Lara Henriques

Fundação Oswaldo Cruz

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Raquel Seruca

Universidade do Porto

Gianpaolo Suriano

Universidade do Porto

Fátima Carneiro

i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto

Ana Rita Soares Mateus

Universidade do Porto

Marc Mareel

Ghent University Hospital

John C. Atherton

University of Nottingham

Patrícia Baptista

Fundação Oswaldo Cruz

Andreia Vasconcelos

Science Oxford

ÂM

Ângela M. Costa

i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto

Maria José Oliveira

i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto

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Universidade do Porto

i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto

IPO Porto

Fundação Oswaldo Cruz

University of Minho

Hospital de São João

Instituto Português de Oncologia Francisco Gentil

University of Coimbra

Instituto Nacional de Engenharia Biomédica

Institute of Mechanical Engineering and Industrial Mangement

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Evaluation of molecular assays for the detection of respiratory viruses with a view to structuring the national Wastewater-based epidemiology program

Lara Henriques Renan Faustino Pedro Henrique Cardoso Marilda Mendonça Siqueira Mônica Arruda

Introduction:Along COVID-19 pandemic, several countries considered wastewater based epidemiology (WBE) as an additional surveillance approach.WBE works out as an early alert system, as viral concentration in wastewater precedes the increase in incidence in about 1-2 weeks.As a populational approach, it is not limited to clinical cases.Due to diversity in concentration methods that preclude detection, qRT-PCR targets, sample complexity and resulting bias in comparative outcomes, standardization is still a global challenge.Objectives: Aligned with the Health Emergency Department, this study aims the development and standardization of laboratory assay for viral detection to support the establishment of a WBE program for outbreaks and health emergencies.Methodology: Two main protocols were compared: (P1) viral concentration with electronegative membrane filtration (100mL) followed by direct manual extraction and (P2) concentration using magnetic nanobeads (10mL) followed by automated extraction.Virus like particle (VLP) containing SARS-CoV-2 (SC2), Influenza A and B (FLUA; FLUB), Syncytial Respiratory Virus (RSV) were inoculated into WW samples and negative control (0.1 to 0.5uL/mL).SC-2 quantification was determined based on a standard curve.Moreover, sets of primers/probes (Bio-manguinhos) for detection and quantification of respiratory viruses by qRT-PCR were optimized for WW samples. Results:No inhibition of the primers/probes set was observed in the 8 samples tested with VPL, even in low concentrations.In polluted river samples (N=4), SC-2 concentration in P2 (10.8gc/uL ± 5.8) was comparable to P1 (4.4gc/uL ± 4.3) despite using a 10x smaller volume.In sewage samples (N=13) there was no significant difference between P1 (5.08gc/uL ± 6.7) and P2 (2.5 ± 2.4).Although, we identified the presence of FLU-A (3) and RSV (3).The P2 was the easiest handled and fastest one; For 20 samples using P2, 2hrs were necessary in comparison with 6.5h for P1. Conclusion:The P2 protocol, when associated with Bio-manguinhos optimized RT-PCR assay, represents a more useful tool for WBE, reducing the volume of samples and processing time without affecting the sensitivity.This preliminary data shows our ability to optimize the assay with a view to scaling up of a national network.

10.35259/isi.biomang.2024_63906

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Identification of signalling pathways influenced by E-cadherin

Lara Henriques

9 Resumo 11 Abbreviations 13 Introduction 15 Cancer 15 Gastric Cancer 16 Pathology of Gastric Cancer 17 Hereditary Diffuse Gastric Cancer (HDGC) 19 E-Cadherin 20 E-Cadherin Protein 20 E-Cadherin’s Role in Malignant Cells 22 E-Cadherin in Gastric Cancer 23 CDH1 Mutational Status 24 E-cadherin and Cellular Signalling 25 Epithelial-Mesenchymal Transition (EMT) 26 E-cadherin and Matrix Metalloproteinases (MMPs) 27 E-cadherin and Integrins 29 E-cadherin and RTKs (Receptor Tyrosine Kinases) 30 RTKs in gastric cancer 33 E-cadherin and PI3K/AKT 33 Aim 35 Material and Methods 37 Cell Culture and Reagents 37 Preparation of Cell Lysates 37 Immunoprecipitation 37 Immunoblot Analysis 39 Immunocytochemistry 39 Antibodies 39 Zymography 39 Matrigel Invasion Assay 40

CDH1

VE-cadherin

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Helicobacter pylori Induces Gastric Epithelial Cell Invasion in a c-Met and Type IV Secretion System-dependent Manner

Journal of Biological Chemistry (2006)

Maria José Oliveira Ana Catarina Costa Ângela M. Costa Lara Henriques Gianpaolo Suriano

CagA

Pathogenicity island

10.1074/jbc.m607067200

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Protein Matrices for Improved Wound Healing: Elastase Inhibition by a Synthetic Peptide Model

Biomacromolecules (2010)

Andreia Vasconcelos Ana Paula Pêgo Lara Henriques Meriem Lamghari Artur Cavaco‐Paulo

The unique properties of silk fibroin were combined with keratin to develop new wound-dressing materials. Silk fibroin/keratin (SF/K) films were prepared to reduce high levels of elastase found on chronic wounds. This improved biological function was achieved by the incorporation of a small peptide synthesized based on the reactive-site loop of the Bowman−Birk Inhibitor (BBI) protein. In vitro degradation and release were evaluated using porcine pancreatic elastase (PPE) solution as a model of wound exudate. It was found that biological degradation and release rate are highly dependent on film composition. Furthermore, the level of PPE activity can be tuned by changing the film composition, thus showing an innovative way of controlling the elastase−antielastase imbalance found on chronic wounds.

10.1021/bm100537b

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Genetics of Hereditary Diffuse Gastric Cancer: Progress and Future Challenges

Future Oncology (2006)

Gianpaolo Suriano Paulo Michel Pinheiro Ferreira Ana Rita Soares Mateus Joana Simões‐Correia Lara Henriques

Hereditary diffuse gastric cancer (HDGC) is a rare cancer susceptibility syndrome. One third of HDGC syndrome families carry germline mutations of the E-cadherin gene. Owing to the limitation of the current endoscopic screening techniques and since no chemoprevention is yet available, total prophylactic gastrectomy is the only option offered to carriers of inactivating mutations in genetic counseling. In this regard, 30% of the E-cadherin germline mutations reported to date are of the missense type, and since their pathogenic significance is not straightforward, the management of carriers of such mutations is suboptimal. In the absence of definitive clinical evidence, functional in vitro studies together with in silico analysis have been used to infer the pathogenic significance of germline missense mutations. Since most of the HDGC families reported to date are negative for E-cadherin germline mutations, the identification of alternative genes underlying the tumorigenesis of diffuse gastric has become an important target for research.

10.2217/14796694.2.3.363

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Citations (13)