atrophy and degeneration.
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http://dx.doi.org/10.1177/0300985817716262DOI Listing
January 2018

Oncogene-induced senescence and its evasion in a mouse model of thyroid neoplasia.

Mol Cell Endocrinol 2018 01 23;460:24-35. Epub 2017 Jun 23.

Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universita' Federico II c/o Istituto di Endocrinologia e Oncologia Sperimentale, CNR, Via S Pansini 5, 80131 Naples, Italy. Electronic address:

Here we describe a conditional doxycycline-dependent mouse model of RET/PTC3 (NCOA4-RET) oncogene-induced thyroid tumorigenesis. In these mice, after 10 days of doxycycline (dox) administration, RET/PTC3 expression induced mitogen activated protein kinase (MAPK) stimulation and a proliferative response which resulted in the formation of hyperplastic thyroid lesions. This was followed, after 2 months, by growth arrest accompanied by typical features of oncogene-induced senescence (OIS), including upregulation of p16INK4A and p21CIP, positivity at the Sudan black B, activation of the DNA damage response (DDR) markers γH2AX and pChk2 T68, and induction of p53 and p19ARF. After 5 months, about half of thyroid lesions escaped OIS and formed tumors that remained dependent on RET/PTC3 expression. This progression was accompanied by activation of AKT-FOXO1/3a pathway and increased serum TSH levels.
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http://dx.doi.org/10.1016/j.mce.2017.06.023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5741508PMC
January 2018

Immunopathological Features of Canine Myocarditis Associated with Leishmania infantum Infection.

Biomed Res Int 2016 19;2016:8016186. Epub 2016 Jun 19.

Unit of Pathology, Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, 80137 Naples, Italy.

Myocarditis associated with infectious diseases may occur in dogs, including those caused by the protozoa Neospora caninum, Trypanosoma cruzi, Babesia canis, and Hepatozoon canis. However, although cardiac disease due to Leishmania infection has also been documented, the immunopathological features of myocarditis have not been reported so far. The aim of this study was to examine the types of cellular infiltrates and expression of MHC classes I and II in myocardial samples obtained at necropsy from 15 dogs with an established intravitam diagnosis of visceral leishmaniasis. Pathological features of myocardium were characterized by hyaline degeneration of cardiomyocytes, necrosis, and infiltration of mononuclear inflammatory cells consisting of lymphocytes and macrophages, sometimes with perivascular pattern; fibrosis was also present in various degrees. Immunophenotyping of inflammatory cells was performed by immunohistochemistry on cryostat sections obtained from the heart of the infected dogs. The predominant leukocyte population was CD8+ with a fewer number of CD4+ cells. Many cardiomyocytes expressed MHC classes I and II on the sarcolemma. Leishmania amastigote forms were not detected within macrophages or any other cell of the examined samples. Our study provided evidence that myocarditis in canine visceral leishmaniasis might be related to immunological alterations associated with Leishmania infection.
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http://dx.doi.org/10.1155/2016/8016186DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4930798PMC
February 2017

AKT1E¹⁷K Is Oncogenic in Mouse Lung and Cooperates with Chemical Carcinogens in Inducing Lung Cancer.

PLoS One 2016 9;11(2):e0147334. Epub 2016 Feb 9.

Dipartimento di Medicina Sperimentale e Clinica, Università Magna Graecia, Catanzaro, Italy.

The hotspot AKT1E17K mutation in the pleckstrin homology domain of AKT1 occurs in approximately 0.6-2% of human lung cancers. Recently, we have demonstrated that AKT1E17K transforms immortalized human bronchial cells. Here by use of a transgenic Cre-inducible murine strain in the wild type Rosa26 (R26) locus (R26-AKT1E17K mice) we demonstrate that AKT1E17K is a bona-fide oncogene and plays a role in the development of lung cancer in vivo. In fact, we report that mutant AKT1E17K induces bronchial and/or bronchiolar hyperplastic lesions in murine lung epithelium, which progress to frank carcinoma at very low frequency, and accelerates tumor formation induced by chemical carcinogens. In conclusion, AKT1E17K induces hyperplasia of mouse lung epithelium in vivo and cooperates with urethane to induce the fully malignant phenotype.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147334PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4747507PMC
July 2016

Age related skeletal muscle atrophy and upregulation of autophagy in dogs.

Vet J 2015 Oct 6;206(1):54-60. Epub 2015 Jul 6.

Department of Veterinary Medicine and Animal Production, Laboratory of Comparative Neuromuscular Diseases, University of Naples Federico II, via Delpino 1, 80137 Napoli, Italy. Electronic address:

Sarcopenia, the age related loss of muscle mass and strength, is a multifactorial condition that occurs in a variety of species and represents a major healthcare concern for older adults in human medicine. In veterinary medicine, skeletal muscle atrophy is often observed in dogs as they reach old age, but the process is not well understood. Autophagy is a mechanism for degradation and recycling of cellular constituents and is potentially involved in sarcopenia. The aim of the present study was to evaluate the expression of three markers of autophagy, Beclin 1, LC3 and p62, in muscle wasting of geriatric dogs, to establish whether the levels of autophagy change with increasing age. Muscle biopsies from 25 geriatric dogs were examined and compared with those from five healthy young dogs. Samples from older dogs, assessed by routine histology, histoenzymatic staining and immunohistochemistry, showed evidence of muscle atrophy, sarcoplasmic vacuolisation and mitochondrial alterations. Furthermore, in 80% of the muscle samples from the older dogs, marked intracytoplasmic staining for Beclin 1 and LC3 was observed. Significantly greater expression of LC3 II and Beclin 1, but lower expression of p62, was found by Western blotting, comparing muscle samples from old vs. young dogs. The results of the study suggest that enhanced autophagy might be one of the factors underlying muscle atrophy in dogs as they age.
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http://dx.doi.org/10.1016/j.tvjl.2015.07.005DOI Listing
October 2015

Ccdc6 knock-in mice develop thyroid hyperplasia associated to an enhanced CREB1 activity.

Oncotarget 2015 Jun;6(17):15628-38

Istituto per l'Endocrinologia ed Oncologia Sperimentale del CNR e/o Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli "Federico II", Naples, Italy.

CCDC6 was originally identified upon rearrangement with RET in human thyroid papillary carcinomas generating the RET/PTC1 oncogene. We have previously reported that CCDC6 interacts with CREB1 and represses its transcriptional activity. Since the function of at least one allele of CCDC6 is lost following RET/PTC1 rearrangements, we aimed at the generation of mice, carrying a CCDC6 mutant gene. Previous studies suggested that the coiled-coil domain of CCDC6, mainly encoded by human exon 2, is required for the protein function. Therefore, we engineered a murine Ccdc6 construct, carrying a deletion of the exon 2, that was able to exert only a mild repression on CREB1 transcriptional activity, with respect to the wild type Ccdc6. Subsequently, we generated Ccdc6-ex2 knock-in mice. These mice developed thyroid hyperplasia associated with an enhanced CREB1 activity and an increased expression of the CREB-1 regulated genes. These results strongly support a CCDC6 promoting role, ascribed to its functional impairment, in the development of thyroid papillary carcinomas harboring the RET/PTC1 oncogene.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4558175PMC
http://dx.doi.org/10.18632/oncotarget.3858DOI Listing
June 2015

Angiostrongylus vasorum: epidemiological, clinical and histopathological insights.

BMC Vet Res 2014 Sep 28;10:236. Epub 2014 Sep 28.

Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via Della Veterinaria 1, Naples, 80137, Italy.

Background: Canine angiostrongylosis is a nematode infection in domestic dogs and wild carnivores. The present report focuses on epidemiological, clinical and histopathological findings in a case of fatal disseminated angiostrongylosis in a dog living in southern Italy and provides data on the extent of the spread of Angiostrongylus vasorum in the same area.

Case Presentation: A 4-year-old female English Setter from the Campania region of southern Italy was referred with a 2-week history of cough and severe respiratory distress that did not respond to antimicrobial therapy. Based on clinical, radiological, echographical and cytological findings (including the presence of larvae), a suspect diagnosis of lungworm infection was performed. After few days the dog died due to progressive clinical aggravation. Complete postmortem examination was conducted within 24 hours from death and samples from lungs, heart, liver, kidney, spleen, stomach and small intestine were fixed in 10% buffered formalin. Grossly, several hemorrhagic foci were observed mostly in the lungs, liver, kidney. Microscopically, the lungs contained numerous, multifocal to coalescing granulomas composed of epitheliod macrophages, multinucleated giant cells and some neutrophils, frequently associated with parasite eggs and larvae. The lungs contained many firm nodules, many adult nematodes approximately 1.5 to 2 cm in length were observed in cut sections and identified as A. vasorum. A subsequent parasitological survey performed with FLOTAC on stray dogs living in the same area showed the presence of A. vasorum larvae in 17 of 1639 stray dogs examined (1.04%).

Conclusion: This survey provides new data on distribution of A. vasorum and underlines that canine angiostrongylosis should be considered as differential diagnosis in dogs.
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http://dx.doi.org/10.1186/s12917-014-0236-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4193975PMC
September 2014

Outbreak of fatal nitrate toxicosis associated with consumption of fennels (Foeniculum vulgare) in cattle farmed in Campania region (southern Italy).

Environ Sci Pollut Res Int 2014 May 23;21(9):6252-7. Epub 2014 Jan 23.

Department of Veterinary Medicine and Animal Production, University of Naples Federico II, via Delpino, 1-80137, Naples, Italy.

Nitrate and nitrite are toxicants that have become increasingly significant environmental chemicals. Increase in environmental distribution of nitrogenous compounds, especially in surface and ground water, has been attributed to the intensive use of nitrate as agricultural fertilizers and to increasing amounts of nitrogenous wastes produced by municipalities, industries, and feedlots. The purpose of this study is to illustrate a fatal nitrate toxicosis in cattle associated with the consumption of fennels (Foeniculum vulgare). Fifteen cows from the same farm suddenly developed weakness, muscular tremors, respiratory distress, and finally convulsions. The affected animals died within 24 to 48 h from the onset of the clinical signs. Five cows underwent a complete post-mortem examination. In all examined animals, gross lesions included presence of dark unclotted blood around the nostrils and the anal region, moderate inflammation of the gastrointestinal mucosa, and brown discoloration of the skeletal muscles and kidneys. The histological examination showed tubular degeneration and congestion of glomerular vessels in the kidney. Toxicological analysis detected nitrates at 4 672.2 ppm in the fennels used to feed the animals. The source of exposure to nitrates was identified in the fennels. The fennels were grown in a polluted area of the Campania region in southern Italy and distributed in a public market for human consumption. The waste from the sale of the fennels was fed to the cows. The accumulation of nitrates in some vegetables poses a risk not only for animal health but also for human and environmental safety.
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http://dx.doi.org/10.1007/s11356-014-2520-9DOI Listing
May 2014

Bovine papillomavirus type 2 infection and a series of mesenchymal tumors of the urinary bladder in cattle.

Biomed Res Int 2013 5;2013:814635. Epub 2013 Jun 5.

Department of Veterinary Medicine and Animal Productions, Naples University Federico II, Naples, Italy.

This report describes the histopathology of two hundred and fifty-three mesenchymal tumors of the urinary bladder in cattle grazing on lands rich in bracken fern. Approximately 80% were hemangiomas and angiosarcomas. Hemangioma (capillary, cavernous, and large vessels) was the most frequent mesenchymal tumor and was more common than angiosarcoma. Although the appearance of endothelial cells can vary remarkably, epithelioid angiosarcomas, often containing multinucleated cells, were the most frequent malignant vascular tumors. Hemangiopericytoma and tumors of muscle and soft connective tissue origin, alone and/or in association with tumor-like lesions, were less frequently seen. Furthermore, forty-five cases of intravascular papillary endothelial hyperplasia (IPEH), a lesion not previously reported in the urinary bladder of cattle, were also described. Bovine papillomavirus type-2 DNA was amplified in tumor samples. Forty vascular tumors were investigated by dual-labeling immunofluorescence, and, for the first time, a coexpression of E5 and platelet-derived growth factor β receptor (PDGF β R) was shown to occur. The results show that the BPV-2 E5 oncoprotein binds to the activated form of the PDGF β receptor thus playing an important role in mesenchymal as well as epithelial carcinogenesis of the urinary bladder. Furthermore, these findings demonstrate that BPV-2 infects both epithelial and mesenchymal cells.
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http://dx.doi.org/10.1155/2013/814635DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3687770PMC
February 2014

Peritoneal morphological changes due to pneumoperitoneum: the effect of intra-abdominal pressure.

Eur J Pediatr Surg 2014 Aug 25;24(4):322-7. Epub 2013 Jun 25.

Division of Veterinary Medicine, Department of Pathology and Animal Health, University of Naples Federico II, Naples, Italy.

Introduction: Carbon dioxide (CO2) used in laparoscopy evokes local and systemic effects. This study was designed to evaluate the histopathologic morphologic changes due to CO2 and air insufflation, at different pressure levels, on visceral and parietal peritoneum in rats.

Materials And Methods: A total of 56 rats were object of the study, randomly divided into five groups. Pneumoperitoneum (PN) was maintained for 30 minutes, at a flow rate of 0.5 L/min and at a pressure of 10 and 6 mm Hg with CO2 (group S1-S2, n = 32) and filtered air (group A1-A2, n = 16). Only anesthesia was performed in the fifth group (group C, n = 8). Peritoneal samples were obtained 24 hours later for blinded histological evaluation. A grading system was adopted to evaluate histological peritoneal changes (0, no change; 1, mild; 2, moderate; and 3, severe) such as mesothelial aspect, inflammatory response, edema, and hemorrhage. The score reflected the severity of damage and was calculated by the sum of the degree evaluated separately. Values were compared with the analysis of variance analysis.

Results: CO2 and air insufflation caused reactive mesothelial cells and peritoneal inflammation of different degrees depending on the level of intra-abdominal pressure (IAP) and type of gas. These modifications were absent in group C and were less evident in low pressure S2 group with respect to S1 and A1-A2 groups. The average values of histopathologic peritoneal score showed significant differences between S2 (11.5) versus S1 groups (16.83) with respect to A groups (A1 = 27.83; A2 = 20.5) and compared with the controls (C = 2.5).

Conclusions: Our data confirm that PN affects the peritoneal integrity. The grades of morphological peritoneal changes are related to the level of IAP. Low CO2 pressure causes minor peritoneal changes with respect to high pressure and air insufflation.
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http://dx.doi.org/10.1055/s-0033-1349057DOI Listing
August 2014

Role of serotonergic system in the pathogenesis of fibrosis in canine idiopathic inflammatory myopathies.

Neuromuscul Disord 2012 Jun 6;22(6):549-57. Epub 2012 Mar 6.

Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Via S. Pansini 5, 80131 Naples, Italy.

Idiopathic inflammatory myopathies are muscle diseases characterized by inflammation, necrosis, and fibrosis. The neurotransmitter serotonin (5-HT) has been shown to promote fibrosis in many tissues and organs by activating TGFβ-1 signaling. In this study, we evaluated the potential role of 5-HT in the pathogenesis of fibrosis in canine idiopathic inflammatory myopathies. Muscle biopsies from dogs affected by masticatory muscle myositis or polymyositis and from healthy dogs were processed for immunohistochemistry and Western blotting. The immunohistochemical analysis showed a strong expression of 5-HT in muscle tissues of affected dogs, whereas the amine was absent in the muscles of healthy dogs. Biochemical analysis showed increased expression levels of the selective 5-HT2A receptor in the muscle specimens of the most severely affected dogs versus controls. Further, increased phosphorylation levels of the TGFβ-1 signaling mediators SMAD2/3 and ERK1/2 were detected in tissue samples from affected dogs as compared to tissues from healthy dogs. Although further studies are needed, our findings highlight for the first time a potential role of 5-HT in the development of fibrosis in canine idiopathic inflammatory myopathies, thus supporting other evidence that 5-HT pro-fibrotic activity occurs via activation of TGFβ-1 signaling pathway.
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http://dx.doi.org/10.1016/j.nmd.2012.01.009DOI Listing
June 2012

Histochemical and immunohistological approach to comparative neuromuscular diseases.

Folia Histochem Cytobiol 2009 ;47(2):143-52

Department of Pathology and Animal Health, University of Naples Federico II, Naples, Italy.

The broad category of neuromuscular diseases covers conditions that involve the weakness or wasting of the body muscles. These problems may occur in the spinal cord, the peripheral nerves or the muscle fibers. Some may be hereditary, while others are acquired. Commonly recognized conditions fall into the categories of myopathies, which are diseases of the muscle like muscular dystrophy, disorders of the junction where the nerve impulses are transmitted to the muscle like myasthenia gravis, and neuropathies, which are diseases of the peripheral nervous system. The diagnosis of most neuromuscular diseases rest on careful clinical evaluation of the patient, electromyography, the muscle biopsy, and in some instances, molecular genetic studies. Muscle biopsy, associated to histochemical and immunohistological techniques, plays a key role in diagnosis of many neuromuscular disorders. A number of morphological abnormalities of muscle can be recognized on histological stains such as haematoxylin and eosin and Engel trichrome. Histochemical techniques are essential for the study of muscle biopsies for four main reasons. First, they demonstrate the non-uniform nature of the muscle highlighting the different biochemical properties of specific fibre type and their selective involvement in certain disease processes. Second, they may show an absences of a particular enzyme. Third, an excess of a particular substrate can be demonstrated. Fourth, they may show structural changes in the muscle which would not be apparent with routine histological stains, such as the enzyme-deficient cores in central core disease "mouth-eaten" fibers, and abnormalities in the distribution of mitochondria. In some neuromuscular disorders there could be only non-specific myopathological features. However, a number of proteins, including sarcolemmal, sarcomeric, and nuclear proteins as well as enzymes with defects responsible for neuromuscular disorders, have been identified during the past two decades, allowing a more specific and firm diagnosis of muscle diseases. Identification of protein defects relies predominantly on immunohistochemical preparations and on Western blot analysis. While immunohistochemistry is very useful in identifying abnormal expression of primary protein abnormalities in recessive conditions, it is less helpful in detecting primary defects in dominantly inherited disorders. Abnormal immunohistochemical expression patterns can be confirmed by Western blot analysis which may also be informative in dominant disorders. Besides identification of specific protein defects, immunohistochemistry is also helpful in the differentiation of inflammatory myopathies by subtyping cellular infiltrates and demonstrating up-regulation of subtle immunological parameters. This review will summarize and describe the impact that histochemistry and immunohistochemistry has had and the possibilities it has opened up in the diagnosis of neuromuscular disorders in human as well as in veterinary myology.
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http://dx.doi.org/10.2478/v10042-009-0066-3DOI Listing
April 2010

Syrian hamster infected with Leishmania infantum: a new experimental model for inflammatory myopathies.

Muscle Nerve 2010 Mar;41(3):355-61

Department of Pathology and Animal Health, University of Naples Federico II, Via Delpino 1, 80137 Naples, Italy.

Idiopathic inflammatory myopathies (IIMs) are inflammatory disorders of unknown origin. On the basis of clinical, histopathological, and immunological features, they can be differentiated into three major and distinct subsets: dermatomyositis; polymyositis; and inclusion-body myositis. Although a few animal models for IIM are currently available, they lack several characteristic aspects of IIMs. The aim of our study was to examine skeletal muscle involvement in an experimental animal model of visceral leishmaniasis, a disseminated infection caused by the protozoan parasite Leishmania infantum, and to compare features of associated inflammation with those of human IIM. Syrian hamsters infected intraperitoneally with amastigotes of L. infantum were killed at 3 or 4 months post-infection, and the skeletal muscles were studied. Focal inflammation was predominantly observed in the endomysium and, to a lesser extent, in perivascular areas. Degenerating muscle fibers were also found, as well as myonecrosis. Immunofluorescence with confocal laser scanning microscopy was used to characterize the phenotype of inflammatory infiltrates and the distribution of MHC class I and II in muscle biopsies. The infiltrating inflammatory cells consisted mainly of T cells, and CD8(+) T cells were found in non-necrotic muscle fibers that expressed MHC class I on the sarcolemma. In addition to T cells, several macrophages were present. The model we are proposing closely resembles polymyositis and may be useful in studying certain aspects of this disease such as the role of T cells in muscle inflammation and myocytotoxicity, while also providing novel therapeutic targets.
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http://dx.doi.org/10.1002/mus.21502DOI Listing
March 2010

Isolated lepromatous conjunctivo-corneal granuloma in a cat from Italy.

Vet Ophthalmol 2009 Mar-Apr;12(2):97-101

Department of Clinical Sciences, Unit of Surgery, University of Naples Federico II, via Delpino 1, 80137 Naples, Italy.

Objective: To describe a case of a conjunctivo-corneal mass in a cat associated with acid-fast bacilli.

Methods: A 2-year-old female black European Short-Hair cat, living outdoors in a suburban environment in Italy, was referred for evaluation of a nodular, vascularized mass of 2 weeks duration. The mass involved the dorsal bulbar conjunctiva at the temporal canthus of OS and invaded the sclera and cornea. Routine ophthalmic and systemic examination, serologic testing, cytology and histology of the mass were performed. Mycobacterium specific polymerase chain reaction (PCR) of variable regions 1, 2 and 3 of the 16S ribosomal RNA (rRNA) gene was also performed.

Results: Neutrophils, lymphocytes, macrophages and giant cells with intracytoplasmic acid-fast bacilli were seen on cytological examination. The histological examination confirmed the presence of a granulomatous lesion with acid-fast bacilli within macrophages. Bacteriological culture of the material from the lesion was negative for Mycobacterium spp. Mycobacterium 16S rRNA gene specific PCR was positive. A diagnosis of feline leprosy was made. The owners refused any treatment, and 1 year later the lesion was still present.

Conclusions: Veterinary ophthalmologists should be aware of conjunctivo-corneal leproma as an unusual symptom of leprosy.
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http://dx.doi.org/10.1111/j.1463-5224.2008.00676.xDOI Listing
July 2009

Canine inflammatory myopathy associated with Leishmania Infantum infection.

Neuromuscul Disord 2009 Feb 11;19(2):124-30. Epub 2008 Dec 11.

Department of Pathology and Animal Health, University of Naples Federico II, Naples, Italy.

Inflammatory myopathy associated with several infectious diseases occurs in dogs including those caused by Toxoplasma gondii, Neospora caninum, Ehrlichia canis and Hepatozoon canis. However, muscle disease due to Leishmania infection has been poorly documented. The aim of this study was to examine the distribution and types of cellular infiltrates and expression of MHC class I and II in muscle biopsies obtained from 15 male beagle dogs from a breeder group with an established diagnosis of leishmaniasis. Myopathic features were characterized by necrosis, regeneration, fibrosis and infiltration of mononuclear inflammatory cells consisting of lymphocytes, plasma cells and histiocytes. The predominant leukocyte populations were CD3+, CD8+ and CD45RA+ with lesser numbers of CD4+ cells. Many muscle fibers had MHC class I and II positivity on the sarcolemma. There was a direct correlation between the severity of pathological changes, clinical signs, and the numbers of Leishmania amastigotes. Our studies provided evidence that: 1) Leishmania should be considered as a cause of IM in dogs; 2) Leishmania is not present within muscle fibers but in macrophages, and that 3) the muscle damage might be related to immunological alterations associated with Leishmania infection. Leishmania spp. should also be considered as a possible cause in the pathogenesis of human myositis.
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http://dx.doi.org/10.1016/j.nmd.2008.10.013DOI Listing
February 2009

Veterinary myology.

Neuromuscul Disord 2007 Aug 11;17(8):662. Epub 2007 Jun 11.

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http://dx.doi.org/10.1016/j.nmd.2007.04.007DOI Listing
August 2007

Expression of major histocompatibility complex class I and class II antigens in canine masticatory muscle myositis.

Neuromuscul Disord 2007 Apr 13;17(4):313-20. Epub 2007 Mar 13.

Department of Pathology and Animal Health-Unit of Anatomic Pathology, University of Naples Federico II, 80137 Naples, Italy.

Studies in human immune-mediated inflammatory myopathies have documented expression of major histocompatibility complex class I (MHC class I) and class II (MHC class II) antigens on muscle fiber membranes in the presence or absence of cellular infiltration. Here we evaluate the presence and distribution of these antigens in canine masticatory muscle myositis, an immune-mediated inflammatory myopathy. Twelve samples of temporalis and masseter muscles from dogs with a clinical diagnosis of canine masticatory muscle myositis were examined by immunohistochemistry and double-immunofluorescence confocal microscopy. MHC class I and class II antigens were expressed in muscle fibers independent of inflammatory cell infiltration. Furthermore MHC class I and class II antigens were expressed on the sarcolemma and co-localized with dystrophin. Our results suggest that MHC class I and class II expression in canine masticatory muscle myositis may play a role in the initiation and maintenance of the pathological condition, rather than just a consequence of a preceding local inflammation.
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http://dx.doi.org/10.1016/j.nmd.2007.01.012DOI Listing
April 2007

Local and systemic impact of pneumoperitoneum on prepuberal rats.

Pediatr Surg Int 2007 May;23(5):453-7

Department of Pediatric, Pediatric Surgery Unit, Second University of Naples, Via Pansini 5, 80131 Naples, Italy.

Pneumoperitoneum (PN) and the gas used to insufflate the abdominal cavity during laparoscopy seem to be responsible for local and systemic modifications. The aim of this study was to verify the effects of intra-abdominal carbon dioxide (CO(2)) and air insufflation on the peritoneum, as well as the cortico-surrenal response in prepuberal rats. Sixty prepuberal rats were divided into three groups: in the first (S, n = 36), PN was induced with CO(2), whereas in the second (A, n = 14), it was induced with filtered room air; in both conditions, insufflation lasted 30 min at a pressure of 10-12 mmHg. The third group (C, n = 10), underwent general anesthesia only. Two hours after inducing anaesthesia, 12 rats in group S, 6 in group A and 6 in group C were killed and the remaining, after 24 h; specimens of the visceral and the parietal peritoneum were obtained for histological examination, blood sample was taken for cortisol and DHEA-S assays at the different study periods. At the histological examination performed 2 h later, the groups S and A presented inflammatory cell infiltrate in the parietal and visceral peritoneum; this finding was even more marked in group A, which presented also congestion, hemorrhage and disruption of the cell line. Twenty-four hours after the experiment, the peritoneum of the two insufflated groups presented chronic infiltrate and reactive mesothelial cells with congestion, which was more evident in group A, but totally absent in group C. Cortisol levels were significantly higher in groups S and A (2.15:1 ratio) killed 2 h later compared to those killed 24 h later and to the control group. DHEA-S levels were not significantly different between the groups. Our results demonstrate that the chemical, physical and molecular impact of CO(2) on the peritoneum causes inflammation and tissue damage, this was even more evident 24 h after our experiment and in the air insufflated group. PN induced a significant variation in blood cortisol levels at 2 h. The CO(2) insufflation should be limited in patients with pre-existing peritoneal damage.
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http://dx.doi.org/10.1007/s00383-006-1860-zDOI Listing
May 2007

Evaluation of static telepathology in veterinary diagnostic cytology.

Vet Clin Pathol 2006 Sep;35(3):303-6

Facoltà di Medicina Veterinaria, Università degli Studi di Napoli Federico II, Napoli, Italy.

Background: Telepathology is the process of diagnostic pathology performed on digital images viewed on a display screen rather than by conventional light microscopy with glass slides. At present, electronic mail (e-mail) attachment is the most common communication medium for telepathology, because it is relatively simple and incurs minimal cost.

Objective: Our objective was to test whether telediagnosis through Internet e-mail could be applied to veterinary diagnostic cytology.

Methods: Using 20 cytology cases, on which a consensus diagnosis was reached by 3 experienced pathologists, a total of 130 images were digitized and sent as e-mail attachments from the Unit of Anatomic Pathology of the Faculty of Veterinary Medicine of Naples to the Unit of Pathology of the Faculty of Veterinary Medicine of Messina, Italy. The images were reviewed at the Unit of Pathology of Messina, where the consulting pathologist formulated his diagnoses by visualizing the images on the monitor of a computer. The telediagnoses were compared with the consensus diagnoses and with the consulting pathologist's interpretation using conventional light microscopy at a later date.

Results: The median time to capture images was approximately 30 minutes for each case. The median time to make a telecytodiagnosis was a few seconds for each case. Overall, there was good agreement (85%) between the consensus diagnosis and the consultant's telediagnosis. In 100% of the cases there was agreement between the consulting pathologist's telediagnosis and conventional glass slide diagnosis.

Conclusion: Telepathology by e-mail provides acceptable efficacy and a faster turnaround time than post and can be applied to veterinary diagnostic cytology.
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http://dx.doi.org/10.1111/j.1939-165x.2006.tb00135.xDOI Listing
September 2006