Impact of microwave-assisted extraction on roasted coffee carbohydrates, caffeine, chlorogenic acids and coloured compounds

Supplementary data to this article can be found online at https://doi.org/10.1016/j.foodres.2019.108864.Microwave-assisted extraction (MAE) allows to quickly achieve soluble compounds from solid matrices due to the promotion of temperatures higher than the solvent (atmospheric) boiling point, once a closed-vessel system is used for operating at high pressure. In this study, the feasibility of MAE for producing high yield coffee extracts with properties that allow their commercial application was tested through a quality by design approach. It was studied the influence of time of extraction (1, 5.5, 10 min), temperature (120, 150, 180 °C) and the mass-to-volume (m/V) ratio (2, 4, 6 g/60 mL) in the overall extraction yield (2447%, w/w), carbohydrates content (1843%, w/w), sugars composition, caffeine (47%, w/w), 5-caffeoylquinic acid (12%, w/w), colour and antioxidant activity of the extracts. FTIR analysis was used to study the resemblance of coffee extracts and commercial instant coffee. MAE allowed overall extraction yields considerably higher than the home brewing methods, mainly when performed at 180 °C, with a substantial increase in arabinogalactans (AG) extraction associated to higher temperatures. Temperature exerted a crucial role in coffee extracts differentiation, although time and m/V ratio also lead to different values in the responses. Under a circular economy concept, MAE was able to produce extracts that can be used as defined food/brew ingredients and provides a galactomannan and cellulose rich residue that can also be valued as a source of dietary fibre.Thanks are due to the University of Aveiro and FCT/MCT for the financial support for the QOPNA research Unit (FCT UID/QUI/00062/2019) through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement, and to the Portuguese NMR Network. Guido R. Lopes was supported by individual doc grant (SFRH/BD/104855/2014) by FCT. Cláudia P. Passos contract (CEECIND/01873/2017) was supported by FCT. This work was also funded by national funds (OE), through FCT, in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19.info:eu-repo/semantics/publishedVersio

Modulation of infusion processes to obtain coffee-derived food ingredients with distinct composition

Coffee infusion experiments were conducted to infer how operational variables (time, temperature, mass to volume ratio, and grinding) might affect the efficiency and/or selectivity of compounds extraction. Although the different variables have extensively been reported independently, to the best of our knowledge, no experimental design was yet delineated to study the simultaneous effect of variables in coffee composition. This study fulfills this gap by constructing surface models that reflect the responses in a wide-ranging design space. The freeze-dried extracts were compared regarding the overall yield of extraction, carbohydrate content and composition, caffeine, chlorogenic acid (5-CQA) content, color, and viscosity. Temperature was the major factor for coffee extracts differentiation, regarding both overall and carbohydrates yield and composition. The extraction process efficiency is more related to galactomannans extraction than arabinogalactans. Varying operational conditions, coffee extracts with distinct chemical properties are obtained from the same roasted coffee, broadening their applications in food formulations.Thanks are due to the University of Aveiro and FCT/MCT for the financial support for the QOPNA research Unit (FCT UID/QUI/00062/2019) through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement, and to the Portuguese NMR Network. Guido R. Lopes and Cláudia P. Passos were supported by individual doc (SFRH/BD/104855/2014) and post-doc (SFRH/BDP/107881/2015) grants by FCT, respectiveinfo:eu-repo/semantics/publishedVersio

Comparison of chromosomal and array-based comparative genomic hybridization for the detection of genomic imbalances in primary prostate carcinomas

BACKGROUND: In order to gain new insights into the molecular mechanisms involved in prostate cancer, we performed array-based comparative genomic hybridization (aCGH) on a series of 46 primary prostate carcinomas using a 1 Mbp whole-genome coverage platform. As chromosomal comparative genomic hybridization (cCGH) data was available for these samples, we compared the sensitivity and overall concordance of the two methodologies, and used the combined information to infer the best of three different aCGH scoring approaches. RESULTS: Our data demonstrate that the reliability of aCGH in the analysis of primary prostate carcinomas depends to some extent on the scoring approach used, with the breakpoint estimation method being the most sensitive and reliable. The pattern of copy number changes detected by aCGH was concordant with that of cCGH, but the higher resolution technique detected 2.7 times more aberrations and 15.2% more carcinomas with genomic imbalances. We additionally show that several aberrations were consistently overlooked using cCGH, such as small deletions at 5q, 6q, 12p, and 17p. The latter were validated by fluorescence in situ hybridization targeting TP53, although only one carcinoma harbored a point mutation in this gene. Strikingly, homozygous deletions at 10q23.31, encompassing the PTEN locus, were seen in 58% of the cases with 10q loss. CONCLUSION: We conclude that aCGH can significantly improve the detection of genomic aberrations in cancer cells as compared to previously established whole-genome methodologies, although contamination with normal cells may influence the sensitivity and specificity of some scoring approaches. Our work delineated recurrent copy number changes and revealed novel amplified loci and frequent homozygous deletions in primary prostate carcinomas, which may guide future work aimed at identifying the relevant target genes. In particular, biallelic loss seems to be a frequent mechanism of inactivation of the PTEN gene in prostate carcinogenesis

Carbohydrates as targeting compounds to produce infusions resembling espresso coffee brews using quality by design approach

All coffee brews are prepared with roasted coffee and water, giving origin to espresso, instant, or filtered coffee, exhibiting distinct physicochemical properties, depending on the extraction conditions. The different relative content of compounds in the brews modulates coffee body, aroma, and colour. In this study it was hypothesized that a coffee infusion allows to obtain extracts that resemble espresso coffee (EC) physicochemical properties. Carbohydrates (content and composition) were the target compounds as they are organoleptically important for EC due to their association to foam stability and viscosity. The freeze-drying of the extracts allowed better dissolution properties than spray-drying. Instant coffee powders were obtained with chemical overall composition resembling espresso, although with lower lipids content. The extracts were able to produce the characteristic foam through CO2 injection or salts addition. Their redissolution at espresso concentration allowed a viscosity, foamability and volatile profile representative of an espresso coffee, opening new exploitation possibilities.Thanks are due to the University of Aveiro and FCT/MCT for the financial support for the QOPNA research Unit (FCT UID/QUI/00062/2019) and the LAQV-REQUIMTE (UIDB/50006/2020) through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement. The authors thank FCT co-financed by Programa Operacional Competitividade e Internacionalização, Portugal 2020 and União Europeia by the FEDER (FCT - Compete2020 - Portugal 2020 – FEDER/EU) N° POCI-01-0145-FEDER-029560, project “PulManCar”. Guido R. Lopes was supported by an individual doctoral grant by FCT (SFRH/BD/104855/2014). Sílvia Petronilho (SFRH/BPD/117213/2016) and Cláudia P. Passos (CEECIND/00813/2017) also thanked FCT for the Post-doc grant and the assistant research contract, respectively. The authors thank Prof. Margarida Almeida for the use of spray-drying facilities and Ana Bastos for the help in the equipment operation.info:eu-repo/semantics/publishedVersio

The initial journey of patients with metastatic pancreatic cancer (PaCTO project): a nationwide survey among Portuguese specialist physicians

This work was supported by AstraZeneca. The funders had no role in study design, data collection, and analysis nor in the preparation of the manuscript.ABSTRACT - Introduction: We aimed to characterize the initial healthcare journey of metastatic pancreatic ductal adenocarcinoma (mPDAC) patients in Portugal, including healthcare provision and factors affecting therapeutic decisions, namely BRCA mutation testing. Methods: This is a descriptive cross-sectional, web-based survey using a convenience sampling approach. Portuguese oncologists and pathologists who routinely work with mPDAC patients from different geographical regions and settings were invited to participate in the study via email (December 2020). Descriptive statistical analyses were performed, with categorical variables reported as absolute and relative frequencies, and continuous variables with non-normal distribution as median and interquartile range (IQR) (Stata v.15.0). Results: Seventy physicians participated in the study (43 oncologists, 27 pathologists). According to the responses, a median of 28 patients per center (IQR 12–70) was diagnosed with PDAC in the previous year; 22 of them were referred (IQR 8–70) to mPDAC. The pointed median time from patients’ first hospital admission until disease diagnosis/staging is between 2 and 4 weeks. Endoscopic ultrasound with fine-needle biopsy is available in most hospitals (86%). Around 50% of physicians request BRCA testing; the assessment of additional biomarkers besides BRCA is requested by 40% of professionals. Half of them stated that BRCA testing should be requested earlier–upon histological diagnosis, especially because the median time for results is of 4.0 weeks (IQR 4–8). PARP inhibitors such as olaparib, when available, would be the therapy of choice for most oncologists (71%) if no disease progression occurs after 4 months. Treatment selection is usually grounded on clinical criteria (e.g., performance status, liver function). Around 45% of patients use FOLFIRINOX/mFOLFIRINOX as the first-line therapy. Gemcitabine + nab-paclitaxel is used by 35% of patients as the second-line therapy. Conclusions: Physicians in Portugal support the increasing role of patient-tailored treatments in mPDAC, whose selection should be grounded on tumoral subtyping and molecular profiling. Further efforts to develop multidisciplinary teams, standardize clinical practice, and optimize the implementation of new target therapies are needed.RESUMO - Introdução: Este estudo teve como objetivo caracterizar o percurso inicial dos doentes com adenocarcinoma ductal pancreático metastático (ACDPm) em Portugal, incluindo a prestação de cuidados de saúde e determinação de fatores que afetam as decisões terapêuticas, nomeadamente o teste de mutações BRCA.Métodos: Trata-se de um estudo descritivo transversal (web-based) usando uma abordagem de amostragem por conveniência. Médicos oncologistas e anatomopatologistas portugueses dedicados ao ACDPm e de diferentes regiões geográficas e instituições foram convidados a participar do estudo por email (Dez-2020). Foram realizadas análises estatísticas descritivas, com variáveis categóricas relatadas como frequências absolutas e relativas, e variáveis contínuas com distribuição não-normal como mediana e intervalo interquartil (IIQ) (Stata v.15.0).Resultados: Setenta médicos participaram do estudo (43 oncologistas, 27 patologistas). De acordo com as respostas, uma mediana de 28 doentes por centro (IIQ 12–70) foi diagnosticada com ACDP no ano anterior; 22 deles (IIQ 8–70) referentes a ACDPm. O tempo médio desde a primeira admissão hospitalar dos doentes até o diagnóstico/estadiamento da doença foi entre 2–4 semanas. A ultrassonografia endoscópica com biópsia por agulha fina é realizada pela maioria dos hospitais (86%). Aproximadamente 50% dos médicos referem solicitar o teste BRCA; a avaliação de biomarcadores adicionais além do BRCA é solicitada por 40% dos profissionais. Metade dos médicos assume que o teste BRCA deveria ser solicitado mais precocemente – durante o diagnóstico histológico, principalmente porque o tempo médio para obtenção do resultado é de 4,0 semanas (IIQ 4–8). Os inibidores PARP, como o olaparibe, quando disponíveis, seriam a terapia de escolha para a maioria dos oncologistas (71%) caso não haja progressão da doença após quatro meses. A seleção dos tratamentos é usualmente baseada em critérios clínicos (por exemplo, performance status, função hepática). Em cerca de 45% dos doentes é utilizado FOLFIRINOX/mFOLFIRINOX como terapia de primeira linha. Um esquema com Gemcitabina + nab-paclitaxel é usado em 35% dos doentes como terapia de segunda linha.Conclusões: Os médicos em Portugal apoiam o papel crescente do tratamento individualizado no ACDPm, cuja seleção deve ser baseada na subtipagem tumoral e no perfil molecular. São necessários mais esforços para capacitar as equipas multidisciplinares, desenvolver práticas clínicas padronizadas e otimizar a implementação de novas terapias-alvo.info:eu-repo/semantics/publishedVersio

Distinção entre incidências numa mamografia

O cancro de mama é o mais comum nas mulheres sendo a segunda principal causa de morte por cancro entre as mesmas. A mamografia tem mostrado ser uma ferramenta eficaz na deteção do cancro da mama em fases precoces e consequentemente mais facilmente tratável, continuando assim a ser a principal modalidade de imagiologia usada para o diagnosticar. O diagnóstico assistido por computador (CAD) tem sido visto como primordial para a deteçãoprecoce de regiões suspeitas em mamogramas permitindo, assim, reduzir as taxas de morte. Este artigo consiste na utilização das características de imagens mamográficas por um algoritmo computacional capaz de fazer a distinção entre as diferentes incidências possíveis: mama esquerda/direita e projeções Crânio-caudal/Médio-lateral Oblíqua. Deste modo, o algoritmo desenvolvido contribui para a automatização da fase de identificação mamária em sistemas de diagnóstico assistido por computador

Sustainable exopolysaccharide production by rhizobium viscosum CECT908 using corn steep liquor and sugarcane molasses as sole substrates

Microbial exopolysaccharides (EPS) are promising alternatives to synthetic polymers in a variety of applications. Their high production costs, however, limit their use despite their outstanding properties. The use of low-cost substrates such as agro-industrial wastes in their production, can help to boost their market competitiveness. In this work, an alternative low-cost culture medium (CSLM) was developed for EPS production by Rhizobium viscosum CECT908, containing sugarcane molasses (60 g/L) and corn steep liquor (10 mL/L) as sole ingredients. This medium allowed the production of 6.1 ± 0.2 g EPS/L, twice the amount produced in the standard medium (Syn), whose main ingredients were glucose and yeast extract. This is the first report of EPS production by R.viscosum using agro-industrial residues as sole substrates. EPSCSLM and EPSSyn exhibited a similar carbohydrate composition, mainly 4-linked galactose, glucose and mannuronic acid. Although both EPS showed a good fit to the HerschelBulkley model, EPSCSLM displayed a higher yield stress and flow consistency index when compared with EPSSyn, due to its higher apparent viscosity. EPSCSLM demonstrated its potential use in Microbial Enhanced Oil Recovery by enabling the recovery of nearly 50% of the trapped oil in sand-pack column experiments using a heavy crude oil.info:eu-repo/semantics/publishedVersio

Guiding stem cell tenogenesis by modulation of growth factor signaling and cell-scale biophysical cues in bioengineered constructs

Tendon injuries and tendinopathies are increasingly prevalent health problems currently lacking effective treatments. Tissue engineering offers promising strategies to boost the low innate regenerative ability of tendons. Within this context, the simultaneous leveraging of both physical and biochemical cues by engineered scaffolding systems can be explored to promote a stronger tenogenic response from stem cells. Here, molecularly imprinted polymeric nanoparticles (MINPs) against transforming growth factor (TGF)-β3 are combined with bioinspired anisotropic hydrogels to produce tenogenesis-inductive constructs. MINPs are first solid phase-imprinted against a TGF-β3 epitope, achieving an affinity comparable to monoclonal antibodies. MINPs and magnetically-responsive microfibers are then encapsulated together with adipose-derived stem cells within gelatin-based hydrogels, applying a magnetostatic field during gelation to align the microfibers. The created anisotropic microstructure guides cell growth and elongation unidirectionally, while MINPs act as artificial receptors for TGF-β3, potentiating its paracrine action in the cellular microenvironment. The combination of both stimuli proves effective at increasing TGF-β signaling, which promotes the expression of tendon-associated genes and corresponding protein synthesis, suggesting that microstructural cues and biomolecule sequestration act in tandem to direct cell fate commitment. Overall, this system recapitulates several elements of tendon development, constituting a promising strategy for the regeneration of this tissue

Combined germline and tumor mutation signature testing identifies new families with NTHL1 tumor syndrome

NTHL1 tumor syndrome is an autosomal recessive rare disease caused by biallelic inactivating variants in the NTHL1 gene and which presents a broad tumor spectrum. To contribute to the characterization of the phenotype of this syndrome, we studied 467 index patients by KASP assay or next-generation sequencing, including 228 patients with colorectal polyposis and 239 patients with familial/personal history of multiple tumors (excluding multiple breast/ovarian/polyposis). Three NTHL1 tumor syndrome families were identified in the group of patients with polyposis and none in patients with familial/personal history of multiple tumors. Altogether, we identified nine affected patients with polyposis (two of them diagnosed after initiating colorectal cancer surveillance) with biallelic pathogenic or likely pathogenic NTHL1 variants, as well as two index patients with one pathogenic or likely pathogenic NTHL1 variant in concomitance with a missense variant of uncertain significance. Here we identified a novel inframe deletion classified as likely pathogenic using the ACMG criteria, supported also by tumor mutational signature analysis. Our findings indicate that the NTHL1 tumor syndrome is a multi-tumor syndrome strongly associated with polyposis and not with multiple tumors without polyposis.info:eu-repo/semantics/publishedVersio

A novel spliced fusion of MLL with CT45A2 in a pediatric biphenotypic acute leukemia

Background: Abnormalities of 11q23 involving the MLL gene are found in approximately 10% of human leukemias. To date, nearly 100 different chromosome bands have been described in rearrangements involving 11q23 and 64 fusion genes have been cloned and characterized at the molecular level. In this work we present the identification of a novel MLL fusion partner in a pediatric patient with de novo biphenotypic acute leukemia. Methods: Cytogenetics, fluorescence in situ hybridization (FISH), molecular studies (RT-PCR and LDI-PCR), and bioinformatic sequence analysis were used to characterize the CT45A2 gene as novel MLL fusion partner in pediatric acute leukemia. Results: Fluorescence in situ hybridization of the patient G-banded metaphases demonstrated a cryptic insertion of 11q23 in Xq26.3 involving the MLL gene. Breakpoint fusion analysis revealed that a DNA fragment of 653 kb from 11q23, containing MLL exons 1-9 in addition to 16 other 11q23 genes, was inserted into the upstream region of the CT45A2 gene located at Xq26.3. In addition, a deletion at Xq26.3 encompassing the 3' region of the DDX26B gene (exons 9-16) and the entire CT45A1 gene was identified. RNA analysis revealed the presence of a novel MLL-CT45A2 fusion transcript in which the first 9 exons of the MLL gene were fused in-frame to exon 2 of the CT45A2 gene, resulting in a spliced MLL fusion transcript with an intact open reading frame. The resulting chimeric transcript predicts a fusion protein where the N-terminus of MLL is fused to the entire open reading frame of CT45A2. Finally, we demonstrate that all breakpoint regions are rich in long repetitive motifs, namely LINE/L1 and SINE/Alu sequences, but all breakpoints were exclusively identified outside these repetitive DNA sequences. Conclusion: We have identified CT45A2 as a novel spliced MLL fusion partner in a pediatric patient with de novo biphenotypic acute leukemia, as a result of a cryptic insertion of 11q23 in Xq26.3. Since CT45A2 is the first Cancer/Testis antigen family gene found fused with MLL in acute leukemia, future studies addressing its biologic relevance for leukemogenesis are warranted