Telomerase promoter mutations in cancer: an emerging molecular biomarker?

João Vinagre, Vasco Pinto and Ricardo Celestino contributed equally to the manuscript.

Immunohistochemical molecular phenotypes of gastric cancer based on SOX2 and CDX2 predict patient outcome

Background Gastric cancer remains a serious health concern worldwide. Patients would greatly benefit from the discovery of new biomarkers that predict outcome more accurately and allow better treatment and follow-up decisions. Here, we used a retrospective, observational study to assess the expression and prognostic value of the transcription factors SOX2 and CDX2 in gastric cancer. Methods SOX2, CDX2, MUC5AC and MUC2 expression were assessed in 201 gastric tumors by immunohistochemistry. SOX2 and CDX2 expression were crossed with clinicopathological and follow-up data to determine their impact on tumor behavior and outcome. Moreover, SOX2 locus copy number status was assessed by FISH (N = 21) and Copy Number Variation Assay (N = 62). Results SOX2 was expressed in 52% of the gastric tumors and was significantly associated with male gender, T stage and N stage. Moreover, SOX2 expression predicted poorer patient survival, and the combination with CDX2 defined two molecular phenotypes, SOX2+CDX2- versus SOX2-CDX2+, that predict the worst and the best long-term patients’ outcome. These profiles combined with clinicopathological parameters stratify the prognosis of patients with intestinal and expanding tumors and in those without signs of venous invasion. Finally, SOX2 locus copy number gains were found in 93% of the samples reaching the amplification threshold in 14% and significantly associating with protein expression. Conclusions We showed, for the first time, that SOX2 combined with CDX2 expression profile in gastric cancer segregate patients into different prognostic groups, complementing the clinicopathological information. We further demonstrate a molecular mechanism for SOX2 expression in a subset of gastric cancer cases.

A reconstrução tomográfica em SPECT com utilização dos novos métodos iterativos “WBR”: comparação com métodos analíticos

Os métodos utilizados pela Medicina moderna no âmbito da Imagem Molecular e na sua capacidade de diagnosticar a partir da “Função do Orgão” em vez da “Morfologia do Orgão”, vieram trazer á componente fundamental desta modalidade da Imagiologia Médica – A Medicina Nuclear – uma importância acrescida, que se tem traduzido num aumento significativo no recurso á sua utilização nas diferentes formas das suas aplicações clínicas. Para além dos aspectos meramente clínicos, que só por si seriam suficientes para ocupar várias dissertações como a presente; a própria natureza desta técnica de imagem, com a sua inerente baixa resolução e tempos longos de aquisição, vieram trazer preocupações acrescidas quanto ás questões relacionadas com a produtividade (nº de estudos a realizar por unidade de tempo); com a qualidade (aumento da resolução da imagem obtida) e, com os níveis de actividade radioactiva injectada nos pacientes (dose de radiação efectiva sobre as populações). Conhecidas que são então as limitações tecnológicas associadas ao desenho dos equipamentos destinados á aquisição de dados em Medicina Nuclear, que apesar dos avanços introduzidos, mantêm mais ou menos inalteráveis os conceitos base de funcionamento de uma Câmara Gama, imaginou-se a alteração significativa dos parâmetros de aquisição (tempo, resolução, actividade), actuando não ao nível das condições técnico-mecânicas dessa aquisição, mas essencialmente ao nível do pós-processamento dos dados adquiridos segundo os métodos tradicionais e que ainda constituem o estado da arte desta modalidade. Este trabalho tem então como objectivo explicar por um lado, com algum pormenor, as bases tecnológicas que desde sempre têm suportado o funcionamento dos sistemas destinados á realização de exames de Medicina Nuclear, mas sobretudo, apresentar as diferenças com os inovadores métodos, que aplicando essencialmente conhecimento (software), permitiram responder ás questões acima levantadas.

Engineering Temporal and Spatial Aspects in OWL using Patterns

WWW is a huge, open, heterogeneous system, however its contents data is mainly human oriented. The Semantic Web needs to assure that data is readable and “understandable” to intelligent software agents, though the use of explicit and formal semantics. Ontologies constitute a privileged artifact for capturing the semantic of the WWW data. Temporal and spatial dimensions are transversal to the generality of knowledge domains and therefore are fundamental for the reasoning process of software agents. Representing temporal/spatial evolution of concepts and their relations in OWL (W3C standard for ontologies) it is not straightforward. Although proposed several strategies to tackle this problem but there is still no formal and standard approach. This work main goal consists of development of methods/tools to support the engineering of temporal and spatial aspects in intelligent systems through the use of OWL ontologies. An existing method for ontology engineering, Fonte was used as framework for the development of this work. As main contributions of this work Fonte was re-engineered in order to: i) support the spatial dimension; ii) work with OWL Ontologies; iii) and support the application of Ontology Design Patterns. Finally, the capabilities of the proposed approach were demonstrated by engineering time and space in a demo ontology about football.

Evolution of Drug Resistance: Insight on TEM β-Lactamases Structure and Activity and β-Lactam Antibiotics

Since the discovery of the first penicillin bacterial resistance to β-lactam antibiotics has spread and evolved promoting new resistances to pathogens. The most common mechanism of resistance is the production of β-lactamases that have spread thorough nature and evolve to complex phenotypes like CMT type enzymes. New antibiotics have been introduced in clinical practice, and therefore it becomes necessary a concise summary about their molecular targets, specific use and other properties. β-lactamases are still a major medical concern and they have been extensively studied and described in the scientific literature. Several authors agree that Glu166 should be the general base and Ser70 should perform the nucleophilic attack to the carbon of the carbonyl group of the β-lactam ring. Nevertheless there still is controversy on their catalytic mechanism. TEMs evolve at incredible pace presenting more complex phenotypes due to their tolerance to mutations. These mutations lead to an increasing need of novel, stronger and more specific and stable antibiotics. The present review summarizes key structural, molecular and functional aspects of ESBL, IRT and CMT TEM β-lactamases properties and up to date diagrams of the TEM variants with defined phenotype. The activity and structural characteristics of several available TEMs in the NCBI-PDB are presented, as well as the relation of the various mutated residues and their specific properties and some previously proposed catalytic mechanisms.

The role of molecular imaging in modern drug development

Drug development represents a highly complex, inefficient and costly process. Over the past decade, the widespread use of nuclear imaging, owing to its functional and molecular nature, has proven to be a determinant in improving the efficiency in selecting the candidate drugs that should either be abandoned or moved forward into clinical trials. This helps not only with the development of safer and effective drugs but also with the shortening of time-to-market. The modern concept and future trends concerning molecular imaging will assumedly be hybrid or multimodality imaging, including combinations between high sensitivity and functional (molecular) modalities with high spatial resolution and morphological techniques.

Synthesis of molecular biomimetics

Biomimetics has paved the way toward new materials and technologies inspired in Nature. Biomolecules and their supramolecular organization have today a leading role in biomimetics, benefiting from the recent advances in nanotechnology. The production of biomimetic materials may be however a difficult task, because Nature does it very well. The use of several building blocks assembled in bottom-up arrangement is without doubt at the core of this process. Such building blocks include different molecules or molecular arrangements, of synthetic or natural origin, such as amino acids, lipids, carbohydrates, nucleic acids, carbon allotropes, dendrimers, or organosilanes, among others. The most common approaches to produce synthetic biomimetic materials are reported herein, with special emphasis to building blocks and their supramolecular arrangement.

Myoglobin-biomimetic electroactive materials made by surface molecular imprinting on silica beads and their use as ionophores in polymeric membranes for potentiometric transduction

Myoglobin (Mb) is among the cardiac biomarkers playing a major role in urgent diagnosis of cardiovascular diseases. Its monitoring in point-of-care is therefore fundamental. Pursuing this goal, a novel biomimetic ionophore for the potentiometric transduction of Mb is presented. It was synthesized by surface molecular imprinting (SMI) with the purpose of developing highly efficient sensor layers for near-stereochemical recognition of Mb. The template (Mb) was imprinted on a silane surface that was covalently attached to silica beads by means of self-assembled monolayers. First the silica was modified with an external layer of aldehyde groups. Then, Mb was attached by reaction with its amine groups (on the external surface) and subsequent formation of imine bonds. The vacant places surrounding Mb were filled by polymerization of the silane monomers 3-aminopropyltrimethoxysilane (APTMS) and propyltrimethoxysilane (PTMS). Finally, the template was removed by imine cleavage after treatment with oxalic acid. The results materials were finely dispersed in plasticized PVC selective membranes and used as ionophores in potentiometric transduction. The best analytical features were found in HEPES buffer of pH 4. Under this condition, the limits of detection were of 1.3 × 10−6 mol/L for a linear response after 8.0 × 10−7 mol/L with an anionic slope of −65.9 mV/decade. The imprinting effect was tested by preparing non-imprinted (NI) particles and employing these materials as ionophores. The resulting membranes showed no ability to detect Mb. Good selectivity was observed towards creatinine, sacarose, fructose, galactose, sodium glutamate, and alanine. The analytical application was conducted successfully and showed accurate and precise results.

The potential of molecular imprinting as a biosensing devices for monitoring the CEA cancer biomarker

6th Graduate Student Symposium on Molecular Imprinting

Carnitine tailored Sensors on Surface Molecular Imprinting based on Graphene layers

III Jornadas de Electroquímica e Inovação (Electroquímica e Nanomateriais), na Universidade de Trás-os-Montes e Alto Douro, Vila Real, 16 a 17 de Setembro de 2013

Host-Tailored Sensors for Carnitine Potentiometric Measurements based on Surface Molecular Imprinting

Graduate Student Symposium on Molecular Imprinting 2013, na Queen’s University, Belfast, United Kingdom, 15 a 17 de Agosto de 2013

Role of oxidative stress-induced systemic and cavernosal molecular alterations in the progression of diabetic erectile dysfunction

Background Erectile dysfunction (ED) is a prevalent complication of diabetes, and oxidative stress is an important feature of diabetic ED. Oxidative stress-induced damage plays a pivotal role in the development of tissue alterations. However, the deleterious effects of oxidative stress in the corpus cavernosum with the progression of diabetes remain unclear. The aim of this study was to evaluate systemic and penile oxidative stress status in the early and late stages of diabetes. Methods Male Wistar streptozotocin-diabetic rats (and age-matched controls) were examined 2 (early) and 8 weeks (late) after the induction of diabetes. Systemic oxidative stress was evaluated by urinary H2O2 and the ratio of circulating reduced/oxidized glutathione (GSH/GSSG). Penile oxidative status was assessed by H2O2 production and 3-nitrotyrosine (3-NT) formation. Cavernosal endothelial nitric oxide synthase (eNOS) was analyzed by quantitative immunohistochemistry. Dual immunofluorescence was also performed for 3-NT and α-smooth muscle actin (α-SMA) and eNOS–α-SMA. Results There was a significant increase in urinary H2O2 levels in both diabetic groups. The plasma GSH/GSSG ratio was significantly augmented in late diabetes. In cavernosal tissue, H2O2 production was significantly increased in late diabetes. Reactivity for 3-NT was located predominantly in cavernosal smooth muscle (SM) and was significantly reduced in late diabetes. Quantitative immunohistochemistry revealed a significant decrease in eNOS levels in cavernosal SM and endothelium in late diabetes. Conclusions The findings indicate that the noxious effects of oxidative stress are more prominent in late diabetes. Increased penile protein oxidative modifications and decreased eNOS expression may be responsible for structural and/or functional deregulation, contributing to the progression of diabetes-associated ED.