Psychological and social interventions for the promotion of mental health in people living in low- and middle-income countries affected by humanitarian crises.

BACKGROUND: Because of wars, conflicts, persecutions, human rights violations, and humanitarian crises, about 84 million people are forcibly displaced around the world; the great majority of them live in low- and middle-income countries (LMICs). People living in humanitarian settings are affected by a constellation of stressors that threaten their mental health. Psychosocial interventions for people affected by humanitarian crises may be helpful to promote positive aspects of mental health, such as mental well-being, psychosocial functioning, coping, and quality of life. Previous reviews have focused on treatment and mixed promotion and prevention interventions. In this review, we focused on promotion of positive aspects of mental health. OBJECTIVES: To assess the effects of psychosocial interventions aimed at promoting mental health versus control conditions (no intervention, intervention as usual, or waiting list) in people living in LMICs affected by humanitarian crises. SEARCH METHODS: We searched CENTRAL, MEDLINE, Embase, and seven other databases to January 2023. We also searched the World Health Organization's (WHO) International Clinical Trials Registry Platform and ClinicalTrials.gov to identify unpublished or ongoing studies, and checked the reference lists of relevant studies and reviews. SELECTION CRITERIA: Randomised controlled trials (RCTs) comparing psychosocial interventions versus control conditions (no intervention, intervention as usual, or waiting list) to promote positive aspects of mental health in adults and children living in LMICs affected by humanitarian crises. We excluded studies that enrolled participants based on a positive diagnosis of mental disorder (or based on a proxy of scoring above a cut-off score on a screening measure). DATA COLLECTION AND ANALYSIS: We used standard Cochrane methods. Our primary outcomes were mental well-being, functioning, quality of life, resilience, coping, hope, and prosocial behaviour. The secondary outcome was acceptability, defined as the number of participants who dropped out of the trial for any reason. We used GRADE to assess the certainty of evidence for the outcomes of mental well-being, functioning, and prosocial behaviour. MAIN RESULTS: We included 13 RCTs with 7917 participants. Nine RCTs were conducted on children/adolescents, and four on adults. All included interventions were delivered to groups of participants, mainly by paraprofessionals. Paraprofessional is defined as an individual who is not a mental or behavioural health service professional, but works at the first stage of contact with people who are seeking mental health care. Four RCTs were carried out in Lebanon; two in India; and single RCTs in the Democratic Republic of the Congo, Jordan, Haiti, Bosnia and Herzegovina, the occupied Palestinian Territories (oPT), Nepal, and Tanzania. The mean study duration was 18 weeks (minimum 10, maximum 32 weeks). Trials were generally funded by grants from academic institutions or non-governmental organisations. For children and adolescents, there was no clear difference between psychosocial interventions and control conditions in improving mental well-being and prosocial behaviour at study endpoint (mental well-being: standardised mean difference (SMD) 0.06, 95% confidence interval (CI) -0.17 to 0.29; 3 RCTs, 3378 participants; very low-certainty evidence; prosocial behaviour: SMD -0.25, 95% CI -0.60 to 0.10; 5 RCTs, 1633 participants; low-certainty evidence), or at medium-term follow-up (mental well-being: mean difference (MD) -0.70, 95% CI -2.39 to 0.99; 1 RCT, 258 participants; prosocial behaviour: SMD -0.48, 95% CI -1.80 to 0.83; 2 RCT, 483 participants; both very low-certainty evidence). Interventions may improve functioning (MD -2.18, 95% CI -3.86 to -0.50; 1 RCT, 183 participants), with sustained effects at follow-up (MD -3.33, 95% CI -5.03 to -1.63; 1 RCT, 183 participants), but evidence is very uncertain as the data came from one RCT (both very low-certainty evidence). Psychosocial interventions may improve mental well-being slightly in adults at study endpoint (SMD -0.29, 95% CI -0.44 to -0.14; 3 RCTs, 674 participants; low-certainty evidence), but they may have little to no effect at follow-up, as the evidence is uncertain and future RCTs might either confirm or disprove this finding. No RCTs measured the outcomes of functioning and prosocial behaviour in adults. AUTHORS' CONCLUSIONS: To date, there is scant and inconclusive randomised evidence on the potential benefits of psychological and social interventions to promote mental health in people living in LMICs affected by humanitarian crises. Confidence in the findings is hampered by the scarcity of studies included in the review, the small number of participants analysed, the risk of bias in the studies, and the substantial level of heterogeneity. Evidence on the efficacy of interventions on positive mental health outcomes is too scant to determine firm practice and policy implications. This review has identified a large gap between what is known and what still needs to be addressed in the research area of mental health promotion in humanitarian settings.

Mycotoxins' Prevalence in Food Industry By-Products: A Systematic Review.

The recovery of biomolecules from food industry by-products is of major relevance for a circular economy strategy. However, by-products' contamination with mycotoxins represents a drawback for their reliable valorization for food and feed, hampering their application range, especially as food ingredients. Mycotoxin contamination occurs even in dried matrices. There is a need for the implantation of monitoring programs, even for by-products used as animal feed, since very high levels can be reached. This systematic review aims to identify the food by-products that have been studied from 2000 until 2022 (22 years) concerning mycotoxins' contamination, distribution, and prevalence in those by-products. PRISMA ("Preferred Reporting Items for Systematic Reviews and MetaAnalyses") protocol was performed via two databases (PubMed and SCOPUS) to summarize the research findings. After the screening and selection process, the full texts of eligible articles (32 studies) were evaluated, and data from 16 studies were considered. A total of 6 by-products were assessed concerning mycotoxin content; these include distiller dried grain with solubles, brewer's spent grain, brewer's spent yeast, cocoa shell, grape pomace, and sugar beet pulp. Frequent mycotoxins in these by-products are AFB1, OTA, FBs, DON, and ZEA. The high prevalence of contaminated samples, which surpasses the limits established for human consumption, thus limiting their valorization as ingredients in the food industry. Co-contamination is frequent, which can cause synergistic interactions and amplify their toxicity.

Drug Repurposing for Cystic Fibrosis: Identification of Drugs That Induce CFTR-Independent Fluid Secretion in Nasal Organoids.

Individuals with cystic fibrosis (CF) suffer from severe respiratory disease due to a genetic defect in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which impairs airway epithelial ion and fluid secretion. New CFTR modulators that restore mutant CFTR function have been recently approved for a large group of people with CF (pwCF), but ~19% of pwCF cannot benefit from CFTR modulators Restoration of epithelial fluid secretion through non-CFTR pathways might be an effective treatment for all pwCF. Here, we developed a medium-throughput 384-well screening assay using nasal CF airway epithelial organoids, with the aim to repurpose FDA-approved drugs as modulators of non-CFTR-dependent epithelial fluid secretion. From a ~1400 FDA-approved drug library, we identified and validated 12 FDA-approved drugs that induced CFTR-independent fluid secretion. Among the hits were several cAMP-mediating drugs, including ß2-adrenergic agonists. The hits displayed no effects on chloride conductance measured in the Ussing chamber, and fluid secretion was not affected by TMEM16A, as demonstrated by knockout (KO) experiments in primary nasal epithelial cells. Altogether, our results demonstrate the use of primary nasal airway cells for medium-scale drug screening, target validation with a highly efficient protocol for generating CRISPR-Cas9 KO cells and identification of compounds which induce fluid secretion in a CFTR- and TMEM16A-indepent manner.

Fat Oxidation of Fatty Fish vs. Meat Meal Diets Under in vitro Standardized Semi-Dynamic Gastric Digestion.

Meat and fish are introduced into the diet as a source of protein, but these muscle foods present different fatty acid (FA) compositions and different lipid stabilities. Fatty fish is expected to oxidize due to its higher content of polyunsaturated FA (PUFA), whereas the higher heme-Fe content of red meat will also affect lipid stability. Combining other food ingredients within a meal also influences lipid oxidation, which will not stop after meals intake. This is due to the acidic environment of the stomach together with the presence of metallic ions, a process that is scarcely understood. The goal of this study was to evaluate the oxidation of fatty fish vs. meat meal diets under in vitro standardized semi-dynamic gastric conditions and FA release from the stomach to the duodenum. Meal diets composed by 25% beef meal (BM) or fatty fish meal (FM), 25% fried potatoes, and 50% sugar soft drink were prepared. Proximate composition, FA and amino acid profiles, and meals quality indices were evaluated. Their differences in composition led to different total gastric digestion time of 242.74 (BM) and 175.20 (FM) minutes. Using the INFOGEST semi-dynamic gastric model, 4 gastric emptying (GE) were simulated in both meals. In each GE, FA profile and lipid oxidation products (LOPs) formation were assessed. As a result, more than 50% FA release to the duodenum occurred in GE1, whose percentage decreased with the time of digestion. FM exhibited the highest LOPs formation, which corroborates the high peroxidizability index measured for this meal diet. Higher LOPs formation occurred in the later GEs, which released less FA. This suggests that higher times of residence in the stomach increase FA oxidation. This study shows a higher formation of LOPs during digestion of FM using a whole meal approach. These results relate to its richness in PUFAs compared to BM. Despite higher LOPs formation, FM digests that reached duodenum still contain higher content of unoxidized PUFAs compared with BM and a desirable ω3/ω6 PUFAs ratio of ~0.43. LOPs formation in PUFA-rich meals could be reduced if those meals have a low caloric value, avoiding large times of residence in the stomach and consequently high levels of oxidation.

Exploring YAP1-centered networks linking dysfunctional CFTR to epithelial-mesenchymal transition.

Mutations in the CFTR anion channel cause cystic fibrosis (CF) and have also been related to higher cancer incidence. Previously we proposed that this is linked to an emerging role of functional CFTR in protecting against epithelial-mesenchymal transition (EMT). However, the pathways bridging dysfunctional CFTR to EMT remain elusive. Here, we applied systems biology to address this question. Our data show that YAP1 is aberrantly active in the presence of mutant CFTR, interacting with F508del, but not with wt-CFTR, and that YAP1 knockdown rescues F508del-CFTR processing and function. Subsequent analysis of YAP1 interactors and roles in cells expressing either wt- or F508del-CFTR reveal that YAP1 is an important mediator of the fibrotic/EMT processes in CF. Alongside, five main pathways emerge here as key in linking mutant CFTR to EMT, namely, (1) the Hippo pathway; (2) the Wnt pathway; (3) the TGFß pathway; (4) the p53 pathway; and (5) MYC signaling. Several potential hub proteins which mediate the crosstalk among these pathways were also identified, appearing as potential therapeutic targets for both CF and cancer.

CFTR interactome mapping using the mammalian membrane two-hybrid high-throughput screening system.

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride and bicarbonate channel in secretory epithelia with a critical role in maintaining fluid homeostasis. Mutations in CFTR are associated with Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder in Caucasians. While remarkable treatment advances have been made recently in the form of modulator drugs directly rescuing CFTR dysfunction, there is still considerable scope for improvement of therapeutic effectiveness. Here, we report the application of a high-throughput screening variant of the Mammalian Membrane Two-Hybrid (MaMTH-HTS) to map the protein-protein interactions of wild-type (wt) and mutant CFTR (F508del), in an effort to better understand CF cellular effects and identify new drug targets for patient-specific treatments. Combined with functional validation in multiple disease models, we have uncovered candidate proteins with potential roles in CFTR function/CF pathophysiology, including Fibrinogen Like 2 (FGL2), which we demonstrate in patient-derived intestinal organoids has a significant effect on CFTR functional expression.

Systems Approaches to Unravel Molecular Function: High-content siRNA Screen Identifies TMEM16A Traffic Regulators as Potential Drug Targets for Cystic Fibrosis.

An attractive approach to treat people with Cystic Fibrosis (CF), a life-shortening disease caused by mutant CFTR, is to compensate for the absence of this chloride/bicarbonate channel by activating alternative (non-CFTR) chloride channels. One obvious target for such "mutation-agnostic" therapeutic approach is TMEM16A (anoctamin-1/ANO1), a calcium-activated chloride channel (CaCC) which is also expressed in the airways of people with CF, albeit at low levels. To find novel TMEM16A regulators of both traffic and function, with the main goal of identifying candidate CF drug targets, we performed a fluorescence cell-based high-throughput siRNA microscopy screen for TMEM16A trafficking using a double-tagged construct expressed in human airway cells. About 700 genes were screened (2 siRNAs per gene) of which 262 were identified as candidate TMEM16A modulators (179 siRNAs enhanced and 83 decreased TMEM16A traffic), being G-protein coupled receptors (GPCRs) enriched on the primary hit list. Among the 179 TMEM16A traffic enhancer siRNAs subjected to secondary screening 20 were functionally validated. Further hit validation revealed that siRNAs targeting two GPCRs - ADRA2C and CXCR3 - increased TMEM16A-mediated chloride secretion in human airway cells, while their overexpression strongly diminished calcium-activated chloride currents in the same cell model. The knockdown, and likely also the inhibition, of these two TMEM16A modulators is therefore an attractive potential therapeutic strategy to increase chloride secretion in CF.

In vitro gastric bioaccessibility of avocado peel extract in beef and soy-based burgers and its impact on Helicobacter pylori risk factors.

The objective of the present study was to investigate the impact of phenolic-rich avocado peel extract (APE) as an ingredient in beef and soy-based burgers to increase their antioxidant activity, reduce lipid and protein oxidation during gastric digestion, and inhibit urease and anhydrase carbonic activity, which are considered as key factors in the main steps of Helicobacter pylori adhesion in the stomach. The gastric bioaccessible fraction of soy and beef burgers with added 0.5% APE obtained by in vitro digestion exhibited a higher content of phenolic compounds, including monomeric and oligomeric (epi)catechin forms and quercetin, and reduced levels of thiobarbituric acid-reactive substances (TBARS) and carbonyls (49% to 73% and 57% to 60%, respectively) when compared with control burgers. Moreover, the burgers with APE inhibited urease and carbonic anhydrase activity. Results generally showed that including APE reduces the primary risk factors associated with H. pylori infection.

Synergy in Cystic Fibrosis Therapies: Targeting SLC26A9.

SLC26A9, a constitutively active Cl- transporter, has gained interest over the past years as a relevant disease modifier in several respiratory disorders including Cystic Fibrosis (CF), asthma, and non-CF bronchiectasis. SLC26A9 contributes to epithelial Cl- secretion, thus preventing mucus obstruction under inflammatory conditions. Additionally, SLC26A9 was identified as a CF gene modifier, and its polymorphisms were shown to correlate with the response to drugs modulating CFTR, the defective protein in CF. Here, we aimed to investigate the relationship between SLC26A9 and CFTR, and its role in CF pathogenesis. Our data show that SLC26A9 expression contributes to enhanced CFTR expression and function. While knocking-down SLC26A9 in human bronchial cells leads to lower wt- and F508del-CFTR expression, function, and response to CFTR correctors, the opposite occurs upon its overexpression, highlighting SLC26A9 relevance for CF. Accordingly, F508del-CFTR rescue by the most efficient correctors available is further enhanced by increasing SLC26A9 expression. Interestingly, SLC26A9 overexpression does not increase the PM expression of non-F508del CFTR traffic mutants, namely those unresponsive to corrector drugs. Altogether, our data indicate that SLC26A9 stabilizes CFTR at the ER level and that the efficacy of CFTR modulator drugs may be further enhanced by increasing its expression.

Explore Gastric Lipolysis and Lipid Oxidation of Conventional versus Pasture-Based Milk by a Semi-dynamic In Vitro Digestion Model.

Research on gastric lipolysis of commercial cow's milk with different fatty acid (FA) compositions is scarce. Gastric lipase exhibits specificity for the sn-3 chain position of triacylglycerols, whose structure is influenced by milk FA composition. Therefore, during gastric digestion of conventional (C) vs pasture-based (P) milk, differences may occur on lipolysis, which has impact on free FA available, influencing their absorption/metabolism rate and physiological hormonal responses. Those two milk types were subjected to the INFOGEST semi-dynamic digestion model. Five gastric emptying points were analyzed for oxidative degradation of polyunsaturated fatty acids (PUFA) and individual free FA. The relative release of medium-chain FA (C8:0-C12:0) was higher than that of longer-chain FA (C14:0-C18:0), and a linear increase in markers of PUFA oxidative degradation occurred along gastric digestion. Quantitatively, C8:0, C18:2n-6, C18:3n-3, and CLAc9t11 were higher (P < 0.001) in P milk when compared with C milk.

Incorporation of avocado peel extract to reduce cooking-induced hazards in beef and soy burgers: A clean label ingredient.

In this study, the benefits of using avocado peel extract (APE), rich in phenolic compounds, to reduce the oxidation and formation of harmful compounds resulting from cooking, were investigated. Beef and soy-based burgers with the addition of APE (0.5% and 1%) were studied after pan frying concerning proximate and physicochemical characteristics, inhibition of protein and lipid oxidation products (thiobarbituric acid reactive substances [TBARS], hexanal, and carbonyls), heterocyclic aromatic amines (HAs) and acrylamide formation. Sensory analysis was additionally performed. APE-affected proximate composition, protein, fat, and ash contents (%) were found to be markedly higher in APE-incorporated burgers (~28.32 ± 0.29, ~14.00 ± 0.01, and ~1.57 ± 0.05, respectively), compared with the controls (~26.55 ± 0.51, ~12.77 ± 0.32, and ~1.48 ± 0.16, respectively). Lower concentrations of TBARS, hexanal, and carbonyls were observed in APE-treated burgers on Days 1 and 10, post-cooking, compared to controls. Overall, it was found that APE had a greater protective effect than the positive control (sodium ascorbate incorporated) in beef burgers. In soy burgers, the positive control demonstrated pro-oxidant activity. The addition of 0.5% APE was found to inhibit HAs and acrylamide formation in beef and soy burgers. Although the addition of APE affected the color of both meat and soy burgers, it did not impact consumer preference. It was therefore concluded that APE extract might be a suitable clean-label alternative to synthetic antioxidants, and that it can protect and increase the nutritional value of meat and meat-free burgers.

Pharmacological Modulation of Ion Channels for the Treatment of Cystic Fibrosis.

Cystic fibrosis (CF) is a life-shortening monogenic disease caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR) protein, an anion channel that transports chloride and bicarbonate across epithelia. Despite clinical progress in delaying disease progression with symptomatic therapies, these individuals still develop various chronic complications in lungs and other organs, which significantly restricts their life expectancy and quality of life. The development of high-throughput assays to screen drug-like compound libraries have enabled the discovery of highly effective CFTR modulator therapies. These novel therapies target the primary defect underlying CF and are now approved for clinical use for individuals with specific CF genotypes. However, the clinically approved modulators only partially reverse CFTR dysfunction and there is still a considerable number of individuals with CF carrying rare CFTR mutations who remain without any effective CFTR modulator therapy. Accordingly, additional efforts have been pursued to identify novel and more potent CFTR modulators that may benefit a larger CF population. The use of ex vivo individual-derived specimens has also become a powerful tool to evaluate novel drugs and predict their effectiveness in a personalized medicine approach. In addition to CFTR modulators, pro-drugs aiming at modulating alternative ion channels/transporters are under development to compensate for the lack of CFTR function. These therapies may restore normal mucociliary clearance through a mutation-agnostic approach (ie, independent of CFTR mutation) and include inhibitors of the epithelial sodium channel (ENaC), modulators of the calcium-activated channel transmembrane 16A (TMEM16, or anoctamin 1) or of the solute carrier family 26A member 9 (SLC26A9), and anionophores. The present review focuses on recent progress and challenges for the development of ion channel/transporter-modulating drugs for the treatment of CF.

Western Dietary Pattern Antioxidant Intakes and Oxidative Stress: Importance During the SARS-CoV-2/COVID-19 Pandemic.

The importance of balanced dietary habits, which include appropriate amounts of antioxidants to maintain the immune system, has become increasingly relevant during the current SARS-CoV-2/COVID-19 pandemic, because viral infections are characterized by high oxidative stress. Furthermore, the measures taken by governments to control the pandemic have led to increased anxiety, stress, and depression, which affect physical and mental health, all of which are influenced by nutritional status, diet, and lifestyle. The Mediterranean diet (MD), Atlantic diet (AD), and the Dietary Guidelines for Americans all provide the essential vitamins, minerals, and phenolic compounds needed to activate enzymatic and nonenzymatic antioxidant responses. However, viral pandemics such as the current COVID-19 crisis entail high oxidative damage caused by both the infection and the resultant social stresses within populations, which increases the probability and severity of infection. Balanced dietary patterns such as the MD and the AD are characterized by the consumption of fruit, vegetables, legumes, olive oil, and whole grains with low intakes of processed foods and red meat. For a healthy lifestyle in young adults, the MD in particular provides the required amount of antioxidants per day for vitamins D (0.3-3.8 µg), E (17.0 mg), C (137.2-269.8 mg), A (1273.3 µg), B-12 (1.5-2.0 µg), and folate (455.1-561.3 µg), the minerals Se (120.0 µg), Zn (11.0 mg), Fe (15.0-18.8 mg), and Mn (5.2-12.5 mg), and polyphenols (1171.00 mg) needed to maintain an active immune response. However, all of these diets are deficient in the recommended amount of vitamin D (20 µg/d). Therefore, vulnerable populations such as elders and obese individuals could benefit from antioxidant supplementation to improve their antioxidant response. Although evidence remains scarce, there is some indication that a healthy diet, along with supplemental antioxidant intake, is beneficial to COVID-19 patients.

Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation.

Casein kinase 2 (CK2) is a highly ubiquitous and conserved serine/threonine kinase that forms a tetramer consisting of a catalytic subunit (CK2α) and a regulatory subunit (CK2ß). Despite being ubiquitous, CK2 is commonly found at higher expression levels in cancer cells, where it inhibits apoptosis, and supports cell migration and proliferation. The Ca2+-activated chloride channel TMEM16A shows similar effects in cancer cells: TMEM16A increases cell proliferation and migration and is highly expressed in squamous cell carcinoma of the head and neck (HNSCC) as well as other malignant tumors. A microscopy-based high-throughput screening was performed to identify proteins that regulate TMEM16A. Within this screen, CK2 was found to be required for proper membrane expression of TMEM16A. small interfering (si) RNA-knockdown of CK2 reduced plasma membrane expression of TMEM16A and inhibited TMEM16A whole cell currents in (cystic fibrosis bronchial epithelial) CFBE airway epithelial cells and in the head and neck cancer cell lines Cal33 and BHY. Inhibitors of CK2, such as TBB and the preclinical compound CX4549 (silmitasertib), also blocked membrane expression of TMEM16A and Ca2+-activated whole cell currents. siRNA-knockout of CK2 and its pharmacological inhibition, as well as knockdown or inhibition of TMEM16A by either niclosamide or Ani9, attenuated cell proliferation. Simultaneous inhibition of CK2 and TMEM16A strongly potentiated inhibition of cell proliferation. Although membrane expression of TMEM16A is reduced by inhibition of CK2, our data suggest that the antiproliferative effects by inhibition of CK2 are mostly independent of TMEM16A. Simultaneous inhibition of TMEM16A by niclosamide and inhibition of CK2 by silmitasertib was additive with respect to blocking cell proliferation, while cytotoxicity was reduced when compared to solely blockade of CK2. Therefore, parallel blockade TMEM16A by niclosamide may assist with anticancer therapy by silmitasertib.

Influence of culinary practices on protein and lipid oxidation of chicken meat burgers during cooking and in vitro gastrointestinal digestion.

The impact of six culinary practices - oven/microwave combined with/without seasoning with oregano/beer - on lipid and protein oxidation of chicken burgers after cooking and after in vitro digestion was assessed. Five oxidation markers - malondialdehyde (MDA), 4-hydroxy-2-nonenal (HNE), hexanal (HEX), carbonyls, and Schiff bases - as well as free amino acids and total fatty acids content were measured. Oregano prevented MDA, HEX, and HNE formation during cooking, while beer seems not to influence their formation. After in vitro digestion, MDA, carbonyls, and Schiff bases increased, regardless of the culinary practice, while HNE and HEX values were reduced. Globally, cooking with oregano exhibited the lowest losses of PUFAs and formation of all oxidation markers, thus it should be used as a mitigation strategy to avoid the formation of oxidation products during cooking, as well as to prevent their formation during in vitro digestion.

Stability of antibacterial and coccidiostat drugs on chicken meat burgers upon cooking and in vitro digestion.

The impact of culinary practices - oven or microwave cooking combined with herbs and/or beer - on antibacterial and coccidiostat drugs stability and bioaccessibility in chicken meat was evaluated. Fourteen compounds from 6 classes (ß-lactams, tetracyclines, sulfonamides, fluoroquinolones, macrolides, and coccidiostats) were monitored after cooking and in vitro digestion (INFOGEST protocol) at two fortification levels. Depending on their reduction, the presence of transformation products derived from cooking or digestion was investigated. In general, compounds were stable during cooking except amoxicillin, chlortetracycline and tylosin (reductions > 50%). Molecular rearrangement and dechlorination reactions are the most probable transformations derived from cooking. Adding herbs/beer does not benefit their reductions. During in vitro digestion, maximum bioaccessibilities of 60% were observed for all quantified compounds. As drugs and bile salts interact, increasing the absorption of lipophilic drugs, their bioaccessibility predictions must not be based only on the determination of their free form using LC-MS/MS.

Transport of mycotoxins across human gastric NCI-N87 and intestinal Caco-2 cell models.

Aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FB1), and ochratoxin A (OTA) are prevalent mycotoxins co-occurring in food, and their oral intake is conceivable to occur in the gastrointestinal epithelium. The intestinal absorption of some mycotoxins has been studied but only considering their isolated intake, while their gastric absorption in humans has not been explored. This study evaluated the bidirectional in vitro transport of four mycotoxins, isolated and in mixture, across gastric NCI-N87 and intestinal Caco-2 monolayers. AFB1 and DON were bidirectionally transported, more rapidly for AFB1; whereas OTA and FB1 were only transported in the absorptive direction, the first in both monolayers, and the second only in the gastric epithelium. The mixture of four mycotoxins exhibited some differences in cell uptake/excretion ratios. AFB1 presented the highest fraction absorbed (>96%) isolated and in mixture, followed by DON (72.8 and 82.9%); and OTA (11 and 66%) when transported isolated and in mixture, respectively. Different absorptive patterns on both epithelia were found when mycotoxins are transported isolated or in mixture. Further investigation on combined ingestion of toxins and their mixed transport should be considered for the proper evaluation of human absorption and toxicity of those mycotoxins considering their frequent co-occurrence and consequent co-exposure.

Influence of oven and microwave cooking with the addition of herbs on the exposure to multi-mycotoxins from chicken breast muscle.

Mycotoxins can contaminate poultry production via fungal infection of feeds. The impact of different cooking methods on mycotoxins stability and bioaccessibility is scarcely studied. Recent cooking practices such as the use of roasting bags along with some seasonings to cook chicken have become a trend to maintain the tenderness of the meat. This study evaluated the impact of oven roasting and microwaving of chicken breast muscles, with/without roasting bags and/or herbs addition on the stability and bioaccessibility of the 10 prevalent mycotoxins. Cooking itself reduced mycotoxins content, while using bags did not confer any advantage; adding herbs reduced mycotoxins content by up to 60%, but also appeared to increase their bioaccessibility. Thus, strategies to reduce their increased bioaccessibility should be the subject of future research. Notwithstanding, adding herbs prior to cooking could be used as a mitigation strategy to reduce mycotoxins in meat.

Toxicological interactions between mycotoxins from ubiquitous fungi: Impact on hepatic and intestinal human epithelial cells.

Aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FB1) and ochratoxin A (OTA) are toxic fungal metabolites co-occurring naturally in the environment. This study aimed to evaluate the toxicological interactions of these mycotoxins concerning additive, antagonistic and synergistic toxicity towards human cells. The theoretical biology-based Combination index-isobologram method was used to evaluate the individual and binary effect of these toxins and determine the type of the interaction using as models Caco-2 (intestinal) and HepG2 (hepatic) cells. Cytotoxicity was assessed using the MTT test at the concentrations of 0.625-20 µM for all the compounds. DON exerted the highest toxicity toward both cells, OTA and AFB1 also showed a dose-effect response, whereas no toxicity was verified for FB1. Synergism or antagonism effects occurred when exposing AFB1-DON and AFB1-OTA on Caco-2 cells at higher or lower concentrations, respectively; while DON-OTA showed synergism throughout all inhibition levels. Concerning HepG2, AFB1-DON exerted a strong synergism, regardless of the level; whereas AFB1-OTA had slight synergism/nearly additive effect; and, OTA-DON had a moderate antagonism/nearly additive effect. Synergistic strengths as high as a dose reduction index of 10 for AFB1-DON were observed in hepatic cells. Taken together our findings indicate that the toxicological effects differ regarding the type of mycotoxins used for combinations and the stronger synergistic effect was observed for mixtures containing DON in both cells. Therefore, even though DON has not been classified as to its carcinogenicity to humans, this mycotoxin may present a serious threat to health, mainly when co-occurring in the environment.

A novel microscopy-based assay identifies extended synaptotagmin-1 (ESYT1) as a positive regulator of anoctamin 1 traffic.

An attractive possibility to treat Cystic Fibrosis (CF), a severe condition caused by dysfunctional CFTR, an epithelial anion channel, is through the activation of alternative (non-CFTR) anion channels. Anoctamin 1 (ANO1) was demonstrated to be a Ca2+-activated chloride channel (CaCC) and thus of high potential to replace CFTR. Despite that ANO1 is expressed in human lung CF tissue, it is present at the cell surface at very low levels. In addition, little is known about regulation of ANO1 traffic, namely which factors promote its plasma membrane (PM) localization. Here, we generated a novel cellular model, expressing an inducible 3HA-ANO1-eGFP construct, and validated its usage as a microscopy tool to monitor for ANO1 traffic. We demonstrate the robustness and specificity of this cell-based assay, by the identification of siRNAs acting both as ANO1 traffic enhancer and inhibitor, targeting respectively COPB1 and ESYT1 (extended synaptotagmin-1), the latter involved in coupling of the endoplasmic reticulum to the PM at specific microdomains. We further show that knockdown of ESYT1 (and family members ESYT2 and ESYT3) significantly decreased ANO1 current density. This ANO1 cell-based assay constitutes an important tool to be further used in high-throughput screens and drug discovery of high relevance for CF and cancer.