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Persimmon fruit processing-derived waste and by-products, such as peels and pomace, are important sources of dietary fiber and phytochemicals. Revalorizing these by-products could help promote circular nutrition and agricultural sustainability while tackling dietary deficiencies and chronic diseases. In this study, fiber-rich fractions were prepared from the by-products of Sharoni and Brilliant Red persimmon varieties. These fractions were quantified for their phenolic composition and assessed for their ability to promote the growth of beneficial human colonic Firmicutes species and for their in vitro anti-inflammatory potential. Gallic and protocatechuic acids, delphinidin, and cyanidin were the main phenolics identified. Faecalibacterium prausnitzii strains showed significantly higher growth rates in the presence of the Brilliant Red fraction, generating more than double butyrate as a proportion of the total short-chain fatty acids (39.5% vs. 17.8%) when compared to glucose. The fiber-rich fractions significantly decreased the inflammatory effect of interleukin-1ß in Caco-2 cells, and the fermented fractions (both from Sharoni and Brilliant Red) significantly decreased the inflammatory effect of interleukin-6 and tumor necrosis factor-α in the RAW 264.7 cells. Therefore, fiber-rich fractions from persimmon by-products could be part of nutritional therapies as they reduce systemic inflammation, promote the growth of beneficial human gut bacteria, and increase the production of beneficial microbial metabolites such as butyrate.
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Anti-Inflamatórios , Colo , Fibras na Dieta , Diospyros , Humanos , Fibras na Dieta/farmacologia , Fibras na Dieta/análise , Diospyros/química , Camundongos , Anti-Inflamatórios/farmacologia , Colo/microbiologia , Colo/efeitos dos fármacos , Colo/metabolismo , Animais , Células RAW 264.7 , Células CACO-2 , Microbioma Gastrointestinal/efeitos dos fármacos , Firmicutes , Faecalibacterium prausnitzii , Frutas/química , Fator de Necrose Tumoral alfa/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Hidroxibenzoatos/farmacologia , Hidroxibenzoatos/análise , Fenóis/farmacologia , Fenóis/análise , Fermentação , Ácido Gálico/farmacologia , Antocianinas/farmacologia , Antocianinas/análiseRESUMO
Pseudo-cereals such as buckwheat (Fagopyrum esculentum) are valid candidates to promote diet biodiversity and nutrition security in an era of global climate change. Buckwheat hulls (BHs) are currently an unexplored source of dietary fibre and bioactive phytochemicals. This study assessed the effects of several bioprocessing treatments (using enzymes, yeast, and combinations of both) on BHs' nutrient and phytochemical content, their digestion and metabolism in vitro (using a gastrointestinal digestion model and mixed microbiota from human faeces). The metabolites were measured using targeted LC-MS/MS and GC analysis and 16S rRNA gene sequencing was used to detect the impact on microbiota composition. BHs are rich in insoluble fibre (31.09 ± 0.22% as non-starch polysaccharides), protocatechuic acid (390.71 ± 31.72 mg/kg), and syringaresinol (125.60 ± 6.76 mg/kg). The bioprocessing treatments significantly increased the extractability of gallic acid, vanillic acid, p-hydroxybenzoic acid, syringic acid, vanillin, syringaldehyde, p-coumaric acid, ferulic acid, caffeic acid, and syringaresinol in the alkaline-labile bound form, suggesting the bioaccessibility of these phytochemicals to the colon. Furthermore, one of the treatments, EC_2 treatment, increased significantly the in vitro upper gastrointestinal release of bioactive phytochemicals, especially for protocatechuic acid (p < 0.01). The BH fibre was fermentable, promoting the formation mainly of propionate and, to a lesser extent, butyrate formation. The EM_1 and EC_2 treatments effectively increased the content of insoluble fibre but had no effect on dietary fibre fermentation (p > 0.05). These findings promote the use of buckwheat hulls as a source of dietary fibre and phytochemicals to help meet dietary recommendations and needs.
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Fagopyrum , Humanos , Fagopyrum/metabolismo , Cromatografia Líquida , RNA Ribossômico 16S/metabolismo , Espectrometria de Massas em Tandem , Fibras na Dieta/metabolismo , Compostos Fitoquímicos/metabolismoRESUMO
Introduction Safe and effective management of agitated patients poses multiple challenges for healthcare professionals. Patients placed in restraints because of agitated behavior are at a higher risk of complications, including death. This intervention was designed to provide emergency department staff a framework for de-escalation, improve teamwork, and reduce the use of violent physical restraints. Methods Emergency medicine nurses, patient support associates, and protective services officers underwent a 90-minute educational intervention in 2017. A 30-minute lecture focusing on communication and early use of medication for agitation was followed by a simulation using standardized participants, then a structured debriefing. A standardized return-on-learning tool determined participants' reactions to and application of the educational intervention. Additionally, data was collected and reported as a ratio of number of restraints applied each month compared to total emergency department visits that month. Data were analyzed comparing the six months before the education and the subsequent six months after the education. Results A pilot group of 30 emergency department staff members completed the educational intervention. The intervention contributed to the overall decrease in restraint use in the department. Most participants (86%) felt more confident in their ability to manage agitated patients. Conclusion An interdisciplinary simulation-enhanced educational intervention successfully reduced use of restraints in the emergency department and improved staff attitudes toward de-escalation techniques for agitated patients.
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Small grain cereals are frequently infected with mycotoxigenic Fusarium fungi. Oats have a particularly high risk of contamination with type A trichothecene mycotoxins; their glucoside conjugates have also been reported. Agronomy practices, cereal variety and weather conditions have been suggested to play a role in Fusarium infection in oats. The current study investigates concentrations of free and conjugated Fusarium mycotoxins in organic and conventional oats grown in Scotland. In 2019, 33 milling oat samples (12 organic, 21 conventional) were collected from farmers across Scotland, together with sample questionnaires. Samples were analysed for 12 mycotoxins (type A trichothecenes T-2-toxin, HT-2-toxin, diacetoxyscirpenol; type B trichothecenes deoxynivalenol, nivalenol; zearalenone and their respective glucosides) using LC-MS/MS. The prevalence of type A trichothecenes T-2/HT-2 was very high (100% of conventional oats, 83% of organic oats), whereas type B trichothecenes were less prevalent, and zearalenone was rarely found. T-2-glucoside and deoxynivalenol-glucoside were the most prevalent conjugated mycotoxins (36 and 33%), and co-occurrence between type A and B trichothecenes were frequently observed (66% of samples). Organic oats were contaminated at significantly lower average concentrations than conventional oats, whereas the effect of weather parameters were not statistically significant. Our results clearly indicate that free and conjugated T-2- and HT-2-toxins pose a major risk to Scottish oat production and that organic production and crop rotation offer potential mitigation strategies.
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Fusarium , Micotoxinas , Toxina T-2 , Tricotecenos do Tipo B , Zearalenona , Micotoxinas/análise , Avena/microbiologia , Grão Comestível/química , Zearalenona/análise , Cromatografia Líquida , Contaminação de Alimentos/análise , Espectrometria de Massas em Tandem , Toxina T-2/análise , Escócia , GlucosídeosRESUMO
Only 6 to 8 % of the UK adults meet the daily recommendation for dietary fibre. Fava bean processing lead to vast amounts of high-fibre by-products such as hulls. Bean hull fortified bread was formulated to increase and diversify dietary fibre while reducing waste. This study assessed the bean hull: suitability as a source of dietary fibre; the systemic and microbial metabolism of its components and postprandial events following bean hull bread rolls. Nine healthy participants (53·9 ± 16·7 years) were recruited for a randomised controlled crossover study attending two 3 days intervention sessions, involving the consumption of two bread rolls per day (control or bean hull rolls). Blood and faecal samples were collected before and after each session and analysed for systemic and microbial metabolites of bread roll components using targeted LC-MS/MS and GC analysis. Satiety, gut hormones, glucose, insulin and gastric emptying biomarkers were also measured. Two bean hull rolls provided over 85 % of the daily recommendation for dietary fibre; but despite being a rich source of plant metabolites (P = 0·04 v. control bread), these had poor systemic bioavailability. Consumption of bean hull rolls for 3 days significantly increased plasma concentration of indole-3-propionic acid (P = 0·009) and decreased faecal concentration of putrescine (P = 0·035) and deoxycholic acid (P = 0·046). However, it had no effect on postprandial plasma gut hormones, bacterial composition and faecal short chain fatty acids amount. Therefore, bean hulls require further processing to improve their bioactives systemic availability and fibre fermentation.
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Fabaceae , Hormônios Gastrointestinais , Adulto , Humanos , Voluntários Saudáveis , Putrescina , Pão/análise , Cromatografia Líquida , Estudos Cross-Over , Espectrometria de Massas em Tandem , Fibras na Dieta/análise , Fabaceae/metabolismo , Ácido Desoxicólico , Glicemia/análiseRESUMO
INTRODUCTION: Hepatocellular adenomas are a rare but serious cause of bleeding, which is further complicated by pregnancy. Interprofessional cooperation is a key component of residency education, thus simulations designed to integrate multiple programs are mutually beneficial. This simulation details surgical and obstetric management of a pregnant patient in hemorrhagic shock from a bleeding hepatocellular adenoma. Objectives for the study were to evaluate learners' confidence to 1) prioritize the care of a pregnant patient with hemoperitoneum and hemorrhagic shock, 2) demonstrate interdisciplinary collaboration with other specialties, 3) apply massive transfusion protocol (MTP) in the appropriate clinical setting, and 4) analyze critical decisions for evaluating pregnant females with severe abdominal pain. METHODS: Obstetric, general surgery, and anesthesia residents, along with labor and delivery nurses participated in a simulated clinical scenario that focused on the management of a pregnant patient in hemorrhagic shock. The learners evaluated the educational session using a standard Return on Investment in Learning survey immediately following the session. RESULTS: A total of 23 residents and medical students gave feedback on the experience. The main learning objectives were met with increased confidence in the four learning objectives by 77.3-95.4% of responders. Overall, greater than 90% of participants felt the simulation was relevant to their training and realistic, with 100% responding that the course provided new, or clarified existing information for them. CONCLUSION: A multidisciplinary simulation-based educational intervention was successful in improving learner confidence in managing a complicated surgical emergency in a pregnant patient with inter-residency cooperation.
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OBJECTIVE: The novel coronavirus-19 (COVID-19) has taken an immense physical, social, and emotional toll on frontline healthcare workers. Research has documented higher levels of anxiety, depression, and burnout among healthcare workers during the pandemic. Thus, creative interventions are needed now more than ever to provide brief, accessible support to frontline workers. Virtual reality is a rapidly growing technology with potential psychological applications. In this study, we piloted a three-minute Tranquil Cinematic-VR simulation of a nature scene to lower subjective stress among frontline healthcare workers in COVID-19 treatment units. We chose to film a nature scene because of the extensive empirical literature documenting the benefits of nature exposure and health. METHODS: A convenience sample of frontline healthcare workers, including direct care providers, indirect care providers, and support or administrative services, were recruited from three COVID-19 units located in the United States. Inclusion criteria for participation included adults aged 18 years and older who could read and speak in English and were currently employed by the healthcare system. Participants viewed a 360-degree video capture of a lush, green nature preserve in an Oculus Go or Pico G2 4K head-mounted display. Prior to viewing the simulation, participants completed a brief demographic questionnaire and the visual analogue scale to rate their subjective stress on a 10-point scale, with 1 = 'Not at all stressed' to 10 = 'Extremely stressed.' We conducted paired t-tests to examine pre- and post-simulation changes in subjective stress as well as Kruskal-Wallis tests and Mann-Whitney U tests to examine differences by demographic variables. All analyses were conducted in SPSS statistical software version 28.0. We defined statistical significance as a p-value less than .05. RESULTS: A total of 102 individuals consented to participate in the study. Eighty-four (82.4%) participants reported providing direct patient care, 73 (71.6%) identified as women, 49 (48.0%) were between the ages of 25-34 years old, and 35 (34.3%) had prior experience with VR. The pre-simulation mean stress score was 5.5±2.2, with a range of 1 to 10. Thirty-three (32.4%) participants met the 6.8 cutoff for high stress pre-simulation. Pre-simulation stress scores did not differ by any demographic variables. Post-simulation, we observed a significant reduction in subjective stress scores from pre- to post-simulation (mean change = -2.2±1.7, t = 12.749, p < .001), with a Cohen's d of 1.08, indicating a very large effect. Further, only four (3.9%) participants met the cutoff for high stress after the simulation. Post-simulations scores did not differ by provider type, age range, gender, or prior experience with virtual reality. CONCLUSIONS: Findings from this pilot study suggest that the application of this Tranquil Cinematic-VR simulation was effective in reducing subjective stress among frontline healthcare workers in the short-term. More research is needed to compare the Tranquil Cinematic-VR simulation to a control condition and assess subjective and objective measures of stress over time.
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Esgotamento Profissional/terapia , COVID-19 , Pessoal de Saúde/psicologia , Realidade Virtual , Adulto , Ansiedade , Esgotamento Profissional/diagnóstico , COVID-19/epidemiologia , Terapias Complementares , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Adulto JovemRESUMO
Background Studies have demonstrated the use of resuscitative endovascular balloon occlusion catheters of the aorta (REBOA) in the setting of postpartum hemorrhage and traumatic hemorrhagic shock. However, REBOA is infrequently utilized leading to a lack of clinician comfort. This study's aim was to demonstrate the utility of REBOA in a hemorrhaging pregnant trauma patient and improve clinician comfort with the placement of REBOA while emphasizing collaboration between medical specialties. Methods A multidisciplinary in-situ simulation was developed for the management of a pregnant patient with an abdominal gunshot wound evaluated by obstetrics and surgery teams. A trauma survey, emergency c-section, massive transfusion protocol (MTP), and evaluation for and placement of REBOA were indicated during the simulation. A standardized Return on Learning questionnaire was utilized to determine participants' reactions and confidence gained during the simulation. Results A total of 32 of 41 participants completed the survey (78%). A statistically significant increase in confidence was reported in the ability to prioritize the care of a pregnant patient with hemorrhagic shock (p = 0.016), apply MTP to the appropriate clinical setting (p = 0.03), and analyze critical decisions made for abdominal trauma in pregnant patients (p = 0.006). Specifically for physicians, a significant increase in confidence in the ability to identify indications/contraindications for REBOA placement in hemorrhaging patients was observed (p = 0.021). Conclusions A multidisciplinary simulation for the management of a pregnant patient in hemorrhagic shock secondary to penetrating abdominal trauma improved learner confidence in MTP, care of pregnant patients in hemorrhagic shock, and abdominal trauma in pregnancy. Physician learners gained confidence in indications for REBOA placement in abdominal trauma. This simulation was highly relevant to all participants.
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PURPOSE: This study evaluated the postprandial effects following consumption of buckwheat, fava bean, pea, hemp and lupin compared to meat (beef); focussing on biomarkers of satiety, gut hormones, aminoacids and plant metabolites bioavailability and metabolism. METHODS: Ten subjects (n = 3 men; n = 7 women; 42 ± 11.8 years of age; BMI 26 ± 5.8 kg/m2) participated in six 1-day independent acute interventions, each meal containing 30 g of protein from buckwheat, fava bean, pea, hemp, lupin and meat (beef). Blood samples were collected during 24-h and VAS questionnaires over 5-h. RESULTS: Volunteers consumed significantly higher amounts of most amino acids from the meat meal, and with few exceptions, postprandial composition of plasma amino acids was not significantly different after consuming the plant-based meals. Buckwheat meal was the most satious (300 min hunger scores, p < 0.05).Significant increase in GLP-1 plasma (AUC, iAUC p = 0.01) found after hemp compared with the other plant-based meals. Decreased plasma ghrelin concentrations (iAUC p < 0.05) found on plant (hemp) vs. meat meal. Several plasma metabolites after hemp meal consumption were associated with hormone trends (partial least squares-discriminant analysis (PLS-DA): 4-hydroxyphenylpyruvic acid, indole 3-pyruvic acid, 5-hydoxytryptophan, genistein and biochanin A with GLP-1, PYY and insulin; 3-hydroxymandelic acid and luteolidin with GLP-1 and ghrelin and 4-hydroxymandelic acid, benzoic acid and secoisolariciresinol with insulin and ghrelin. Plasma branched-chain amino acids (BCAAs), (iAUC, p < 0.001); and phenylalanine and tyrosine (iAUC, p < 0.05) were lower after buckwheat comparison with meat meal. CONCLUSION: Plants are valuable sources of amino acids which are promoting satiety. The impact of hemp and buckwheat on GLP-1 and, respectively, BCAAs should be explored further as could be relevant for aid and prevention of chronic diseases such as type 2 diabetes. Study registered with clinicaltrial.gov on 12th July 2013, study ID number: NCT01898351.
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Cannabis , Diabetes Mellitus Tipo 2 , Fagopyrum , Hormônios Gastrointestinais , Aminoácidos , Glicemia/metabolismo , Cannabis/metabolismo , Estudos Cross-Over , Fagopyrum/metabolismo , Feminino , Grelina , Voluntários Saudáveis , Humanos , Insulina , Masculino , Refeições , Período Pós-PrandialRESUMO
NEW FINDINGS: What is the central question of this study? Does exercise affect vitamin D receptor expression in T lymphocytes in young, middle-aged and older adults? What is the main finding and its importance? Moderate-intensity cycling exercise increases vitamin D receptor expression in vitamin D-deficient men, independent of age, presenting a strategy to combat the prevalence of vitamin D deficiency. ABSTRACT: Vitamin D plays a key role in the modulation of the immune system, mediated through the intracellular vitamin D receptor (VDR). Exercise has been shown to influence the activity and availability of the VDR. The aim of this study was to investigate the effect of age on basal immune cell (T-lymphocyte) VDR expression and the subsequent effect of acute aerobic exercise to modulate VDR expression in peripheral T cells. Thirty-five men were included in the study (mean ± SD: age 44 ± 17 years and body mass index 25.7 ± 3.1 kg/m2 ), separated into three age groups: 18-30 (n = 12), 31-45 (n = 11) and 60-75 years (n = 12). Participants completed two trials [control (CON) and aerobic exercise (AE)], with blood samples collected pre- and postexercise (0, 1 and 3 h). Peripheral blood T cells were isolated and analysed for VDR expression by flow cytometry. The results show that advanced age is associated with lower VDR expression in T cells (882 ± 274, 796 ± 243 and 594 ± 174 geomean in the 18-30, 31-45 and 60-75 year age groups, respectively). Acute AE was successful at acutely increasing VDR expression in T cells, irrespective of age. Advanced age corresponds to a lower T-cell VDR expression, which might be responsible for age-associated development of chronic conditions and autoimmunity. Exercise was successful in increasing VDR expression in T cells irrespective of age and independent of exercise-induced T-cell mobilization.
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Exercício Físico , Receptores de Calcitriol , Deficiência de Vitamina D , Adolescente , Adulto , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Receptores de Calcitriol/metabolismo , Linfócitos T/metabolismo , Vitamina D/metabolismo , Deficiência de Vitamina D/metabolismo , Adulto JovemRESUMO
Background Agitated or aggressive patients pose a high risk of emotional and physical harm to hospital staff. Healthcare associates have the highest rate of workplace violence among studied fields. Learning to effectively de-escalate a patient who is a danger to self or others is key to reducing these incidents. This course was designed to improve education and communication among staff on a hospital surgical floor when verbal de-escalation is needed. Methods A ninety-minute simulation-based multidisciplinary curriculum was developed. This included a 30-minute didactic discussion, 10-minute simulation, and 50-minute debrief. Learners included nurses, patient service associates, and protective services officers from a medical/surgical unit. Data were collected using a validated return on investment in learning protocol and the Management of Aggression and Violence Attitude Scale (MAVAS) tool. Results Our return on investment in learning showed that more than 97% of learners felt safer in managing agitated patients after participating in the training. The MAVAS tool was used in pre- and post-format and showed a significant trend toward the importance of clear communication and role clarity when de-escalating a patient. Conclusions A combination of didactic teaching and simulated experience allowed for greater confidence, communication, and teamwork in de-escalating an agitated patient in a community hospital surgical unit.
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Mycotoxins are important food contaminants that commonly co-occur with modified mycotoxins such as mycotoxin-glucosides in contaminated cereal grains. These masked mycotoxins are less toxic, but their breakdown and release of unconjugated mycotoxins has been shown by mixed gut microbiota of humans and animals. The role of different bacteria in hydrolysing mycotoxin-glucosides is unknown, and this study therefore investigated fourteen strains of human gut bacteria for their ability to break down masked mycotoxins. Individual bacterial strains were incubated anaerobically with masked mycotoxins (deoxynivalenol-3-ß-glucoside, DON-Glc; nivalenol-3-ß-glucoside, NIV-Glc; HT-2-ß-glucoside, HT-2-Glc; diacetoxyscirpenol-α-glucoside, DAS-Glc), or unconjugated mycotoxins (DON, NIV, HT-2, T-2, and DAS) for up to 48 h. Bacterial growth, hydrolysis of mycotoxin-glucosides and further metabolism of mycotoxins were assessed. We found no impact of any mycotoxin on bacterial growth. We have demonstrated that Butyrivibrio fibrisolvens, Roseburia intestinalis and Eubacterium rectale hydrolyse DON-Glc, HT-2 Glc, and NIV-Glc efficiently and have confirmed this activity in Bifidobacterium adolescentis and Lactiplantibacillus plantarum (DON-Glc only). Prevotella copri and B. fibrisolvens efficiently de-acetylated T-2 and DAS, but none of the bacteria were capable of de-epoxydation or hydrolysis of α-glucosides. In summary we have identified key bacteria involved in hydrolysing mycotoxin-glucosides and de-acetylating type A trichothecenes in the human gut.
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Bactérias/enzimologia , Proteínas de Bactérias/metabolismo , Microbioma Gastrointestinal , Intestinos/microbiologia , Micotoxinas/metabolismo , Acetilação , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Microbiologia de Alimentos , Glucosídeos/metabolismo , Humanos , Hidrólise , Micotoxinas/efeitos adversos , Medição de Risco , Especificidade por Substrato , Fatores de Tempo , Tricotecenos/metabolismoRESUMO
Benefits from micronutrients within fruit juice and smoothies are well documented, but fewer studies research the role of phytochemicals. Well-controlled human studies are essential to evaluate their impact, particularly on glucose and lipid regulation but also gastrointestinal health. Planning these studies requires data on the potential molecular targets. Here we report a comprehensive metabolomic (LC-MS) analysis of the phytochemical composition of four commonly consumed beverages, including data on whether they are free to be absorbed early in the gastrointestinal tract or bound to other plant components. Smoothies contained a wide range of phenolics (free and bound), whereas the fruit juices contained higher amounts of fewer compounds. Orange juice was rich in bound hesperidin (1.97 ± 0.39 mg/100 mL) and hydroxycinnamic acids, likely to be delivered to the colon with the potential to have an impact on gut health. Apple juice contained free chlorogenic acid (3.11 ± 1.03 mg/100 mL), phloridzin (0.40 ± 0.03 mg/100 mL), catechin (0.090 ± 0.005 mg/100 mL), and epicatechin (0.38 ± 0.02 mg/100 mL), suggesting potential roles in glucose uptake reduction or positive effects on systemic blood flow. Redox screening established that differences in chemical composition impacted on bioactivity, highlighting the importance of availability from the matrix. This suggests that fruit-based beverage interventions should target specific mechanisms depending on the fruits from which they are comprised and in particular, the availability of the individual constituents.
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Legumes are a source of health-promoting macro- and micronutrients, but also contain numerous phytochemicals with useful biological activities, an example of which are saponins. Epidemiological studies suggest that saponins may play a role in protection from cancer and benefit human health by lowering cholesterol. Therefore, they could represent good candidates for specialised functional foods. Following the consumption of a soya-rich high-protein weight-loss diet (SOYA HP WL), the concentrations of Soyasaponin I (SSI) and soyasapogenol B (SSB) were determined in faecal samples from human volunteers (n = 10) and found to be between 1.4 and 17.5 mg per 100 g fresh faecal sample. SSB was the major metabolite identified in volunteers' plasma (n = 10) after consumption of the soya test meal (SOYA MEAL); the postprandial (3 h after meal) plasma concentration for SSB varied between 48.5 ng/mL to 103.2 ng/mL. The metabolism of SSI by the gut microbiota (in vitro) was also confirmed. This study shows that the main systemic metabolites of soyasaponin are absorbed from the gut and that they are bioavailable in plasma predominantly as conjugates of sapogenol. The metabolism and bioavailability of biologically active molecules represent key information necessary for the efficient development of functional foods.
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The hypoglycaemic effects of two Ribes sp. i.e., anthocyanin-rich black currants (BC) were compared to green currants (GC), which are low in anthocyanins to establish which compounds are involved in the regulation of postprandial glycaemia. We determined the effect of the currants on inhibiting carbohydrate digestive enzymes (α-amylase, α-glucosidase), intestinal sugar absorption and transport across CaCo-2 cells. The digestion of these currants was modelled using in vitro gastrointestinal digestion (IVGD) to identify the metabolites present in the digested extracts by LC-MS/MS. Freeze-dried BC and IVDG extracts inhibited yeast α-glucosidase activity (P<.0001) at lower concentrations than acarbose, whereas GC and IVDG GC at the same concentrations showed no inhibition. BC and GC both showed significant inhibitory effects on salivary α-amylase (P<.0001), glucose uptake (P<.0001) and the mRNA expression of sugar transporters (P<.0001). Taken together this suggests that the anthocyanins which are high in BC have their greatest effect on postprandial hyperglycaemia by inhibiting α-glucosidase activity. Phytochemical analysis identified the phenolics in the currants and confirmed that freeze-dried BC contained higher concentrations of anthocyanins compared to GC (39.80 vs. 9.85 g/kg dry weight). Specific phenolics were also shown to inhibit salivary α-amylase, α-glucosidase, and glucose uptake. However, specific anthocyanins identified in BC which were low in GC were shown to inhibit α-glucosidase. In conclusion the anthocyanins in BC appear to regulate postprandial hyperglycaemia primarily but not solely by inhibiting α-glucosidase while other phenolics modulate salivary α-amylase, glucose uptake and sugar transporters which together could lower the associated risk of developing type-2 diabetes.
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Antocianinas/química , Glucose/metabolismo , Inibidores de Glicosídeo Hidrolases/farmacologia , Hiperglicemia/metabolismo , Ribes/química , alfa-Amilases Salivares/metabolismo , Glicemia/análise , Células CACO-2 , Sobrevivência Celular , Cromatografia Líquida , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Frutas/química , Trato Gastrointestinal/metabolismo , Humanos , Fenol/química , Fenol/metabolismo , Fenóis/química , Período Pós-Prandial , Açúcares/química , Espectrometria de Massas em Tandem , alfa-Glucosidases/metabolismoRESUMO
Fusarium mycotoxins are common contaminants in cereals and often co-occur with plant-derived mycotoxin sugar conjugates. Several of these modified mycotoxins are not degraded in the small intestine and hence carried through to the large intestine where microbial transformation may occur. This study aims to assess the gastrointestinal stability of the trichothecenes HT-2 toxin (HT-2), HT-2-ß-glucoside (HT-2-Glc), diacetoxyscirpenol (DAS), DAS-α-glucoside (DAS-Glc) and fumonisin B1 (FB1), N-(1-deoxy-d-fructos-1-yl) fumonisin-B1 (NDF-FB1). All tested modified mycotoxins were stable under upper gastrointestinal (GI) conditions. In faecal batch culture experiments, HT-2-Glc was hydrolysed efficiently and no further microbial biotransformation of HT-2 was observed. DAS-Glc hydrolysis was slow and DAS was de-acetylated to 15-monoacetoxyscripenol. NDF-FB1 was hydrolysed at the slowest rate and FB1 accumulation varied between donor samples. Our results demonstrate that all tested modified mycotoxins are stable in the upper GI tract and efficiently hydrolysed by human gut microbiota, thus potentially contributing to colonic toxicity. Hence the microbial biotransformation of any novel modified mycotoxins needs to be carefully evaluated.
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Grão Comestível/química , Fumonisinas/metabolismo , Fusarium , Microbioma Gastrointestinal , Glucosídeos/metabolismo , Intestino Grosso , Tricotecenos/metabolismo , Adulto , Biotransformação , Feminino , Contaminação de Alimentos , Trânsito Gastrointestinal , Humanos , Hidrólise , Intestino Grosso/metabolismo , Intestino Grosso/microbiologia , Intestino Delgado/metabolismo , Masculino , Micotoxinas Mascaradas/metabolismo , Pessoa de Meia-Idade , Micotoxinas/metabolismo , Poaceae , Toxina T-2/análogos & derivados , Toxina T-2/metabolismo , Trato Gastrointestinal Superior/metabolismoRESUMO
Cereal foods are commonly contaminated with multiple mycotoxins resulting in frequent human mycotoxin exposure. Children are at risk of high-level exposure because of their high cereal intake relative to body weight. Hence, this study aims to assess multimycotoxin exposure in UK children using urinary biomarkers. Spot urines (n = 21) were analyzed for multimycotoxins (deoxynivalenol, DON; nivalenol, NIV; ochratoxin A, OTA; zearalenone, ZEN; α-zearalenol, α-ZEL; ß-zearalenol, ß-ZEL; T-2 toxin, T-2; HT-2 toxin, HT-2; and aflatoxin B1 and M1, AFB1, AFM1) using liquid chromatography-coupled tandem mass spectrometry. Urine samples frequently contained DON (13.10 ± 12.69 ng/mL), NIV (0.36 ± 0.16 ng/mL), OTA (0.05 ± 0.02 ng/mL), and ZEN (0.09 ± 0.07 ng/mL). Some samples (1-3) contained T-2, HT-2, α-ZEL, and ß-ZEL but not aflatoxins. Dietary mycotoxin estimation showed that children were frequently exposed to levels exceeding the tolerable daily intake (52 and 95% of cases for DON and OTA). This demonstrates that UK children are exposed to multiple mycotoxins through their habitual diet.
Assuntos
Biomarcadores/urina , Micotoxinas/urina , Aflatoxinas/urina , Criança , Pré-Escolar , Dieta/efeitos adversos , Exposição Ambiental/efeitos adversos , Feminino , Contaminação de Alimentos/estatística & dados numéricos , Humanos , Masculino , Ocratoxinas/urina , Inquéritos e Questionários , Toxina T-2/análogos & derivados , Toxina T-2/urina , Tricotecenos/urina , Reino Unido , Zearalenona/urina , Zeranol/análogos & derivados , Zeranol/urinaRESUMO
Using simple solvent extraction and enzymatic hydrolysis, a rapid LC-MS/MS method for quantification of free and conjugated forms of anthocyanidins and anthocyanins in plasma and urine samples was developed and validated. A mixed enzymatic treatment containing ß-glucuronidase (100â¯Uâ¯mL-1) and sulfatase (2.5â¯Uâ¯mL-1) for 5â¯min (37⯰C; pH 6) was optimal condition for deconjugation of anthocyanidins and anthocyanins in urine and plasma samples. The LC-MS/MS allowed quantifying thirteen different anthocyanidins and anthocyanins simultaneously. The developed LC-MS/MS method was precise and accurate over multiple days and nominal concentrations. The stability assessment study confirmed that the long-term storage and/or periodic use of plasma and urine samples might have a considerable impact on the stability of some anthocyanidins. The method was successfully applied to measure anthocyanidins and anthocyanins in plasma and urine samples following consumption of acute blueberry test meals.
Assuntos
Antocianinas/sangue , Antocianinas/urina , Análise Química do Sangue/métodos , Urinálise/métodos , Mirtilos Azuis (Planta)/química , Cromatografia Líquida , Humanos , Espectrometria de Massas em Tandem , Fatores de TempoRESUMO
Modeling disease with human pluripotent stem cells (hPSCs) is hindered because the impact on cell phenotype from genetic variability between individuals can be greater than from the pathogenic mutation. While "footprint-free" Cas9/CRISPR editing solves this issue, existing approaches are inefficient or lengthy. In this study, a simplified PiggyBac strategy shortened hPSC editing by 2 weeks and required one round of clonal expansion and genotyping rather than two, with similar efficiencies to the longer conventional process. Success was shown across four cardiac-associated loci (ADRB2, GRK5, RYR2, and ACTC1) by genomic cleavage and editing efficiencies of 8%-93% and 8%-67%, respectively, including mono- and/or biallelic events. Pluripotency was retained, as was differentiation into high-purity cardiomyocytes (CMs; 88%-99%). Using the GRK5 isogenic lines as an exemplar, chronic stimulation with the ß-adrenoceptor agonist, isoprenaline, reduced beat rate in hPSC-CMs expressing GRK5-Q41 but not GRK5-L41; this was reversed by the ß-blocker, propranolol. This shortened, footprint-free approach will be useful for mechanistic studies.
Assuntos
Sistemas CRISPR-Cas/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Células-Tronco Pluripotentes Induzidas/citologia , Miócitos Cardíacos/citologia , Diferenciação Celular/genética , Linhagem Celular , Edição de Genes/métodos , HumanosRESUMO
Mycotoxin contamination of cereal grains causes well-recognized toxicities in animals and humans, but the fate of plant-bound masked mycotoxins in the gut is less well understood. Masked mycotoxins have been found to be stable under conditions prevailing in the small intestine but are rapidly hydrolyzed by fecal microbiota. This study aims to assess the hydrolysis of the masked mycotoxin deoxynivalenol-3-glucoside (DON3Glc) by the microbiota of different regions of the porcine intestinal tract. Intestinal digesta samples were collected from the jejunum, ileum, cecum, colon, and feces of 5 pigs and immediately frozen under anaerobic conditions. Sample slurries were prepared in M2 culture medium, spiked with DON3Glc or free deoxynivalenol (DON; 2 nmol/ml), and incubated anaerobically for up to 72 h. Mycotoxin concentrations were determined using liquid chromatography-tandem mass spectrometry, and the microbiota composition was determined using a quantitative PCR methodology. The jejunal microbiota hydrolyzed DON3Glc very slowly, while samples from the ileum, cecum, colon, and feces rapidly and efficiently hydrolyzed DON3Glc. No further metabolism of DON was observed in any sample. The microbial load and microbiota composition in the ileum were significantly different from those in the distal intestinal regions, whereas those in the cecum, colon and feces did not differ.IMPORTANCE Results from this study clearly demonstrate that the masked mycotoxin DON3Glc is hydrolyzed efficiently in the distal small intestine and large intestine of pigs. Once DON is released, toxicity and absorption in the distal intestinal tract likely occur in vivo This study further supports the need to include masked metabolites in mycotoxin risk assessments and regulatory actions for feed and food.