Processo inflamatório crônico granulomatoso experimental em peixes: um estudo morfológico, ultraestrutural e imunohistoquímico

O objetivo do presente estudo foi avaliar o processo inflamatório crônico granulomatoso induzido experimentalmente em Oreochromis niloticus através da inoculação de BCG e elucidar aspectos da reação inflamatória em peixes para uma melhor compreensão da filogenia do processo. Os resultados obtidos por microscopia de luz comum e ultra-estrutural demonstraram a participação de macrófagos, trombócitos, linfócitos, eosinófilos, células plasmáticas e células gigante tipo corpo estranho no processo inflamatório. Além desses tipos celulares, uma reação granulomatosa constituída predominantemente de células epitelióides também foram observadas ultraestruturalmente. Essas células epitelióides desenvolveram desmossomos ao longo do experimento, e também passaram a expressar receptores para citoqueratina, características estas de células epiteliais. Células pigmentares (melanomacrófagos), envolvendo de maneira crescente toda a formação granulomatóide e, participando ativamente da reação inflamatória crônica granulomatosa

Processo inflamatório crônico granulomatoso experimental em peixes: um estudo morfológico, ultraestrutural e imunohistoquímico

The purpose of the present study was to evaluate the chronic granulomatous inflammatory process experimentally induced in Oreochromis niloticus by the inoculation of BCG and to clarify aspects of the inflammatory reaction in fish for a better understanding of the phylogeny of the process. Results obtained by light microscopy and ultrastructural analysis showed the participation of macrophages, thrombocytes, lymphocytes, eosinophils, plasma cells and foreign type giant cells in the inflammatory process. Besides these cell types, a granulomatous reaction mainly consisting of epithelioid cells was also observed ultrastructurally. These epithelioid cells developed desmosomes throughout the experiment, and also began to express receptors for cytokeratin. Both findings are immunocytochemically characteristic of epithelial cells. Pigment cells (melanomacrophages) could also be seen surrounding all the granulomatoid formation in a crescent fashion and participating in the chronic granulomatous inflammatory reaction.O objetivo do presente estudo foi avaliar o processo inflamatório crônico granulomatoso induzido experimentalmente em Oreochromis niloticus através da inoculação de BCG e elucidar aspectos da reação inflamatória em peixes para uma melhor compreensão da filogenia do processo. Os resultados obtidos por microscopia de luz comum e ultra-estrutural demonstraram a participação de macrófagos, trombócitos, linfócitos, eosinófilos, células plasmáticas e células gigante tipo corpo estranho no processo inflamatório. Além desses tipos celulares, uma reação granulomatosa constituída predominantemente de células epitelióides também foram observadas ultraestruturalmente. Essas células epitelióides desenvolveram desmossomos ao longo do experimento, e também passaram a expressar receptores para citoqueratina, características estas de células epiteliais. Células pigmentares (melanomacrófagos), envolvendo de maneira crescente toda a formação granulomatóide e, participando ativamente da reação inflamatória crônica granulomatosa

Utilização da classificação de Dorfman-Warnke (modificada por Burke) em patologia veterinária na avaliação da linfadenite induzida por parvovírus canino em cobaias (Cavia porcellus )

Em patologia humana, a análise e interpretação das linfadenopatias reacionais baseia-se na caracterização morfológica das várias regiões do linfonodo. Em patologia veterinária não há essa padronização. No presente estudo, utilizamos as bases da classificação de Dorfman-Warnke (modificada por Burke), utilizada na área médica humana para avaliarmos uma linfadenite viral experimental em cobaias. Parvovírus canino foi inoculado em coxim plantar de cobaias e a resposta dos linfonodos poplíteos avaliada. No fim de cada período experimental, linfonodos foram excisados, pesados e processados histologicamente em cortes corados pelos métodos de Giemsa, Mallory, Gordon & Sweets e Hematoxilina-Eosina e por meio de imunoistoquímica com anticorpos pan-T e pan-B. Foi observado aumento de peso significante (p<0,05) dos linfonodos ipsilaterais ao sítio de inoculação em comparação com os contralaterais. A resposta folicular nos primeiros linfonodos foi caracterizada por reação do centro germinativo, apresentando perda parcial da delimitação da zona do manto. Em região paracortical aumentou o número de células blásticas, ocorrendo hipertrofia de vênulas pós-capilares. Associou-se discreta hiperplasia de cordões medulares e densificação do estroma reticular. A reação observada apresenta semelhanças com a identificada em humanos frente a estímulo viral, sugerindo a adequação do uso dessa classificação também em medicina veterinária.In human pathology the adequate system used to analyze and interprete reactive lymphadenopathies is based on the morphological characterization of the various areas within the lymphoid node. In veterinary pathology there is no standardization of histopathological evaluation of lymph nodes. Therefore, in this study, we applied the Dorfman-Warnke (modified by Burke) human classification of reactive lymphadenopathies to evaluate virus-induced lymphadenitis in guinea-pigs. Canine parvovirus was inoculated into the footpads of guinea-pigs and the response of popliteal lymph nodes was histologically studied. The footpads were excised after different periods of time, processed, and stained with HE, Mallory, Gordon & Sweets, Giemsa and by immunohistochemical methods. A significant increase in the weight of ipsilateral lymph nodes to the inoculation site (p<0.05) was observed. The follicular response in the lymph nodes was characterized by the reaction of the germinative center, which showed partial loss of delimitation in the mantle zone. Increased numbers of blast cells and hypertrophy of post-capillary venules were evidenced in the paracortical area of the lymph nodes. There was a discrete hyperplasia of the medullary cords and a general enlargement of the reticular net associated with those alterations. The reactional response observed was similar to viral lymphadenitis identified in humans, justifying the attempts to systematize this classification for veterinary use just like it is currently done in human medicine

Main neoplasic processes in psittacines birds kept in captivity

Um estudo anatomopatológico envolvendo 130 psitacídeos oriundos de centros de triagens, zoológicos e criadouros, visando a determinação da causa de morte destes animais, revelou a presença de neoplasias em sete animais, totalizando 5,4% dos casos. Os processos neoplasicos foram caracterizados como colangiocarcinoma, colangioma, linfoma, hemangiossarcoma e carcinoma renal primário, sendo o colangiocarcinoma o mais freqüentemente encontrado, presente em três animais. Este trabalho descreve estas neoplasias, levando em consideração os aspectos macro e microscópicos, meios de diagnósticos e espécies acometidas.An anatomopathologic study of 130 captivy psittacines from zoos, breeders and rehabilitation centers showed the presence of neoplastic disease in seven birds, totalizing 5.4% of the cases. The neoplasms were identified like cholangiocarcinoma, cholangioma, lymphoma, haemangiosarcoma and renal cell carcinoma. The cholangiocarcinoma was the tumor most commonly observed. it was present in birds. This study describes the macro and microscopic findings of these tumors; also reports diagnostic methods employed to identify these neoplasms.Fundação de Amparo à Pesquis

Immunoexpression of Her-2 protein in fine-needle aspirates of breast carcinoma: correlation with surgical specimens

PURPOSE: to determine the expression of HER-2 protein in fine-needle aspirates obtained from infiltrating breast duct carcinoma and in the corresponding surgical specimens and to correlate HER-2 expression with histological grade. Methods: forty-eight smears fixed in alcohol and previously stained by the Papanicolaou technique and histological sections of corresponding paraffin-embedded tissue blocks were submitted to immunohistochemistry by the avidin-biotin-peroxidase method using a polyclonal antibody (A0485-Dako). The reactions were carried out simultaneously with antigen recovery. The results were analyzed by a semiquantitative method according to the HercepTest score. Scores of 3+ were considered to be positive. RESULTS: positivity was observed in 39.6% of the smears and in 35.4% of the respective histological sections, with almost perfect cytohistologic concordance (kappa 0.82). The method of evaluation of histological sections was found to be reproducible regarding the smears and standardized the interpretation of the results. The correlation between HER-2 positivity and histological grade was not statistically significant but there was a tendency towards HER-2 expression in less differentiated tumors. CONCLUSION: the positivity of the immunocytochemical reactions observed in smears was comparable to that obtained in the histological sections, indicating that the material obtained by fine-needle aspiration biopsies previously stained by the Papanicolaou technique can be used for the evaluation of HER-2 expression.OBJETIVO: avaliar a imunoexpressão da proteína HER-2 em punção biópsia aspirativa de carcinoma de ductos infiltrantes da mama e nos espécimes cirúrgicos correspondentes. Correlacionar a imunoexpressão da proteína HER-2 com o grau histológico. MÉTODO: foram examinados 48 esfregaços fixados em álcool, previamente corados pela técnica de Papanicolaou, e cortes histológicos de blocos de parafina do tecido correspondente. Todos foram submetidos à técnica de imuno-histoquímica, utilizando o método da avidina-biotina-peroxidase com um anticorpo policlonal (A0485-Dako). As reações foram realizadas simultaneamente com recuperação antigênica. A avaliação dos resultados foi feita por método semiquantitativo, segundo a graduação do HercepTest. Foram considerados positivos apenas os casos graduados em 3+. RESULTADOS: observou-se positividade de 39,6% nos esfregaços e 35,4% nos cortes histológicos respectivos, sendo a concordância cito-histológica nos casos positivos quase perfeita (kappa 0,82). A utilização do método de avaliação dos cortes histológicos mostrou-se reprodutível nos esfregaços e padronizou a interpretação dos resultados. A correlação entre a positividade para proteína HER-2 e o grau histológico não foi significante, mas houve tendência a imunoexpressão mais freqüente em tumores menos diferenciados. CONCLUSÃO: a positividade das reações imunocitoquímicas nos esfregaços foi comparável à obtida nos cortes histológicos, podendo portanto o material de punção aspirativa por agulha fina, previamente corado pela técnica de Papanicolaou, ser utilizado na avaliação da imunoexpressão da proteína HER-2.Universidade Federal de São Paulo (UNIFESP) Departamento de Patologia Escola Paulista de MedicinaInstituto Adolfo LutzUNIFESP, Depto. de Patologia Escola Paulista de MedicinaSciEL

Papilomas cutâneos em tartarugas marinhas: um estudo morfológico, ultra-estrutural e imunoistoquímico em espécies brasileiras

Onze tartarugas verdes juvenis (Chelonia mydas) originárias do Oceano Atlântico, Brasil, com múltiplas lesões cutâneas papilomatosas foram examinadas. Histologicamente, os papilomas exibiam proliferação estromal hiperplásica e proliferação epidermal. As células epiteliais apresentavam alterações nucleares sugestivas de infecção viral e pleomorfismo nuclear severo. Um halo grande nuclear estava presente nos casos de proliferação epitelial; nessas células, características nucleares foram freqüentemente discarióticos, sem inclusão. Todos os fibropapilomas examinados foram negativos para antígenos grupo-específico para papilomavírus (BPV) e antígenos grupo-específico para herpesvírus (HSV1/HSV2) pela técnica peroxidase-antiperoxidase. Investigação por microscopia eletrônica foi negativa para partículas virais de papilomavírus e herpesvírus.Eleven juvenile green turtles (Chelonia mydas) from Atlantic Ocean, Brazil, with multiple cutaneous papillomatosis were examined. Histologically, the papillomas exhibit stromal hyperplasia proliferation and epithelial proliferation. The epithelial cells had nuclear changes suggestive of viral infection and severe nuclear pleomorphism. A large nuclear halo was present in the cases of epithelial proliferation; in these cells, nuclear features were frequently dyscariotic, without inclusion. All fibropapillomas examined were negative for papillomavirus group-specific antigens (BPV) and herpesvirus group-specific antigens (HSV1 / HSV2) by the peroxidase-antiperoxidase technique. Electronic microscopy investigation was negative for papillomaviruses and herpes-viruses particles

The association of p16INK4A and fragile histidine triad gene expression and cervical lesions

Objective. This cross-sectional study was intended to assess the association between immunohistochemical analysis of p16INK4A and fragile histidine triad (FHIT) and the presence of precancerous cervical lesions. Materials and Methods. Women seen at Pe´ rola Byington Hospital, São Paulo, Brazil, with histologically confirmed cervicitis (n = 31), cervical intraepithelial neoplasia (CIN) 1 (n = 30), CIN 2,3 (n = 30), and cervical cancer (n = 7) had also cervical material collected for liquid-based cytology, human papillomavirus Hybrid Capture 2 (HC2) test, and p16 and FHIT immunohistochemical reactions. Results. p16 and FHIT reactions were scored as the following: G1%, 1% to 5%, 95% to 25%, and 925%. Receiver operating curve analysis was used to select p16 and FHIT score cutoffs for further categorical analyses. All but one of the 37 CIN 2,3/cancer cases had a p16 score of greater than 1% to 5%. Among the 61 cervicitis/CIN 1 cases, 46 (75%) had a p16 score lower than 1% to 5%. In contrast, no association of FHIT expression and severity of cervical lesions could be demonstrated in this data set. Receiver operating curve analyses suggested the score of 1% to 5% for p16 as the cutoff that best discriminates CIN 2,3/cancer from cervicitis/CIN 1. No cutoff for FHIT scores could be suggested with data set. Conclusions. p16, but not FHIT expression, has the potential to be used as complementary diagnostic tool to investigate human papillomavirusYinduced cervical lesions, if these results are confirmed in larger studies.(undefined

Histopathological Characterization and Whole Exome Sequencing of Ectopic Thyroid: Fetal Architecture in a Functional Ectopic Gland from Adult Patient

Ectopic thyroid results from a migration defect of the developing gland during embryogenesis causing congenital hypothyroidism. But it has also been detected in asymptomatic individuals. This study aimed to investigate the histopathological, functional, and genetic features of human ectopic thyroids. Six samples were histologically examined, and the expression of the specific thyroid proteins was assessed by immunohistochemistry. Two samples were submitted to whole exome sequencing. An oropharynx sample showed immature fetal architecture tissue with clusters or cords of oval thyrocytes and small folliclesone sample exhibited a normal thyroid pattern while four showed colloid goiter. All ectopic thyroids expressed the specific thyroid genes and T4 at similar locations to those observed in normal thyroid. No somatic mutations associated with ectopic thyroid were found. This is the first immature thyroid fetal tissue observed in an ectopic thyroid due to the arrest of structural differentiation early in the colloid stage of development that proved able to synthesize thyroid hormone but not to respond to TSH. Despite the ability of all ectopic thyroids to synthetize specific thyroid proteins and T4, at some point in life, it may be insufficient to support body growth leading to hypothyroidism, as observed in some of the patients.FAPESP Grant [2009/53840-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Sao Paulo, Brazil [2010/12005-9, 2014/24549-4]Instituto da TiroideUniv Sao Paulo FMUSP, Fac Med, Cellular & Mol Endocrine Lab, Thyroid Unit,LIM 25, Ave Doutor Arnaldo 455, BR-01246904 Sao Paulo, SP, BrazilSao Paulo Publ Hlth Serv, Adolfo Lutz Inst, Av Dr Arnaldo 355, BR-01246000 Sao Paulo, SP, BrazilHead & Neck Surg Santa Catarina Hosp, Av Paulista 200, BR-01310000 Sao Paulo, SP, BrazilUNESP, Botucatu Sch Med, Dept Internal Med, Av Prof Montenegro,S-N Dist Rubiao Jr, BR-18618687 Botucatu, SP, BrazilHosp Pediat Dr Juan Garrahan, Serv Endocrinol, Combate Pozos 1881,C1245AAM, Buenos Aires, DF, ArgentinaUniv Estadual Campinas, Fac Ciencias Med, Dept Cirurgia, Disciplina Cirurgia Cabeca & Pescoco, R Tessalia Vieira Camargo 126, BR-13083887 Campinas, SP, BrazilUniv Fortaleza Unifor, Med Sch, Av Washington Soares 1321, BR-60811905 Fortaleza, CE, BrazilUniv Fed Sao Paulo UNIFESP, Postgrad Program Biotechnol, Pedro Toledo 669, BR-04039903 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Postgrad Programs Biotechnol & Struct & Funct Bio, Dept Ciencias Biol, Thyroid Mol Sci Lab,UNIFESP, Pedro Toledo 669, BR-04039903 Sao Paulo, SP, BrazilHosp Sirio Libanes, Mol Oncol Ctr, Rua Prof Daher Cutait 69, BR-01308060 Sao Paulo, SP, BrazilUniv Fed Sao Paulo UNIFESP, Postgrad Program Biotechnol, Pedro Toledo 669, BR-04039903 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Postgrad Programs Biotechnol & Struct & Funct Bio, Dept Ciencias Biol, Thyroid Mol Sci Lab,UNIFESP, Pedro Toledo 669, BR-04039903 Sao Paulo, SP, BrazilFAPESP [2009/53840-0]FAPESP[2010/12005-9, 2014/24549-4]Web of Scienc

Immunocytochemical expression of p16 INK4A and Ki-67 in cytologically negative and equivocal pap smears positive for oncogenic human papillomavirus

This study was designed to analyze the cross-sectional comparison of the p16 INK4A and Ki-67 immunocytochemical expression in negative and equivocal (atyp-ical squamous cells of undetermined significance (ASC-US)) liquid-based cytology (LBC) samples testing positive for high-risk human papillomavirus (HPV) types with HC2 assay or polymerase-chain reaction (PCR). A series of 199 consecutive LBC speci-mens derived from the same number of women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, São Paulo, were analyzed using immunocytochemistry for expression of p16 INK4A and Ki-67 in negative and equivocal LBC samples testing positive for high-risk HPV types with hybrid capture II test (HC2) or PCR. All patients with at least one test positive (cytology, PCR, and/or HC2) were followed each 6 months for 3 years. The follow-up procedure consisted of visual examination, colposcopic inspection, cytology, and HC2 assay. Among the neg-ative cytologic samples, 101 were HPV-positive and 55 HPV-negative. Of the HPV-pos-itive group, 59 of 101 cases (58.4%) were positive for both p16 and Ki67 immunostaining, and 17 of 101 (16.8%) were negative for both. The proportion of Ki-67-positivity increased almost in parallel with the increasing grade of p16-positivity (p = 0.0001 for linear trend). In the HPV-negative group, both markers were negative in 41 of 55 cases (74.5%), and no statistical relationship was observed between the two markers (Pearson, p = 0.595). HPV-positive ASC-US samples demonstrated a simulta-neous positive immunoreaction for p16 and Ki67 in 11 of 16 cases (68.7%), whereas 3 (18.7%) were concurrently negative. The relationship between the two markers was of borderline significance (Pearson, p = 0.053), but no linear relationship was found be-tween the graded p16 and Ki-67 expression (p = 0.065 for linear trend). In the HPV-negative ASC-US group, there was no statistical association between the graded p16 and Ki-67 positivity (Pearson, p = 0.281). After 36 months of follow-up of the ASC-US patients, 6 women still displayed ASC-US smear, of which 4 of 6 were HPV-positive and expressed both p16 and Ki-67 markers. Two of 43 ASC-US smears had high-grade squamous intraepithelial lesions diagnosed (4.6%), and 1 had low-grade squamous intraepithelial lesion (2.3%). All of those were positive for HPV, p16 and Ki-67. Patients with ASC-US diagnosis and positive high-risk HPV status and positive for p16 INK4A Ki67 should be carefully observed to exclude occurrence of a squamous intraepithelial lesion. The combination of these two markers can be a useful implement for manage-ment of women with equivocal cytology.European Commission (EC) - INCO-DEV Programme - Project #ICA4-CT-2001-10013