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The use of a low-cost split-ring resonator (SRR) passive sensor for the real-time permittivity characterization of hydrocarbon fluids is proposed in this paper. The characterization of the sensor is performed through both full-wave simulation and measurements. Thanks to the analysis of several crude samples, the possibility of discrimination between different types of crude and the estimation of several of their properties are demonstrated. Between them, the estimation of sulfur, aromatic hydrocarbons, and salt-water concentrations either in normal ambient conditions or in a high-pressure and high-temperature environment can be mentioned. Experiments were run both at normal ambient conditions and pressures up to 970 bar and temperatures up to 200 °C.
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BACKGROUND: Grapevine reproductive development has direct implications on yield. It also impacts on berry and wine quality by affecting traits like seedlessness, berry and bunch size, cluster compactness and berry skin to pulp ratio. Seasonal fluctuations in yield, fruit composition and wine attributes, which are largely driven by climatic factors, are major challenges for worldwide table grape and wine industry. Accordingly, a better understanding of reproductive processes such as gamete development, fertilization, seed and fruit set is of paramount relevance for managing yield and quality. With the aim of providing new insights into this field, we searched for clones with contrasting seed content in two germplasm collections. RESULTS: We identified eight variant pairs that seemingly differ only in seed-related characteristics while showing identical genotype when tested with the GrapeReSeq_Illumina_20K_SNP_chip and several microsatellites. We performed multi-year observations on seed and fruit set deriving from different pollination treatments, with special emphasis on the pair composed by Sangiovese and its seedless variant locally named Corinto Nero. The pollen of Corinto Nero failed to germinate in vitro and gave poor berry set when used to pollinate other varieties. Most berries from both open- and cross-pollinated Corinto Nero inflorescences did not contain seeds. The genetic analysis of seedlings derived from occasional Corinto Nero normal seeds revealed that the few Corinto Nero functional gametes are mostly unreduced. Moreover, three genotypes, including Sangiovese and Corinto Nero, were unexpectedly found to develop fruits without pollen contribution and occasionally showed normal-like seeds. Five missense single nucleotide polymorphisms were identified between Corinto Nero and Sangiovese from transcriptomic data. CONCLUSIONS: Our observations allowed us to attribute a seedlessness type to some variants for which it was not documented in the literature. Interestingly, the VvAGL11 mutation responsible for Sultanina stenospermocarpy was also discovered in a seedless mutant of Gouais Blanc. We suggest that Corinto Nero parthenocarpy is driven by pollen and/or embryo sac defects, and both events likely arise from meiotic anomalies. The single nucleotide polymorphisms identified between Sangiovese and Corinto Nero are suitable for testing as traceability markers for propagated material and as functional candidates for the seedless phenotype.
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Frutas/crescimento & desenvolvimento , Frutas/genética , Reprodução/genética , Sementes/crescimento & desenvolvimento , Sementes/genética , Vitis/crescimento & desenvolvimento , Vitis/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Genes de Plantas , Variação Genética , Genótipo , Hibridização Genética , Polimorfismo de Nucleotídeo ÚnicoRESUMO
RNA can be modified in over 100 distinct ways, and these modifications are critical for function. Pseudouridine synthases catalyse pseudouridylation, one of the most prevalent RNA modifications. Pseudouridine synthase 7 modifies a variety of substrates in Saccharomyces cerevisiae including tRNA, rRNA, snRNA, and mRNA, but the substrates for other budding yeast Pus7 homologues are not known. We used CRISPR-mediated genome editing to disrupt Candida albicans PUS7 and find absence leads to defects in rRNA processing and a decrease in cell surface hydrophobicity. Furthermore, C. albicans Pus7 absence causes temperature sensitivity, defects in filamentation, altered sensitivity to antifungal drugs, and decreased virulence in a wax moth model. In addition, we find C. albicans Pus7 modifies tRNA residues, but does not modify a number of other S. cerevisiae Pus7 substrates. Our data suggests C. albicans Pus7 is important for fungal vigour and may play distinct biological roles than those ascribed to S. cerevisiae Pus7.
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Candida albicans/genética , Transferases Intramoleculares/metabolismo , RNA Fúngico/genética , RNA Ribossômico/genética , Animais , Antifúngicos/farmacologia , Sistemas CRISPR-Cas , Candida albicans/efeitos dos fármacos , Candida albicans/enzimologia , Candidíase/microbiologia , Deleção de Genes , Edição de Genes , Humanos , Interações Hidrofóbicas e Hidrofílicas , Transferases Intramoleculares/genética , Larva/microbiologia , Mariposas/microbiologia , Processamento Pós-Transcricional do RNA , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Especificidade por SubstratoRESUMO
Grapevines (Vitis vinifera L.) form the basis of viticulture, and are susceptible to diseases such as downy mildew (Plasmopara viticola) and powdery mildew (Erysiphe necator). Therefore, successful viticulture programs require the use of pesticides. Breeding for resistance is the only eco-friendly solution. Marker-assisted selection is currently widely used for grapevine breeding. Consequently, traits of interest must be tagged with molecular markers linked to quantitative trait loci (QTL). We herein present our findings regarding genetic mapping and QTL analysis of resistance to downy and powdery mildew diseases in the progenies of the GF.GA-47-42 ('Bacchus' × 'Seyval') × 'Villard blanc' cross. Simple sequence repeats and single nucleotide polymorphisms of 151 individuals were analyzed. A map consisting of 543 loci was screened for QTL analyses based on phenotypic variations observed in plants grown in the field or under controlled conditions. A major QTL for downy mildew resistance was detected on chromosome 18. For powdery mildew resistance, a QTL was identified on chromosome 15. This QTL was replaced by a novel QTL on chromosome 18 in 2003 (abnormally high temperatures) and 2004. Subsequently, both QTLs functioned together. Additionally, variations in the timing of the onset of veraison, which is a crucial step during grape ripening, were studied to identify genomic regions affecting this trait. A major QTL was detected on linkage group 16, which was supplemented by a minor QTL on linkage group 18. This study provides useful information regarding novel QTL-linked markers relevant for the breeding of disease-resistant grapevines adapted to current climatic conditions.
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Resistência à Doença , Proteínas de Plantas/genética , Locos de Características Quantitativas , Vitis/crescimento & desenvolvimento , Mapeamento Cromossômico/métodos , Cromossomos de Plantas/genética , Ligação Genética , Repetições de Microssatélites , Melhoramento Vegetal , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Polimorfismo de Nucleotídeo Único , Vitis/genética , Vitis/microbiologiaRESUMO
Introduction: Introduction: among the groups more affected by the COVID-19 pandemic were patients undergoing chronic hemodialysis (HD) treatment due to their comorbidities, advanced age, impaired innate and adaptive immune function, and increased nutritional risk due to their underlying inflammatory state. All of these factors contribute to a higher risk of severe complications and worse outcomes compared to the general population when infected with SARS-CoV-2. Objective: the objective of this study was to describe the nutritional characteristics of and their potential association with the prognosis of COVID-19 in patients undergoing chronic HD treatment. Method: a descriptive, retrospective, observational design. All cases of COVID-19 in patients undergoing chronic treatment at the Hemodialysis Unit of Hospital de Manises, Valencia, Spain, from the start of the pandemic to before vaccination were included. Results: for that, 189 patients were studied, who received chronic HD treatment in the hospital unit, 22 patients were diagnosed with COVID-19 (12 %) in that period. The mean age was 71 years, 10 were women, the Charlson index was 6.59 points, diabetes mellitus 10, vintage HD 51.6 months, 2 patients had previously received a currently non-functioning kidney transplant, 16 had arteriovenous fistula as vascular access, and 6 had central vascular access. The mean dialysis session time was 220.14 minutes and the initial value of the single dose of the Kt/V pool was 1.7. 16 patients had body composition measurement, a strong association (p < 0.05) was identified between mortality and BMI, as well as mortality and FTI. Furthermore, the differences between deceased and surviving groups in the serum levels of various variables related to nutritional status were analyzed, finding significant differences with p < 0.05 in the value of triglycerides and ferritin. Conclusions: higher body mass index and higher body fat content, along with lower baseline levels of triglycerides and ferritin, were significantly associated with higher COVID-19 mortality in patients on chronic hemodialysis. These findings suggest that the initial nutritional status of these patients can significantly influence the prognosis of SARS-CoV-2 infection.
Introducción: Introducción: entre los grupos más afectados por la pandemia de COVID-19 se encuentran los pacientes en tratamiento crónico de hemodiálisis (HD) por sus comorbilidades, edad avanzada, deterioro de la función inmune innata y adaptativa, y mayor riesgo nutricional por su estado inflamatorio de base. Todos estos factores contribuyen a un mayor riesgo de complicaciones graves y peores resultados en comparación con la población general cuando se infectan con SARS-CoV-2. Objetivo: el objetivo de este estudio es describir las características nutricionales y su potencial asociación con el pronóstico de COVID-19 en pacientes en tratamiento crónico de HD. Método: diseño observacional retrospectivo y descriptivo. Se incluyeron todos los casos de COVID-19 en pacientes en tratamiento crónico en la Unidad de Hemodiálisis del Hospital de Manises, Valencia, desde el inicio de la pandemia hasta antes de la vacunación. Resultados: de 189 pacientes que recibieron tratamiento de HD crónica en la unidad hospitalaria, 22 pacientes fueron diagnosticados con COVID-19 (12 %) en ese período. La edad media fue de 71 años, 10 eran mujeres, índice de Charlson de 6,59 puntos, diabetes mellitus 10, tiempo en diálisis 51,6 meses, 2 pacientes habían recibido previamente un trasplante renal actualmente no funcionante, 16 tenían fístula arteriovenosa como acceso vascular, y 6 tenían acceso vascular central. El tiempo medio de la sesión de diálisis fue de 220,14 minutos y el valor inicial de la dosis única del pool de Kt/V fue de 1,7. Tenían medición de la composición corporal 16 pacientes, se identificó una fuerte asociación (p < 0,05) entre mortalidad e IMC, así como mortalidad y FTI. Además las diferencias entre los grupos de fallecidos y sobrevivientes en los niveles séricos de diversas variables relacionadas con el estado nutricional fueron analizados, encontrando diferencias significativas con p < 0,05 en el valor de triglicéridos y ferritina. Conclusiones: un índice de masa corporal más alto y un mayor contenido de grasa corporal, junto con niveles basales más bajos de triglicéridos y ferritina, se asocian significativamente a una mayor mortalidad por COVID-19 entre los pacientes en hemodiálisis crónica. Estos hallazgos sugieren que el estado nutricional inicial de estos pacientes puede influir significativamente en el pronóstico de la infección por SARS-CoV-2.
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COVID-19 , Estado Nutricional , Diálise Renal , Humanos , COVID-19/terapia , COVID-19/mortalidade , COVID-19/complicações , COVID-19/epidemiologia , Masculino , Feminino , Estudos Retrospectivos , Idoso , Prognóstico , Pessoa de Meia-Idade , Espanha/epidemiologia , Idoso de 80 Anos ou mais , Falência Renal Crônica/terapia , Falência Renal Crônica/complicações , Falência Renal Crônica/mortalidade , Índice de Massa CorporalRESUMO
Introduction: MISTRG mice have been genetically modified to allow development of a human myeloid compartment from engrafted human CD34+ haemopoietic stem cells, making them particularly suited to study the human innate immune system in vivo. Here, we characterized the human neutrophil population in these mice to establish a model that can be used to study the biology and contribution in immune processes of these cells in vivo. Methods and results: We could isolate human bone marrow neutrophils from humanized MISTRG mice and confirmed that all neutrophil maturation stages from promyelocytes (CD11b-CD16-) to end-stage segmented cells (CD11b+CD16+) were present. We documented that these cells possessed normal functional properties, including degranulation, reactive oxygen species production, adhesion, and antibody-dependent cellular cytotoxicity towards antibody-opsonized tumor cells ex vivo. The acquisition of functional capacities positively correlated with the maturation state of the cell. We found that human neutrophils were retained in the bone marrow of humanized MISTRG mice during steady state. However, the mature segmented CD11b+CD16+ human neutrophils were released from the bone marrow in response to two well-established neutrophil-mobilizing agents (i.e., G-CSF and/or CXCR4 antagonist Plerixafor). Moreover, the neutrophil population in the humanized MISTRG mice actively reacted to thioglycolate-induced peritonitis and could infiltrate implanted human tumors, as shown by flow cytometry and fluorescent microscopy. Discussion: These results show that functional human neutrophils are generated and can be studied in vivo using the humanized MISTRG mice, providing a model to study the various functions of neutrophils in inflammation and in tumors.
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Compostos Heterocíclicos , Neutrófilos , Humanos , Camundongos , Animais , Mobilização de Células-Tronco Hematopoéticas , Medula Óssea , ImunidadeRESUMO
The use of non-Saccharomyces yeasts in brewing is a useful tool for developing new products to meet the growing consumer demand for innovative products. Non-Saccharomyces yeasts can be used both in single and in mixed fermentations with Saccharomyces cerevisiae, as they are able to improve the sensory profile of beers, and they can be used to obtain functional beers (with a low ethanol content and melatonin production). The aim of this study was to evaluate this capacity in eight non-Saccharomyces strains isolated from Madrid agriculture. For this purpose, single fermentations were carried out with non-Saccharomyces strains and sequential fermentations with non-Saccharomyces and the commercial strain SafAle S-04. The Wickerhamomyces anomalus strain CLI 1028 was selected in pure culture for brewing beer with a low ethanol content (1.25% (v/v)) for its fruity and phenolic flavours and the absence of wort flavours. The best-evaluated strains in sequential fermentation were CLI 3 (Hanseniaspora vineae) and CLI 457 (Metschnikowia pulcherrima), due to their fruity notes as well as their superior bitterness, body, and balance. Volatile compounds and melatonin production were analysed by GC and HPLC, respectively. The beers were sensory-analysed by a trained panel. The results of the study show the potential of non-Saccharomyces strains in the production of low-alcohol beers, and as a flavour enhancement in sequential fermentation.
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The IFAPA research center "Rancho de la Merced" (Jerez, Spain) hosts one of the oldest and most diverse grapevine germplasm repositories in Europe, and is aimed at providing feasible solutions to deal with any agronomic trait by exploring its genetic variability and by means of association and Deoxyribonucleic Acid (DNA) editing studies. In this work, we focused on a wine and dual-use grapevine subcollection that consists of 930 accessions. Genetic analysis allowed to identify 521 unique genotypes. After comparing them with several databases, matches were found for 476 genetic profiles while the remaining 45 have not been previously described. Combination with clustering analysis suggested a total pool of 481 Vitis vinifera accessions that included some table cultivars. Several synonymies, homonymies and mislabeling have also been detected. Structure analysis allowed identifying six clusters according to eco-geographic cultivation areas and one additional group including non-vinifera accessions. Diversity analysis pointed out that Spanish Mediterranean varieties are genetically closer to oriental genotypes than to European varieties typical of oceanic and continental climates. The origin of Spanish varieties is discussed in depth considering our data and previous studies. Analysis of molecular variance partition confirmed a well-structured germplasm, although differentiation among groups had a much lower effect on genetic variability than differences within groups, which are strongly related to a very high heterozygosity. A core collection that covers all allele richness is proposed. It is constituted of about 13% of total accessions, and each cluster inferred by structure analysis is represented.
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BACKGROUND: Current immunotherapy for patients with high-risk neuroblastoma involves the therapeutic antibody dinutuximab that targets GD2, a ganglioside expressed on the majority of neuroblastoma tumors. Opsonized tumor cells are killed through antibody-dependent cellular cytotoxicity (ADCC), a process mediated by various immune cells, including neutrophils. The capacity of neutrophils to kill dinutuximab-opsonized tumor cells can be further enhanced by granulocyte-macrophage colony-stimulating factor (GM-CSF), which has been shown in the past to improve responses to anti-GD2 immunotherapy. However, access to GM-CSF (sargramostim) is limited outside of Northern America, creating a high clinical need for an alternative method to stimulate dinutuximab responsiveness in the treatment of neuroblastoma. In this in vitro study, we have investigated whether clinically well-established granulocyte colony-stimulating factor (G-CSF) can be a potentially suitable alternative for GM-CSF in the dinutuximab immunotherapy regimen of patients with neuroblastoma. METHODS: We compared the capacity of neutrophils stimulated either in vitro or in vivo with GM-CSF or G-CSF to kill dinutuximab-opsonized GD2-positive neuroblastoma cell lines and primary patient tumor material. Blocking experiments with antibodies inhibiting either respective Fc gamma receptors (FcγR) or neutrophil integrin CD11b/CD18 demonstrated the involvement of these receptors in the process of ADCC. Flow cytometry and live cell microscopy were used to quantify and visualize neutrophil-neuroblastoma interactions. RESULTS: We found that G-CSF was as potent as GM-CSF in enhancing the killing capacity of neutrophils towards neuroblastoma cells. This was observed with in vitro stimulated neutrophils, and with in vivo stimulated neutrophils from both patients with neuroblastoma and healthy donors. Enhanced killing due to GM-CSF or G-CSF stimulation was consistent regardless of dinutuximab concentration, tumor-to-neutrophil ratio and concentration of the stimulating cytokine. Both GM-CSF and G-CSF stimulated neutrophils required FcγRIIa and CD11b/CD18 integrin to perform ADCC, and this was accompanied by trogocytosis of tumor material by neutrophils and tumor cell death in both stimulation conditions. CONCLUSIONS: Our preclinical data support the use of G-CSF as an alternative stimulating cytokine to GM-CSF in the treatment of high-risk neuroblastoma with dinutuximab, warranting further testing of G-CSF in a clinical setting.
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Adjuvantes Imunológicos/farmacologia , Anticorpos Monoclonais/farmacologia , Antineoplásicos Imunológicos/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Citotoxicidade Imunológica/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Neuroblastoma/tratamento farmacológico , Neutrófilos/efeitos dos fármacos , Antígeno CD11b/metabolismo , Antígenos CD18/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Técnicas de Cocultura , Humanos , Neuroblastoma/imunologia , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Neutrófilos/imunologia , Neutrófilos/metabolismo , Neutrófilos/patologia , Receptores de IgG/metabolismo , Trogocitose/efeitos dos fármacos , Microambiente TumoralRESUMO
High-risk neuroblastoma, especially after recurrence, still has a very low survival rate. Immune checkpoint inhibitors targeting T cells have shown remarkable clinical efficacy in adult solid tumors, but their effects in pediatric cancers have been limited so far. On the other hand, targeting myeloid immune checkpoints, such as CD47-SIPRα, provide the opportunity to enhance antitumor effects of myeloid cells, including that of neutrophils, especially in the presence of cancer-opsonizing antibodies. Disialoganglioside (GD2)-expressing neuroblastoma cells targeted with anti-GD2 antibody dinutuximab are in part eradicated by neutrophils, as they recognize and bind the antibody targeted tumor cells through their Fc receptors. Therapeutic targeting of the innate immune checkpoint CD47-SIRPα has been shown to promote the potential of neutrophils as cytotoxic cells in different solid tumor indications using different cancer-targeting antibodies. Here, we demonstrate that the capacity of neutrophils to kill dinutuximab-opsonized neuroblastoma cells is also controlled by the CD47-SIRPα axis and can be further enhanced by antagonizing CD47-SIRPα interactions. In particular, CD47-SIRPa checkpoint inhibition enhanced neutrophil-mediated ADCC of dinutuximab-opsonized adrenergic neuroblastoma cells, whereas mesenchymal neuroblastoma cells may evade immune recognition by a reduction of GD2 expression. These findings provide a rational basis for targeting CD47-SIRPα interactions to potentiate dinutuximab responsiveness in neuroblastomas with adrenergic phenotype.
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Over the last decades, cancer immunotherapies such as checkpoint blockade and adoptive T cell transfer have been a game changer in many aspects and have improved the treatment for various malignancies considerably. Despite the clinical success of harnessing the adaptive immunity to combat the tumor, the benefits of immunotherapy are still limited to a subset of patients and cancer types. In recent years, neutrophils, the most abundant circulating leukocytes, have emerged as promising targets for anti-cancer therapies. Traditionally regarded as the first line of defense against infections, neutrophils are increasingly recognized as critical players during cancer progression. Evidence shows the functional plasticity of neutrophils in the tumor microenvironment, allowing neutrophils to exert either pro-tumor or anti-tumor effects. This review describes the tumor-promoting roles of neutrophils, focusing on their myeloid-derived suppressor cell activity, as well as their role in tumor elimination, exerted mainly via antibody-dependent cellular cytotoxicity. We will discuss potential approaches to therapeutically target neutrophils in cancer. These include strategies in humans to either silence the pro-tumor activity of neutrophils, or to activate or enhance their anti-tumor functions. Redirecting neutrophils seems a promising approach to harness innate immunity to improve treatment for cancer patients.
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Neoplasias/imunologia , Ativação de Neutrófilo , Neutrófilos/imunologia , Imunidade Adaptativa , Animais , Citotoxicidade Celular Dependente de Anticorpos , Divisão Celular/imunologia , Movimento Celular/imunologia , Humanos , Imunidade Inata , Imunoterapia/métodos , Camundongos , Células Supressoras Mieloides/imunologia , Invasividade Neoplásica/imunologia , Metástase Neoplásica/imunologia , Neoplasias/irrigação sanguínea , Neoplasias/terapia , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/terapia , Neovascularização Patológica/imunologia , Microambiente Tumoral/imunologiaRESUMO
The area of the Garda Lake within the Trentino province (north of Italy) is the northernmost part of Europe where the Mediterranean species Olea europaea L. is traditionally cultivated. 'Casaliva' is claimed as the main variety traditionally grown in the Garda Trentino area (GT) from which a world renowned niche extra virgin olive oil is produced. Since a dominant presence of 'Casaliva' would link the fruit set success and yield to a self-pollination compatibility system, a deep genetic survey of the olive tree population in the GT has been performed with the aim of establishing the actual varietal composition and of understanding from which pollen donor the 'Casaliva' olives originate. Forty-four different genetic profiles were observed among the 205 leaf samples collected from 106 ancient trees through the analysis of 20 nuclear microsatellite markers. The varietal composition in modern orchards was also explored and the vast majority of the additional 151 trees analyzed showed the same genotype as the ancient accessions of 'Casaliva'. The results support the long historical link of 'Casaliva' with the GT and, besides a high varietal homogeneity, they also revealed the presence of olive genetic resources essential to fruit production. In fact, the parentage analysis of 550 embryos from drupes of 'Casaliva' evidenced that a cross-fertilization system is favored and a list of candidate cultivars most suitable as local pollinizers of 'Casaliva' was identified.
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Frutas/genética , Genótipo , Repetições de Microssatélites , Olea/genética , Árvores/genética , Frutas/embriologia , Olea/embriologia , PolinizaçãoRESUMO
Terpenoids, especially monoterpenes, are major aroma-impact compounds in grape and wine. Previous studies highlighted a key regulatory role for grapevine 1-deoxy-D-xylulose 5-phosphate synthase 1 (VvDXS1), the first enzyme of the methylerythritol phosphate pathway for isoprenoid precursor biosynthesis. Here, the parallel analysis of VvDXS1 genotype and terpene concentration in a germplasm collection demonstrated that VvDXS1 sequence has a very high predictive value for the accumulation of monoterpenes and also has an influence on sesquiterpene levels. A metabolic engineering approach was applied by expressing distinct VvDXS1 alleles in the grapevine model system "microvine" and assessing the effects on downstream pathways at transcriptional and metabolic level in different organs and fruit developmental stages. The underlying goal was to investigate two potential perturbation mechanisms, the former based on a significant over-expression of the wild-type (neutral) VvDXS1 allele and the latter on the ex-novo expression of an enzyme with increased catalytic efficiency from the mutated (muscat) VvDXS1 allele. The integration of the two VvDXS1 alleles in distinct microvine lines was found to alter the expression of several terpenoid biosynthetic genes, as assayed through an ad hoc developed TaqMan array based on cDNA libraries of four aromatic cultivars. In particular, enhanced transcription of monoterpene, sesquiterpene and carotenoid pathway genes was observed. The accumulation of monoterpenes in ripe berries was higher in the transformed microvines compared to control plants. This effect is predominantly attributed to the improved activity of the VvDXS1 enzyme coded by the muscat allele, whereas the up-regulation of VvDXS1 plays a secondary role in the increase of monoterpenes.
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The genetic varietal authentication of wine was investigated according to DNA isolation procedures reported for enological matrices and also by testing 11 commercial extraction kits and various protocol modifications. Samples were collected at different stages of the winemaking process of renowned Italian wines Brunello di Montalcino, Lambruschi Modenesi, and Trento DOC. Results demonstrated not only that grape DNA loss is produced by the fermentation process but also that clarification and stabilization operations contribute to the reduction of double-stranded DNA content on wine. Despite the presence of inhibitors, downstream PCR genotyping yielded reliable nuclear and chloroplast SSR markers for must samples, whereas no amplification or inconsistent results were obtained at later stages of the vinification. In addition, a TaqMan genotyping assay based on cultivar-specific single-nucleotide polymorphisms (SNPs) was designed, which allowed assessment of grapevine DNA mixtures. Once the wine matrix limitations are overcome, this sensitive tool may be implemented for the relative quantification of cultivars used for blend wines or frauds.
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Vitis/genética , Vinho/análise , Fermentação , Contaminação de Alimentos/análise , Genótipo , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Palestine has a wide range of agro-ecological concerns and hosts a large variety of plants. Grapes are part of the cultural heritage and provide an indispensable food ingredient. Local cultivars have been traditionally identified on the basis of morphological traits, geographical origin, or names of the vineyard owner; therefore, the occurrence of homonymy, synonymy, and misnaming significantly prevents their valorization. DNA profiling by 22 common SSR markers was used to characterize 43 putative cultivars grown mainly for local table grape consumption at the southern highland regions of West-Bank, to further evaluate genetic diversity and relationships of the population. Consistent matching of SSR markers with grapevines cultivated in neighboring countries or maintained in European germplasm collections was found for 8 of the 21 different non-redundant genotypes discovered, suggesting possible synonyms as well as the occurrence of breeding selections formerly developed in the USA. Genetic relationships inferred from SSR markers clearly assigned Palestinian cultivars to the Proles orientalis subpr. Antasiatica ancestral population, and they even remarked the connection between local resources and cultivars generated from international table grape breeding. This study supports the value of collection and conservation of vines endemic to a region of immense historical importance for viticulture.
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DNA de Plantas/genética , Repetições de Microssatélites/genética , Filogenia , Vitis/genética , Variação Genética , Genótipo , Humanos , Oriente MédioRESUMO
RESUMEN: Los dispositivos de administración electrónica de nicotina aparecieron con el objetivo de reemplazar el uso del tabaco y contribuir al reciente crecimiento de las políticas antitabaco. Actualmente, los efectos que producen los químicos que contiene son desconocidos. El objetivo de este trabajo consistió en describir la información encontrada en la literatura sobre los posibles daños generados por los dispositivos de administración electrónica de nicotina, en la cavidad bucal y los tejidos periodontales. Se realizó una búsqueda electrónica y manual, sin límite de idioma ni año. Se excluyeron, opiniones de expertos y artículos que estudiaran otros dispositivos de administración de nicotina. Se seleccionaron 18 artículos, 7 estudios in-vitro, 2 estudios longitudinales, 2 ensayos clínicos y 7 reportes de caso. Se encontró que existen niveles de citotoxicidad elevados, observando cambios a nivel de morfología y metabolismo celular. Sin embargo, si lo comparamos con el tabaco, se observa que los niveles de toxicidad son menores en los dispositivos de administración electrónica de nicotina. En los estudios longitudinales y ensayos clínicos se observó un aumento del sangramiento al sondaje y de la circulación sanguínea, medida con láser Doppler, al cambiar de cigarro convencional a cigarro electrónico. Los reportes de caso informan de importantes traumatismos en el territorio maxilofacial, causados por la explosión de estos dispositivos, durante su uso. Los dispositivos de administración electrónica de nicotina son tóxicos a nivel de las células periodontales, generando necrosis y daños al ADN celular. Presentan riesgos de uso, reportándose traumatismos graves a nivel oral y maxilofacial, por sobrecalentamiento de las baterías. Los ensayos clínicos y estudios longitudinales no fueron concluyentes, por lo que se debe seguir investigando en la materia.
ABSTRACT: Electronic nicotine delivery systems were introduced to replace tobacco use and contribute to the anti-smoking policies. Today, the effect the chemicals it contains produce in the human body, are unknown. The aim of this work was to describe the information found in the literature about the possible damage produced in the periodontal tissue and the oral cavity, by the electronic nicotine delivery systems. An electronic and manual search was carried by a single reviewer, without year or language limit, excluding expert's opinions and articles that studied other nicotine delivery systems. Eighteen articles were selected; 7 in vitro studies, 2 longitudinal studies, 2 clinical trials and 7 case reports. Toxicity levels were found to be high, showing changes in cellular morphology and metabolism. Comparing with conventional cigarette, toxicity levels were lower in the electronic nicotine delivery systems. Longitudinal studies and clinical trials observed an increase in bleeding on probing and in blood circulation, by using laser doppler velocimetry, when changing from conventional cigarette to electronic cigarette. Also reports about explosions while using these electronic devises were encountered in the literature, with grave consequences in the maxillofacial territory. Electronic nicotine delivery systems show toxic levels, generating necrosis and DNA damage in periodontal ligament and gingival fibroblast cells. The risk of using these devices is high, due to possible explosion following overheating of the lithium battery, causing facial and oral trauma. Clinical trials and longitudinal studies were not conclusive, so investigations should continue in this matter.
Assuntos
Humanos , Doenças Periodontais/induzido quimicamente , Nicotina/efeitos adversos , Nicotina/farmacologia , Sistemas Eletrônicos de Liberação de Nicotina , BocaRESUMO
Located in the center of the Mediterranean landscape and with an extensive coastal line, the territory of what is today Italy has played an important role in the history of human settlements and movements of Southern Europe and the Mediterranean Basin. Populated since Paleolithic times, the complexity of human movements during the Neolithic, the Metal Ages and the most recent history of the two last millennia (involving the overlapping of different cultural and demic strata) has shaped the pattern of the modern Italian genetic structure. With the aim of disentangling this pattern and understanding which processes more importantly shaped the distribution of diversity, we have analyzed the uniparentally-inherited markers in â¼900 individuals from an extensive sampling across the Italian peninsula, Sardinia and Sicily. Spatial PCAs and DAPCs revealed a sex-biased pattern indicating different demographic histories for males and females. Besides the genetic outlier position of Sardinians, a North West-South East Y-chromosome structure is found in continental Italy. Such structure is in agreement with recent archeological syntheses indicating two independent and parallel processes of Neolithisation. In addition, date estimates pinpoint the importance of the cultural and demographic events during the late Neolithic and Metal Ages. On the other hand, mitochondrial diversity is distributed more homogeneously in agreement with older population events that might be related to the presence of an Italian Refugium during the last glacial period in Europe.
Assuntos
Cromossomos Humanos Y/genética , DNA Mitocondrial/genética , Variação Genética , Genética Populacional/métodos , Cromossomos Humanos Y/classificação , Análise por Conglomerados , DNA Mitocondrial/classificação , Feminino , Geografia , Haplótipos/genética , Humanos , Itália , Masculino , Filogenia , Análise de Componente Principal , Fatores de TempoRESUMO
Cultivars used for wine and table grape have self-fertile hermaphrodite flowers whereas wild European vines and American and Asian species are dioecious, having either male or female flowers. Consistent with previous studies, the flower sex trait was mapped as a single major locus on chromosome 2 based on a pure Vitis vinifera population segregating for hermaphrodite and female progeny, and a hybrid population producing all three flower sex types. The sex locus was placed between the same SSR and SNP markers on both genetic maps, although abnormal segregation hampered to fine map the genomic region. From a total of 55 possible haplotypes inferred for three SSR markers around the sex locus, in a population of 132 V. sylvestris accessions and 171 V. vinifera cultivars, one of them accounted for 66 % of the hermaphrodite individuals and may be the result of domestication. Specific size variants of the VVIB23 microsatellite sequence within the 3'-UTR of a putative YABBY1 gene were found to be statistically significantly associated with the sex alleles M, H and f; these markers can provide assistance in defining the status of wild grapevine germplasm.
Assuntos
Mapeamento Cromossômico/métodos , Flores/genética , Haplótipos/genética , Organismos Hermafroditas/genética , Repetições de Microssatélites , Vitis/genética , Embaralhamento de DNA , DNA de Plantas/análise , DNA de Plantas/genética , Marcadores Genéticos/genéticaRESUMO
Vlad III The Impaler, also known as Dracula, descended from the dynasty of Basarab, the first rulers of independent Wallachia, in present Romania. Whether this dynasty is of Cuman (an admixed Turkic people that reached Wallachia from the East in the 11(th) century) or of local Romanian (Vlach) origin is debated among historians. Earlier studies have demonstrated the value of investigating the Y chromosome of men bearing a historical name, in order to identify their genetic origin. We sampled 29 Romanian men carrying the surname Basarab, in addition to four Romanian populations (from counties Dolj, Nâ=â38; Mehedinti, Nâ=â11; Cluj, Nâ=â50; and Brasov, Nâ=â50), and compared the data with the surrounding populations. We typed 131 SNPs and 19 STRs in the non-recombinant part of the Y-chromosome in all the individuals. We computed a PCA to situate the Basarab individuals in the context of Romania and its neighboring populations. Different Y-chromosome haplogroups were found within the individuals bearing the Basarab name. All haplogroups are common in Romania and other Central and Eastern European populations. In a PCA, the Basarab group clusters within other Romanian populations. We found several clusters of Basarab individuals having a common ancestor within the period of the last 600 years. The diversity of haplogroups found shows that not all individuals carrying the surname Basarab can be direct biological descendants of the Basarab dynasty. The absence of Eastern Asian lineages in the Basarab men can be interpreted as a lack of evidence for a Cuman origin of the Basarab dynasty, although it cannot be positively ruled out. It can be therefore concluded that the Basarab dynasty was successful in spreading its name beyond the spread of its genes.