Metabolic Adaptations Correlated with Antibody Response after Immunization with Inactivated SARS-CoV-2 in Brazilian Subjects.

The adsorbed vaccine SARS-CoV-2 (inactivated) produced by Sinovac (SV) was the first vaccine against COVID-19 to be used in Brazil. To understand the metabolic effects of SV in Brazilian subjects, NMR-based metabolomics was used, and the immune response was studied in Brazilian subjects. Forty adults without (group-, n = 23) and with previous COVID-19 infection (group+, n = 17) were followed-up for 90 days postcompletion of the vaccine regimen. After 90 days, our results showed that subjects had increased levels of lipoproteins, lipids, and N-acetylation of glycoproteins (NAG) as well as decreased levels of amino acids, lactate, citrate, and 3-hydroxypropionate. NAG and threonine were the highest correlated metabolites with N and S proteins, and neutralizing Ab levels. This study sheds light on the immunometabolism associated with the use of SV in Brazilian subjects from Rio de Janeiro and identifies potential metabolic markers associated with the immune status.

Microbiota associated with peri-implantitis-A systematic review with meta-analyses.

AIM: To answer the following PECO question: "In systemically healthy human subjects (P), which are the differences between peri-implantitis (E) and peri-implant health/mucositis (C) in terms of bacterial presence/count (O)?" MATERIALS AND METHODS: Cross-sectional studies fulfilling specific inclusion criteria established to answer the PECO question were included. Two review authors independently searched for studies, screened the titles and abstracts, did full-text analysis, extracted the data from the included reports, and performed the risk of bias assessment through an adaptation of the Newcastle/Ottawa tool for cross-sectional studies and of the JBI critical appraisal checklist. In case of disagreement, a third reviewer author took the final decision. Study results were summarized using random effects meta-analyses. RESULTS: A total of 12 studies were included, involving 1233 participants and 1513 implants. Peri-implantitis was associated with the presence of S. epidermidis (Odds ratio, OR = 10.28 [95% Confidence interval, CI: 1.26-83.98]), F. nucleatum (OR = 7.83 [95% CI: 2.24-27.36]), T. denticola (OR = 6.11 [95% CI: 2.72-13.76]), T. forsythia (OR = 4.25 [95% CI: 1.71-10.57]), P. intermedia (OR = 3.79 [95% CI: 1.07-13.35]), and P. gingivalis (OR = 2.46 [95% CI: 1.21-5.00]). Conversely, the presence of A. actinomycetemcomitans (OR = 3.82 [95% CI: 0.59-24.68]), S. aureus (OR = 1.05 [95% CI: 0.06-17.08]), and C. rectus (OR = 1.48 [95% CI: 0.69-3.17]) was not associated with peri-implantitis. CONCLUSIONS: Peri-implantitis is associated with the presence of S. epidermidis and specific periodontopathogens (P. gingivalis, T. forsythia, T. denticola, F. nucleatum, and P. intermedia). (CRD42021254589).

Optical coherence tomography findings in fungal uveitis.

PURPOSE: The purpose of the study was to describe the vitreal, retinal, and choroidal features of eyes affected by endogenous endophthalmitis (EE) and evaluate the effects of systemic antifungal drug treatment and pars plana vitrectomy by using spectral domain optical coherence tomography (SD-OCT). METHODS: Medical records and SD-OCT images of eyes diagnosed with EE at a single uveitis tertiary referral center in Brazil were acquired at the time of diagnosis, after 7 days of high-dose antifungal drug treatment, and at follow-up assessments performed 30 days after resolution. RESULTS: Thirteen eyes were enrolled in the study. All patients showed hyperreflective round-shaped lesions on SD-OCT and pre-retinal aggregates. Five eyes responded to antifungal systemic oral drugs despite showing vitreous opacity. The response to treatment was observable on optical coherence tomography (OCT) images. CONCLUSION: Fungal endophthalmitis showed typical features on SD-OCT, facilitating early diagnosis and treatment despite the absence of vitreous culture or biopsy. This study suggests that OCT images can be used to support diagnosis by physicians who do not have access to vitreoretinal surgery.

Generation of a triple-fluorescent mouse strain allows a dynamic and spatial visualization of different liver phagocytes in vivo.

Resident and circulating immune cells have been extensively studied due to their almost ubiquitous role in cell biology. Despite their classification under the "immune cell department", it is becoming increasingly clear that these cells are involved in many different non-immune related phenomena, including fetus development, vascular formation, memory, social behavior and many other phenotypes. There is a huge potential in combining high-throughput assays - including flow cytometry and gene analysis - with in vivo imaging. This can improve our knowledge in both basic and clinical cell biology, and accessing the expression of markers that are relevant in the context of both homeostasis and disease conditions might be instrumental. Here we describe how we generated a novel mouse strain that spontaneously express three different fluorescence markers under control of well-studied receptors (CX3CR1, CCR2 and CD11c) that are involved in a plethora of stages of cell ontogenesis, maturation, migration and behavior. Also, we assess the percentage of the expression and co-expression of each marker under homeostasis conditions, and how these cells behave when a local inflammation is induced in the liver applying a cutting-edge technology to image cells by confocal intravital microscopy.

Immune and metabolic shifts during neonatal development reprogram liver identity and function.

BACKGROUND & AIMS: The liver is the main hematopoietic site in embryos, becoming a crucial organ in both immunity and metabolism in adults. However, how the liver adapts both the immune system and enzymatic profile to challenges in the postnatal period remains elusive. We aimed to identify the mechanisms underlying this adaptation. METHODS: We analyzed liver samples from mice on day 0 after birth until adulthood. Human biopsies from newborns and adults were also examined. Liver immune cells were phenotyped using mass cytometry (CyTOF) and expression of several genes belonging to immune and metabolic pathways were measured. Mortality rate, bacteremia and hepatic bacterial retention after E. coli challenge were analyzed using intravital and in vitro approaches. In a set of experiments, mice were prematurely weaned and the impact on gene expression of metabolic pathways was evaluated. RESULTS: Human and mouse newborns have a sharply different hepatic cellular composition and arrangement compared to adults. We also found that myeloid cells and immature B cells primarily compose the neonatal hepatic immune system. Although neonatal mice were more susceptible to infections, a rapid evolution to an efficient immune response was observed. Concomitantly, newborns displayed a reduction of several macronutrient metabolic functions and the normal expression level of enzymes belonging to lipid and carbohydrate metabolism was reached around the weaning period. Interestingly, early weaning profoundly disturbed the expression of several hepatic metabolic pathways, providing novel insights into how dietary schemes affect the metabolic maturation of the liver. CONCLUSION: In newborns, the immune and metabolic profiles of the liver are dramatically different to those of the adult liver, which can be explained by the differences in the liver cell repertoire and phenotype. Also, dietary and antigen cues may be crucial to guide liver development during the postnatal phase. LAY SUMMARY: Newborns face major challenges in the extra-uterine life. In fact, organs need to modify their cellular composition and gene expression profile in order to adapt to changes in both microbiota and diet throughout life. The liver is interposed between the gastrointestinal system and the systemic circulation, being the destination of all macronutrients and microbial products from the gut. Therefore, it is expected that delicately balanced mechanisms govern the transformation of a neonatal liver to a key organ in adults.

Protease-activated receptor 2 (PAR2) upregulates granulocyte colony stimulating factor (G-CSF) expression in breast cancer cells.

Protease-activated receptor 2 (PAR2) is a G-protein coupled receptor which is activated upon cleavage of its N-terminal region. PAR2 has been associated with many aspects regarding tumor progression, such as the production of pro-tumoral cytokines. Granulocyte colony-stimulating factor (G-CSF) is a cytokine essential to neutrophil production and maturation, and it is often overexpressed in tumors. In this study, we evaluated the ability of PAR2 to modulate G-CSF expression. PAR2 and G-CSF were significantly more expressed in metastatic (4T1 and MDA-MB-231) as compared to non-metastatic (67NR and MCF7) breast cancer cell lines. In addition, PAR2 stimulation by a synthetic agonist peptide significantly increased G-CSF gene expression in the metastatic cell lines. Knockdown of PAR2 in 4T1 cells decreased G-CSF expression and secretion. In addition, treatment of 4T1 with the commercial PAR2 antagonist, ENMD-1068, significantly decreased G-CSF expression. cBioPortal analyses of the TCGA database showed a significant co-occurrence of G-CSF and PAR2 gene overexpression in breast cancer samples. In conclusion, our data suggest that PAR2 contributes to G-CSF expression in breast cancer cells, possibly favoring tumor progression.

IL-33 signalling in liver immune cells enhances drug-induced liver injury and inflammation.

OBJECTIVE AND DESIGN: The aim of this study was to investigate the contribution of IL-33/ST2 axis in the onset and progression of acute liver injury using a mice model of drug-induced liver injury (DILI). MATERIAL AND TREATMENTS: DILI was induced by overdose administration of acetaminophen (APAP) by oral gavage in wild-type BALB/c, ST2-deficient mice and in different bone marrow chimeras. Neutrophils were depleted by anti-Ly6G and macrophages with clodronate liposomes (CLL). METHODS: Blood and liver were collected for biochemical, immunologic and genetic analyses. Mice were imaged by confocal intravital microscopy and liver non-parenchymal cells and hepatocytes were isolated for flow cytometry, genetic and immunofluorescence studies. RESULTS: Acetaminophen overdose caused a massive necrosis and accumulation of immune cells within the liver, concomitantly with IL-33 and chemokine release. Liver non-parenchymal cells were the major sensors for IL-33, and amongst them, neutrophils were the major players in amplification of the inflammatory response triggered by IL-33/ST2 signalling pathway. CONCLUSION: Blockage of IL-33/ST2 axis reduces APAP-mediated organ injury by dampening liver chemokine release and activation of resident and infiltrating liver non-parenchymal cells.

Reciprocal modulation of histone deacetylase inhibitors sodium butyrate and trichostatin A on the energy metabolism of breast cancer cells.

Tumor cells display different bioenergetic profiles when compared to normal cells. In the present work we showed metabolic reprogramming by means of inhibitors of histone deacetylase (HDACis), sodium butyrate and trichostatin A in breast cancer cells representing different stages of aggressiveness and metabolic profile. When testing the effect of NaB and TSA on viability of cells, it was shown that non-tumorigenic MCF-10A cells were less affected by increasing doses of the drugs than the tumorigenic, hormone dependent, tightly cohesive MCF-7, T-47D and the highly metastatic triple-negative MDA-MB 231 cells. T-47D cells were the most sensitive to treatment with both, NaB and TSA. Experiments measuring anchorage- independent growth of tumor cells showed that MCF-7, T-47D, and MDA-MB-231 cells were equally sensitive to the treatment with NaB. The NaB induced an attenuation of glycolysis, reflected by a decrease in lactate release in MCF-7 and T47D lines. Pyruvate kinase activity was significantly enhanced by NaB in MDA-MB-231 cells only. In contrast, the inhibitor enhanced lactate dehydrogenase activity specifically in T-47 D cells. Glucose-6-phosphate dehydrogenase activity was shown to be differentially modulated by NaB in the cell lines investigated: the enzyme was inhibited in MCF-7 cells, whereas in T-47D and MDA-MB-231 cells, G6PDH was activated. NaB and TSA were able to significantly increase the oxygen consumption by MDA-MB-231 and T-47D cells. Collectively the results show that epigenetic changes associated to acetylation of proteins in general affect the energy metabolism in all cancer cell lines and that mitochondria may occupy a central role in metastasis.

Monkeypox Infection Causing Conjunctival Vesicles and Anterior Uveitis.

Human monkeypox is a zoonosis caused by an orthopoxvirus and the clinical presentation resembles that of smallpox and chickenpox. The disease may start with a prodrome that includes lymphadenopathy, headache, fatigue, and fever, followed by a vesiculo-pustular rash. Ocular manifestations such as conjunctivitis and edema are present in approximately 20% of affected people, with a greater incidence among unvaccinated patients. Corneal involvement has also been reported and can result in corneal scarring and severe forms of keratitis. The natural course of the disease is most often benign and self-limiting, however, in some individuals, especially immunocompromised patients, there is a risk of complications such as bronchopneumonia, encephalitis, and vision loss. Herein, we present a case of a patient with monkeypox which caused conjunctival vesicles and anterior uveitis.

Ophthalmic Manifestations of Monkeypox Virus Infection: A Cases Series.

This retrospective case series aims to describe the ophthalmic manifestations of the Monkeypox virus infection in seven patients evaluated in two countries of South America (Colombia and Brazil). Two had skin lesions in the eyelid, and five had conjunctivitis. None had intraocular involvement. Three of seven patients had a history of Human Immunodeficiency Virus infection, and all patients had lesions in the genital region, suggesting sexual-contact transmission. In 6 of 7 cases, conjunctival RT-PCR was positive for the Monkeypox virus, including one case without conjunctival vesicles. In all cases, lesions resolved without complications, and just two required antiviral treatment. All patients demonstrated improvement without complications. RT-PCR positivity in conjunctiva demonstrated the presence of the Monkeypox virus, suggesting that ocular-mediated transmission could be plausible. Ophthalmologists should be aware of this ophthalmic manifestation.

Method optimization for the extraction of chlorogenic acids from coffee parchment: An ecofriendly alternative.

One of the principal byproducts of coffee roasting is the coffee parchment. It is abundant in bioactive substances, including derivatives of chlorogenic acids, which are well-known for their exceptional antioxidant effects. It is advantageous to use environmentally friendly extraction techniques on such residues since it adds value to the entire coffee production process supply chain. The aim of this work was to assess and enhance the ability of non-conventional extraction techniques to extract derivatives of chlorogenic acid from coffee parchment. A central composite design was used to maximize the recovery of those phenolic compounds. The optimized extraction conditions were with 5 min extraction period, at a temperature of 70 °C, and 80% ethanol in the extractor solvent. In this conditions extraction recovery of chlorogenic acids was of 0.8% by the use of microwave-aided extraction (MAE). The optimized conditions are practical, economical, and ecologically friendly method to extract phenolic compounds and, consequently, underscores the potential for sustainable utilization of coffee parchment, offering a valuable contribution to the development of environmentally conscious strategies within the coffee industry.

3-Ishwarone, a rare ishwarane sesquiterpene from Peperomia scandens Ruiz & Pavon: structural elucidation through a joint experimental and theoretical study.

3-Ishwarone, (1), a sesquiterpene with a rare ishwarane skeleton, was isolated from Peperomia scandens Ruiz & Pavon (Piperaceae). Its structure was unambiguously determined by 1D- and 2D-NMR and infrared analyses, as well as by comparative theoretical studies which involved calculations of 13C-NMR chemical shifts, using the Density Functional Theory (DFT) with the mPW1PW91 hybrid functional and Pople's 6-31G(d) basis set, and of vibrational frequencies, using the B3LYP hybrid functional and triple ζ Dunning's correlation consistent basis set (cc-pVTZ), of (1) and three of its possible diastereomers, compounds 2-4.

Bilateral Ampiginous Choroiditis following Confirmed SARS-CoV-2 Infection.

INTRODUCTION: Ampiginous Choroiditis is a rare posterior uveitis that combines clinical features of Acute Multifocal Posterior Placoid Pigment Epitheliopathy and Serpiginous Chorioretinitis. Its pathophysiology is poorly understood and further studies are necessary to understand which mechanisms start the immunologic reaction. CASE REPORT: The purpose of this article is to report a well-documented case of Ampiginous Choroiditis following in seven days a RT-PCR confirmed SARS-CoV-2 infection, suggesting that the infection might have contributed as a trigger. CONCLUSION: Timely diagnosis and correct treatment are paramount to improve the visual outcomes, and the patient had successful response to systemic steroids.

The ADAM17 sheddase complex regulator iTAP/Frmd8 modulates inflammation and tumor growth.

The metalloprotease ADAM17 is a sheddase of key molecules, including TNF and epidermal growth factor receptor ligands. ADAM17 exists within an assemblage, the "sheddase complex," containing a rhomboid pseudoprotease (iRhom1 or iRhom2). iRhoms control multiple aspects of ADAM17 biology. The FERM domain-containing protein iTAP/Frmd8 is an iRhom-binding protein that prevents the precocious shunting of ADAM17 and iRhom2 to lysosomes and their consequent degradation. As pathophysiological role(s) of iTAP/Frmd8 have not been addressed, we characterized the impact of iTAP/Frmd8 loss on ADAM17-associated phenotypes in mice. We show that iTAP/Frmd8 KO mice exhibit defects in inflammatory and intestinal epithelial barrier repair functions, but not the collateral defects associated with global ADAM17 loss. Furthermore, we show that iTAP/Frmd8 regulates cancer cell growth in a cell-autonomous manner and by modulating the tumor microenvironment. Our work suggests that pharmacological intervention at the level of iTAP/Frmd8 may be beneficial to target ADAM17 activity in specific compartments during chronic inflammatory diseases or cancer, while avoiding the collateral impact on the vital functions associated with the widespread inhibition of ADAM17.

Semaphorin 4B is an ADAM17-cleaved adipokine that inhibits adipocyte differentiation and thermogenesis.

OBJECTIVE: The metalloprotease ADAM17 (also called TACE) plays fundamental roles in homeostasis by shedding key signaling molecules from the cell surface. Although its importance for the immune system and epithelial tissues is well-documented, little is known about the role of ADAM17 in metabolic homeostasis. The purpose of this study was to determine the impact of ADAM17 expression, specifically in adipose tissues, on metabolic homeostasis. METHODS: We used histopathology, molecular, proteomic, transcriptomic, in vivo integrative physiological and ex vivo biochemical approaches to determine the impact of adipose tissue-specific deletion of ADAM17 upon adipocyte and whole organism metabolic physiology. RESULTS: ADAM17adipoq-creΔ/Δ mice exhibited a hypermetabolic phenotype characterized by elevated energy consumption and increased levels of adipocyte thermogenic gene expression. On a high fat diet, these mice were more thermogenic, while exhibiting elevated expression levels of genes associated with lipid oxidation and lipolysis. This hypermetabolic phenotype protected mutant mice from obesogenic challenge, limiting weight gain, hepatosteatosis and insulin resistance. Activation of beta-adrenoceptors by the neurotransmitter norepinephrine, a key regulator of adipocyte physiology, triggered the shedding of ADAM17 substrates, and regulated ADAM17 expression at the mRNA and protein levels, hence identifying a functional connection between thermogenic licensing and the regulation of ADAM17. Proteomic studies identified Semaphorin 4B (SEMA4B), as a novel ADAM17-shed adipokine, whose expression is regulated by physiological thermogenic cues, that acts to inhibit adipocyte differentiation and dampen thermogenic responses in adipocytes. Transcriptomic data showed that cleaved SEMA4B acts in an autocrine manner in brown adipocytes to repress the expression of genes involved in adipogenesis, thermogenesis, and lipid uptake, storage and catabolism. CONCLUSIONS: Our findings identify a novel ADAM17-dependent axis, regulated by beta-adrenoceptors and mediated by the ADAM17-cleaved form of SEMA4B, that modulates energy balance in adipocytes by inhibiting adipocyte differentiation, thermogenesis and lipid catabolism.

Risk factors for recurrences and visual impairment in patients with ocular toxoplasmosis: A systematic review and meta-analysis.

BACKGROUND: Ocular toxoplasmosis (OT) is caused by the parasite Toxoplasma gondii. OT is the leading cause of posterior uveitis globally; it is a recurrent disease that may result in visual impairment and blindness. This systematic review and meta-analysis aim to summarize and evaluate the risk factors for recurrences, visual impairment, and blindness described in the literature worldwide. METHODS AND FINDINGS: We performed a systematic literature search in PubMed, Embase, VHL, Cochrane Library, Scopus, and DANS EASY Archive. All studies reporting patients with clinically and serologically confirmed OT presenting any clinical or paraclinical factor influencing recurrences, visual impairment, and blindness were included. Studies presenting secondary data, case reports, and case series were excluded. An initial selection was made by title and abstract, and then the studies were reviewed by full text where the eligible studies were selected. Then, the risk of bias was assessed through validated tools. Data were extracted using a validated extraction format. Qualitative synthesis and quantitative analysis were done. This study was registered on PROSPERO (CRD42022327836). RESULTS: Seventy two studies met the inclusion criteria. Fifty-three were summarized in the qualitative synthesis in three sections: clinical and environmental factors, parasite and host factors, and treatment-related factors. Of the 72 articles, 39 were included in the meta-analysis, of which 14 were conducted in South America, 13 in Europe, four in Asia, three multinational, two in North America and Central America, respectively, and only one in Africa. A total of 4,200 patients with OT were analyzed, mean age ranged from 7.3 to 65.1 year of age, with similar distribution by sex. The frequency of recurrences in patients with OT was 49% (95% CI 40%-58%), being more frequent in the South American population than in Europeans. Additionally, visual impairment was presented in 35% (95% CI 25%-48%) and blindness in 20% (95% CI 13%-30%) of eyes, with a similar predominance in South Americans than in Europeans. On the other hand, having lesions near the macula or adjacent to the optic nerve had an OR of 4.83 (95% CI; 2.72-8.59) for blindness, similar to having more than one recurrence that had an OR of 3.18 (95% CI; 1.59-6.38). Finally, the prophylactic therapy with Trimethoprim/Sulfamethoxazole versus the placebo showed a protective factor of 83% during the first year and 87% in the second year after treatment. CONCLUSION: Our Systematic Review showed that clinical factors such as being older than 40 years, patients with de novo OT lesions or with less than one year after the first episode, macular area involvement, lesions greater than 1 disc diameter, congenital toxoplasmosis, and bilateral compromise had more risk of recurrences. Also, environmental and parasite factors such as precipitations, geographical region where the infection is acquired, and more virulent strains confer greater risk of recurrences. Therefore, patients with the above mentioned clinical, environmental, and parasite factors could benefit from using prophylactic therapy.

[Information Systems on violence against women: an integrative review]. / Sistemas de Informação sobre violência contra as mulheres: uma revisão integrativa.

Violence against women, enhanced by issues of race/ethnicity, class and generation, constitutes one of the main obstacles to overcoming gender inequalities in all spheres of life, including the private domain. Over the past decades, actions to confront the phenomenon have advanced significantly in Brazil, particularly the growing output and the systematization of information on the topic. In this sense, Information Systems (IS) are important tools in the collection, processing/organization and dissemination of official statistics on this type of violence, the objective of which is to assist in the planning and implementation of intersectoral public policies. It is noteworthy that these devices are characterized as a space for disputes of diverse interests. That is, they are not neutral instruments, but they bring with them a worldview that guides them. Thus, a critical analysis of information systems on violence against women is necessary, dealing with how they characterize the phenomenon, as well as feeding decisions and planning actions. Based on an integrative review, this paper aims to analyze how information systems on violence against women are portrayed in national and international academic production.

A violência contra as mulheres, potencializada por questões de raça/etnia, classe e geração, constitui-se um dos principais obstáculos para a superação das desigualdades de gênero em todas as esferas da vida, incluindo o espaço privado. Ao longo das últimas décadas, ações de enfrentamento do fenômeno avançaram de forma significativa no Brasil, particularmente a produção crescente e a sistematização de informação sobre o mesmo. Nessa direção, os Sistemas de Informação (SI) constituem-se ferramentas importantes na coleta, processamento/organização e difusão de estatísticas oficiais sobre esse tipo de violência, cujo objetivo é auxiliar o planejamento e implementação de políticas públicas intersetoriais. Ressalta-se que esses dispositivos se caracterizam como um espaço de disputas de interesses diversos. Isto é, não são instrumentos neutros, mas trazem em si uma visão de mundo que os orienta. Assim, faz-se necessário uma análise crítica dos sistemas de informação sobre violência contra as mulheres, tratando como caracterizam o fenômeno, bem como têm alimentado as decisões e planejamento de ações. A partir de uma revisão integrativa, o presente trabalho tem como objetivo analisar como os sistemas de informação sobre violência contra a mulher são retratados pela produção acadêmica nacional e internacional.

Quantitative 1H NMR method for analyzing primaquine diphosphate in active pharmaceutical ingredients.

Thermal analysis, Fourier Transform IR, the isotropic chemical shift of 1H NMR in different solvents, their temperature dependence and spin-lattice relaxation time constant (T1), solution 1D and 2D NMR, and solid-state 13C and 31P NMR (magic angle spinning NMR) were employed to obtain full information and elucidate the structures of primaquine diphosphate (PQD) samples used for quality controlling malaria medicines. Additionally, a simple, rapid, specific, and reliable quantitative method (qNMR) was developed to determine the PQD level in the raw material of active pharmaceutical ingredients (APIs). The method was developed using ethylene carbonate (EC) as the internal standard and dimethylsulfoxide-d6 (DMSO-d6) as the NMR solvent. For the API qNMR, 1H NMR signals at 3.82 and 1.22 ppm were used. The qNMR methodology, through the linearity, range, LOD and LOQ, stability, precision, robustness, and accuracy, was validated within the requirements of guidelines. The accuracy of the qNMR was evaluated by comparing it to a pharmacopeial HPLC technique and there were no statistical differences (p > 0.05). The proposed qNMR method authentically supports and endorses the current pharmacopoeial methods used for determining the PQD content.

1H NMR and UPLC-HRMS-based metabolomic approach for evaluation of the grape maturity and maceration time of Touriga Nacional wines and their correlation with the chemical stability.

Touriga Nacional is a well-adapted Portuguese grape variety in São Francisco River Valley (northeastern Brazil). Nevertheless, it has only been indicated to short-term consumption because of the lack of chemical stability, which is attributed to low grape acidity and incomplete phenolic maturity. Therefore, we used Ultra-Performance Liquid Chromatography coupled High-resolution Mass Spectrometry, Nuclear Magnetic Resonance and chemometrics (PCA and PLS-DA) to evaluate the grape maturity and maceration time on chemical composition of wines from two harvest seasons. Moreover, we investigated how these experimental factors could affect their chemical stability. Grapes maturity showed to be the main effect. Overall, phenolic acids and short-chain organic acids were found to be at higher levels in wines produced with unripe grapes from February and shorter maceration time (p < 0.05). Proanthocyanidins and other flavonoids were increased in wines macerated for longer time using overripe grapes harvested in July. Furthermore, stable wines were made from overripe grapes, which contained more galacturonic acid.