Crystal violet stain as a selective stain for the assessment of mitotic figures in oral epithelial dysplasia and oral squamous cell carcinoma.

Background: Assessment of mitotic figures (MFs) is routinely practiced as prognostic indicator in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC), but identification of MFs poses a problem in terms of staining characteristics. Aim: To evaluate effectiveness of crystal violet stain for staining of MFs and its comparison with hematoxylin and eosin (H and E) stain. Materials and Methods: Study sample includes archival tissues embedded in paraffin blocks diagnosed as OED (n = 30) and OSCC (n = 30). The control group comprised of tissue specimen from oral mucosa of healthy volunteers (n = 30). Two serial sections of each tissue specimen were stained separately with H and E stain and 1% crystal violet stain. The stained sections were observed under microscope for identification and counting of MFs. Data obtained was statistically analyzed by using the Man-Whitney U test. Results: A significant increase in number of MFs was observed in OED and OSCC in comparison with normal oral mucosa. There was a highly significant increase in number of MFs in crystal violet stained tissue sections when compared with H and E stain. Metaphase is the most commonly observed phase of mitosis in crystal violet stain when compared with H and E stain for all three groups. Conclusion: Crystal violet stain can be considered as selective stain for mitotic figures.

Study of parameters to ensure quality control in histopathology reporting: A meta-analysis at a tertiary care center.

Context : When surgical pathology reports are dispatched to patients and clinicians, sometimes they are discovered to have errors, and it is a common practice for the pathologists to issue amended reports. Measuring the rate at which surgical pathology reports are amended can be used as a tool for assuring quality control in histopathology. Aim : The aim of this study was determine the parameters that can be used as an assessment tool to minimize errors in histopathology. Materials and Methods : This study was carried out at a major histopathology center. The duration of this study was from January 2001 through January 2011(ten years). Following parameters were looked for: Interpretational errors, permanent and frozen section correlation, intradepartmental consultation and cases sent for second opinion, cases brought in tissue committee meetings, audits, and cases discussed in hospital meetings. Results : A total of 28,1931 surgical pathology cases were signed out during the ten-year period. On these, addendums were issued on 5730 cases (2.0%). Additional report issued on 3521 (1.3%). Addendum/corrected report issued for 2209 cases, which was 0.7%, representing the true interpretational error. And out of this number, a second opinion was taken for 5980 cases, and 78 were sent abroad for second opinion. Conclusion : Review by a second pathologist is a strong tool to minimize errors in surgical pathology reporting. This may be done prior to or after the report is dispatched and the case is discussed in the hospital for treatment purposes. This analysis concludes that true interpretational error occurred only in 0.7% of cases, which is an attribute to the strong peer review in the department.

Primary cutaneous amyloidosis: A clinico-pathological study with emphasis on polarized microscopy.

Background: Primary localized cutaneous amyloidosis (PCA) is a relatively rare condition characterized by amyloid deposition in dermis without systemic involvement. Although, histopathological examination of the lesion reveals amorphous eosinophilic deposits in papillary dermis examination of congo red stained slides under polarized light will give definitive diagnosis Aims: To study the clinicopathological features of cutaneous amyloidosis emphasizing the utility of polarized light in diagnosis. Materials and Methods : A clinicopathological study of primary cutaneous amyloidosis over a period of 8 years was undertaken. All the cases, clinically diagnosed and histopathologically proven as cutaneous amyloidosis were stained with congo red and studied under polarized light. Results and Conclusions: Of the 45 cases of clinically suspected amyloidosis, 32 cases were proven histopathologically as primary cutaneous amyloidosis and confirmed by congo red stain under polarized light which showed apple green birefringence. Among the two types of PCA, lichen amyloidosis was the most common variant accounting to 65.63% with pure cases of macular amyloidosis accounting for only 15.63%. Biphasic amyloidosis was seen in 18.75%. Knee was the commonest site of involvement with pruritis being the most common symptom. Histopathologically, the most common findings were hyperkeratosis, irregular acanthosis and expansion of dermal papillae by amyloid deposits showing apple green birefringence under polarized microscope with congo red staining. Although, H and E stain gives a clue for the diagnosis of amyloid nevertheless congo red staining under polarized light forms a very sensitive and definitive method for confirmation.

Core biopsies of the breast: Diagnostic pitfalls.

The incidence of breast cancer is increasing worldwide. In this review article, the authors compare and contrast the incidence of breast cancer, and the inherent differences in the United States (US) and India in screening techniques used for diagnosing breast cancer. In spite of these differences, core biopsies of the breast are common for diagnosis of breast cancer in both countries. The authors describe "Best Practices" in the reporting and processing of core biopsies and in the analysis of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor Receptor 2 (Her2/neu). The pitfalls in the diagnosis of fibroepithelial lesions of the breast on core biopsy are discussed, as also the significance of pseudoangiomatous stromal hyperplasia of the breast (PASH) is discussed in core biopsy. In this review, the management and diagnosis of flat epithelial atypia and radiation atypia are elaborated and the use of immunohistochemistry (IHC) in papillary lesions, phyllodes tumor, and complex sclerosing lesions (radial scars) is illustrated. Rarer lesions such as mucinous and histiocytoid carcinoma are also discussed.

Evaluation of the effect of presence of blood in the stomach on endoscopic diagnostic tests for Helicobacter pylori infection.

Introduction: Presence of blood in the stomach has been thought to affect the performance of diagnostic tests used in detecting Helicobacter pylori (H. pylori) in the stomach. This study evaluated the effect of blood on the efficacy of rapid urease test (RUT) and microscopic appearance of the biopsy after staining with Giemsa stain. Materials and Methods: Patients with bleeding oesophageal varices who met the inclusion criteria were tested for H. pylori by RUT and microscopic examination of the biopsy. A repeat endoscopy, RUT and histology were done one month following initial presentation. The performance of the diagnostic tests was evaluated with and without the presence of intraluminal blood. A combined result of the two tests, RUT and histology, carried out in presence or absence of blood for the diagnosis of H. pylori, when considered together was considered as the gold standard. Results: Thirty six patients included in the study were in the ages ranging between 15-60 years (mean age = 44.14 years ±2.1). The combination of tests at both visits showed 20/36 (55.6%) patients were positive for H. pylori. The decrease in H. pylori positivity in the presence of blood was significant for RUT (8.3% vs. 38.9%; P=0.005) and combined test (19.4% vs. 47.2%; P=0.02) but the decrease in positivity for histology (11.1% vs 30.6%) was not significant (P=0.08). In the presence of blood, the sensitivity of RUT, histology and combined tests were 15%, 20% and 35%, respectively. In the absence of blood, the sensitivity of RUT, histology and combination of tests was 70%, 55% and 85%, respectively. Conclusion: Blood in the stomach significantly decreased the sensitivity of RUT, histology and the combination of both. Negative results of these tests in acute upper gastro intestinal (GI) bleeding should therefore be interpreted carefully.

Histochemistry and morphometric analysis of muscle fibers from patients with duchenne muscular dystrophy (DMD) / Histoquímica y análisis morfométrico de las fibras musculares de p0acientes con distrofia muscular de duchenne (DMD)

The aim of the study was to analyze the muscle fibers by histochemistry and morphometric methods from patients with Duchenne muscular dystrophy (DMD). Muscle biopsies were taken from the vastus lateralis muscle of five boys between 13 and 15-years of age, with clinical diagnosis of DMD. The histochemistry was performed using myofibrillar ATPases (9.6, 4.6 and 4.3). To morphometrical analysis a computerized semiautomatic system and software Image-Lab was used. ATPase staining showed atrophy of muscle fibers. Fibrosis and adipose deposition occurred in variable degree depending of muscular involvement. The morphometrical analysis showed an increase of size (percentage) to type I fiber than other types in all patients. Furthermore, the type I fiber had a larger cross-sectional area and mean diameter than type IIa and IIb fibers. Both histochemistry and morphometric analysis could be important tools for qualitative and quantitative diagnostics of muscle fibers attacked in this type of disease.

El objetivo del estudio fue analizar las fibras musculares mediante histoquímica y métodos morfométricos en pacientes con distrofia muscular de Duchenne (DMD). Se tomaron biopsias musculares del músculo vasto lateral de cinco niños entre 13 y 15 años de edad, con diagnóstico clínico de DMD. La histoquímica se realizó mediante ATPasa miofibrilar (9.6, 4.6 y 4.3). Para el análisis morfométrico se utilizó un sistema semiautomático computarizado y software de imagen de laboratorio. La tinción de ATPasa mostró una atrofia de las fibras musculares. La fibrosis y depósito adiposo se observó en grado variable dependiendo del compromiso muscular. El análisis morfométrico mostró un aumento de tamaño (porcentaje) de fibras tipo I en todos los pacientes. Además, la fibra tipo I tuvo un área de sección transversal y diámetro medio mayor que las fibras tipos IIa y IIb. Tanto la histoquímica y el análisis morfométrico pueden ser herramientas importantes para el diagnóstico cualitativo y cuantitativo de las fibras musculares comprometidas en este tipo de enfermedad.

Localization of sugar residues in the stomach of three species of monkeys (Tupaiidae glis, Nycticebus cocang and Callithrix jacchus) by lectin histochemistry / Localización de los residuos de azúcar en el estómago de tres especies de monos (Tupaiidae glis, Nycticebus cocang y Callithrix jacchus) por histoquímica de lectina

The stomach of three species of non-human primates was investigated by lectin histochemistry to clarify the staining affinity and distribution patterns of their sugar residues. All gastric regions, with little differences between the deep and superficial parts of the same region, were rich in. in N-acetylglucosamine and/or neuraminic acid. Although, the superficial regions of the gastric mucosa were scanty in N-acetylgalactosamine, a- D-glucose and a -D-mannose, the deep parts of the gastric mucosa were rich in these sugars. In conclusion, there is a difference among the mucosubstances of surface and foveolar mucous cells, mucous neck cells, and gastric gland cells. This indicates heterogeneous composition of gastric mucus, or mucus molecules with variations in the degree of glycosylation of their oligosaccharide chains in the different cells which suggest that lectin binding affinity in the gastric mucosa correlated mostly to the degree of cellular differentiation.

El estómago de tres especies de primates no humanos fue investigado por histoquímica de lectinas para determinar la afinidad de tinción y los patrones de distribución de sus residuos de azúcar. Todas las regiones gástricas, con pequeñas diferencias entre las partes profundas y superficiales de la misma región, eran ricas en N-acetilglucosamina y/o ácido neuramínico. Si bien, las regiones superficiales de la mucosa gástrica eran escasas en N-acetilgalactosamina, a-D-glucosa y a-D-manosa, las partes profundas de la mucosa gástrica eran ricas en estos azúcares. En conclusión, existe una diferencia entre las mucosustancias de la superficie y células mucosas foveolares, células mucosas del cuello y células de las glándulas gástricas. Esto indica una composición heterogénea de la mucosa gástrica, o moléculas de moco con variaciones en el grado de glicosilación de sus cadenas de oligosacáridos en las diferentes células, sugieriendo que la afinidad de union de lectinas en la mucosa gástrica se relacionada principalmente con el grado de diferenciación celular.

Diferencias en la morfología de las células prostáticas en la circulación sanguínea (cpcs), células cancerosas aisladas (CPCS) en la medula ósea y células cancerosas infiltrando microgramos en la medula ósea de pacientes con cáncer prostático, detectadas utilizando métodos de inmumocitoquímica / Morphological differences between circulating prostate cells in blood and bone marrow and those infiltrating bone marrow fragments in patients with prostate cancer, detected using immunocytochemical methods

Objetivo: Describir y determinar diferencias en la morfología celular entre células prostáticas en la circulación sanguínea y muestras de la medula ósea en pacientes con cáncer prostático. Métodos y Pacientes: Después de un consentimiento informado por escrito, muestras de sangre venosa y médula ósea fueron extraídas de pacientes con cáncer prostático histológicamente confirmado. Células mononucleares fueron obtenidas por centrifugación diferencial y una impronta preparada de la biopsia de la médula ósea. Las células prostáticas fueron identificadas con inmunocitoquímica, definida como anfígeno prostático especifico positivo y CD45 (pan-Ieucocito) negativo. Resultados: De los 71 sujetos estudiados, las células obtenidas en sangre y aspiración de la medula ósea, fueron redondas u ovaladas, a diferencia de las células en los microgramos o fragmentos que tuvieron las características típicas de las células malignas: irregulares, con diferencias en tamaño y con largos procesos citoplasmáticos. Conclusiones: Las diferencias en la morfología pueden ser explicadas debido a que habrían 2 tipos de células, unas circulando o en transito entre la sangre y otros tejidos -como médula ósea- con apariencia redonda u ovalada vías células en los microgrumos o fragmentos, que se han infiltrado la membrana endoostial y representan verdadera micro- metástasis.

Describe and determine morphological differences between prostate cells found in blood and bone marrow in men with prostate cancer. Methods and Patients: After written informed consent, samples of blood and bone marrow were obtained from men with histological confirmed prostate cancer. Mononuclear cells were obtained by differential centrifugation and a touch-prep made from the biopsy sample. Prostate cells were identified using immunocytochemistry and defined as cells positive for anti-prostate specific antigen and negative for anti-CD45 (pan-leucocyte). Results: From the 71 patients studied, cells found in blood and bone marrow aspirates were round or oval, whereas the cells detected in microfragments of the biopsy showed the characters of a typical cancer cell, irregular in shape and size with long cytoplasmic processes. Conclusions: These morphological differences could be explained in that there are 2 populations of cells, those in blood or bone marrow aspirates with a round or oval appearance are in transit, circulating between the blood and other tissues such as bone marrow, and those found in microfragments which represent true micrometastasis.

Biópsia muscular com estudo histoquímico: experiência inicial da Faculdade de Medicina de São José do Rio Preto / Muscle histochemistry: a study of the first 55 biopsies in São José do Rio Preto

São apresentados os primeiros 55 casos de biópsia muscular com estudo histoquímico da Faculdade de Medicina de São José do Rio Preto. A histoquímica muscular permite: 1. Revelar a natureza não uniforme dotecido muscular demonstrando as propriedades bioquímicas de diferentes tipos de fibra e sua participação seletiva em determinadas doenças; 2. Detectar ausência de enzimas específicas; 3. Detectar excesso ou acúmulo de substratos específicos; 4. Detectar anormalidades estruturais musculares que não aparecem nas reações histológicas rotineiras. A média de idade foi de 28,6 anos, variando de 25 dias a 76 anos, 56,4% eram do sexo masculino e 43,6% feminino. Foram rotineiramente utilizadas as seguintes técnicas: Hematoxilina &Eosina, Tricrômio de Gomori modificado, Citocromo-C-Oxidase, Dinucleotídeo Adenina Nicotinamida Desidrogenase reduzida pelo tetrazólio, Succinato-Desidrogenase, Fosfatase Alcalina, Fosfatase Ácida, Adenosina Trifosfato miofibrilar pré-incubadas a pH 9,4, 4,6 e 4,3, Ácido Periódico de Schiff e Oil-Red-O. Anormalidades foram encontradas em 72,8% das biópsias, sendo de padrão distrófico em 17 casos, inflamatório em 7 casos, mitocondrial em 6 casos, reinervação crônica em 4 casos, glicogenose em 2 casos e outros diagnósticos em 4 casos. Anormalidades mínimas ou inespecíficas foram encontradas em 5 casos (9%) e em10 casos (18,2%) foram normais. Nossos achados foram semelhantes aos de outros descritos na literatura nacional. A biópsia muscular como estudo histoquímico, ainda inexistente na região Noroeste do estado deSão Paulo, é uma das armas diagnósticas mais importantes para as doenças neuromusculares.

We present the muscle histochemistry from the first 55 biopsies at the State Medical School of São José doRio Preto, São Paulo, Brazil. Muscle histochemistry first, demonstrate the non-uniform nature of muscular tissue showing biochemical properties from different fiber types and their selective involvement in particular diseases; second, demonstrate the absence of specific enzymes; third, demonstrate abnormal storage of specific substrates, and fourth, demonstrate structural abnormalities that are not seen on ordinary histological stainings. The mean age was 28.6 years, ranging from 25 days to 76 years, 56.4% male and 43.6% female. Ourroutine techniques, done in all biopsies, included: Hematoxylin and Eosin, Gomori Trichrome, CytochromeOxidase, NADH dehydrogenase, Succinic Dehydrogenase, Alkaline Phosphatase, Acid Phosphatase, ATPase pH 9.4, 4.6 and 4.3, Periodic-acid Schiff and Oil-Red-O. Abnormalities were found in 72.8%, being dystrophicin 17 cases, inflammatory in 7 cases, mitochondrial in 6 cases, chronic reinnervation in 4 cases, glycogen storage disorder in 2 cases and other diagnostics in 4 cases. Minimal or unspecific abnormalities were found in 5 cases (9%) and were normal in 10 cases (18.2%). Our findings were similar to others described in Brazil. Muscle histochemistry is one of the most significant diagnostic tools for neuromuscular disorders and was not done before in this region of Brazil.

Gleason scoring of prostatic carcinoma: impact of a web-based tutorial on inter- and intra-observer variability.

A total of 40 cases of prostatic adenocarcinomas were scored independently by four pathologists using the Gleason scoring system. After attending a web-based tutorial, the scoring was repeated by all. Consensus scores were obtained by simultaneous viewing of each case in a multihead microscope by all four pathologists. The scores were then compared. The pretutorial kappa (kappa) values ranged from 0.36 to 0.64 with an average of 0.459. After the tutorial, the kappa values ranged from 0.44 to 0.678 with the average kappa value increasing to 0.538, thus indicating an improvement in the agreement. The intraobserver agreement ranged from 0.435 to 0.788. We conclude that web-based tutorials with emphasis on images developed by experts serve to achieve an uniformity in reporting.

O corpo gorduroso de Aedes (Stegomyia) aegypti (Diptera; Nematocera) (Linnaeus, 1762): estudo morfológico do órgão em diferentes idades e condições alimentares, isolamento, cultivo primário e transcriptoma dos enócitos

O corpo gorduroso é o principal órgão do metabolismo intermediário dos insetos e a principal fonte de componentes da hemolinfa. Dois tipos celulares estão presentes no corpo gorduroso dos dípteros: os trofócitos e os enócitos. O presente trabalho teve como objetivos testar as seguintes hipóteses: (a) o corpo gorduroso de A. aegypti é capaz de se reorganizar do ponto de vista ultra-estrutural, histoquímico e morfométrico de acordo com a idade e com o tipo de alimentação; (b) os enócitos de A. aegypti podem ser cultivados e (c) os mesmos expressam transcritos relacionados com os processos de desintoxicação, síntese de lipídios e imunidade inata. Para tanto, foram utilizadas fêmeas recém-emergidas, fêmeas com 18d de idade, sendo um grupo 18h e outro quatro dias após a alimentação sangüínea e fêmeas com 18d alimentadas com açúcar. Nas fêmeas alimentadas com açúcar, o corpo gorduroso está mais desenvolvido do que nas pós-mergidas e nas alimentadas com sangue. Nessas mesmas fêmeas, os trofócitos apresentam o citoplasma preenchido basicamente por gotículas de lipídio devido à lipogênese. Após a alimentação sangüínea, os lóbulos do corpo gorduroso estão achatados, o que pode ser explicado pelo esvaziamento do órgão que exporta nutrientes para os ovários. As alterações dos trofócitos após a alimentação sangüínea incluem o aumento do diâmetro nuclear, a diminuição das gotículas de lipídios, o alargamento das mitocôndrias e a presença de lisossomos nos trofócitos. Os enócitos puderam ser mantidos viáveis por 57 dias em cultivo primário e o citoplasma dessas células é quase todo preenchido pelo retículo endoplasmático liso (REL). Os transcritos mais abundantes encontrados nos enócitos correspondem à citocromo P450, responsável pela desintoxicação e às outras proteínas relacionadas com o metabolismo de lipídios.

Também são encontrados transcritos. relacionados com o reconhecimento e de destruição de patógenos. Provavelmente, a presença do REL e a localização periférica dos enócitos no corpo gorduroso de A. aegypti facilita o processo de desintoxicação, a produção e a secreção de lipídios para a hemolinfa, além do reconhecimento de patógenos e da secreção de componentes antimicrobianos. A presença do REL desenvolvido e a expressão de várias enzimas comprometidas com a desintoxicação e o metabolismo de lipídios sugerem que os enócitos participam da homeostasia e síntese lipídica em A. aegypti. Além disso, nossos resultados confirmam o corpo gorduroso de A. aegypti como um órgão capaz de reorganizar sua microanatomia, componentes citoplasmáticos e aspectos subcelulares de acordo com a idade e o tipo de dieta. Também é possível cultivar os enócitos e sua purificação, cultivo primário e transcriptoma constituem importantes ferramentas para que futuros estudos possam investigar a atuação dessas células na fisiologia e na imunidade inata de mosquitos

Morphoquantitative analysis of the intraepithelial neoplasic uterine cells by computer image system / Análisis morfocuantitativo de las células neoplásicas intraepiteliales uterinas por sistema computarizado

The purpose of the present research was to detect the DNA variations by Feulgen stained chromatin and the packing degree of the cervical chromatin in the clinical cases of human uterine carcinomas. Scanning microphotometry was carried out on slides prepared from block Feulgen-stained. Accurate control procedures were performed using normal human cell nuclei. The cervical cells was morphometric studied using the Image Lab System for citologic classification. The overall diagnostic accuracy for neoplasia type I (CIN I) and type II (CIN II), demonstrated by stain interpetration values of a sensitive detection of DNA neuroploidy and a significant difference between the two groups in scanning static cytophotometry.

La investigación tuvo como objetivo detectar las posibles variaciones del DNA en el núcleo de las células marcadas por la reacción histoquímica de Feulgen en individuos con diagnóstico de neoplasias intraepiteliales de grados I y II del cuello uterino. La microcitometría fue realizada en láminas provenientes de hospitales universitarios y estatales, sometidas a reacción de Feulgen y analizadas en el sistema de imagen (Image Lab) para cuantificación. Los resultados encontrados entre las células neoplásicas de grado I (NIC I) y del grado II (NIC II) demostraron que el método es sensible y eficiente en la cuantificación entre las diferentes clasificaciones, ayudando así en los diagnósticos de las lesiones del cuello uterino en su fase inicial.

O corpo gorduroso de Aedes (Stegomyia) aegypti (Diptera; Nematocera) (Linnaeus, 1762): estudo morfológico do órgão em diferentes idades e condições alimentares, isolamento, cultivo primário e transcriptoma dos enócitos / The fat body Aedes (Stegomyia) aegypti (Dipetra; Nematocera) (Linnaeus, 1762): Morphological study of the organ in different ages and diets, isolation, primary culture and the transcriptome of the oenocytes

O corpo gorduroso é o principal órgão do metabolismo intermediário dos insetos e a principal fonte de componentes da hemolinfa. Dois tipos celulares estão presentes no corpo gorduroso dos dípteros: os trofócitos e os enócitos. O presente trabalho teve como objetivos testar as seguintes hipóteses: (a) o corpo gorduroso de A. aegypti é capaz de se reorganizar do ponto de vista ultra-estrutural, histoquímico e morfométrico de acordo com a idade e com o tipo de alimentação; (b) os enócitos de A. aegypti podem ser cultivados e (c) os mesmos expressam transcritos relacionados com os processos de desintoxicação, síntese de lipídios e imunidade inata. Para tanto, foram utilizadas fêmeas recém-emergidas, fêmeas com 18d de idade, sendo um grupo 18h e outro quatro dias após a alimentação sangüínea e fêmeas com 18d alimentadas com açúcar. Nas fêmeas alimentadas com açúcar, o corpo gorduroso está mais desenvolvido do que nas pós-mergidas e nas alimentadas com sangue. Nessas mesmas fêmeas, os trofócitos apresentam o citoplasma preenchido basicamente por gotículas de lipídio devido à lipogênese. Após a alimentação sangüínea, os lóbulos do corpo gorduroso estão achatados, o que pode ser explicado pelo esvaziamento do órgão que exporta nutrientes para os ovários. As alterações dos trofócitos após a alimentação sangüínea incluem o aumento do diâmetro nuclear, a diminuição das gotículas de lipídios, o alargamento das mitocôndrias e a presença de lisossomos nos trofócitos. Os enócitos puderam ser mantidos viáveis por 57 dias em cultivo primário e o citoplasma dessas células é quase todo preenchido pelo retículo endoplasmático liso (REL). Os transcritos mais abundantes encontrados nos enócitos correspondem à citocromo P450, responsável pela desintoxicação e às outras proteínas relacionadas com o metabolismo de lipídios.

Também são encontrados transcritos. relacionados com o reconhecimento e de destruição de patógenos. Provavelmente, a presença do REL e a localização periférica dos enócitos no corpo gorduroso de A. aegypti facilita o processo de desintoxicação, a produção e a secreção de lipídios para a hemolinfa, além do reconhecimento de patógenos e da secreção de componentes antimicrobianos. A presença do REL desenvolvido e a expressão de várias enzimas comprometidas com a desintoxicação e o metabolismo de lipídios sugerem que os enócitos participam da homeostasia e síntese lipídica em A. aegypti. Além disso, nossos resultados confirmam o corpo gorduroso de A. aegypti como um órgão capaz de reorganizar sua microanatomia, componentes citoplasmáticos e aspectos subcelulares de acordo com a idade e o tipo de dieta. Também é possível cultivar os enócitos e sua purificação, cultivo primário e transcriptoma constituem importantes ferramentas para que futuros estudos possam investigar a atuação dessas células na fisiologia e na imunidade inata de mosquitos

Caracterización histoquímica y morfométrica de la musculatura esquelética del miembro pelviano del caprino (Capra hircus) / Histochemical and morphometric characterization of the skeletal musculature of the hind limb of the caprine (Capra hircus)

Se estudiaron los músculos de la región de la cadera, del muslo y del pie del miembro pelviano del caprino, en condiciones de explotación semi-intensiva con técnicas histoquímicas, con la finalidad de conocer la distribución porcentual de los distintos tipos de fibras. Además, contribuye a la caracterización del músculo normal, condición indispensable para el estudio posterior de la patología, la genética y eventualmente, para determinar algunas características organolépticas de la carne. Se tomaron muestras in vivo de los músculos bíceps femoral, glúteo medio, glúteo profundo, vasto intermedio, vasto lateral, gastrocnemios, soleo, semimembranoso, semitendinoso y tensor de la fascia lata, en cinco (5) cabras de 21-30 Kg. de peso. Las muestras se congelaron en isopentano enfriado en nitrógeno líquido. Los cortes se realizaron en un críostato a -20°C, con un espesor de 10 µm. Se realizaron las reacciones de laAdenosina Trifosfatasa (ATP-asa) con un pH de preincubación 4,33 y 4,37 para determinar la distribución porcentual de tipos de fibras. El músculo vasto intermedio presentó 100 por ciento de fibras tipo I. Los tres músculos glúteos mostraron alrededor de 45 por ciento fibras tipo I, 22 por ciento de tipo IIa y 33 por ciento tipo IIb. El resto de los músculos tuvo 29 por ciento de las fibras tipo I, 31por ciento del tipo IIa y 40 por ciento del tipo IIb. El promedio de área de fibra más alto correspondió al músculo vasto intermedio con 4.070 µm² y el promedio de área por tipo de fibra reveló que la fibra más grande fue la fibra tipo I con 2.682 µm² y la más pequeña fue la tipo IIb con 2.046 µm². En conclusión, se caracterizó la composición miofibrilar de diez (10) músculos del miembro pelviano de caprino de la raza Alpino Francés.

The muscles of the hip, high and leg region of the goats were studied under conditions of explotation semi intensive with technical histochemical, with the purpose of knowing the percentage distribution of the different types of fibers, that which contributes to the characterization of the normal muscle, indispensable condition for the later study of the pathology, the genetic one and possibly to determine some organoleptics characteristic of the meat. They took samples in alive of the muscles Biceps femoris, Middle gluteal, Deep gluteal, Vastus intermedius, Vastus lateralis, Gastrocnemius, Soleus, Semimembronosus, Semitendinosus and Tensor fasciae latae, in five (5) goats of 21-30 Kg. of weight. The samples froze in isopentano cooled in liquid nitrogen. The cuts were carried out in a cryostat to -20°C, with a thickness of 10 µm. They were carried out the reactions of the Adenosine Trifosfatase (ATP-handle) with a pH of preincubation 4.33 and 4.37 to determine the percentage distribution of types of fibers. The muscle Vastus intermedius presents 100% of fibers type I, almost all of high capacity oxidative. The three muscles Gluteal showed around 45% fibers type I, 22% typeIIa and 33% type IIb. The rest of the muscles had 29% of the fibers type I, 31% of the type IIa and 40% of the type IIb. The highest average of fiber area was that of the m. Vastus intermedius with 4,070 µm² and the area average for fiber type revealed that the biggest fiber was the fiber type I with 2,682 µm² and the smallest was the type IIb with 2,046 µm². In conclusion, the composition myofibril of ten was characterized (10) muscles of the later member of goats of the French Alpine beed.

Intestinal Auerbach plexus structures with NADH histochemistry in three strains of spontaneous diabetic rats / Estructuras del plexo de Auerbach intestinal con histoquímica del NADH en tres líneas de ratas espontáneamente diabéticas

The enteric nervous system comprises two major systems: the submucosal and the myenteric plexus. Theaim of this study was to describe the myenteric plexusfrom three strains of spontaneous diabetic rats from thehistological point of view. Samples of small intestineand of proximal and distal colon were obtained fromthree spontaneous diabetic rats i.e., eSS, eSMT, βstrains and 1-year old Wistar rats. Specimens werestained with NADH (β-nicotinamide adenine dinucleotide,reduced form) histochemical technique andexamined with light microscope. Microscopically little modifications in mesh-like structure of intestinal Auerbach’s plexus from eSS were detected in comparisonwith Wistar rats samples. Intestinal plexus of eSMT and β rats showed disruption of mesh-like structures, modifications in the slightly colored background (smooth muscle) and augmented vascularization. Small intestine and colon are affected. In short: In our spontaneously diabetic rat models, mesh-like structure of Auerbach’s plexus is strain dependent.

El sistema nerviso entérico comprende dos sistemas mayores: el plexo submucoso y el plexo mientérico. Este estudio describe la estructura histológica del plexo mientérico en tres líneas de ratas espontáneamente diabéticas. Especímenes de intestino delgado, colon proximal y colon distal fueron obtenidos de tres líneas de ratas espontáneamente diabéticas: eSS, eSMT, β y Wistar de 1 año de edad. Los materiales obtenidos fueron procesados con la técnica histoquímica del NADH (β-nicotinamida adenina dinucleotido, forma reducida) y observados en un microscopio óptico. Pequeñas modificaciones histológicas en la estructura reticular del plexo de Auerbach intestinal pueden ser detectados en las ratas eSS cuando son comparadas con las ratas Wistar. La estructura reticular del plexo de Auerbach de las ratass eSMT y β muestran una desaparición de dicha estructura reticular, disminución de la coloración de base (músculo liso) y un aumento de la vascularización. Tanto el intestino delgado como el colon están afectados. Resumiendo: en nuestros modelos experimentales de ratas diabéticas la estructura reticular del plexo de Auerbach es dependiente de la línea de rata estudiada.

Avaliação histoquímica da mucosa gastrointestinal de ratos expostos ao álcool / Histochemical avaliation of the gastrointestinl mucosa of rats exposed to alcohol

Objetivo: avaliar através da histoquímica com lectinas a expressão de glicosaminoglicanos (mucinas) nas células do trato gastrointestinal de ratos expostos ao etanol. Método: ratos foram expostos ao etanol (3 g/kg do peso) durante 15, 30 e 45 dias. Após a perfusão, o estômago e o intestino fixados em formalina tamponada a 10% e fragmentos desses órgãos emblocados em parafina. Os cortes histológicos (4µm) foram incubados com lectil1as (PNA, WGA e ConA) conjugadas à peroxidase. A marcação das lectinas se revelou com o DAB-H202. seguida pela contra-coloração com hematoxilina. Os métodos Periodic acid-shiff (PAS) e Alcian blue foram usados para avaliar a expressão de glicosaminoglicanos (mucinas). Resultados: verificou-se que na mucosa gástrica a expressão das mucinas aumentou progressivamente durante a exposição ao etanol, conforme o aumento do número de células PAS e Alcian blue positivas. Por outro lado, na mucosa intestinal nenhuma diferença considerável se observou no número dessas células. Todas as lectinas testadas apresentaram um aumento no padrão de marcação relacionado ao maior período de exposição ao etanol. A PNA foi a mais seletiva das lectinas, reconhecendo, exclusivamente, células do cólon e das glândulas gástricas. A Con A apresentou uma intensa marcação nas regiões apicais das glândulas do cólon e do estômago, enquanto que a WGA reagiu, intensamente, nas células caliciformes e células de Paneth nas glândulas do epitélio intestinal. Conclusão: esses achados demonstram que a exposição ao etanol, leva a uma alteração dos padrões de reconhecimento das lectinas nas células do tecido gastrointestinal, bem como, um aumento a expressão gástrica das mucinas.

Objectives: in this work, lectin histochemistry was used to evaluate the expression of glycosaminoglycan (mucin) in gastrointestinal cells of rats exposed to ethanol. Method: Rats were exposed to ethanol (3g/kg of weight) during 15, 30 and 45 days. After perfusion, the stomach and intestine were formalin 10% fixed and the tissues were paraffin embedded. Slices (4mm) were incubated with peroxidase-conjugated lectins (PNA, WGA and Con A). Lectin staining was revealed with DAB-H2O 2followed by haematoxylin counterstaining. Periodic acid-Shiff (PAS)and Alcian Blue staining methods were used to evaluate glycosaminoglycan expression. Results: indicated that, in gastric mucosae, acid and neutral mucin expression rose progressively during ethanol exposition while in intestinal mucosae no considerable difference was observed in the number of PAS- and Alcian Blue-positive staining cells. All lectins presented an increasing binding pattern related to the period of ethanol exposure. PNA was the most selective lectin, recognizing exclusively colon cells and gastric glands. Con A presented an intense binding to the bottom region of the colon stomach glands while WGA intensely bound to the caliceform and Paneth cells in the intestinal glands. Conclusion: These jindings demonstrate that ethanol exposure led to a different pattern of lectin recognition of the cells of the gastrointestinal tissue.

Role of FAB classification of acute leukemias in era of immunophenotyping.

French-American-British classification for leukemias had been widely accepted due to its objectiveness and good reproducibility. WHO classification of leukemias was formulated in 1997 with a purpose of further enhancing the objectivity. However, the requirement of cytogenetics and immunophenotyping makes it difficult for many countries like India to put WHO classification in routine use. This study was carried to know the effectiveness of FAB classification in an era of technical advancement. A retrospective analysis of all acute leukemias over a period of 2 years was done. Out of total of 469 cases of acute leukemias, 193 were diagnosed as Acute Lymphoblastic Leukemia (ALL), 200 as Acute Myeloid Leukemia (AML), and 76 cases diagnosed as Acute Leukemia, cytochemically undifferentiated. Hence, only 16% of all leukemias remained unclassifiable. Subclassification of AML cases revealed a much higher percentage of AML-M3, as compared to western literature. In conclusion, FAB classification, based on morphology and simple cytochemical stains, remains effective enough, although cytogenetics and immunophenotyping can add to diagnostic accuracy in some cases.

Técnica histoquímica aplicada ao tecido ósseo desmineralizado e parafinado para o estudo do osteócito e suas conexões / Histochemical tecnique for osteocyte and its connections study in descalcified and paraffined bone tissue

O osteócito vem sendo alvo de pesquisas recentes. A avaliação in situ de sua morfologia, da atividade e das características de suas conexões é difícil, portanto é realizada apenas com técnicas avançadas. Devido à importância desse tipo celular na manutenção da matriz óssea, este estudo propõe uma técnica de coloração pela prata como alternativa para o estudo do osteócito e suas conexões em tecido ósseo desmineralizado e parafinado. Cortes de 4æm do fêmur de ratas foram desmineralizados, desparafinados em xilol e hidratados em concentrações decrescentes de álcool etílico (ETOH) e água miliQ. Para a impregnação foram utilizadas soluções de nitrato de prata a 50 por cento e de ácido fórmico a 1 por cento com 2 por cento de gelatina microbiológica em estufa a 40ºC. Essa técnica permite visualizar facilmente as bordas lacunares dos osteócitos e suas conexões, proporcionando uma alternativa simples e eficaz para o estudo da morfologia desse tipo celular até então ainda não proposta com essa finalidade.

Significance of AgNOR score in benign and malignant soft tissue tumours.

AgNOR staining was employed on FNAC and histopathological sections obtained from patients with soft tissue tumours. The study comprised of 20 normal appearing soft tissues, 74 benign and 36 malignant soft tissue tumours. The slides were stained with AgNOR in order to differentiate between benign and malignant soft tissue tumours. The mean AgNOR count in normal appearing soft tissues, benign lesions and malignant lesions was 1.04+/-0.10 (0.94-1.14), 1.51+/-0.21 (1.1-2.1) and 4.96+/-1.33 (2.57-7.21) respectively. The mean AgNOR count was found to be higher in benign soft tissue tumours as compared to normal appearing soft tissues and the difference was found to be statistically significant. The mean AgNOR count in soft tissue sarcomas was found to be higher as compared to both normal appearing soft tissues and benign soft tissue tumours and the results were found to be statistically significant. The increased AgNOR score in both benign and malignant soft tissue tumours as compared to normal appearing soft tissues indicates high proliferative activity. Thus AgNOR staining is a simple and useful method for estimating tumour cell proliferation thereby differentiating normal appearing soft tissues from benign and malignant soft tissue tumours.