The wine sector is one of the most economically important agro-food businesses. The wine market value is largely associated to terroir, in some cases resulting in highly expensive wines that attract ...fraudulent practices. The existent wine traceability system has some limitations that can be overcome with the development of new technological approaches that can tackle this problem with several means. This review aims to call attention to the problem and to present several strategies that can assure a more reliable and authentic wine system, identifying existent technologies developed for the sector, which can be incorporated into the current traceability system.
Since the last decade of the last century, it is known that many old grapevine varieties are descendants of other varieties through natural crossing. Portugal has an important program for the ...conservation of representative samples of intra-varietal variability of all autochthonous varieties, managed by the Portuguese Association for Grapevine Diversity (PORVID), which makes looking for genotypes with dubious identification an important activity from a perspective of its valorisation. This communication presents the results of the molecular analysis of 5,000 samples (accessions) from the PORVID’s collection, using nine microsatellite
loci
currently recommended by the International Organization of Vine and Wine (OIV) for genetic grapevine identification. The results obtained confirmed the molecular identity of 4,220 samples corresponding to 214 varieties present in the official list of Portuguese varieties. In 780 samples, 95 profiles with a plural number of accessions revealed not to be listed in the Vitis International Variety Catalogue (VIVC) database, corresponding to possible varieties either descendent from natural crossing from at least one known parental variety, or from undetermined origin. Furthermore, the need for a comprehensive strategy aimed at uncovering other hidden varieties is discussed to prevent their imminent loss, deepen understanding of their origin, and add economic value and sustainability to the vine and wine sector.
The accurate varietal identification is an essential requirement for every process involved in the exploitation of grapevine resources and derived products. The advancements achieved during the last ...years allowed the simultaneous analysis of multiple molecular markers capable of identifying grapevine varieties. Despite the establishment of a recommended set of nine microsatellite (SSR) markers for this purpose, their effective application with DNA extracted from must and wine samples remains a challenging task. This work aimed to develop High Resolution Melting (HRM) assays based on SSR markers applicable for grapevine varietal identification using leaf, must and wine samples. The grapevine varieties used were Cabernet Sauvignon, Touriga Franca, Touriga Nacional and Rufete. A total of 12 SSR markers were used to screen the varieties: nine markers recommended by the OIV (VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VVS2, VrZAG62 and VrZAG79) and three markers selected based on their repeat motif and length (VvIv35, VChr5c and VChr9a). The results from multiplex PCR amplification of DNA from wine samples revealed that these three markers performed better than the nine established SSR markers. HRM assays were developed targeting markers VvIv35, VChr5c and VChr9a, successfully discriminating the varietal composition in must DNA samples. Promising results were obtained using wine DNA, where assay HRM-VChr9a proved to have the highest discriminant power. The HRM-SSR assays need to be applied in a larger number of varieties, to explore its suitability for grapevine fingerprinting applications throughout the wine chain. Overall, the proposed small SSR makers can be more suitable for wine DNA analysis. The HRM-SSR approach presented here provides fast results, allowing the complete discrimination of varietal composition in must DNA. It also shows to be a promising tool to discriminate the varieties using wine DNA, a task usually hampered by the inherent complexity of wine samples.
•PCR amplification of SSR markers is widely used for grapevine identification.•Must and wine DNA degradation restricts the application of SSR markers.•Small allele size SSRs are amplified in leaf, must and wine DNA from 4 varieties.•Amplification success of small SSRs is compared to recommended OIV SSR markers.•HRM assays targeting three SSRs suitable for varietal discrimination are optimized.
The increasing incidence of fraud demands for new robust methods to assess food authenticity. Over the past decades, biosensors have emerged as practical testing devices with exponential growth in ...diverse research fields. Their numerous advantages have contributed to their implementation in the food sector, with applications ranging from the identification of pathogens, chemical compounds and allergens to spoilage detection and Genetically Modified Organisms (GMO) identification in various food products.
This review explores the development of DNA-based biosensors for food authenticity assessment with a focus on species identification. The role and versatility of nucleic acids as analytes and biorecognition elements are discussed, and the available conventional methods are presented. The main transducing principles involved in biosensing, and the use of nanomaterials are briefly introduced. The application of various DNA-based biosensors over the last decade is shown, highlighting the main innovations and how these have contributed to the improvement of their performance. The final section addresses how different technologies can influence biosensor manufacturing and optimization, so these can become established rapid on-site testing devices used to assess food authenticity.
In the food sector, little research has been carried out in food authenticity regarding biosensors. The development of these devices is mainly aimed at species identification in meat and derived products, although other equally relevant products should be targeted. Several recent technological advances have been successfully integrated into biosensors and must be further explored to promote the establishment of these devices in food authenticity assessment.
•DNA biosensors for food authenticity focused on species identification are explored.•The potential of DNA as biorecognition element and analyte is discussed.•Different biosensors are presented to show progress from the last decade.•New technological advances are being implemented into improved DNA biosensors.•DNA biosensors to authenticate high value food items are currently lacking.
Lung cancer is the main malignant cancer reported worldwide, with one of the lowest survival rates. Deletions in the Epidermal Growth Factor Receptor (EGFR) gene are often associated with non-small ...cell lung cancer (NSCLC), a common subtype of lung cancer. The detection of such mutations provides key information for the diagnosis and treatment of the disease; therefore, the early screening of such biomarkers is of vital importance. The need for fast, reliable, and early detection means applied to NSCLC has led to the development of highly sensitive devices that can detect cancer-associated mutations. Such devices, known as biosensors, are a promising alternative to more conventional detection methods and can potentially alter the way cancer is diagnosed and treated. In this study, we report the development of a DNA-based biosensor, namely a quartz crystal microbalance (QCM), applied to the detection of NSCLC, from liquid biopsies samples. The detection, as is the case of most DNA biosensors, is based on the hybridization between the NSCLC-specific probe and the sample DNA (containing specific mutations associated with NSCLC). The surface functionalization was performed with a blocking agent (dithiothreitol) and thiolated-ssDNA strands. The biosensor was able to detect specific DNA sequences in both synthetic and real samples. Aspects such as reutilization and regeneration of the QCM electrode were also studied.
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•DNA based QCM biosensor applied to Non-Small Cell Lung Cancer.•Detection from cfDNA present in plasma.•Detection of EGFR deletion in exon 19.
•HRM can be used as an alternative approach to SSR genotyping.•A new somatic variant derived from Cerceal Branco was identified.•A new variety unidentified in the VIVC database was found.
Precise ...identification is crucial to study and for the valorisation of ancient grapevine varieties, with many synonyms and misidentifications hampering this task. SSR-Multiplex PCR and SNP genotyping combined with HRM were used, to identify 19 varieties present in an ancient Portuguese vineyard. The combination of the methodologies allowed the identification of 18 grapevines, from which 10 were correctly identified by ampelography and the remaining were misidentified. SSR and SNP data retrieved for one sample did not match any profile publicly available, however pedigree analysis suggests it is a new variety. One sample identified as Cerceal Branco presented an unexpected red berry. A sample identified as Malvasia Preta/Preta Roxa also presented a berry colour non coincident with its molecular identification. This suggests they might represent new somatic variants of existent varieties. Three HRM assays were performed to test its robustness in comparison to SSR/SNP results. Molecular identification was successful, revealing possible new variants. HRM revealed to be useful in identifying varieties by melting curve profile comparison against reference material. The results provide a better knowledge of a valuable pool of genetic patrimony: ancient varieties that represent a piece of history and heritage from an important winegrowing region.
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Olive oil and wine production have a worldwide economic impact. Their market reliability is under great concern because of the increasing number of fraud and adulteration attempts. The need for a ...traceability system in all its extension is crucial particularly for the cases of olive oils and wines with certified labels, in which only a limited number of olives and grapevine varieties, respectively, are allowed in a restricted well-defined geographical area. Molecular markers have been vastly applied to the food sector, and in particular High-Resolution DNA Melting technology has been successfully applied for olive oil and wine authentication, as part of the traceability system. In this review, the applications of HRM and their usefulness for this sector considering, Safety, Security and Authenticity will be reviewed. A broad overview of the HRM technique will be presented, focusing on the aspects that are crucial for its success, in particular the new generation of fluorescent dsDNA dyes used for amplicon detection and quantification, and the data analysis. A brief outlook on the olive oil and wine authenticity procedures, based on new DNA technology advances, and in which way this may influence the future establishment of a traceability system will be discussed.
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•High Resolution Melting DNA as a technique for olive oil and wine authenticity•The HRM performance with saturating and non-saturating dyes are summarized.•Methods of extracting data information and their limitations
•Label-free DNA-based biosensor applied to authenticity.•Varietal identification in different matrices (leaf, must and wine samples)•PCR-free detection.
The diversity found among the Vitis vinifera ...L. species allows the production of wines with very different characteristics. The development of platforms suitable for food composition analysis is currently an emerging area. Among these, DNA biosensors have been developed for a wide variety of applications, ranging from food safety to authenticity. The main aim of this work was to study the detection capacity of the DNA-based optical biosensor using different V. vinifera matrices (leaf, must and wine). Genomic DNA was extracted from leaf, must and wine of three V. vinifera varieties and was tested on the long-period grating (LPG) DNA-based biosensor developed within our group. The biosensor was able to distinguish the varieties even using DNA extracted from complex matrices, revealing its potential to be applied in wine authenticity.
The wine sector requires quick and reliable methods for Vitis vinifera L. varietal identification. The number of V. vinifera varieties is estimated in about 5,000 worldwide. Single Nucleotide ...Polymorphisms (SNPs) represent the most basic and abundant form of genetic sequence variation, being adequate for varietal discrimination. The aim of this work was to develop DNA-based assays suitable to detect SNP variation in V. vinifera, allowing varietal discrimination. Genotyping by sequencing allowed the detection of eleven SNPs on two genes of the anthocyanin pathway, the flavanone 3-hydroxylase (F3H, EC: 1.14.11.9), and the leucoanthocyanidin dioxygenase (LDOX, EC 1.14.11.19; synonym anthocyanidin synthase, ANS) in twenty V. vinifera varieties. Three High Resolution Melting (HRM) assays were designed based on the sequencing information, discriminating five of the 20 varieties: Alicante Bouschet, Donzelinho Tinto, Merlot, Moscatel Galego and Tinta Roriz. Sanger sequencing of the HRM assay products confirmed the HRM profiles. Three probes, with different lengths and sequences, were used as bio-recognition elements in an optical biosensor platform based on a long period grating (LPG) fiber optic sensor. The label free platform detected a difference of a single SNP using genomic DNA samples. The two different platforms were successfully applied for grapevine varietal identification.
Leveling Up Hydrogels: Hybrid Systems in Tissue Engineering Neves, Sara C.; Moroni, Lorenzo; Barrias, Cristina C. ...
Trends in biotechnology (Regular ed.),
March 2020, 2020-Mar, 2020-03-00, 20200301, Letnik:
38, Številka:
3
Journal Article
Recenzirano
Odprti dostop
Hydrogels can mimic several features of the cell native microenvironment and have been widely used as synthetic extracellular matrices (ECMs) in tissue engineering and regenerative medicine (TERM). ...However, some applications have specifications that hydrogels cannot efficiently fulfill on their own. Incorporating reinforcing structures like fibrous scaffolds or particles into hydrogels, as hybrid systems, is a promising strategy to improve their functionality. We describe recent advances in the fabrication and application of these hybrid systems, where structural properties and stimuli responsiveness of hydrogels are enhanced while their ECM-like features are preserved. Furthermore, we discuss how these systems can contribute to the development of more complex tissue engineered structures in the rapidly evolving field of TERM.
Native tissue mimicry is still a major challenge in TERM. Extracellular matrix (ECM) analogs should ideally provide biologically relevant biochemical/biophysical cues, in an adequate spatial-temporal manner, to properly stimulate cultured and/or recruited cells towards functional tissue formation.Hydrogels are the most attractive ECM analogs, but it is difficult to tune them to mimic most of the native tissue-specific features alone without concomitantly hindering key biological processes. One promising strategy to overcome this relies on the use of hybrid systems, combining hydrogels with other structures.Providing structural/mechanical reinforcement, spatial patterning/guidance, and/or stimuli responsiveness to hydrogel-based systems emerged as effective methodologies to more closely replicate the plethora of features and signals that native ECM presents to cells.
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