Purification, biochemical characterization and fibrinolytic potential of proteases produced by bacteria of the genus Bacillus: a systematic literature review.

Thrombosis is a hematological disorder characterized by the formation of intravascular thrombi, which contributes to the development of cardiovascular diseases. Fibrinolytic enzymes are proteases that promote the hydrolysis of fibrin, promoting the dissolution of thrombi, contributing to the maintenance of adequate blood flow. The characterization of new effective, safe and low-cost fibrinolytic agents is an important strategy for the prevention and treatment of thrombosis. However, the development of new fibrinolytics requires the use of complex methodologies for purification, physicochemical characterization and evaluation of the action potential and toxicity of these enzymes. In this context, microbial enzymes produced by bacteria of the Bacillus genus are promising and widely researched sources to produce new fibrinolytics, with high thrombolytic potential and reduced toxicity. Thus, this review aims to provide a current and comprehensive understanding of the different Bacillus species used for the production of fibrinolytic proteases, highlighting the purification techniques, biochemical characteristics, enzymatic activity and toxicological evaluations used.

Whole body diffusion for metastatic disease assessment in neuroendocrine carcinomas: comparison with OctreoScan® in two cases.

Neuroendocrine tumor (NET) patients must be adequately staged in order to improve a multidisciplinary approach and optimal management for metastatic disease. Currently available imaging studies include somatostatin receptor scintigraphy, like OctreoScan®, computed tomography (CT), scans and magnetic resonance imaging (MRI), which analyze vascular concentration and intravenous contrast enhancement for anatomic tumor localization. However, these techniques require high degree of expertise for interpretation and are limited by their availability, cost, reproducibility, and follow-up imaging comparisons. NETs significantly reduce water diffusion as compared to normal tissue. Diffusion-weighted imaging (DWI) in MRI has an advantageous contrast difference: the tumor is represented with high signal over a black normal surrounding background. The whole-body diffusion (WBD) technique has been suggested to be a useful test for detecting metastasis from various anatomic sites. In this article we report the use of DWI in MRI and WBD in two cases of metastatic pulmonary NET staging in comparison with OctreoScan® in order to illustrate the potential advantage of DWI and WBD in staging NETs.

Upfront Radiosurgery for Treatment of Symptomatic Obstructive Hydrocephalus due to Brain Tumors.

Introduction Hydrocephalus is a build-up of cerebrospinal fluid (CSF) in the brain and is characterized by abnormal dilatation of the cerebral ventricles. Patients can be either asymptomatic, have symptoms related to primary tumors, or have hydrocephalus-related symptoms. Generally, symptomatic patients are candidates for ventriculoperitoneal (VP) shunt placement to reduce acute symptoms. Little evidence exists regarding the resolution of symptomatic hydrocephalus secondary to brain tumors using stereotactic radiosurgery (SRS) alone as a primary treatment option. Methods The present study is a retrospective series of eight patients (six men and two women) diagnosed with obstructive hydrocephalus due to brain tumors treated with radiosurgery between April 2013 and February 2021. The primary endpoint of the present study is to report our institutional experience regarding the control of symptomatic obstructive hydrocephalus due to brain tumors treated with upfront radiosurgery. Results The mean age was 52 years (range, 5-79). The most common presenting symptoms included headache (100%), vision-related symptoms (75%), and ataxia (37.5%). All patients showed symptom improvement after radiosurgery, five (62.5%) patients showed resolution in less than three days and the rest of the patients resolved hydrocephalus in a longer timeframe (more than three days). All patients lowered their Evans index compared to the index documented before radiosurgery, in a range from 0.02 to 0.17. Conclusion Radiosurgery is a non-invasive alternative treatment for primary and secondary brain tumors that debut with obstructive hydrocephalus, tumors expected to have a high alpha/beta ratio might be suitable to attempt radiosurgery to avoid permanently implanted devices such as VP shunts or other invasive procedures such as a third ventriculostomy. The present study demonstrated that in selected cases SRS can lead to hydrocephalus symptom resolution along with a decrease in ventricular size in a relatively short time frame. Little evidence exists regarding the effect of SRS on symptomatic hydrocephalus resolution and further histology-specific studies are required. We acknowledge that this approach requires immediate access to radiosurgery and close clinical follow-up to ensure success.

Environmental health indicators and a case study of air pollution in Latin American cities.

Environmental health indicators (EHIs) are applied in a variety of research and decision-making settings to gauge the health consequences of environmental hazards, to summarize complex information, or to compare policy impacts across locations or time periods. While EHIs can provide a useful means of conveying information, they also can be misused. Additional research is needed to help researchers and policy-makers understand categories of indicators and their appropriate application. In this article, we review current frameworks for environmental health indicators and discuss the advantages and limitations of various forms. A case study EHI system was developed for air pollution and health for urban Latin American centers in order to explore how underlying assumptions affect indicator results. Sixteen cities were ranked according to five indicators that considered: population exposed, children exposed, comparison to health-based guidelines, and overall PM(10) levels. Results indicate that although some overall patterns in rankings were observed, cities' relative rankings were highly dependent on the indicator used. In fact, a city that was ranked best under one indicator was ranked worst with another. The sensitivity of rankings, even when considering a simple case of a single pollutant, highlights the need for clear understanding of EHIs and how they may be affected by underlying assumptions. Careful consideration should be given to the purpose, assumptions, and limitations of EHIs used individually or in combination in order to minimize misinterpretation of their implications and enhance their usefulness.

Experimental schistosomiasis in the common marmoset Callithrix jacchus.

In order to evaluate Callithrix jacchus as an animal model for mansoni schistosomiasis, a group of 10 male animals were once percutaneously exposed to 250 cercariae of the Schistosoma mansoni SLM (São Lourenço da Mata) strain. Animals were periodically bled for measuring serum level of enzymes and proteins and for blood cell counting. When comparing pre-infection to post-infection values, a significant increase was found for alkaline phosphatase at 15 to 120 days p.i., differential counts of eosinophil at 45 and 60 days, and total protein and global eosinophil counts at 120 days. No Schistosoma mansoni eggs were found in stools. Adult worms of small size were recovered from five animals. At day 120, the number of Schistosoma mansoni eggs/g of tissue was 0-289.7 (liver), 0-30.1 (large intestine) and 0-171.4 (small intestine). These findings lead us to classify Callithrix jacchus as a non-permissive host to the SLM strain of Schistosoma mansoni.

Web-based versus traditional lecture: are they equally effective as a flexible bronchoscopy teaching method?

This study compares the traditional live lecture to a web-based approach in the teaching of bronchoscopy and evaluates the positive and negative aspects of both methods. We developed a web-based bronchoscopy curriculum, which integrates texts, images and animations. It was applied to first-year interns, who were later administered a multiple-choice test. Another group of eight first-year interns received the traditional teaching method and the same test. The two groups were compared using the Student's t-test. The mean scores (± SD) of students who used the website were 14.63 ± 1.41 (range 13-17). The test scores of the other group had the same range, with a mean score of 14.75 ± 1. The Student's t-test showed no difference between the test results. The common positive point noted was the presence of multimedia content. The web group cited as positive the ability to review the pages, and the other one the role of the teacher. Web-based bronchoscopy education showed results similar to the traditional live lecture in effectiveness.

Histomorfometria e índice proliferativo (Ki-67) no carcinoma escamocelular in situ de pregas vocais / Histomorphometry and proliferative index in squamous cell carcinoma in situ of the vocal cords

INTRODUÇÃO: As displasias da laringe são precursoras do carcinoma escamocelular invasivo e constituem lesões pouco comuns, pois a maior parte dos casos é detectada como carcinoma invasor. OBJETIVO: Caracterizar as displasias acentuadas/carcinoma in situ da prega vocal quanto a área do epitélio, diâmetro dos núcleos e índice proliferativo, comparando esses dados com os obtidos no epitélio escamoso normal e de transição. MATERIAL E MÉTODO: Entre as 1.400 biópsias e peças cirúrgicas de lesões intraepiteliais e carcinomas invasivos de laringe (1994-2006), foram selecionados cinco casos (0,35%) pareados com dois controles de epitélio normal e com a idade; todos foram submetidos a análise morfométrica e imuno--histoquímica (Ki-67). RESULTADOS: Comparando-se o epitélio displásico com o normal e de transição observou-se maior área e diâmetro nuclear no epitélio displásico; não houve diferenças significativas no diâmetro dos núcleos por camada entre o epitélio displásico e de transição; e o índice proliferativo foi maior no epitélio displásico com núcleos corados em todas camadas e menor com núcleos limitados a camada basal e parabasal no epitélio normal e de transição. CONCLUSÃO: verifica-se maior área no epitélio displásico. O diâmetro dos núcleos do epitélio displásico e do de transição são semelhantes, mas este apresenta núcleos uniformes em toda extensão em contraste com o pleomorfismo do epitélio displásico. O índice proliferativo (Ki-67) contribui para o diagnóstico diferencial das lesões escamosas da prega vocal, pois no carcinoma in situ/displasia acentuada observa-se maior número de núcleos marcados, os quais são vistos em todas as camadas no epitélio.

INTRODUCTION: Laryngeal dysplasias, precursors of squamous cell carcinomas, are uncommon lesions, inasmuch as most cases are diagnosed as invasive carcinomas. OBJECTIVE: To characterize severe dysplasia/in situ carcinoma of the vocal cords by comparing the area of involved epithelium, nuclear diameter, and proliferative index with transition and normal squamous epithelia. MATERIAL AND METHOD: Among 1,400 surgical and biopsy specimens of laryngeal lesions (1994-2006), five cases (0.35%) were selected and compared according to age with two controls with normal epithelium. Furthermore, all of them were identified and submitted to morphometric and immunohistochemical analysis (Ki-67). RESULTS: After comparing dysplastic with transition and normal squamous epithelia, it was observed an increased nuclear diameter in dysplastic epithelium. Additionally, there was no significant difference in nuclear diameter per layer between dysplastic and transition epithelia. Moreover, the proliferative index was higher in dysplastic epithelium with stained nuclei in all layers and lower in normal and transition epithelia with nuclei concentrated on the basal and parabasal layers. CONCLUSION: Dysplastic epithelium occupies larger areas. Nuclear diameter is similar in dysplastic and transition epithelia. However, the latter has uniform nuclei, in contrast with pleomorphism in dysplastic epithelium. The proliferative index (Ki-67) contributes to the differential diagnosis of intraepithelial lesions of the vocal cords, inasmuch as in severe dysplasia/in situ carcinoma there is a higher number of stained nuclei, which are seen in all epithelial layers.

Perfil imunoistoquímico das proteínas p53 e Ki67 em pacientes com retocolite ulcerativa inespecífica / Imunohistochemical profile of p53 and Ki67 from patients with unspecific ulcerative colitis

Objetivos: O presente estudo avaliou, através da imunohistoquímica e estudo histopatológico, as principais alterações na mucosa intestinal de pacientes acometidos pela retocolite ulcerativa crônica idiopática (RCUI) apenas tratada clinicamente (n=30) ou subseqüentemente tratada com Proctocolectomia total e construção de bolsa ileal (n=30). Métodos: Desta forma, foram selecionados fragmentos de tecido intestinal submetidos à imunomarcação para as proteínas p53 e Ki67 e coloração por hematoxilina-eosina para análise histopatológica comparativa. Resultados: Os resultados obtidos indicam importantes diferenças no perfil inflamatório e presença de áreas de erosão/desgate da mucosa colônica de ambos os grupos estudados. Quanto a imunoexpressão, observou-se uma maior reatividade de padrão nuclear principalmente nos tecidos inflamados dos pacientes com RCUI tratados clinicamente. Conclusões: Estes resultados sugerem que, de acordo com o perfil histopatológico e imunohistoquímico, a mucosa colônica de pacientes submetidos ao tratamento cirúrgico apresenta menos alterações que os pacientes tratados clinicamente. Assim, os dados sugerem que estes métodos podem auxiliar no diagnóstico e monitoramento de pacientes sob diferentes rotinas terapêuticas.

Objectives: The present study evaluated, trough immunohistochemistry and histopatologic analysis, the main alterations in intestinal mucous from patients with unspecific ulcerative colitis (IUC) clinically (n=30) or surgically treated (n=30) with ileal J-pouch and permanent ileostomy. Methods: Thus, tissue slices from intestinal mucous were submitted to imunostains for p53 and Ki67 proteins, and Haematoxilin-eosin staining for comparative histopathological evaluation. Results: The results indicated important differences in inflammatory profile and erosion/necrosis sites on the both investigated groups. Related to imunoexpression was observed a higher reactivity with nuclear pattern, mainly in inflammatory tissues from IUC patients clinically treated. Conclusions: Those results showed that, according to histopathological and imunohistochemical parameters, the colonic mucous from IUC patients submitted to surgical treatment exhibited less alterations than IUC patients clinically treated. The data suggest that those methods could be helpful to diagnosis and assist the accompaniment of patients under different therapeutic routines.

Avaliação histomorfométrica de micronúcleos e colágeno como métodos adicionais no diagnóstico diferencial de neoplasias cutâneas / Histomorphometrical evaluation of the micronucleous and collagen how additional methods in skin neoplasms diagnosis

Este estudo avaliou a densidade de micronúcleos e a distribuição do colágeno intersticial em tumores benignos e malignos de pele. Foram selecionados casos de ceratose seborréica, ceratose actínica, ceratoacantoma e carcinoma basocelular. Os resultados demonstraram um aumento significante do número de micronúcleos nas lesões de carcinoma basocelular e ceratoacantoma. Por outro lado, houve uma redução da área média de colágeno intersticial nas lesões dos mesmos, em relação ao tecido normal. Conclui-se então, que este tipo de análise poderá ser utilizado como parâmetro complementar na diferenciação de lesões cutâneas difíceis de serem diagnosticadas apenas pelos métodos convencionais de análise histopatológica.

This study established the density of micronuclei and distribution of the interstitial collagen in benign andmalign skin tumors. Skin samples from basal cell carcinoma, keratoacanthoma, seborreic keratosis and actinic keratosis were used. The results showed a significant variation in the micronucleus number in the basal cell carcinoma and keratoacanthoma. On the other hand, the mean area of collagen was lower in these ones in relation to the normal tissue. It was concluded that this analysis can be used as complementar parameter in the differentiation of cutaneous lesions with hard diagnosis carried out by means of conventional methods of histopathological analysis.

Análise histológica e histoquímica de fatores prognósticos em pacientes com retocolite ulcerativa / Histological and histochemical analysis of prognostic factors in patients with ulcerative colitis

OBJETIVO: Visando oferecer um método que auxilie o diagnóstico diferencial ou mesmo que seja indicador da transformação neoplásica, a galectina-3 surge como um potencial marcador da fisiopatologia de diversos tipos de câncer, entre eles o câncer colorretal. MÉTODOS: o perfil imuno-histoquímico foi obtido do tecido intestinal de pacientes com retocolite ulcerativa (n=20) e sua respectiva contraparte normal, de ambos os sexos e idade média de 55 anos. Os tecidos foram submetidos à histoquímica da reação de Schiff (PAS) para visualizações das inclusões de glicosaminoglicanos e tricrômico de Masson para visualização das fibras colágenas. Fragmentos de tecido (4 µm) foram submetidos à técnica de imuno-histoquímica para a proteína galectina-3. A marcação tecidual foi analisada através de uma estação de trabalho, contendo um microscópio óptico equipado com uma câmera digital, ambos acoplados a um computador contendo o software OPTIMAS®. RESULTADOS: Os resultados demonstram uma diminuição da expressão da galectina-3 no sitio inflamatório da retocolite ulcerativa. Houve um aumento significativo (p<0.022) no número de células caliciformes dos pacientes tratados clinicamente, porém com redução do conteúdo glandular. CONCLUSÕES: Os dados sugerem que a atividade glandular juntamente com a redução da expressão da galectina-3 possam auxiliar o diagnóstico e o monitoramento desses pacientes.

OBJECTIVES: Aiming to offer a diagnosis method or a prognostic marker, galectin-3 appears as a potential marker of the physiopathology of diverse diseases and some types of cancer, between them the colorectal cancer. METHODS: In our study, the immunohistochemical and histological profiles had been taken from intestinal biopsies of patients with ulcerative colitis (n=20) and its respective normal counterpart, from both males and females and average age of 55 years-old. The biopsies had been submitted to Periodic Acid of Schiff (PAS) for visualization of glycosaminoglycans inclusions, and Masson's tricromic for visualization of collagen fibers. Fragments (4µm) had been submitted to the technique of immunohistochemistry for galectin-3 protein. The profiles of tecidual coloration had been analyzed through a work station contend an optic microscope copulated with a digital camera, both connected a computer contend software OPTIMAS®. RESULTS: Our results demonstrate a reduction of galectin-3 expression in ulcerative colitis inflammatory site from surgically and clinically treated patients. The goblet cells number from the surgically treated patients had suffered a significant reduction (p<0.022), however with increase of its glandular content. CONCLUSIONS: The data suggest that the glandular activity together with the reduction of the galectin-3 expression can assist the accompaniment of these patients.

Análise morfométrica de micronúcleos e colágeno intersticial em tumores de mama / Morphometric analysis of micronucleuses and interstitial collagen in breast tumors

O presente trabalho estabeleceu a densidade de micronúcleos e a distribuição do colágeno intersticial no tecido tumoral benigno e maligno de mama humana. Foram utilizadas a reação histoquímica de Feulgen e tricrômico de Masson, e a análise digital de imagens. Foram selecionados fragmentos de tecido mamário, diagnosticados como Fibroadenoma (FA), Doença fibrocística (DF) e Carcinoma Ductal Infiltrante (CDI). Para controle, foram utilizadas amostras de plástica mamária com tecido normal. A reação histoquímica foi realizada através da reação de Feulgen, para evidenciação dos micronúcleos, e, para a análise de colágeno intersticial, foi utilizada a coloração tricrômico de Masson. O número médio de micronúcleos exibiu valores semelhantes em amostra de FA (90,8 ± 4,5), DF (80,3 ± 4,0) e tecido normal (83,0 ± 4,1). Entretanto, valores significativamente menores que aqueles foram encontrados no CDI (176,5 ± 88,3). Quanto à distribuição do colágeno intersticial, constatou-se um aumento estatisticamente significante (p< 0,001) nos casos de FA (8101,4 ± 405,07) e DF (7046,78 ± 352,33), quando comparados ao tecido mamário normal (1733,13 ± 86,65) e ao CDI (2165,94 ± 108,29). Pode-se concluir que tanto o método de análise de imagem como a reação de Feulgen mostraram-se eficientes para evidenciar as alterações do núcleo celular e da matriz extracelular de tumores de mama.

Experimental schistosomiasis in the Common Marmoset Callithrix jacchus

Com o objetivo de avaliar o Callithrix jacchus (sagüi) como modelo experimental para a esquistossomose, um grupo de 10 animais, machos, adultos jovens, foram expostos a 250 cercárias da cepa SLM (São Lourenço da Mata) do Schistosoma mansoni, pela via percutânea. A intervalos de 15 dias até 120 dias apos infecção foram dosados os níveis de proteínas totais e enzimas séricas, além de realizados exames parasitológico e hematológico. Aumento significativo foi observado para: fosfatase alcalina a partir do 150 dia; contagem global de eosinófilos aos 120 dias, e diferencial de eosinófilos aos 45 e 60 dias p.i. Nao se observou ovos de Schistosoma nas fezes. A perfusão cinco animais apresentaram vermes adultos. Todos os vermes encontrados eram diminutos. O número de ovos foi 0-289,7 ovos/g de tecido no fígado, 0-30,1 no intestino grosso, 0-171,4 no intestino delgado. Esta observaçoes levam-nos a concluir que o Callithrix jacchus seja não-permissivo ao Schistosoma.

Altered lectin-binding sites in normal colon and ulcerative colitis

O presente trabalho objetivou, através de histoquímica com lectinas e análise digital de imagens, avaliar a expressão de carboidratos em amostras de colo normal e com colite ulcerativa. A partir de fragmentos de mucosa intestinal foram obtidos cortes histológicos (4mm) que foram incubados com lectinas (Con A, WGA, LTA e PNA), e os resultados das marcações foram avaliados através de microscopia óptica e sistema de análise de imagens. Os resultados obtidos revelaram uma intensa marcação para as células inflamatórias, principalmente neutrófilos infiltrados no tecido de reto e sigmóide, bem como células das glândulas intestinais. As lectinas WGA e LTA exibiram padrões distintos de marcação entre o epitélio normal e os casos de colite ulcerativa. As lectinas PNA e Con A falharam em reconhecer os carboidratos celulares nos casos estudados em ambos os grupos. Os resultados obtidos foram confirmados pela análise de imagem. As observações obtidas sugerem que as lectinas WGA e LTA são marcadores promissores para diferenciar o epitélio normal do padrão inflamatório da colite ulcerativa, indicando uma expressão distinta de N-acetilglicosamina e L-fucose nos respectivos casos estudados.

Complete acute occlusion of the portal vein in dogs: II. A study of acid-base balance aimed at its application in liver transplantation

Como etapa inicial de um plano geral de pesquisa sobre avaliacao de derivacoes portossistemicas descompressivas, habitualmente usadas em transplante hepatico ortotopico experimental, os autores estudaram 20 caes mesticos, adultos, de ambos os sexos e com peso corporal de 15,2 ñ 1,7 kg. Em dez animais procedeu-se a oclusao total e aguda da veia porta durante 2 h (Grupo Oclusao Portal). Nos dez animais restantes, praticou-se a operacao simulada (Grupo Controle). Foram investigados, de modo sistematico, dados do equilibrio acido-basico no sangue arterial e no sangue portal desses animais. Tais dados foram analisados estatisticamente, permitindo concluir com um limite de probabilidade de erro de 5 por cento que a oclusao total e aguda da veia porta praticada em caes, nas condicoes de experimentacao usadas no presente estudo, produziu nos referidos animais: 1. Acidose metabolica no sangue arterial, caracterizada por aumento da concentracao do ion ('cH POT. +') e diminuicoes da pressao parcial de gas carbonico ('pCO IND. 2') e da concentracao de bases totais (cBT); 2. Acidose mista no sangue portal, evidenciada por elevacao da concentracao do ion hidrogenio ('cH POT. +'), da pressao parcial de gas carbonico ('pCO IND. 2') e de diminuicao da concentracao de bases totais (cBT), assim como por hipoxemia...