Annurca apple polyphenol extract selectively kills MDA-MB-231 cells through ROS generation, sustained JNK activation and cell growth and survival inhibition

Polyphenols represent the most studied class of nutraceuticals that can be therapeutics for a large spectrum of diseases, including cancer. In this study, we investigated for the first time the antitumor activities of polyphenol extract from Annurca apple (APE) in MDA-MB-231 triple negative breast cancer cells, and we explored the underlying mechanisms. APE selectively inhibited MDA-MB-231 cell viability and caused G2/M phase arrest associated with p27 and phospho-cdc25C upregulation and with p21 downregulation. APE promoted reactive oxygen species (ROS) generation in MDA-MB-231 cells while it acted as antioxidant in non-tumorigenic MCF10A cells. We demonstrated that ROS generation represented the primary step of APE antitumor activity as pretreatment with antioxidant N-acetylcysteine (NAC) prevented APE-induced G2/M phase arrest, apoptosis, and autophagy. APE downregulated Dusp-1 and induced a significant increase in JNK/c-Jun phosphorylation that were both prevented by NAC. Moreover, downregulation of JNK by its specific inhibitor SP600125 significantly diminished the anticancer activity of APE indicating that ROS generation and sustained JNK activation represented the main underlying mechanism of APE-induced cell death. APE also inhibited AKT activation and downregulated several oncoproteins, such as NF-kB, c-myc, and beta-catenin. In light of these results, APE may be an attractive candidate for drug development against triple negative breast cancer

Crushed Bricks: Demolition Waste as a Sustainable Raw Material for Geopolymers

Demolition activity plays an important role in the total energy consumption of the construction industry in the European Union. The indiscriminate use of non-renewable raw materials, energy consumption, and unsustainable design has led to a redefinition of the criteria to ensure environmental protection. This article introduces an experimental plan that determines the viability of a new type of construction material, obtained from crushed brick waste, to be introduced into the construction market. The potential of crushed brick waste as a raw material in the production of building precast products, obtained by curing a geopolymeric blend at 60 °C for 3 days, has been exploited. Geopolymers represent an important alternative in reducing emissions and energy consumption, whilst, at the same time, achieving a considerable mechanical performance. The results obtained from this study show that the geopolymers produced from crushed brick were characterized by good properties in terms of open porosity, water absorption, mechanical strength, and surface resistance values when compared to building materials produced using traditional technologies

Identification of tumor-associated cassette exons in human cancer through EST-based computational prediction and experimental validation

Background: Many evidences report that alternative splicing, the mechanism which produces mRNAs and proteins with different structures and functions from the same gene, is altered in cancer cells. Thus, the identification and characterization of cancer-specific splice variants may give large impulse to the discovery of novel diagnostic and prognostic tumour biomarkers, as well as of new targets for more selective and effective therapies. Results: We present here a genome-wide analysis of the alternative splicing pattern of human genes through a computational analysis of normal and cancer-specific ESTs from seventeen anatomical groups, using data available in AspicDB, a database resource for the analysis of alternative splicing in human. By using a statistical methodology, normal and cancer-specific genes, splice sites and cassette exons were predicted in silico. The condition association of some of the novel normal/tumoral cassette exons was experimentally verified by RT-qPCR assays in the same anatomical system where they were predicted. Remarkably, the presence in vivo of the predicted alternative transcripts, specific for the nervous system, was confirmed in patients affected by glioblastoma. Conclusion: This study presents a novel computational methodology for the identification of tumor-associated transcript variants to be used as cancer molecular biomarkers, provides its experimental validation, and reports specific biomarkers for glioblastoma

Matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in diagnosis of pleural effusion of malignant origin

OBJECTIVES The aim of the present study was to evaluate the diagnostic accuracy of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in differentiating benign from malignant exudative pleural effusions. METHODS This is a unicentre observational study including 97 consecutive patients with exudative pleural effusions. Metalloproteinase-9, tissue inhibitor of metalloproteinase-1, lactate dehydrogenase, ferritin, carcinoembryonic antigen and carbohydrate antigen 15-3 were measured in pleural effusion and serum by enzyme-linked immunosorbent assay. The activity of metalloproteinase-9 was also evaluated by substrate zymography. The data were correlated with final diagnosis of pleural effusions to evaluate the diagnostic accuracy. RESULTS Of the 97 eligible patients, 6 were excluded. Of the 91 patients included in the study, 70 had malignant pleural effusions and 21 had benign pleural effusions. Both in sera and pleural effusions, matrix metalloproteinase-9 (P < 0.0001), tissue inhibitor of metalloproteinase-1 (P < 0.0001) and carcinoembryonic antigen (P < 0.0001) levels were higher in neoplastic patients than in benign group. Zymography analysis showed a most prominent band at a molecular weight of 92 kDa (metalloproteinase-9) whereas a less intense band was observed at 72 kDa (metalloproteinase-2). A significant correlation was found between metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels in pleural effusion (P < 0.0001; r = 0.8) and serum (P < 0.03; r = 0.2). Pleural effusion metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels showed higher value of sensitivity (97 and 91%, respectively) and specificity (90 and 95%, respectively) compared with other standard markers. Serum metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels showed similar results. Among 70 neoplastic patients, 29 had negative pleural cytology. Of these, 25 presented elevated levels of metalloproteinase-9 and tissue inhibitor of metalloproteinase-1, whereas 4 patients had elevated levels of one of the two markers. CONCLUSIONS Our results showed that metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 might be valuable markers in differentiating benign from malignant pleural effusions. Their levels are neither influenced by the histology and tumour origin nor by the presence of tumour cells in pleural effusions. Thus, their use in clinical practice could help in the selection of patients needing more invasive procedures, such as thoracoscopic biopsy.OBJECTIVES The aim of the present study was to evaluate the diagnostic accuracy of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in differentiating benign from malignant exudative pleural effusions. METHODS This is a unicentre observational study including 97 consecutive patients with exudative pleural effusions. Metalloproteinase-9, tissue inhibitor of metalloproteinase-1, lactate dehydrogenase, ferritin, carcinoembryonic antigen and carbohydrate antigen 15-3 were measured in pleural effusion and serum by enzyme-linked immunosorbent assay. The activity of metalloproteinase-9 was also evaluated by substrate zymography. The data were correlated with final diagnosis of pleural effusions to evaluate the diagnostic accuracy. RESULTS Of the 97 eligible patients, 6 were excluded. Of the 91 patients included in the study, 70 had malignant pleural effusions and 21 had benign pleural effusions. Both in sera and pleural effusions, matrix metalloproteinase-9 (P < 0.0001), tissue inhibitor of metalloproteinase-1 (P < 0.0001) and carcinoembryonic antigen (P < 0.0001) levels were higher in neoplastic patients than in benign group. Zymography analysis showed a most prominent band at a molecular weight of 92 kDa (metalloproteinase-9) whereas a less intense band was observed at 72 kDa (metalloproteinase-2). A significant correlation was found between metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels in pleural effusion (P < 0.0001; r = 0.8) and serum (P < 0.03; r = 0.2). Pleural effusion metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels showed higher value of sensitivity (97 and 91%, respectively) and specificity (90 and 95%, respectively) compared with other standard markers. Serum metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels showed similar results. Among 70 neoplastic patients, 29 had negative pleural cytology. Of these, 25 presented elevated levels of metalloproteinase-9 and tissue inhibitor of metalloproteinase-1, whereas 4 patients had elevated levels of one of the two markers. CONCLUSIONS Our results showed that metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 might be valuable markers in differentiating benign from malignant pleural effusions. Their levels are neither influenced by the histology and tumour origin nor by the presence of tumour cells in pleural effusions. Thus, their use in clinical practice could help in the selection of patients needing more invasive procedures, such as thoracoscopic biopsy

O CONCEITO DE INCLUSÃO DE DISCENTES DE EDUCAÇÃO FÍSICA DE UNIVERSIDADES PÚBLICAS DO ESTADO DE SÃO PAULO NO CONTEXTO SOCIAL DA SUA PRÁTICA

Initial training in Physical Education has been rebuilt according to inclusive paradigms being established in society. This study looks into the concept of inclusion presented by senior Physical Education college students at public institutions of the state of São Paulo. Qualitative research was conducted through questionnaires answered by 322 students. Data were analyzed using Iramuteq software by Reinert Classification analysis. The results showed that students built a concept of inclusion compatible with the principle of complexity, considering legal, social, psychological and affective characteristics included in that proposition.La formación inicial en Educación Física se ha reconstruido según los paradigmas inclusivos en implantación en la sociedad. Así, ese estudio objetivó verificar cuál es el concepto de Inclusión presentado por los estudiantes universitarios que concluyen el curso de Educación Física en instituciones públicas del estado de São Paulo. Así, se desarrolló una investigación cualitativa a través de la aplicación de cuestionario a 322 académicos. Los datos fueron analizados con el programa Iramuteq, empleando los análisis de Clasificación por el Método de Reinert, Análisis de Similitud y Nube de Palabras. Como resultado, se ha identificado que los alumnos participantes han alcanzado la construcción de un concepto de Inclusión de acuerdo con la complejidad del principio, considerando aspectos legales, sociales, psicológicos y afectivos incluidos en esa propuesta. A formação inicial em Educação Física tem se reconstruído segundo os paradigmas inclusivos em implantação na sociedade. Assim, este estudo objetivou verificar o conceito de inclusão apresentado por estudantes universitários concluintes do curso de Educação Física em instituições públicas do estado de São Paulo. Para isso, foi desenvolvida uma pesquisa qualitativa por meio da aplicação de questionário a 322 acadêmicos. Os dados foram analisados pelo programa Iramuteq, por meio das análises de Classificação pelo Método de Reinert, Análise de Similitude e Nuvem de Palavras. Como resultado, foi identificado que os discentes respondentes têm alcançado a construção de um conceito de inclusão condizente com a complexidade do princípio, considerando aspectos legais, sociais, psicológicos e afetivos imbuídos nessa proposta.

Recognition and behavioral assessment of acute pain in cats: literature review

Embora a dor seja considerada o quarto sinal vital e uma das manifestações mais comumente encontradas na prática médica veterinária dos animais domésticos, seu tratamento ainda é inadequado. A dor aguda pós-operatória tem suscitado grande interesse por seu potencial risco de cronificação caso não adequadamente tratada, podendo piorar a recuperação e a qualidade de vida do paciente. O gato é uma das espécies domésticas menos estudadas no que diz respeito ao reconhecimento e controle da dor, e algumas das dificuldades residem na avaliação e na percepção da dor. O consenso sobre os sinais comportamentais da dor nesta espécie publicado em fevereiro de 2016 considerou alguns sinais como confiáveis e sensíveis para a avaliação da dor em gatos, em toda uma gama de diferentes condições clínicas, porém afirma a necessidade da realização de estudos que analisem a sua validade e aplicabilidade clínica, especialmente em relação a diferentes intensidades de dor. Na tentativa de se quantificar a dor são utilizados vários tipos de escalas subjetivas tradicionais e outras que facilitam a avaliação da efetividade da analgesia, a partir da observação de sinais comportamentais espontâneos indicativos de dor, combinada a uma resposta qualitativa à palpação da ferida cirúrgica. Faz-se necessária a utilização de escalas específicas para o tipo de dor (aguda ou crônica) e para a espécie, de modo a minimizar a subjetividade e a parcialidade dos observadores e possibilitando uma melhor assistência ao paciente.Although pain is considered the 4th vital sign and one of the most frequently observed clinical signs in domestic animals’ clinical practice, its treatment is still inadequate despite significant improvement in the last few years. Acute post-operative pain has aroused great interest due to its potential risk of developing into chronic pain, and if not treated properly, it might worsen the recovery and the patient’s quality of life. Cats are one of the least studied species of domestic animals regarding pain recognition and control. Some of the difficulties lie in pain assessment and perception. The consensus published in February 2016 about behavioral signs of pain in cats considered some signs to be reliable and sensitive for the assessment of pain in this species in many different clinical conditions, however it still states that more studies will be necessary in order to evaluate its clinical validity and applicability, especially considering the various pain intensities. As an attempt to quantify pain intensity in cats, several types of traditional subjective scales and others that facilitate pain assessment by combining the observation of spontaneous behavioral signals of pain and qualitative response to palpation of surgical wound are used as tools. It is necessary to use specific scales for each type of pain and for each specific animal species so to minimize the subjectivity and the partiality of the observers, reducing bias and improving efficacy, thus leading to a better patient care

Pheochromocytomas and paragangliomas in children: Data from the Italian Cooperative Study (TREP)

Pheochromocytomas (PCs) are neuroendocrine tumors arising from the chromaffin cells of the adrenal gland, and paragangliomas (PGLs) are their extra-adrenal counterparts arising from ganglia along the sympathetic/parasympathetic chain. Surgery is the cornerstone of treatment. A sporatic or inherited germline mutation is commonly associated

Orexins and receptor OX2R in the gastroenteric apparatus of two teleostean species: Dicentrarchus labrax and Carassius auratus

Orexin A and B peptides and the receptor OX2R were studied in sea bass and goldfish gastroenteric tract by immunoblotting combined with densitometric analysis using NIH Image J software and immunohistochemical techniques. These teleost species present a different gut organization and diverse feeding habits. Immunoblotting experiments showed one band of 16 kDa corresponding to prepro-orexin, and one band of 38 kDa corresponding to the OX2R receptor. Immunohistochemical localization of OXA and OXB was observed in the enteric nervous system throughout the gastroenteric tract of both species. OXA and OXB immunoreactive cells were found in the gastric and intestinal regions of sea bass, and were mainly found in the basal region of folds in intestinal bulb, and in the midgut and hindgut of goldfish. The distribution of OX2R was mainly detected in the mucosa of the gastroenteric tract of sea bass and goldfish. This distribution suggests an endocrine action of OXA and OXB in the gastrointestinal tract as well as involvement in the peripheral control of food intake and digestive processes in both species. This study might also serve to determine the productive factors in breeding and as a baseline for future experimental studies on the regulation of the gastroenteric functions in non-mammalian vertebrates

MEK blockade converts AML differentiating response to retinoids into extensive apoptosis

: The aberrant function of transcription factors and/or kinase-based signaling pathways that regulate the ability of hematopoietic cells to proliferate, differentiate, and escape apoptosis accounts for the leukemic transformation of myeloid progenitors. Here, we demonstrate that simultaneous retinoid receptor ligation and blockade of the MEK/ERK signaling module, using the small-molecule inhibitor CI-1040, result in a strikingly synergistic induction of apoptosis in both acute myeloid leukemia (AML) and acute promyelocytic leukemia (APL) cells with constitutive ERK activation. This proapoptotic synergism requires functional RAR and RXR retinoid receptors, as demonstrated using RAR- and RXR-selective ligands and RAR-defective cells. In the presence of MEK inhibitors, however, retinoid-induced chromatin remodeling, target-gene transcription, and granulocytic differentiation are strikingly inhibited and apoptosis induction becomes independent of death-inducing ligand/receptor pairs; this suggests that apoptosis induction by combined retinoids and MEK inhibitors is entirely distinct from the classical "postmaturation" apoptosis induced by retinoids alone. Finally, we identify disruption of Bcl-2-dependent mitochondrial homeostasis as a possible point of convergence for the proapoptotic synergism observed with retinoids and MEK inhibitors. Taken together, these results indicate that combined retinoid treatment and MEK blockade exert powerful antileukemic effects and could be developed into a novel therapeutic strategy for both AML and APL