Avaliação da utilização de biópsia transbrônquica em pacientes com suspeita clínica de doença pulmonar intersticial

OBJETIVO: Estudar os padrões clínicos, radiológicos e histopatológicos da biópsia transbrônquica (BTB) utilizados para a confirmação diagnóstica em pacientes com suspeita clinica de doença pulmonar intersticial (DPI) atendidos em um hospital universitário de nível terciário. MÉTODOS: Os prontuários, laudos radiológicos e de biópsias transbrônquicas de todos os pacientes com suspeita de DPI submetidos a BTB entre janeiro de 1999 e dezembro de 2006 no Hospital das Clínicas de Botucatu, localizado na cidade de Botucatu (SP), foram revisados. RESULTADOS: Foram incluídos no estudo 56 pacientes. Desses, 11 (19,6%) apresentaram o diagnóstico definitivo de fibrose pulmonar idiopática (FPI), que foi significativamente maior nos casos nos quais DPI era uma possibilidade diagnóstica em comparação com aqueles nos quais DPI era a principal suspeita (p = 0,011), demonstrando a contribuição da BTB para a definição diagnóstica dessas doenças. O exame histopatológico dessas biópsias revelou que 27,3% dos pacientes com FPI apresentavam o padrão de pneumonia organizante, o que pode sugerir doença mais avançada. O padrão histológico indeterminado foi o mais frequente, refletindo a característica periférica da FPI. Entretanto, o padrão fibrose apresentou alta especificidade e alto valor preditivo negativo. Para os padrões sugestivos de FPI em TC, a curva ROC indicou que a melhor relação entre sensibilidade e especificidade ocorreu com a presença de cinco alterações radiológicas, sendo o aspecto de favo de mel fortemente sugestivo de FPI (p = 0,01). CONCLUSÕES: Nas DPIs, a TC de tórax deve ser sempre realizada e a BTB usada em situações individualizadas, conforme a suspeita e distribuição das lesões.

Ativação das células hepáticas estreladas e fibrose hepática em crianças portadoras de hepatite autoimune tipo 1: estudo imuno-histoquímico de biópsias hepáticas pareadas antes do tratamento e após a remissão clínica

The activation of hepatic stellate cells (HSC) is considered the most important event in hepatic fibrogenesis. The precise mechanism of this process is unknown in autoimmune hepatitis (AIH), and more evidence is needed on the evolution of fibrosis. The aim of this study was to assess these aspects in children with type 1 AIH. We analyzed 16 liver biopsy samples from eight patients, paired before treatment and after clinical remission, performed an immunohistochemical study with anti-actin smooth muscle antibody and graded fibrosisand inflammation on a scale of 0:4 (Batts and Ludwig scoring system). We observedthere was no significant reduction in fibrosis scores after 24± 18 months (2.5 ± 0.93 vs. 2.0± 0.53, P = 0.2012). There was an important decrease in inflammation: portal (2.6 ±0.74 vs. 1.3± 0.89, P = 0.0277), periportal/periseptal (3.0 ±0.76 vs. 1.4 ± 1.06, P = 0.0277), and lobular (2.8 ± 1.04 vs. 0.9± 0.99, P =0.0179). Anti-actin smooth muscle antibodies were expressed in the HSC of the initial biopsies (3491.93 ±2051.48 lm2), showing a significant reduction after remission (377.91 ±439.47 lm2) (P = 0.0117). HSC activation was demonstrated in the AIH of children. The reduction of this activation after clinical remission, which may precede a decrease in fibrosis, opens important perspectives in the follow-up of AIH.

Aspectos morfológicos de biópsias musculares em equinos com miopatia sob forma de surto

As miopatias em equinos são classificadas de acordo com aspectos clínicos, morfológicos e moleculares, em três grandes grupos: não associadas ao exercício, associadas ao exercício e devido alteração da condução elétrica do sarcolema. Apesar dos avanços no diagnóstico, a literatura ainda relata surtos de miopatia em equinos sem etiologia esclarecida em diversos países. O objetivo desse estudo foi descrever as alterações histológicas e histoquímicas de biópsias musculares de equinos acometidos por miopatia. Sete equinos da raça Quarto de Milha, com 18-24 meses de idade, apresentaram sinais clínicos de miopatia. Dentre esses animais, cinco apresentaram sinais subagudos leves a moderados e dois apresentaram sinais hiperagudos severos e decúbito lateral. Foram realizadas biópsias musculares utilizando a técnica percutânea, por agulha tipo Bergström, no músculo glúteo médio em todos os animais acometidos. As amostras foram processadas por meio de técnicas histológicas (HE, Tricrômio de Gomori modificado) e histoquímicas (PAS, NADH, ATPase). Nos quadros clínicos mais leves, a principal alteração encontrada foi a presença de fibras vermelhas rajadas do tipo I e IIA, que estão associadas às alterações do metabolismo oxidativo e das funções mitocondriais, como ocorrem nas miopatias mitocondriais. Também foram observadas fibras atróficas do tipo I e IIA, além da presença de agregados subsarcolemais. Nos quadros mais severos o tecido muscular apresentou infiltrado inflamatório, aumento de colágeno, fagocitose, necrose, calcificação e regeneração muscular. Diante dos achados morfológicos, da resposta à terapia com vitamina E e Se e da baixa mortalidade quando comparado aos relatos de miopatia atípica, conclui-se que esse surto foi desencadeado por lesões mitocondriais, caracterizadas pelas fibras musculares vermelhas rajadas, possivelmente devido uma quebra da homeostase de vitamina E e Se, sendo compatível com o diagnóstico de miopatia nutricional.

Histopathologic Changes Induced by Intense Pulsed Light in the Treatment of Poikiloderma of Civatte

Background The clinical efficacy of intense pulsed light (IPL) in the treatment of poikiloderma of Civatte (PC) is well documented, but little is known about microscopic changes. Objective To analyze histopathologic findings on the necks of individuals with PC after IPL therapy. Materials and Methods Fourteen patients with PC on the neck underwent three monthly sessions of IPL. Biopsies and clinical photographs were taken before and 60 similar to days after treatment. A dermatopathologist analyzed histopathologic slides stained with hematoxylin and eosin, Masson's trichrome, Verhoeff-van Gieson and Fontana-Masson or processed for CD-34 immunohistochemistry. The slides also underwent digital image analysis. Clinical results were based on the analysis of the pictures by three dermatologists and on patient satisfaction. Results Intense pulsed light treatment resulted in more-homogeneous melanin distribution; a greater number of fibroblasts and nonfragmented elastic fibers; and greater density (p similar to=similar to.01), color intensity (p similar to=similar to.02), number and thickness of the collagen bundles. No significant changes in vessels number or diameters were observed. Clinical results were positive in 92.9% of the cases. Conclusion IPL treatment of PC induced a more-homogeneous distribution of melanin and increased nonfragmented elastic fibers, collagen density, and intensity. These changes were related to clinical improvement.

Imuno-histoquímica para o diagnóstico precoce de vitiligo

O vitiligo é uma doença de pele freqüente que acomete 1% da população e é caracterizada por máculas despigmentadas conseqüentes à perda progressiva e localizada dos melanócitos da epiderme. Na maioria dos pacientes, o diagnóstico é feito por exame clínico. A biópsia da pele é realizada quando há necessidade de diagnóstico diferencial com doenças hipocromiantes. O diagnóstico histopatológico de vitiligo é difícil nos preparados corados por hematoxilina e eosina (HE). Há poucos estudos sobre a melhoria da qualidade diagnóstica no vitiligo. OBJETIVO: Avaliar a utilidade dos marcadores imuno-histoquímicos proteína S-100, human melanoma black-45 (HMB-45) e Melan-A para o diagnóstico precoce em casos clinicamente suspeitos ou duvidosos de vitiligo. Material e métodos: Lâminas histológicas de biópsias de pele sã e lesada de 10 pacientes com suspeita clínica de vitiligo coradas pelos métodos de HE, proteína S-100, HMB-45 e Melan-A. Utilizou-se contracoloração com Giemsa como modificação técnica para diferenciar a melanina da imunomarcação. RESULTADOS: Seis casos, com manifestação clínica recente, apresentaram infiltrado linfocitário, do tipo dermatite de interface, na pele lesada na HE. As colorações por S-100, HMB-45 e Melan-A marcaram os melanócitos da camada basal da pele sã, e a proteína S-100 evidenciou as células de Langerhans. Na pele lesada, os melanócitos estavam ausentes ou diminuídos quando comparados com a pele normal. A proteína S-100 demonstrou maior número de células de Langerhans, o que é característico das lesões de vitiligo. CONCLUSÃO: A imuno-histoquímica pode ser utilizada como método auxiliar no diagnóstico dos casos duvidosos de vitiligo.

Bone marrow necrosis related to paracoccidioidomycosis: the first eight cases identified at autopsy

To report the first eight bone marrow necrosis (BMN) cases related to paracoccidioidomycosis (PCM) from patient autopsies with well-documented bone marrow (BM) histology and cytology.A retrospective evaluation was performed on BM specimens from eight autopsied patients from Botucatu University Hospital with PCM-related BMN. Relevant BMN literature was searched and analysed.All eight patients had acute PCM. Six had histological only (biopsies) and two cytological only (smears) specimens. Five biopsy specimens revealed severe and one mild coagulation patterned necrotic areas. Five had osteonecrosis. The cytological specimens also showed typical BMN patterns. Paracoccidioides brasiliensis yeast forms were visible within necrotic areas in all cases.

Borderline leprosy: in situ and cytokine profile in supernatant of mononuclear of cell culture

In order to contribute to a better understanding of cytokine participation in borderline leprosy, in the present study we determined - by in vitro and in situ examinations - the production of these cytokine mediation in non-treated borderline tuberculoid (BT) patients and borderline lepromatous (BL) patients. Seven non-treated BT patients, 12 non-treated BL patients, besides 19 healthy individuals (control group), were evaluated. Peripheral blood mononuclear cells (PBMC) were stimulated or not with specific-M. leprae stimulus (whole and sonicated M. leprae antigens) and a non-specific stimulus. After 48 hours, supernatant was collected for TNF-alpha, IFN-gamma, IL-10 and TGF-beta1 cytokine determination by ELISA. Biopsies from cutaneous lesions were submitted to histological analysis and hematoxylin-eosin and Fite-Faraco stainings; the sections then underwent iNOS, IL-10 and TGF-beta1 in situ detection by immunohistochemistry. Cytokine quantification in PBMC supernatants from patients showed that BT patients produced higher levels of IFN-gamma. Compared to healthy individuals, both borderline patient groups produced lower levels of TGF-beta1 while BL patients generated lower IL-10 levels. The in situ iNOS expression was higher in BT patients compared to BL individuals. on the order hand, TGF-beta1 cytokine revealed a higher proportion of immunostained cells in BL patients. There was no significant difference in IL-10 level between BT and BL patients. Regarding cutaneous lesions, in BL patients there was a negative correlation between TGF-beta1 tissue expression and IL-10. Independently of the clinical form, we observed a positive correlation between TGF-beta1 and bacterial index as well as a negative correlation between the TGF-beta1 tissue expression and iNOS. The results even showed a positive correlation between iNOS tissue expression and production of IFN-gamma by PBMC stimulated with M. leprae antigens. Taken together, the histopathological and immunological observations reinforce the notion of immunological instability in borderline leprosy patients and indicating the participation of mixed cytokines profiles in these individuals, specifically a Th1 profile in BT patients and Th2 profile in BL patients, with a possible participation of T-regulatory lymphocytes.

In vitro and skin lesion cytokine profile in Brazilian patients with borderline tuberculoid and borderline lepromatous leprosy

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Vulvar carcinoma in young patients and its relationship with genital warts

We report the occurrence of aggressive vulvar carcinoma associated with condyloma acuminata in three patients: under 33 years old. Discussion of the role of the human papilloma virus (HPV) in the development of vulvar cancer is also presented. Three patients with condyloma associated with aggressive vulvar squamous cell carcinoma, in situ (1 case) and invasive (2 cases), documented by biopsy and/or vulvectomy are presented. In situ hybridization (ISH) was used to characterize the subtypes of HPV. One patient with erythematous systemic lupus developed in situ carcinoma after 5 years. The other two cases also developed aggressive multicentric, invasive squamous cell carcinoma after 10 years of diagnosis of condyloma. In all cases HPV cytological abnormalities were seen throughout the pathological examination. HPV 16 and 18 were present in cells of invasive squamous cell carcinoma in cases 2 and 3. HPV 6 and 11 were detected only in the condyloma area in case 2. HPV 30 was seen only in the condyloma area in case 3. This report emphasizes the need for biopsies of all unusually persistent or treatment-resistant condylomas, particularly in young and/or immunoisuppressed patients.

Development of a real-time PCR method for rapid sexing of human preimplantation embryos

Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), white 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR. (C) 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Prevalence of hysteroscopic findings and histologic diagnoses in patients with abnormal uterine bleeding

Objective: To describe the prevalence of hysteroscopic findings and histologic diagnoses inpatients with abnormal uterine bleeding (AUB).Design: Retrospective series of consecutive patients.Setting: Tertiary care University hospital and private office.Patient(s): Four thousand fifty-four hysteroscopies with biopsy in patients with AUB evaluated between June 1993 and December 2004.Intervention(S): Hysteroscopies were performed using 2.9-mm or 4-mm telescopes with CO2 or saline as the distension media. Biopsies were (lone with a 5-mm grasper or with a Novak's curette.Main Outcome Measure(s): Prevalence of hysteroscopic findings and histologic analysis of biopsies.Result(S): Endometrial polyp was the most frequent hysteroscopic finding, accounting for 1,374 (33.9%) cases. Normal uterine cavity and cervical canal were found in 814 (20.1%) patients. Submucous fibroids were diagnosed in 302 (7.5%) women. Normal endometrium was the most frequent histologic diagnosis, accounting for 1,888 (46.6%) cases. Endometrial polyp was found in 1, 115 (27.5%) patients. Endometrial hyperplasia was diagnosed in 613 (15.1%) and endometrial cancer in 105 (2.6%) women.Conclusion(s): Endometrial polyps are the most frequent hysteroscopic findings in patients with AUB, whereas normal endometrium is the most frequent histologic diagnosis.

High prevalence of aggregative adherent Escherichia coli strains in the mucosa-associated microbiota of patients with inflammatory bowel diseases

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Relationship of IL-1 and TNF-alpha polymorphisms with Helicobacter pylori in gastric diseases in a Brazilian population

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Clinical presentation and salivary gland histopathology of paediatric primary Sjogren's syndrome

ObjectiveExplore the presentation, diagnostic criteria and exocrine gland histopathology of paediatric primary Sjogren's syndrome (PPSjS).MethodsA case series of 8 children is reported and American-European Consensus Group (AECG-2002) criteria were examined, as well as minor labial salivary and lachrymal gland biopsies, which were scored by a pathologist blinded to outcome. For all cases, connective tissue diseases and parotid-related infectious disease were excluded.ResultsAge at onset varied from 5-13 years old; 6 were females, all followed from diagnosis up to the last visit (1-10 years). The main features at presentation were recurrent tender parotid swelling and sialectasis imaging, with decreased salivary function assessed by Tc-99 scintigraphy. Mild sicca symptoms were observed in 4/8 cases. Systemic features, including fatigue, myalgia, arthritis, tenosynovitis, joint contractures, transient Raynaud's and high ESR, were recorded at onset. Autoantibody profile was unremarkable for diagnosis, while lymphocytic infiltration of labial salivary glands and sialectasis were observed in all biopsies (8/8). In lachrymal glands, massive lymphocytic infiltration and lymphocytic gastritis were observed during complementary assessment. Flares were treated with low dose steroids and long-term use of hydroxychloroquine (5/8), although only 318 fulfilled AECG-2002 diagnostic criteria, throughout the disease course.ConclusionPPSjS is rare, slowly progressive and its early presentation is variable. Standardised diagnostic algorithms should include recurrent parotid swelling and early diagnosis should rely mostly on salivary and lachrymal gland histopathology in this age group.

Ação neuro-muscular do veneno crotálico: dados preliminares

Estudamos 6 pacientes, 2 cães e um coelho com intoxicação crotálica. Avaliamos a condução nervosa periférica sensitiva e motora, a transmissão neuromuscular e eletromiografias. As biópsias de músculo foram processadas por histoquímica. Os 6 pacientes apresentaram mononeuropatia sensitiva no nervo periférico adjacente ao local da inoculação do veneno e encontramos evidências histoquímicas de miopatia mitocondrial. Os defeitos da transmissão neuromuscular foram mínimos. A maioria dos autores admite que veneno crotálico determina síndrome miastênica. Nossos achados indicam que ptose palpebral, facies miastênico e fraqueza muscular observados após acidente crotálico, correspondem provavelmente a miopatia mitocondrial, muitas vezes transitória e reversível.