Anthocyanins: Quantification by Tandem Mass Spectrometry.

Mass spectrometry is a high throughput technique widely used for metabolic fingerprinting of plant material. Among the diverse plant metabolites, pigments such as anthocyanins play a determinant role in plant defence mechanisms, protecting them from biotic and abiotic stresses. Anthocyanins are phenolic water-soluble glycosides or acyl-glycosides of anthocyanidins which could be accurately detected and quantified through mass spectrometry. This chapter describes how to extract anthocyanins from higher plant materials and quantify them through a liquid chromatography-mass spectrometry (LC-MS) based method.

Dirofilaria immitis is endemic in rural areas of the Brazilian Amazonas state capital, Manaus.

The canine filarial parasite Dirofilaria immitis has not been reported in Brazil´s Amazonas state capital, Manaus, for over a century. Here, we report one imported and 27 autochthonous D. immitis infections from a microfilarial survey of 766 domestic dog blood samples collected between 2017 and 2021 in Manaus. An Overall prevalence estimate of 15.44% (23/149) was calculated from our two rural collection sites; a prevalence of 1.22% (4/328) was estimated at our periurban collection site, and an overall prevalence of 0.35% (1/289) was calculated from our two urban clinic collections. Our data suggest that in the urban areas of Manaus, where the parasites are very likely vectored by the same species of mosquito that historically vectored Wuchereria bancrofti (Culex quinquefasciatus), prevalence levels are very low and possibly maintained by an influx from rural areas where sylvatic reservoirs and/or more favorable vector transmission dynamics maintain high prevalences.

Intermittent Scanning Glucose Monitoring or Predicted Low Suspend Pump Treatment: Does It Impact Time in Glucose Target and Treatment Preference? The QUEST Randomized Crossover Study.

Objective: To compare glycemic control and treatment preference in children with type 1 diabetes (T1D) using sensor augmented pump (SAP) with predictive low glucose suspend (SmartGuard®) or pump with independent intermittent scanning continuous glucose monitoring (iscCGM, Freestyle libre ®). Methods: In this open label, cross-over study, children 6 to 14 years of age, treated with insulin pump for at least 6 months, were randomized to insulin pump and iscCGM (A) or SAP with SmartGuard® (B) for 5 weeks followed by 5 additional weeks. The difference in percentages of time in glucose target (TIT), (3.9 - 8.0 mmol/l), <3 mmol/l, > 8 and 10 mmol/l, were analyzed using linear mixed models during the final week of each arm and were measured by blinded CGM (IPro2®). Results: 31 children (15 girls) finished the study. With sensor compliance > 60%, no difference in TIT was found, TIT: A 37.86%; 95% CI [33.21; 42.51]; B 37.20%; 95% CI [32.59; 41.82]; < 3 mmol/l A 2.27% 95% CI [0.71; 3.84] B 1.42% 95% CI [-0.13; 2.97]; > 8 mmol/l A 0.60% 95% CI [0.56, 0.67]; B 0.63% [0.56; 0.70]. One year after the study all participants were on CGM compared to 80.7% prior to the study, with a shift of 13/25 participants from iscCGM to SAP. Conclusions: In this study, no significant difference in glycemic control was found whether treated with SAP (SmartGuard®) or pump with iscCGM. The decision of all families to continue with CGM after the study suggests a positive impact, with preference for SmartGuard®. Clinical Trial Registration: [clinicaltrials.gov], identifier NCT03103867.

The Impact of Plant-Based Coatings in "ROCHA" Pear Preservation during Cold Storage: A Metabolomic Approach.

Although new storage technologies have been emerging in recent years, preservation of pear (Pyrus communis L.) remains a challenge for suppliers. Maintenance of desired organoleptic properties throughout cold storage using non-chemical strategies has been investigated and the use of edible coatings has shown potential to delay fruit quality deterioration during cold storage. Thus, the objective of this study is to evaluate the impact of pectin coatings including plant extracts, in "Rocha" pear (Pyrus communis L. cv. Rocha) preservation. A four-month pilot scale assay was performed in both dynamic controlled atmosphere (DCA) (-0.5 °C, 0.5% O2, and 0.4% CO2) and normal atmospheric (NA) conditions (2 °C). For each storage condition, the following three coatings were tested: pectin (3% w/v) (PCT), pectin (3% w/v) + strawberry tree leaves extract (9.5 mg/mL) (CT1), and pectin (3% w/v) + apple pomace extract (16 mg/mL) (CT2). Volatile compounds, potentially related to aroma or ripening status of "Rocha" pear, were monitored alongside with conjugated trienols (CTs) and maturity parameters. The combination of DCA conditions and the application of pectin coatings were able to reduce the release of Rocha pear volatiles associated with ripening status, (particularly esters and sesquiterpenes), as well as reduce CTs, which could contribute to the preservation of Rocha pear for longer periods.

Natural-Based Antioxidant Extracts as Potential Mitigators of Fruit Browning.

Fruit enzymatic browning (EB) inhibition continues to be a challenge in the Food Industry. This physiological disorder results mainly from the oxidation of natural phenolic compounds by polyphenoloxidase (PPO) and peroxidase (POX) leading to the formation of brown pigments. EB can be controlled with the application of antioxidants, reducing/inhibiting the activity of these oxidative enzymes. In this study, strawberry tree (leaves and branches) and apple byproduct were the natural-based extracts (NES) selected, as potential tissue browning inhibitors, within a first screening of fifteen natural-based extracts with antioxidant properties. Phenolic profile, total phenolic content and antioxidant activity of the selected extracts were also performed as well as their depletion effect on the oxidative enzyme's activity and browning inhibiton in fresh-cut pears. Strawberry tree extracts (leaves and branches) revealed higher total phenolic content (207.97 ± 0.01 mg GAE.gNES-1 and 104.07 ± 16.38 mg GAE.gNES-1, respectively), confirmed by the plethora of phenolic compounds identified by LC-ESI-UHR-QqTOF-HRMS and quantified by HPLC. This phytochemical composition was reflected in the low IC50 against PPO and POX obtained. Despite the lower phenolic content (6.76 ± 0.11 mg GAE.gNES-1) and antioxidant activity (IC50 = 45.59 ± 1.34 mg mL-1), apple byproduct extract showed potential in delaying browning. This study highlights the opportunity of byproducts and agricultural wastes extracts as novel anti-browning agents.

Strategies to Preserve Postharvest Quality of Horticultural Crops and Superficial Scald Control: From Diphenylamine Antioxidant Usage to More Recent Approaches.

Horticultural crops are vulnerable to several disorders, which affect their physiological and organoleptic quality. For about forty years, the control of physiological disorders (such as superficial scald) in horticultural crops, particularly in fruit, was achieved through the application of the antioxidant diphenylamine (DPA), usually combined with controlled atmosphere (CA) conditions. However, identification of DPA residues and metabolites in treated fruits, associated with their toxicity, banned the use of this antioxidant in Europe. This triggered the urgent need for novel and, ideally, natural and sustainable alternatives, combined with adequate storage conditions to protect cultivars from harmful agents. This review systematizes the state-of-the-art DPA application on several fresh cultivars, such as apples, pears, and vegetables (potatoes, spinach, etc.), as well as the possible mechanisms of the action and effects of DPA, emphasizing its antioxidant properties. Alternative methods to DPA are also discussed, as well as respective effects and limitations. Recent research on scald development molecular pathways are highlighted to open new non-chemical strategies opportunities. This appraisal shows that most of the current solutions have not lead to satisfactory commercial results; thus, further research aimed to understand the mechanisms underlying postharvest disorders and to design sustainable and safe solutions to improve horticultural products storage is needed.

Biocompatibility and Bioimaging Potential of Fruit-Based Carbon Dots.

Photo-luminescent carbon dots (CD) have become promising nanomaterials and their synthesis from natural products has attracted attention by the possibility of making the most of affordable, sustainable and, readily-available carbon sources. Here, we report on the synthesis, characterization and bioimaging potential of CDs produced from diverse extensively produced fruits: kiwi, avocado and pear. The in vitro cytotoxicity and anticancer potential of those CDs were assessed by comparing human epithelial cells from normal adult kidney and colorectal adenocarcinoma cells. In vivo toxicity was evaluated using zebrafish embryos given their peculiar embryogenesis, with transparent embryos developing ex-utero, allowing a real-time analysis. In vitro and in vivo experiments revealed that the synthesized CD presented toxicity only at concentrations of ≥1.5 mg mL-1. Kiwi CD exhibited the highest toxicity to both cells lines and zebrafish embryos, presenting lower LD50 values. Interestingly, despite inducing lower cytotoxicity in normal cells than the other CDs, black pepper CDs resulted in higher toxicity in vivo. The bio-distribution of CD in zebrafish embryos upon uptake was investigated using fluorescence microscopy. We observed a higher accumulation of CD in the eye and yolk sac, avocado CD being the ones more retained, indicating their potential usefulness in bio-imaging applications. This study shows the action of fruit-based CDs from kiwi, avocado and pear. However the compounds present in these fruit-based CDs and their mechanism of action as a bioimaging agent need to be further explored.

Effect of predicted low suspend pump treatment on improving glycaemic control and quality of sleep in children with type 1 diabetes and their caregivers: the QUEST randomized crossover study.

BACKGROUND: In attempting to achieve optimal metabolic control, the day-to-day management is challenging for a child with type 1 diabetes (T1D) and his family and can have a major negative impact on their quality of life. Augmenting an insulin pump with glucose sensor information leads to improved outcomes: decreased haemoglobin A1c levels, increased time in glucose target and less hypoglycaemia. Fear of nocturnal hypoglycaemia remains pervasive amongst parents, leading to chronic sleep interruption and lack of sleep for the parents and their children. The QUEST study, an open-label, single-centre randomized crossover study, aims to evaluate the impact on time in target, in hypoglycaemia and hyperglycaemia and the effect on sleep and quality of life in children with T1D, comparing a sensor-augmented pump (SAP) with predictive low glucose suspend and alerts to the use of the same insulin pump with a flash glucose measurement (FGM) device not interacting with the pump. METHODS/DESIGN: Subjects meeting the inclusion criteria are randomized to treatment with the SAP or treatment with an insulin pump and independent FGM for 5 weeks. Following a 3-week washout period, the subjects cross over to the other study arm for 5 weeks. During the week before and in the last week of treatment, the subjects and one of their caregivers wear a sleep monitor in order to obtain sleep data. The primary endpoint is the between-arm difference in percentage of time in glucose target during the final 6 days of each treatment arm, measured by a blinded continuous glucose measurement (CGM). Additional endpoints include comparison of quantity and quality of sleep as well as quality of life perception of the subjects and one of their caregivers in the two different treatment arms. Recruitment started in February 2017. A total of 36 patients are planned to be randomized. The study recruitment was completed in April 2018. DISCUSSION: With this study we will provide more information on whether insulin pump treatment combined with more technology (SmartGuard® feature and alerts) leads to better metabolic control. The inclusion of indicators on quality of sleep with less sleep interruption, less lack of sleep and perception of quality of life in both children and their primary caregivers is essential for this study and might help to guide us to further treatment improvement. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03103867 . Registered on 6 April 2017.