Mutagenicity and antimutagenicity of Salacia crassifolia (mart. Ex. Schult.) G. Don. evaluated by Ames test / Mutagenicitade e antimutagenicidade de Salacia crassifolia (mart. Ex. Schult.) G. Don. avaliadas pelo teste de Ames

Abstract Salacia crassifolia (Mart. Ex. Schult.) G. Don. is a bush which belongs to Celastraceae family and occurs specially in Brazilian Cerrado. Its leaves, stem, seeds and fruits are popularly used for several medicinal purposes, such as antitumoral, antirheumatic, anti-inflammatory and antimicrobial. In this study, the mutagenic and antimutagenic activities of S. crassifolia stem bark fractions (hexane, ethyl acetate and hydroalcoholic) were evaluated by the Ames mutagenicity assay in Salmonella typhimurium TA98 and TA100 strains. By the obtained results, all S. crassifolia fractions did not significantly increase the number of prototrophic revertants for histidine (His+) in both S. typhimurium strains tested (p > 0.05), suggesting absence of mutagenicity. Regarding antimutagenicity, the fractions ethyl acetate and hydroalcoholic significantly decreased the number of His+ revertants colonies induced by positive control for strain TA98 (p < 0.05), demonstrating protection against mutagenicity induced by 4-nitroquinolile1-oxide, whereas the hexane fraction did not show antimutagenic effect in this strain. In the TA100 strain, all fractions of S. crassifolia protected DNA against the harmful action of sodium azide, and the hexane fraction exhibited the greatest protection in this work. Thus, it's possible conclude that the fractions of S. crassifolia tested in this study could be used in chemoprevention.

Resumo Salacia crassifolia (Mart. Ex. Schult.) G. Don. é uma árvore que pertence à família Celastraceae e ocorre especialmente no Cerrado Brasileiro. Suas folhas, caule, sementes e frutos são popularmente utilizados para vários fins medicinais, tais como antitumoral, antirreumático, anti-inflamatório e antimicrobiano. Neste estudo, nós avaliamos as atividades mutagênica e antimutagênica de frações da casca do caule de S. crassifolia (hexânica, acetato de etila e hidroalcoólica) pelo ensaio de mutagenicidade de Ames em Salmonella typhimurium, cepas TA98 e TA100. Pelos resultados obtidos todas as frações de S. crassifolia não aumentaram significativamente o número de revertentes prototróficas para histidina (His+) em ambas as cepas de S. typhimurium testadas (p > 0.05), sugerindo ausência de mutagenicidade. Em relação à antimutagenicidade, as frações acetate de etila e hidroalcoólica reduziram significativamente o número de colônias revertentes His+ induzidas pelo controle positive para a cepa TA98 (p < 0.05), demonstrando sua ação protetora contra a mutagenicidade induzida por 4-nitroquinolile1-oxide, enquanto a fração hexânica não demonstrou efeito antimutagênico nesta cepa. Na cepa TA100, todas as frações de S. crassifolia protegeram o DNA contra a ação lesiva de azida sódica, e a fração hexânica exibiu a maior proteção desse trabalho. Assim, concluímos que as frações de S. crassifolia testadas neste estudo poderiam ser utilizadas em quimioprevenção.

Mutagenicity and antimutagenicity of Salacia crassifolia (mart. Ex. Schult.) G. Don. evaluated by Ames test

Abstract Salacia crassifolia (Mart. Ex. Schult.) G. Don. is a bush which belongs to Celastraceae family and occurs specially in Brazilian Cerrado. Its leaves, stem, seeds and fruits are popularly used for several medicinal purposes, such as antitumoral, antirheumatic, anti-inflammatory and antimicrobial. In this study, the mutagenic and antimutagenic activities of S. crassifolia stem bark fractions (hexane, ethyl acetate and hydroalcoholic) were evaluated by the Ames mutagenicity assay in Salmonella typhimurium TA98 and TA100 strains. By the obtained results, all S. crassifolia fractions did not significantly increase the number of prototrophic revertants for histidine (His+) in both S. typhimurium strains tested (p > 0.05), suggesting absence of mutagenicity. Regarding antimutagenicity, the fractions ethyl acetate and hydroalcoholic significantly decreased the number of His+ revertants colonies induced by positive control for strain TA98 (p 0.05), demonstrating protection against mutagenicity induced by 4-nitroquinolile1-oxide, whereas the hexane fraction did not show antimutagenic effect in this strain. In the TA100 strain, all fractions of S. crassifolia protected DNA against the harmful action of sodium azide, and the hexane fraction exhibited the greatest protection in this work. Thus, its possible conclude that the fractions of S. crassifolia tested in this study could be used in chemoprevention.

Resumo Salacia crassifolia (Mart. Ex. Schult.) G. Don. é uma árvore que pertence à família Celastraceae e ocorre especialmente no Cerrado Brasileiro. Suas folhas, caule, sementes e frutos são popularmente utilizados para vários fins medicinais, tais como antitumoral, antirreumático, anti-inflamatório e antimicrobiano. Neste estudo, nós avaliamos as atividades mutagênica e antimutagênica de frações da casca do caule de S. crassifolia (hexânica, acetato de etila e hidroalcoólica) pelo ensaio de mutagenicidade de Ames em Salmonella typhimurium, cepas TA98 e TA100. Pelos resultados obtidos todas as frações de S. crassifolia não aumentaram significativamente o número de revertentes prototróficas para histidina (His+) em ambas as cepas de S. typhimurium testadas (p > 0.05), sugerindo ausência de mutagenicidade. Em relação à antimutagenicidade, as frações acetate de etila e hidroalcoólica reduziram significativamente o número de colônias revertentes His+ induzidas pelo controle positive para a cepa TA98 (p 0.05), demonstrando sua ação protetora contra a mutagenicidade induzida por 4-nitroquinolile1-oxide, enquanto a fração hexânica não demonstrou efeito antimutagênico nesta cepa. Na cepa TA100, todas as frações de S. crassifolia protegeram o DNA contra a ação lesiva de azida sódica, e a fração hexânica exibiu a maior proteção desse trabalho. Assim, concluímos que as frações de S. crassifolia testadas neste estudo poderiam ser utilizadas em quimioprevenção.

Rapidly progressive dementia due to neurosarcoidosis / Demência rapidamente progressiva causada por neurosarcoidose

Rapidly progressive dementia (RPD) is typically defined as a cognitive decline progressing to severe impairment in less than 1-2 years, typically within weeks or months. Accurate and prompt diagnosis is important because many conditions causing RPD are treatable. Neurosarcoidosis is often cited as an unusual reversible cause of RPD. METHODS: We report two cases of neurosarcoidosis presenting as RPD. RESULTS: Case 1: A 61-year-old woman developed a RPD associated with visual loss. In seven months she was dependent for self-care. Magnetic resonance imaging (MRI) revealed temporal and suprasellar brain lesions. Treatment with high-dose intravenous prednisolone was associated with partial improvement. Case 2: A 43-year-old woman who was being treated for diabetes insipidus developed a severe episodic amnesia one year after onset of cognitive symptoms. Previous MRI had shown a hypothalamic lesion and she had been treated with oral prednisone and cyclophosphamide. There was reduction of the MRI findings but no improvement in the cognitive deficits. Brain biopsy disclosed noncaseous granulomas and granulomatous angiitis; treatment was changed to high-dose intravenous methylprednisolone, with poor symptomatic response. CONCLUSION: The diagnosis of RPD due to neurosarcoidosis can be challenging when the disease is restricted to the nervous system. In these cases, clinical presentation of RPD associated with neuroendocrine and visual dysfunction, imaging findings showing hypothalamic lesions and, in some cases, brain biopsy, are the key to a correct diagnosis. It is possible that earlier diagnoses and treatment could have led to a better outcome in these patients.

Demência rapidamente progressiva (DRP) é tipicamente definida como um declínio cognitivo que progride para prejuízo funcional severo em menos de 1-2 anos, geralmente em semanas a meses. O diagnóstico rápido e acurado é fundamental, já que muitas condições que levam a DRP são reversíveis. MÉTODOS: Relatamos dois casos de neurosarcoidose que se apresentaram como DRP. RESULTADOS: Caso 1: Uma mulher de 61 anos desenvolveu uma DRP associada a perda de acuidade visual. Em sete meses evoluiu com dependência para auto-cuidado. A ressonância magnética (RM) revelou lesões encefálicas temporais e supraselares. Evoluiu com melhora parcial após tratamento com metilprednisolona intravenosa em altas doses. Caso 2: Uma mulher de 43 anos que estava em tratamento para diabetes insipidus desenvolveu uma amnésia episódica severa um ano após o início dos sintomas cognitivos. A RM anterior mostrava uma lesão hipotalâmica, e ela recebeu tratamento oral com prednisona e ciclofosfamida. Houve redução dos achados da RM, porem sem melhora dos déficits cognitivos. A biópsia cerebral mostrou granulomas não caseosos e angeíte granulomatosa; o tratamento foi modificado para metilprednisolona intravenosa em altas doses, com resposta sintomática pobre. CONCLUSÃO: O diagnóstico de DRP por neurosarcoidose pode ser desafiador quando a doença está restrita ao sistema nervoso central. Nestes casos, a apresentação clínica da DRP associada a disfunção neuroendócrina e visual, exames de imagem com lesões hipotalâmicas e, em alguns casos, a biópsia cerebral são fundamentais para um diagnóstico correto. é possível que o diagnóstico e tratamento precoces poderiam ter trazido melhores resultados nesses pacientes.

Zeólita na dieta de bovinos de corte / Zeolite in the diet of beef cattle

Avaliou-se o efeito da inclusão de zeólita na dieta de bovinos de corte sobre os parâmetros ruminais, as digestibilidades total e parcial, a produção de nitrogênio, a eficiência microbiana e as características do sangue. Os tratamentos, com base na matéria seca (MS) da dieta, foram: 0; 0,75; 1,5; 2,25 e 3,0% de zeólita. Utilizaram-se cinco bovinos machos mestiços, fistulados no rúmen e abomaso, alimentados com silagem de milho e concentrado, representando 65% de volumoso e 35% de concentrado. O delineamento utilizado foi em quadrado latino 5×5, com cinco períodos de 15 dias. Os animais receberam 15g de dióxido de titânio (TiO2) do terceiro ao 12º dia de cada período. Houve efeito (P<0,05) sobre o consumo diário (kg/dia) de proteína bruta (PB) e efeito (P<0,05) sobre o consumo de MS e de fibra em detergente neutro corrigida para cinza e proteína, expresso em g/kg de peso. A digestibilidade total dos nutrientes digestíveis totais (NDT) foi influenciada pelos tratamentos (P<0,05). A digestibilidade ruminal dos carboidratos não fibrosos (CNF) foi afetada quadraticamente (P<0,05), assim como a intestinal da PB e dos CNF. A ingestão e a excreção fecal de nitrogênio (g/dia) foram influenciadas linearmente (P<0,05) pela inclusão de zeólita. Concluiu-se que a adição de zeólita na dieta, embora tenha aumentado os consumos de MS e de FDN, não melhorou a utilização da ureia em dietas de bovinos de corte.

We evaluated the effect of the inclusion of zeolite in the diet of beef cattle on the ruminal digestibility, total and partial production of nitrogen, microbial efficiency and blood characteristics. Treatments based on the dietary dry matter (DM) were: 0%, 0.75%, 1.5%, 2.25% and 3.0% of zeolite. We used five crossbred steers fistulated in the rumen and abomasum, fed corn silage and concentrate, representing 65% forage and 35% concentrate. The design was a 5x5 latin square with five periods of 15 days. The animals received 15g of titanium dioxide (TiO2) from the third to the 12th day of each period. A significant effect (P<0.05) on the daily consumption (kg/day) of crude protein (CP) was observed (P<0.05) for DM intake and neutral detergent fiber corrected for ash and protein (NDFap) expressed in g/kg. The total digestibility of total digestible nutrients (TDN) was influenced by treatments (P<0.05). Ruminal digestibility of non-fiber carbohydrates (NFC) was affected quadratically (P<0.05) as well as intestinal CP and NFC. Intake and fecal excretion of nitrogen (g/day) were affected linearly (P<0.05) by adding zeolite. It was concluded that the addition of zeolite in the diet, while increasing the intake of DM and NDF, did not improve the use of urea in the diet of beef cattle.

Angiotomografia coronariana multislice na avaliação da origem anômala das artérias coronarianas / Multislice coronary angiotomography in the assessment of coronary artery anomalous origin / Angiotomografía coronaria multislice en la evaluación del origen anómalo de las arterias coronarias

Fundamento: A origem anômala das artérias coronarianas é uma entidade relativamente rara, podendo se apresentar de várias formas clínicas e evoluir de forma adversa. A angiotomografia multislice das artérias coronarianas vem ganhando espaço na prática clínica diária, representando importante método propedêutico, com grande potencial na avaliação anatômica dessas artérias. Objetivo: O objetivo deste estudo é descrever os achados angiotomográficos e a evolução de pacientes com origem anômala das artérias coronarianas. Métodos: Durante o período de janeiro de 2008 a março de 2011, foram avaliados consecutivamente 404 pacientes encaminhados para realização de angiotomografia das coronárias por diversos motivos, com tempo médio de seguimento de 21 meses. Resultados: Nove pacientes (2,2%) apresentaram origem anômala das artérias coronárias, sendo quatro com origem anômala da artéria circunflexa (Cx) em Coronária Direita (CD), dois com origem anômala da CD (um com origem de CD em Cx, e um com origem de CD em seio coronariano esquerdo), um paciente com tronco coronariano único (descendente anterior e Cx saindo do seio coronariano esquerdo), um com trajeto anômalo do tronco da coronária esquerda entre aorta e artéria pulmonar e um paciente apresentando tronco coronariano esquerdo originando-se do seio coronariano direito. Dos pacientes avaliados, um paciente recebeu um cardiodesfibrilador implantável; um paciente evoluiu com morte súbita durante internação hospitalar; e os outros não tiveram intercorrências. Conclusão: A angiotomografia multislice das coronárias representa método propedêutico minimamente invasivo que possibilita detectar a origem, o curso e terminação das anomalias de origem das artérias coronarianas com excelente acurácia, possibilitando o correto diagnóstico e auxiliando no planejamento terapêutico.

Background: Anomalous origin of coronary arteries is a relatively rare entity, which may show different clinical forms and develop an adverse outcome. Multislice coronary angiotomography has gained importance in daily clinical practice, representing an important patient assessment method with great potential in the anatomical evaluation of these arteries. Objective: This study aimed to describe angiotomographic findings and evolution of patients with anomalous origin of coronary arteries. Methods: From January 2008 to March 2011 404 patients were evaluated consecutively, who had been referred for coronary angiotomography for several reasons, with a mean follow-up of 21 months. Results: Nine patients (2.2%) had anomalous origin of coronary arteries, of which four had an anomalous origin of the Circumflex (Cx) artery in the Right Coronary Artery (RCA), two had anomalous origin of the RCA (one originating from the Cx and one in the left coronary sinus), a patient with single coronary trunk (anterior descending and Cx out of the left coronary sinus), one had anomalous left main coronary artery trajectory between the aorta and pulmonary artery and one patient with left coronary trunk originating from the right coronary sinus. Of the evaluated patients, one received an implantable cardiodefibrillator; one died due to sudden death during hospitalization; and the others showed no complications. Conclusion: The multislice coronary angiotomography represents minimally invasive assessment method that allows detecting the origin, course and termination of anomalous coronary arteries with excellent accuracy, allowing the correct diagnosis and aiding therapeutic planning.

RESUMEN FUNDAMENTO: El origen anómalo de las arterias coronarias es una entidad relativamente rara, pudiendo presentarse de varias formas clínicas y evolucionar de forma adversa. La angiotomografía multislice de las arterias coronarias viene ganando espacio en la práctica clínica diaria, representando importante método propedéutico, con gran potencial en la evaluación anatómica de esas arterias. OBJETIVO: El objetivo de este estudio es describir los hallazgos angiotomográficos y la evolución de pacientes con origen anómalo de las arterias coronarias. MÉTODOS: Durante el período de enero de 2008 a marzo de 2011, fueron evaluados consecutivamente 404 pacientes encaminados para realización de angiotomografía de las coronarias por diversos motivos, con tiempo medio de seguimiento de 21 meses. RESULTADOS: Nueve pacientes (2,2%) presentaron origen anómalo de las arterias coronarias, siendo cuatro con origen anómalo de la arteria circunfleja (Cj) en Coronaria Derecha (CD), dos con origen anómalo de la CD (uno con origen de CD en Cj, y uno con origen de CD en seno coronario izquierdo), un paciente con tronco coronario único (descendente anterior y Cj saliendo del seno coronario izquierdo), uno con trayecto anómalo del tronco de la coronaria izquierda entre aorta y arteria pulmonar y un paciente presentando tronco coronario izquierdo originándose del seno coronario derecho. De los pacientes evaluados, un paciente recibió un cardiodesfibrilador implantable; un paciente evolucionó con muerte súbita durante internación hospitalaria; y los otros no tuvieron intercurrencias. CONCLUSÃO: La angiotomografía multislice de las coronarias representa método propedéutico mínimamente invasivo que posibilita detectar el origen, el curso y terminación de las anomalías de origen de las arterias coronarias con excelente precisión, posibilitando el correcto diagnóstico y auxiliando al planeamiento terapéutico.

Cardiac and renal effects induced by different exercise workloads in renovascular hypertensive rats

We examined the effect of exercise training (Ex) without (Ex 0 percent) or with a 3 percent workload (Ex 3 percent) on different cardiac and renal parameters in renovascular hypertensive (2K1C) male Fisher rats weighing 150-200 g. Ex was performed for 5 weeks, 1 h/day, 5 days/week. Ex 0 percent or Ex 3 percent induced similar attenuation of baseline mean arterial pressure (MAP, 119 ± 5 mmHg in 2K1C Ex 0 percent, N = 6, and 118 ± 5 mmHg in 2K1C Ex 3 percent, N = 11, vs 99 ± 4 mmHg in sham sedentary (Sham Sed) controls, N = 10) and heart rate (HR, bpm) (383 ± 13 in 2K1C Ex 0 percent, N = 6, and 390 ± 14 in 2K1C Ex 3 percent, N = 11 vs 371 ± 11 in Sham Sed, N = 10,). Ex 0 percent, but not Ex 3 percent, improved baroreflex bradycardia (0.26 ± 0.06 ms/mmHg, N = 6, vs 0.09 ± 0.03 ms/mmHg in 2K1C Sed, N = 11). Morphometric evaluation suggested concentric left ventricle hypertrophy in sedentary 2K1C rats. Ex 0 percent prevented concentric cardiac hypertrophy, increased cardiomyocyte diameter and decreased cardiac vasculature thickness in 2K1C rats. In contrast, in 2K1C, Ex 3 percent reduced the concentric remodeling and prevented the increase in cardiac vasculature wall thickness, decreased the cardiomyocyte diameter and increased collagen deposition. Renal morphometric analysis showed that Ex 3 percent induced an increase in vasculature wall thickness and collagen deposition in the left kidney of 2K1C rats. These data suggest that Ex 0 percent has more beneficial effects than Ex 3 percent in renovascular hypertensive rats.

Functional differences between two morphologically distinct cell subpopulations within a human colorectal carcinoma cell line

The LISP-I human colorectal adenocarcinoma cell line was isolated from a hepatic metastasis at the Ludwig Institute, Säo Paulo, SP, Brazil. The objective of the present study was to isolate morphologically different subpopulations within the LISP-I cell line, and characterize some of their behavioral aspects such as adhesion to and migration towards extracellular matrix components, expression of intercellular adhesion molecules and tumorigenicity in vitro. Once isolated, the subpopulations were submitted to adhesion and migration assays on laminin and fibronectin (crucial proteins to invasion and metastasis), as well as to anchorage-independent growth. Two morphologically different subpopulations were isolated: LISP-A10 and LISP-E11. LISP-A10 presents a differentiated epithelial pattern, and LISP-E11 is fibroblastoid, suggesting a poorly differentiated pattern. LISP-A10 expressed the two intercellular adhesion molecules tested, carcinoembryonic antigen (CEA) and desmoglein, while LISP-E11 expressed only low amounts of CEA. On the other hand, adhesion to laminin and fibronectin as well as migration towards these extracellular matrix proteins were higher in LISP-E11, as expected from its poorly differentiated phenotype. Both subpopulations showed anchorage-independent growth on a semi-solid substrate. These results raise the possibility that the heterogeneity found in the LISP-I cell line, which might have contributed to its ability to metastasize, was due to at least two different subpopulations herein identified

Evidence for the expression of native Mycobacterium tuberculosis phospholipase C: recognition by immune sera and detection of promoter activity

The genome of Mycobacterium tuberculosis H37Rv contains three contiguous genes (plc-a, plc-b and plc-c) which are similar to the Pseudomonas aeruginosa phospholipase C (PLC) genes. Expression of mycobacterial PLC-a and PLC-b in E. coli and M. smegmatis has been reported, whereas expression of the native proteins in M. tuberculosis H37Rv has not been demonstrated. The objective of the present study was to demonstrate that native PLC-a is expressed in M. tuberculosis H37Rv. Sera from mice immunized with recombinant PLC-a expressed in E. coli were used in immunoblots to evaluate PLC-a expression. The immune serum recognized a 49-kDa protein in immunoblots against M. tuberculosis extracts. No bands were visible in M. tuberculosis culture supernatants or extracts from M. avium, M. bovis and M. smegmatis. A 550-bp DNA fragment upstream of plc-a was cloned in the pJEM12 vector and the existence of a functional promoter was evaluated by detection of ß-galactosidase activity. ß-Galactosidase activity was detected in M. smegmatis transformed with recombinant pJEM12 grown in vitro and inside macrophages. The putative promoter was active both in vitro and in vivo, suggesting that expression is constitutive. In conclusion, expression of non-secreted native PLC-a was demonstrated in M. tuberculosis

Evaluation of an anti-carcinoembryonic monoclonal antibody suitable for immunoscintigraphy

An anti-carcinoembryonic antigen (CEA) monoclonal antibody (mAb 6D1.1) was evaluated in vitro and in vivo to determine its suitability as a tracer for immunoscintigraphy of colorectal carcinomas. Determination of mAb affinity for CEA showed a constant of association of 0.63 + or - 0.11 x 109 M-1. Binding of technetium-99m (99mTc)-6D1.1, labeled by a direct method, to human cultured lineages was highly specific. Binding to only CEA-positive LS-174T cells resulted in a saturable curve inhibited by pre-incubation with unlabeled mAb. No binding at all was observed for the human lineages MeWo (melanoma) or ZR75-30 (breast carcinoma), neither of them expressing CEA cells. Intravenous injection of 99mTc-6D1.1 into nude mice xenografted with human LS-174T tumors resulted in planar images of excellent quality. Localization of an irrelevant mAb labeled with either 99mTc or iodine-125 was never observed in tumor masses. Biodistribution studies on excised tumoral tissue showed retention of 28.48 percent of the injected dose per gram of LS-174T tumor. The tumor-to-blood ratio was 3.46. The same analysis performed on the other three human xenografted tumors studied demonstrated that only the CEA-producing HT-29 (colorectal adenocarcinoma) retained 99mTc-6D1.1 while the other two (ZR75-30 and MeWo) did not. These data demonstrate that this mAb is an adequate tool for targeting CEA-expressing tumors in experimental models

Diameter of the mammalian porin channel in open and "closed" states: direct measurement at the single channel level in planar lipid bilayer

Porin isolated from bovine skeletal muscle was reconstituted in planar lipid bilayers under voltage clamp conditions. A set of non-electrolytes were used as molecular probes for determining the pore diameter. The maximal diameter of the open channel was estimated to be 3.02 + 0.26 nm. As observed for other porin channels, a large transmembrane potential drove the channel into a "closed" state. The channel transition to the low conductance (closed) state was followed by a decrease in the maximal diameter of the channel to 2.4 +- 0.08 nm.

Prevalência de discinesia tardia em pacientes internados nos hospitais psiquiátricos de Salvador, Bahia, Brasil / Prevalence of tardive dyskinesia on hospitalized patients in psychiatric hospitals from Salvador, Bahia, Brazil

Os autores apresentam estudo transversal da prevalência da discinesia tardia (DT) em pacientes internados em unidades psiquiátricas da cidade de Salvador durante o mês de setembro-1992. Após revisäo da literatura, comentam a prevalência média de DT encontrada: 1,65 por cento na populaçäo de 2115 pacientes, usando protocolo baseado na escala de Simpson

Monoclonal antibody-based immunoassay for carcinoembryonic antigen (CEA) measurement in human serum

Clinical oncology requires methods to detect tumor markers in patients sera and tissues Presently, monoclonal antibodies (MAbs based enzyme immunoassays are amongst the most advantageous techniques. Here we present a sensitive and specific double-antibody enzyme immunoassay for serum me surement of carcinoembryonic antigen (CEA). It has been developed with MAbs which recognize nonoverlapping peptide epitopes on the antigen molecule. The capture MAb 6C7 is GOLD 1 highly specific antibody while the tracer MAb 5.D11, labeled with biotin, is GOLD 4 that cross-reacts with nonspecific cross-reacting antigen (NCA) expressed on granulocytes. In addition the biotinylate MAb is shown to be also useful to detect CEA by immunohistochemistry.

Participation of glycosylation sites in the binding of Staphylococcus aureus to laminin

1. Microbial pathogenicity is in many instances associated with the ability to adhere to host surfaces or to extracellular matrix components. 2. Laminin is a major glycoprotein of basement membranes which can promote specific bacterial adhesion. Staphylococcus aureus is a pathogenic bacterium which presents a laminin receptor of about 50-kDa molecular mass (Lopes JD, Reis M per cent Brentani RR (1985). Science, 229: 275-277). 3. Adhesion inhibition assays of [125iodine]-labeled bacteria to laminin demonstrate that the receptor binding site is contained in the pepsin-derived (P1) laminin fragment. 4. Cell adhesion to laminin is unaffected by periodate oxidation of sugars on the surface of bacteria or by removal of divalent cations by ethylenediaminetetraacetic acid (EDTA). In contrast, bacterial adhesion is reduced when laminin is deglycosylated with N-glycosidase F or when bacteria are submitted to controlled trypsin digestion. 5. Laminin binding to the S. aureus 50-kDa band in immunoblotting assays has confirmed all of these results obtained in cell adhesion experiments

Antigen F1 from yersinia pestis forms aqueous channels in lipid bilayer membranes

Antigen F1 is a protein of 17 kDa produced by Yersinia pestis it is cultured at 37 grade C. When incorporated into planar lipid bilayer membranes this protein induces fluctuations on membrane conductance typical of the formation of ionic channels. These fluctuations reveal two distinct unitary conductance sizes, one in the range of 800 to 1400 pS and the other in the range of 140 to 600 pS. Zero current potential measaurements in the presence of a salt gradient show that the channell is not significantly ion selective. The reversal potential measured in the presence of 0.5 MKCl on the cis side and 0.1 MKCl on the trans side was 3.58 ñ 3.98 mV (N=7). The non-selectivity of the channel, in addition to its large conductance, suggests that it forms large aqueous pores. The present results, taken together with other data showing that nantigen F1 inhibits the activity of phagocytic cells, suggest that antigen F1 acts by forming aqueous pores in the membrane of these target cells

Use of monoclonal antibodies against human T cells for clinical diagnosis by immunohistochemical procedures

Monoclonal antibodies (Mabs) were produced against human T cell membrane antigens. Sixteen Mabs were studied and six were selected for immunohistochemical assays on parffin-embedded tonsil section. Two Mabs (2D7 and 1E2) specifically recognized T-lymphocyte areas in sections of pathological tissues originating from lymphoproliferative diseases, and reacted with proteins of approximately 80 KDa. Most of the Mabs produced thus far are only suitable for immunohistochemical assays on frozen section. Only a few Mabs recognize lymphoid markers on paraffin-embedded sections, a procedure which permits a more extensive and practical application of Mabs in clinical diagnosis. These antibodies should be baluable in diagnosing. T. cell-related diseases and their large scale production should reduce laboratory costs because all reahgents currently avaliable are imported

The immunodominant surface antigen of Plasmodium gallinaceum is present in both the salivary gland and oocyst sporozoites

1. Monoclonal antibodies (MAbs) against surface antigens of Plasmodium gallinaceum sporozoites, an avian malaria parasite, were produced using spleen cells from mice immunized with sporozoites from mosquito salivary glands (SGS) or from midadgusts containing oocysts (OoS). 2. All of the 15 MAbs teted (11 anti-SGS and 4 anti-OoS) reacted with SGS and OoS by indirect immunofluorescence and circumsporozoiter precipitation reactions. Fourteen of these MAbs (11 anti-SGS and 3 anti-OoS) produced a Western blot (WB) patten identical to that produced with serum from mice lyperimmunized with viable intacts sporozoites. 3. All MAbs and the immune sera recognized only two polypeptide bands of approximate molecutlar weight 78 and 64KDa. 4. No difference in the WB pattern was observed when-or 12-day SGS or OoS extracts were used as antigens in WB. This antigenic similary was confirmed when the total protein extracts were visualized on silver-stained SDS-PAGE gel

Evolutionary conservation of laminin-binding proteins

The virulence of pathogens and metastatic capacity of cancer cells seems to correlate with the ability to adhere to cells and/or to basement components. A key feature of this mechanism in the expression of specific receptors for the basement membrane protein laminin. There different receptors have been already described in cells phylogenetically very distant, such as human white blood cells, Trichomonas vaginalis and Stapgylococcus aureus, all recognizing laminin with the same range of affinity. We have shown that laminin, which is also found in the circulation, enchances phagocytosis of S. aureus by macrophages in a species-specific fashion. Also, monoclonal antibodies (MAb) raised against the bacterial receptor inhibit the phagocytic enhancement mediated by laminin and recognize laminin-binding proteins in unicellular parasites and mammalian cells. The same Mab 1.H12 elutes a 52-kDa protein from bacterial extracts and a 67-kDa band from cancer cells extracts. Since the MAb is a monospecific reagent, results with 1.H12 strongly suggest and evolutionary conservation of the biding site of phylogenetically different laminin receptors

Production of monoclonal antibodies against Schistosoma mansoni and a trypanosomatid: a methodological report.

Anticorpos monoclonais foram produzidos contra dois diferentes organismos: o tripanosomatideo Herpetomonas muscarum muscarum e Schistosoma mansoni. Verificou-se que a obtencao desses anticorpos depende de variaveis como esquemas de imunizacao, origem e peso molecular do polietilenoglicol, lotes de soro fetal bovino, feederlayers de macrofagos, linhagens de celulas e condicoes de cultura de celulas mielomatosas de camundongo. Aplicando um protocolo aqui descrito, foram obtidos 35 anticorpos monoclonais estaveis contra H.m.muscarum e 10 contra S. mansoni. Com relacao ao tripanosomatideo, um esquema de intervalos menores entre as imunizacoes resultou em anticorpos da classe IgM; ao contrario, intervalos mais longos resultaram em anticorpos predominantemente da classe IgG. Com S. mansoni, infeccoes mais ou menos longas produziram igualmente anticorpos predominantemente da classe IgG.Padroes diferentes de reacao foram observados atraves de imunofluorescencia indireta: alguns anticorpos mostraram predominancia de reacao contra o flagelo do tripanosomatideo. Os anticorpos monoclonais anti-S. mansoni reagiram contra o tegumento ou contra o tubo digestivo em cortes de vermes adultos. A importancia biologica dos dois tipos de anticorpos esta agora sendo investigada

Sindrome de Cushing induzida pelo uso de dexametasona por via nasal sob a forma de spray. Relato de tres casos. / Cushing's syndrome induced by the use of dexamethasone by nasal route under spray form. Report of 3 cases

Sao apresentados tres casos de pacientes com Sindrome de Cushing induzidos pela administracao de Dexametasona sob a forma de spray nasal. Em todos esses casos ha evidencias de inibicao do eixo hipotalamo-pituitaria-adrenal. Em nenhum deles a funcao adrenal foi restaurada ate o momento, pois os pacientes ainda requerem o emprego de administracao oral de corticoide para sua manutencao

Fratura de Pouteau-Colles. Tratamento pela tecnica de Sarmiento (modificada). / Pouteau-Colles' fracture. Treatment by Sarmiento's modified technic

Um estudo em 20 casos utilizando a tecnica de Sarmiento e col. foi realizado, com pequena variacao de tecnica. Os resultados determinam que quando utilizamos a posicao supinada os resultados sao superiores