Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor / Caracterização imunocitoquímica de culturas primárias de tumor venéreo transmissível canino

Immunochemistry with anti-vimentin, anti-lysozyme, anti-alpha 1 antitrypsin, anti-CD3 and anti-CD79α antibodies has been used for characterization of primary cell culture in the transmissible venereal tumor (TVT). Samples for primary cell culture and immunohistochemistry assays were taken from eight dogs with cytological and clinical diagnosis of TVT. To validate the immunochemical results in the primary cell culture of TVT, a chromosome count was performed. For the statistical analysis, the Mann-Whitney test with p<0.05 was used. TVT tissues and culture cells showed intense anti-vimentin immunoreactivity, lightly to moderate immunoreactivity for anti-lysozyme, and mild for anti-alpha-antitrypsin. No marking was achieved for CD3 and CD79α. All culture cells showed chromosomes variable number of 56 to 68. This is the first report on the use of immunocytochemical characterization in cell culture of TVT. Significant statistic difference between immunochemistry in tissue and culture cell was not established, what suggests that the use of this technique may provide greater certainty for the confirmation of tumors in the primary culture. This fact is particularly important because in vitro culture of tumor tissues has been increasingly used to provide quick access to drug efficacy and presents relevant information to identify potential response to anticancer medicine; so it is possible to understand the behavior of the tumor.(AU)

Os anticorpos anti-vimentina, anti-lisozima, anti-alfa 1 antitripsina, anti-CD3 e anti-CD79α foram empregados para a caracterização de culturas primárias de tumor venéreo transmissível canino (TVT). Amostras para cultura primária e imuno-histoquímica foram coletadas de oito cães com diagnóstico clínico e citológico de TVT. Para validar o resultado inmunocitoquímico nas culturas de TVT foi realizada a contagem de cromossomos. Para a análise estatística o teste de Mann-Whitney foi empregado a um nível de significância de p<0.05. As culturas e os tecidos de TVT apresentaram intensa reatividade para vimentina, moderada a leve para Lisozima, moderada para alfa-antitripsina e não houve marcação para CD3 e CD79α. Finalmente, todas as culturas apresentaram números de cromossomos que variaram de 56 a 68. Este é o primeiro relato que apresenta o uso da immunocitoquímica para a caracterização de culturas de TVT. Assim, e devido ao fato de se observar semelhança entre a imunomarcação em células e tecidos, sugere-se que o uso desta técnica possa auxiliar na confirmação de culturas primárias do tumor, fato muito importante porque a utilização da cultura do tumor pode permitir o acesso a informação relevante sobre resposta potencial a um tratamento e conhecimento do comportamento biológico do tumor.(AU)

Proteomic Profiling of Serum from Patients with Tuberculosis

BACKGROUND: Effective treatment and monitoring of tuberculosis (TB) requires biomarkers that can be easily evaluated in blood samples. The aim of this study was to analyze the serum proteome of patients with TB and to identify protein biomarkers for TB. METHODS: Serum samples from 26 TB patients and 31 controls were analyzed by using nano-flow ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in data-independent mode, and protein and peptide amounts were calculated by using a label-free quantitative approach. The generated data were analyzed by using principal component analysis and partial least squares discriminant analysis, a multivariate statistical method. RESULTS: Of more than 500 proteins identified, alpha-1-antitrypsin was the most discriminative, which was 4.4 times higher in TB patients than in controls. Peptides from alpha-1-antitrypsin and antithrombin III increased in TB patients and showed a high variable importance in the projection scores and coefficient in partial least square discriminant analysis. CONCLUSIONS: Sera from patients with TB had higher alpha-1-antitrypsin levels than sera from control participants. Alpha-1-antitrypsin levels may aid in the diagnosis of TB.

A Case of Acute Lymphoblastic Leukemia Presenting with Protein-Losing Enteropathy / 대한소화기학회지

Protein-losing enteropathy (PLE) is a syndrome characterized by excessive gastrointestinal protein loss, resulting in hypoproteinemia and edema. A variety of benign and malignant conditions can be associated with PLE and acute leukemia is a very rare cause of PLE. We report a case of PLE associated with acute lymphoblastic leukemia. A 27-year-old man was admitted due to watery diarrhea, epigastric pain and bilateral leg edema. Laboratory findings showed hypoproteinemia and polycythemia. The diagnosis of PLE and acute lymphoblastic leukemia were confirmed on the measurement of fecal alpha1-antitrypsin clearance and bone marrow examination. After systemic chemotherapy and autologous stem cell transplantation, his clinical symptoms and abnormal laboratory findings were gradually improved.

Enfisema bulloso bilateral gigante por déficit de alfa1-antitripsina / Giant bilateral bullous emphysema due to deficit of alpha1-antritypsin

Se presenta el caso de una paciente de 48 años de edad con un enfisema bulloso bilateral gigante y niveles bajos de alfa1-antitripsina, determinados en el laboratorio Labcel de la Facultad de Ciencias Médicas Dr Miguel Enríquez. La definición de alfa1-antitriptisina es resultante de un defecto genético y aproximadamente el 75 por ciento de los adultos con una carencia grave de esta proteína desarrollan un enfisema que a menudo comienza antes de los 40 años de edad. Con el objetivo de debatir los resultados de los exámenes complementarios realizados, el tratamiento y la evolución de esta paciente, exponemos este trabajo por considerarlo de importancia para la práctica médico-quirúrgica(AU)

The case of a female patient aged 48 with a giant bilateral bullous emphysema and low levels of alpha1-antitripsine determined at the Labcel laboratory of Dr Miguel Enríquez. Faculty of Medical Sciences was reported. The definition of alpha1-antitrypsin is the result of a genetical defect and approximately 75 percent of the adults with a severe lack of this protein develop an emphysema that often begins before the 40 years of age. In order to discuss the results of the complementary tests, the treatment and evolution of this patient, we present this paper for considering it important for the medicosurgical practice(AU)

Instilación intratraqueal de elastasa en la rata: cambios electroforéticos de la alfa1-AT-antitripsina en el lavado broncoalveolar / Intratracheal instillation of elastase in the rat: electrophoretical changes of alphal-antitrypsin in bronchoalveolar lavage fluid

Introduction: Endogenous alphal-antitrypsin alpha is the main inhibitor of the intratracheally instilled elastase in experimental animals. Objective: To evaluate by electrophoresis and immunodetection using western blot analysis, the different forms of alpha1-AT in bronchoalveolar lavage fluid (BALF) of Sprague Dawley rats after intratracheal instillation of elastase, with the hypothesis that the previously observed increment in antielastase activity is due to high levels of active alpha1-AT. Results: In the first hours after elastase instillation the concentration of alpha1-AT increases more than seven times due to an increase in alveolar-capillary permeability. Alpha 1-AT in BAIF is found as the native protein (~ 52 kDa), as complexes of different molecular sizes (> 75 kDa and > 100 kDa) and as a proteolytic product (< 40 kDa). Conclusion: In spite of a high proportion of alpha1-AT in the inactive form as part of different complexes, the increase in alveolar-capillary permeability after elastase treatment contributes to maintain high levels of active alpha. These results could be of importance in other inflammatory lung processes.

Introducción: la antiproteasa alfa 1-antitripsina alfa constituye el principal inhibidor endógeno de la elastasa instilada por vía intratraqueal en modelos experimentales. Objetivo: Evaluar mediante electroforesis e inmunodetección por western blot, las distintas formas en que se encuentra la alfa1-AT en el lavado broncoalveolar (IBA) de ratas Sprague Dawley después de la instilación de elastasa, con la hipótesis de que el aumento en la actividad antielastasa previamente encontrada se acompaña de niveles altos de alfa1-AT activa. Resultados: En las primeras horas post-elastasa la concentración de alfa1-AT en el IBA aumenta más de 7 veces, debido al aumento de la permeabilidad alvéolo-capilar, encontrándose tanto como proteína nativa (~ 52 kDa), como parte de complejos de mayor tamaño (> 75 kDa y > 100 kDa) y como producto de proteólisis (< 40 kDa). Conclusión: A pesar de existir una alta proporción de alfa1-AT inactiva formando complejos, el aumento de la permeabilidad alvéolo-capilar contribuye a mantener niveles altos de alfa1-AT activa. Estos resultados podrían ser extrapolables a distintos procesos inflamatorios pulmonares.

Fecal alpha1--antitrypsin in healthy and intestinal-disorder Thai children.

OBJECTIVE: Determine the normal FA1-AT level in random wet stool of Thai children using RID and NPL, and to study the correlation between RID and NPL methods for measurement of FA1-AT. MATERIAL AND METHOD: Random stool samples were collected from healthy children and intestinal-disorders patients. Alpha1-antitrypsin (FA1-AT) in wet stool samples was measured by nephelometry (NPL) and radial-immunodiffusion (RID) methods. RESULTS: Newborn infants had the highest FA1-AT level during the first day of life and declined to the same level as older children on day 3-4. Median and geometric mean of FA1-AT levels by NPL from healthy children aged 1 month-15 years was 1.23 and 1.11 mg/dL respectively. FA1-AT levels by NPL from children with severe intestinal disorders, displaying median and geometric mean at 6.77 and 12.39 mg/dL respectively, were much higher than healthy children. The RID and NPL methods showed a correlation of r = 0.87 (p < 0.01) and R2 = 0.75. CONCLUSION: Random FA1-AT assay in wet stool is a non-invasive and simple test for supporting diagnosis of protein-losing enteropathy.

Alfa 1 antitrypsin and protease levels in Puerto Rican asthmatics: a pilot study

A cross-sectional pilot study was conducted on a population of 119 asthmatics who had been recruited from the Emergency Room Department of a major hospital in Ponce, Puerto Rico. The purpose of the study was to determine the frequency of the MM, MS, and SS a-i-antiprotease variants. Also, we analyzed the serum levels of the alpha-1-antiprotease inhibitor, quantified the levels of serine proteases in homes of the asthmatic volunteers, and determined whether environmental levels of proteases, regardless of their sources, had any association with either asthma symptoms or alpha-1-antiprotease inhibitor phenotypes. Our results do not support the role of the alpha-1-antiprotease as a risk factor for asthma in the study population as previously reported. Patients who had visited the ED due to asthma on 3 or more occasions had significantly higher trypsin levels than those who had done so 2 or fewer times. Of those asthmatic patients who had daily symptoms, 40had been exposed to high levels of elastase, and 33.3to trypsin. Similarly, 52.9of the patients with 2 or more hospitalizations a year had been exposed to high elastase levels, and 40.5of asthma patients who had nocturnal asthma more than 3 times a week had been exposed to high levels of elastase.

Identification of Proteins Differentially Expressed in the Conventional Renal Cell Carcinoma by Proteomic Analysis

Renal cell carcinoma (RCC) is one of the most malignant tumors in urology, and due to its insidious onset patients frequently have advanced disease at the time of clinical presentation. Thus, early detection is crucial in management of RCC. To identify tumor specific proteins of RCC, we employed proteomic analysis. We prepared proteins from conventional RCC and the corresponding normal kidney tissues from seven patients with conventional RCC. The expression of proteins was determined by silver stain after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The overall protein expression patterns in the RCC and the normal kidney tissues were quite similar except some areas. Of 66 differentially expressed protein spots (p<0.05 by Student t-test), 8 different proteins from 11 spots were identified by MALDI-TOF-MS. The expression of the following proteins was repressed (p<0.05); aminoacylase-1, enoyl-CoA hydratase, aldehyde reductase, tropomyosin alpha-4 chain, agmatinase and ketohexokinase. Two proteins, vimentin and alpha-1 antitrypsin precursor, were dominantly expressed in RCC (p<0.05).

Diagnosis of alpha-1-antitrypsin deficiency by DNA analysis of children with liver disease

BACKGROUND: Alpha-1-antitrypsin deficiency is a genetic disorder which is transmitted in a co-dominant, autosomal form. Alpha-1-antitrypsin deficiency affects mainly the lungs and the liver leading, in the latter case, to neonatal cholestasis, chronic hepatitis or cirrhosis. A precise diagnosis of Alpha-1-antitrypsin deficiency may be obtained by biochemical or molecular analysis. OBJECTIVE: The purpose of this study was to use DNA analysis to examine the presence of an alpha-1-antitrypsin deficiency in 12 children suspected of having this deficiency and who showed laboratory and clinical characteristics of the disease. PATIENTS AND METHODS: Twelve patients, aged 3 months to 19 years, who had serum alpha-1-antitrypsin levels lower than normal and/or had hepatic disease of undefined etiology were studied. The mutant alleles S and Z of the alpha-1-antitrypsin gene were investigated in the 12 children. Alpha-1-antitrypsin gene organization was analyzed by amplification of genoma through the polymerase chain reaction and digestion with the restriction enzymes Xmnl (S allele) and Taq-1 (Z allele). RESULTS: Seven of the 12 patients had chronic liver disease of undefined etiology and the other five patients had low serum levels of alpha-1-antitrypsin as well as a diagnosis of neonatal cholestasis and/or chronic liver disease of undefined etiology. Five of the 12 patients were homozygous for the Z allele (ZZ) and two had the S allele with another allele (*S) different from Z. CONCLUSION: These results show that alpha-1-antitrypsin deficiency is relatively frequent in children with chronic hepatic disease of undefined etiology and/or low alpha-1-antitrypsin levels (41.6 per cent). A correct diagnosis is important for effective clinical follow-up and for genetic counseling.

Determinaçäo quantitativa de alfa-1-antitripsina e suas variaçöes fenotípicas no soro de portadores de infecçöes crônicas / Quantitative determination of alpha-1-antitrypsin and its phenotype variations in the sera of chronics diseases porters

A alfa-1-antitripsina (Ó1-AT) é uma proteína de fase aguda, sendo a mais importante de várias antiproteases, pois atua como protetora contra injúrias nos tecidos. Ela migra eletroforeticamente na regiäo das alfa-globulinas. Neste trabalho foram analisadas amostras de soros de pacientes portadores de infecçöes crônicas por parasitas e bactérias quanto à verificaçäo do fenótipo de Ó1-AT, caracterizado por focalizaçäo isoelétrica, e à sua determinaçäo quantitativa mediante a capacidade de inibiçäo da tripsina. Tanto os fenótipos quanto as dosagens de Ó1-AT inseriram-se dentro dos valores normais estabelecidos para estas metodologias. Estes resultados demonstraram que näo houve correlaçäo entre as patologias analisadas e o nível deste mediador do processo inflamatório. Provavelmnente, outros fatores do hospedeiro, tais como a imunomodulaçäo por citocinas, participaçäo de anticorpos e linfócitos T citotóxicos, estejam envolvidos na cronicidade destas infecçöes.

Colonic proteinase: increased activity in patients in patients with ulcerative colitis

In order to study the colonic intraluminal proteinase-antiproteinase imbalance under inflammatory conditions, we determined proteolytic activity (PA), alpha-1-antitrypsin and the activities of trypsin, chymotrypsin and neutrophil elastase in feces from patients with ulcerative colitis (UC) comparing the results with a control group. A fecal sample was obtained from each of 25 patients with ulcerative colitis and 10 control subjects were studied. The severity of the disease was assessed by the Truelove index. Proteolytic activity was measured lesing azocasein as proteolytic substrate. The fecal concentration of alpha-1-antitrypsin was measured by radial immunodiffusion and the activities of the enzymes were measured using specific substrates. We found an increase in fecal PA, alpha-1-antitrypsin and neutrophil elastase in patients with UC and the correlation between the severity of the disease and the PA was statistically significant (r = 0.62, P < 0.05). We conclude that elevated colonic proteinase activity could contribute to the pathophysiology of ulcerative colitis.

Serum alpha-1-antitrypsin in ischemia and rheumatic heart diseases.

Tissue damage, inflammation and necrosis are hallmarks of myocardial infarction. In the present study significant elevations of serum alpha-1-antitrypsin were noted in coronary artery disease and angina cases. Interestingly chronic rheumatic heart disease which is also characterized by tissue injury. Inflammation revealed normal levels of serum alpha-1-antitrypsin. The level in chronic rheumatic heart disease was 3.37 +/- 0.57 mumol/mt/ml (control level was 3.37 +/- 0.54 mumol/mt/ml). The corollary of these observations is that in heart diseases acute phase response in terms of enhanced levels of alpha-1-antitrypsin differ depending on the causative factors. Except chronic rheumatic heart disease, in all other stressful states studied there is (to a certain degree) an altered systemic homeostasis and haemostasis. On the other hand chronic rheumatic heart disease encompass certain amount of acute phase status in terms of tissue damage and inflammation does exist unaccompanied by altered systemic homeostasis and haemostasis. However, bacteriological etiologies predominate the triggered immune responses. It is hypothesised that serum alpha-1-antitrypsin enhancement will not occur even though acute phase state exists if specific immune responses are also a part of the disease manifestation.

Prenatal development of human major salivary glands. Histological and immunohistochemical characteristics with reference to adult and neoplastic salivary glands

No abstract available.

A study of antitrypsin and macroglobulin levels in serum and saliva of patients with gingivitis.

Periodontal diseases are associated with chronic inflammation. The destruction of connective tissue matrix is responsible for the pathogenesis of chronic inflammatory states. The degradation of matrix is initiated extra and pericellularly by proteinases produced locally at the inflammatory site. The regulation of these proteinases are by inhibitors present in serum and extravascular tissues, and it is the proteinase/proteinase inhibitor balance that determines the progression of chronic inflammatory state. Few contradicting studies are available on changes in the levels of proteinase inhibitors in serum in periodontal disease. The occurrence of these inhibitors in saliva has not been studied in detail. The present study was aimed at measuring the Proteinase inhibitors in serum and saliva of patients with periodontal disease.

Clearance fecal de Ó1 antitripsina: valores normales en niños de la ciudad de Córdoba / Faecal clearence of the Ó1-antitrypsin: values normal in children of the Cordoba city

En las enteropatías perdedoras de proteínas (EPP) el incremento de la permeabilidad de las paredes del TGI causa exudación proteica superior a la normal que se manifiesta por elevada excreción proteica fecal. El estudio del turnover metabólico, mediante la inyección intravenosa de proteínas marcadas con radioisótopos del yodo, provee información sobre el metabolismo proteico, pero no es útil para medir pérdida proteica gastrointestinal pues el yodo libre atraviesa con facilidad la mucosa gástrica en ambos sentidos, lo que conduce a una valoración errónea de la excreción proteíca en la materia fecal (MF). La albúmina marcada con 51Cr permite valorar la pérdida proteíca gastrointestinal caracterizada por un patron de pérdida al bulto. Los mencionados son metodos de referencia, invasores, costosos y su empleo está reglamentado por la legislación especial pues es material radioactivo. El clearance fecal de Ó1 antitripsina es inocuo y accesible a laboratorios clínicos y posee eficacia clínica probada para valorar la excreción proteica gastrointestinal. Se ha implementado la metodología y establecido valores normales de referencia para la población infantil de la ciudad de Córdoba. La muestra estuvo conformoda por 30 niños clínicamente sanos de ambos sexos, cuyas edades estuvieron comprendidas entre los 14 y 120 meses; en el protocolo de estudio de identificación se registró la edad, sexo y análisis de laboratorio. La determinación de Ó1AT sérica y fecal se realizó por inmunodifusión radial simple (IDRS). En las enteropatías puede ocurrir malabsorción proteíca, causante de hipoalbuminemia, o pérdida proteica gastrointestinal (en enteropatías exudativas), la cual afecta las fracciones albúmina y globulinas. El clearance fecal de Ó1AT indica los mililitros de plasma depurados en 24 horas por el tracto gastrointestinal y provee orientación diagnóstica útil para el uso clínico de rutina. Los valores de referencia hallados en la población infantil de Córdoba son: -clearance fecal de Ó1AT= desde no determinable hasta 10.8ml/24h; -concentración de Ó1AT sérica = 118 a 396mg por ciento

Deficiencia de alfa-1 antitripsina: presentación de un estudio familiar

Se informa el caso de un paciente de 65 años de edad quien presentaba un hepatocarcinoma y cirrosis asociados a deficiencia de alfa-1antitripsina (A1-AT). Cuatro de sus seis hermanos, dos hombres y dos mujeres, tenían valor de esta glicoproteína inferiores a los normales, sin manifestación aparente de compromiso clínico hepático ni pulmonar; ninguno tenía antecedentes de hepatitis o colestasis neonatal. La descripción de este primer caso en nuestro medio se propone estimular el interés por esta entidad como causa de hepatopatía crónica, carcinoma hepático primario y enfisema pulmonar, sin factores de riesgo diferentes asociados

Iron, copper and alpha 1-antitrypsin deficiency in the liver of cirrhotic and schistosomotic patients

Há vários estudos sobre depósitos de ferro e cobre e deficiência de alfa 1-antitripsina (A1AT) no fígado em cirróticos, mas nenhum, segundo nossa pesquisa bibliográfica, em esquistossomóticos. Os autores pesquisaram a presença desses metais e a deficiência de A1AT no fígado de 72 cirróticos e 27 esquistossomóticos. Entre esses, cinco eram de forma hepatointestinal e 22 da hepatoesplênica. Entre os cirróticos, 44 eram alcoólatras e 28 näo-alcoólatras. O ferro foi detectado em 23(31,9%) dos cirróticos, dos quais 16(36,3%) eram alcoólatras e 7(25,0%) näo alcoólatras, diferença näo significante. Quanto aos graus, foram observados: 13 casos (56,5%) grau I; cinco casos (21,7%) grau II, e cinco (21,7%) grau III. O cobre foi detectado em 24 cirróticos (33,3%) dos quais 15(34,0%) eram alcoólatras e nove (32,1%) näo alcoólatras, diferença näo significante. A deficiência de A1AT foi observada em dois (2,8%) cirróticos, um dos quais era alcoólatra. HBsAg e HBcAg em tecido hepático foram detectados em cinco cirróticos (6,9%), dos quais três eram alcoólatras. A presença de ferro, e cobre e deficiência de A1AT foi observada em três cirróticos (12,5%) e a de ferro e deficiência de A1AT em dois cirróticos (2,8%). Os autores ressaltam que nenhum paciente esquistossomático apresentou ferro, cobre ou deficiência de A1AT no fígado e discutem aspectos relacionados com a eventual importância dos dados obtidos

"Clearance" fecal da alfa-1-antitripsina / Fecal clearance of alpha 1-antitrypsin

Há enteropatias perdedoras de proteínas (PLE) com e sem sintomas digestivos ou hipoproteinemia. O diagnóstico através da perda fecal de albumina é inviável, exceto por método radioativo. A alternativa da dosagem fecal da alfa-1-antitripsina (AAT) - uma proteína plasmática especialmente resistente à digestao proteolítica - caiu em descrédito: os resultados variavam, indiscriminadamente, com o grau de comprometimento da mucosa e com a concentraçäo plasmática de AAT. O clearance (relaçäo da massa média de AAT fecal por 24 horas com a concentraçäo plasmática da substância) elimina a inespecificidade e evita erros decorrentes do grau de hidrataçäo das fezes ou de seu conteúdo de sólidos.