Effect of Coffee and Chocolate Ingestion on Clozapine Dose and on Plasma Clozapine and Norclozapine Concentrations in Clinical Practice.

BACKGROUND: Some reports point to dietary caffeine intake as a cause of increased plasma clozapine concentrations in certain patients. METHODS: We compared clozapine dose and plasma clozapine and N-desmethylclozapine (norclozapine) concentrations in male and female smokers and nonsmokers in relation to reported (i) coffee (caffeine) and (ii) chocolate (caffeine and theobromine) intake in samples submitted for clozapine therapeutic drug monitoring, 1993-2017. RESULTS: There was information on coffee ingestion for 16,558 samples (8833 patients) from males and 5886 samples (3433 patients) from females and on chocolate ingestion for 12,616 samples (7568 patients) from males and 4677 samples (2939 patients) from females. When smoking was considered, there was no discernible effect of either coffee or chocolate ingestion either on the median dose of clozapine or on the median plasma clozapine and norclozapine concentrations in men and in women. However, cigarette smoking was associated with higher coffee and chocolate consumption. Although male nonsmokers who reported drinking 3 or more cups of coffee daily had significantly higher median plasma clozapine and norclozapine concentrations than those who drank less coffee, they were also prescribed a significantly higher clozapine dose. There was no clear effect of coffee ingestion on plasma clozapine and norclozapine in female nonsmokers. IMPLICATIONS: Inhibition of clozapine metabolism by caffeine at the doses of caffeine normally encountered in those treated with clozapine is unlikely even in male nonsmokers. Measurement of plasma caffeine in an appropriate sample should be considered in any future investigation into a presumed clozapine-caffeine interaction.

Combining fracture mechanics and rheology to investigate the impact of micro-aeration on chocolate oral processing.

This study presents a rigorous mechanical characterisation investigation on milk chocolate with varying porosities, at different temperatures and strain rate levels. Uniaxial compression tests at temperatures varying from 20 °C to 30 °C were performed to measure the bulk properties of chocolate as a function of porosity and temperature. Fracture experiments were also conducted to compute the fracture energy at temperature levels between 20 °C and 30 °C for all tested samples. Additionally, rheological experiments are conducted to compute the viscosity of the different chocolates at 37 °C. This combined experimental analysis of solid mechanics, fracture mechanics, and rheology aims to define the impact of temperature and chocolate's phase change from solid to liquid on its mechanical properties. Moreover, the impact of micro-aeration on the relationship between material properties and temperature is discussed. The results demonstrate a significant impact of both temperature and micro-aeration on the chocolate's material properties; fracture stresses decrease with micro-aeration due to the presence of micro-pores creating weak links in the chocolate matrix, the critical strain energy release rate decreases with micro-aeration at temperatures up to 25 °C and increases at temperatures above 30 °C. Finally, the viscosity at 37 °C increases with increasing porosity due to the obstruction of the flow by micro-pores acting as "solid" particles. The results highlight how the impact of micro-aeration on the material properties of chocolate alters as the testing temperature rises and the material changes phase. The relationships between the micro-aeration and material properties and the dependence of temperature on the different mechanical properties are used to explain the difference in textural attributes as obtained from temporal dominance sensation tests. This study seeks to contribute valuable insights into the field of chocolate technology, emphasizing the need for a combined engineering approach to understand the structural breakdown of chocolate during oral processing as mechanisms such as chewing, melting, mixing and shearing occur.

In-house validation of an LC-MS method for the multiplexed quantitative determination of total allergenic food in chocolate.

Mass spectrometry has been widely accepted as a confirmatory tool for the sensitive detection of undeclared presence of allergenic ingredients. Multiple methods have been developed so far, achieving different levels of sensitivity and robustness, still lacking harmonization of the analytical validation and impairing comparability of results. In this investigation, a quantitative method has been validated in-house for the determination of six allergenic ingredients (cow's milk, hen's egg, peanut, soybean, hazelnut, and almond) in a chocolate-based matrix. The latter has been produced in a food pilot plant to provide a real and well-characterized matrix for proper assessment of method performance characteristics according to official guidelines. In particular, recent considerations issued by the European Committee for Standardization have been followed to guide a rigorous single-laboratory validation and to feature the main method performance, such as selectivity, linearity, and sensitivity. Synthetic surrogates of the peptide markers have been used both in native and labelled forms in matrix-matched calibration curves as external calibrants and internal standards, respectively. A two-order of magnitude range was investigated, focusing on the low concentration range for proper assessment of the detection and quantification limits (LOD and LOQ) by rigorous calibration approach. Conversion factors for all six allergenic ingredients have been determined for the first time to report the final quantitative information as fraction of total allergenic food protein (TAFP) per mass of food (µgTAFP/gfood), since such a reporting unit is exploitable in allergenic risk assessment plans. The method achieved good sensitivity with LOD values ranging between 0.08 and 0.2 µgTAFP/gfood, for all ingredients besides egg and soybean, whose quantitative markers reported a slightly higher limit (1.1 and 1.2 µgTAFP/gfood, respectively). Different samples of chocolate bar incurred at four defined concentration levels close to the currently available threshold doses have been analyzed to test the quantitative performance of the analytical method, with a proper estimate of the measurement uncertainty from different sources of variability. The sensitivity achieved resulted in compliance with the various threshold doses issued or recommended worldwide.

Cocoa Apoplastome Contains Defense Proteins Against Pathogens.

The apoplast performs important functions in the plant, such as defense against stress, and compounds present form the apoplastic washing fluid (AWF). The fungus Moniliophthora perniciosa, the causal agent of witches' broom disease (WBD) in Theobroma cacao, initially colonizes the apoplast in its biotrophic phase. In this period, the fungus can remain for approximately 60 days, until it changes to its second phase, causing tissue death and consequently large loss in the production of beans. To better understand the importance of the apoplast in the T. cacao-M. perniciosa interaction, we performed the first apoplastic proteomic mapping of two contrasting genotypes for WBD resistance (CCN51-resistant and Catongo-susceptible). Based on two-dimensional gel analysis, we identified 36 proteins in CCN-51 and 15 in Catongo. We highlight PR-proteins, such as peroxidases, ß-1,3-glucanases, and chitinases. A possible candidate for a resistance marker of the CCN-51 genotype, osmotin, was identified. The antioxidative metabolism of the superoxide dismutase (SOD) enzyme showed a significant increase (P < 0.05) in the AWF of the two genotypes under field conditions (FD). T. cacao AWF inhibited the germination of M. perniciosa basidiospores (>80%), in addition to causing morphological changes. Our results shed more light on the nature of the plant's defense performed by the apoplast in the T. cacao-M. perniciosa interaction in the initial (biotrophic) phase of fungal infection and therefore make it possible to expand WBD control strategies based on the identification of potential targets for resistance markers and advance scientific knowledge of the disease.

Short-term cocoa bioflavanol supplementation does not improve cold-induced vasodilation in young healthy adults.

PURPOSE: Cold-induced vasodilation (CIVD) is an oscillatory rise in blood flow to glabrous skin that occurs in cold-exposed extremities. Dietary flavanols increase bioavailable nitric oxide, a proposed mediator of CIVD through active vasodilation and/or withdrawal of sympathetic vascular smooth muscle tone. However, no studies have examined the effects of flavanol intake on extremity skin perfusion during cold exposure. We tested the hypothesis that acute and 8-day flavanol supplementation would augment CIVD during single-digit cold water immersion (CWI). METHODS: Eleven healthy adults (24 ± 6 years; 10 M/1F) ingested cocoa flavanols (900 mg/day) or caffeine- and theobromine-matched placebo for 8 days in a double-blind, randomized, crossover design. On Days 1 and 8, CIVD was assessed 2 h post-treatment. Subjects immersed their 3rd finger in warm water (42 °C) for 15 min before CWI (4 °C) for 30 min, during which nail bed and finger pad skin temperature were measured. RESULTS: Flavanol ingestion had no effect on CIVD frequency (Day 1, Flavanol: 3 ± 2 vs. Placebo: 3 ± 2; Day 8, Flavanol: 3 ± 2 vs. Placebo: 3 ± 1) or amplitude (Day 1, Flavanol: 4.3 ± 1.7 vs. Placebo: 4.9 ± 2.6 °C; Day 8, Flavanol: 3.9 ± 1.9 vs. Placebo: 3.9 ± 2.0 °C) in the finger pad following acute or 8-day supplementation (P > 0.05). Furthermore, average, nadir, and apex finger pad temperatures during CWI were not different between treatments on Days 1 or 8 of supplementation (P > 0.05). Similarly, no differences in CIVD parameters were observed in the nail bed following supplementation (P > 0.05). CONCLUSION: These data suggest that cocoa flavanol ingestion does not alter finger CIVD. Clinical Trial Registration Clinicaltrials.gov Identifier: NCT04359082. April 24, 2020.

Vegetarian, vegan, or plant-based? Comparing how different labels influence consumer evaluations of plant-based foods.

Market actors have a role to play in enabling sustainable food transitions. One challenge for these actors is how to promote plant-based foods in ways that appeal to a growing number of consumers. Here we test how different plant-based related labels affect consumer appraisals of a range of foods (cookies, sausages, cheese, chocolate, pasta). In two studies (pre-registered; NUSA = 1148, NGermany = 491), we examined the effects of a 'vegetarian', 'vegan', or 'plant-based' label (compared to no label) on five attributes (healthy, tasty, ethical, pure, environmentally friendly) related to the products. We also measured self-reported likelihood to purchase the products. Overall, the results indicated that the 'plant-based' label was slightly more appealing to participants than the 'vegetarian' and 'vegan' labels. However, contrary to our expectations, neither consumers' information-seeking tendencies nor their pre-existing attitudes toward plant-based foods influenced (i.e., moderated) the effects of the labels. Anticipated taste was a strong and consistent predictor of purchase likelihood for all labeled products, but the ethical and pure attributes also accounted for unique variance in this outcome variable. Taken together, our findings and discussion provide insights into the role of labels and label terminology on consumer appraisals of plant-based foods.

Imagine chocolate: The craving experience questionnaire in the food domain.

Measurement of food craving has gained relevance in the current obesity epidemic. The Craving Experience Questionnaire (CEQ) considers not only craving intensity but also cognitive intrusiveness and imagery vividness as separate craving factors and could thus refine food craving assessment. It is available in two versions with ten items each. The CEQ-F assesses craving frequency across specific time periods and the CEQ-S time-point specific craving strength. Across three independent studies, N = 533 participants completed the German chocolate CEQ-F referenced at the past year to operationalise trait-like craving. Among them, N = 402 also completed the German chocolate CEQ-S referenced at the current moment to operationalise state-like craving. Four-week test-retest reliability was measured. For external validity, we assessed self-reported chocolate consumption, body-mass-index, trait approach motivation, general imagery vividness, and the most widely used food craving questionnaire, namely the Food Cravings Questionnaires in a trait (FCQ-T-r) and state version (FCQ-S), as well as behavioural approach bias (reaction time-based measurement). The three-factor structure was replicated with excellent internal consistency for both CEQ-F and CEQ-S. Test-retest reliability was moderate for both CEQ versions. CEQ-F scores were related to higher levels of chocolate consumption, approach motivation, and FCQ-T-r scores, but not to body-mass-index, imagery vividness, or approach bias. CEQ-S scores were associated with FCQ-S scores and partly with approach bias, but not with approach motivation and imagery vividness. The current results support the factor structure, validity and reliability of the German chocolate CEQ-S and CEQ-F with questions remaining regarding the ability of the CEQ-S to measure state craving. Thus, CEQ-F and CEQ-S usefully contribute to food craving assessment.

Counts of mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic sporeforming bacteria and persistence of Bacillus cereus spores throughout cocoa powder processing chain.

Sporeforming bacteria are a concern in some food raw materials, such as cocoa powder. Samples (n = 618) were collected on two farms and at several stages during cocoa powder manufacture in three commercial processing lines to determine the impact of each stage on bacterial spore populations. Mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic, and Bacillus cereus spore populations were enumerated in all the samples. Genetic diversity in B. cereus strains (n = 110) isolated from the samples was examined by M13 sequence-based PCR typing, partial sequencing of the panC gene, and the presence/absence of ces and cspA genes. The counts of different groups of sporeforming bacteria varied amongst farms and processing lines. For example, the counts of mesophilic aerobic spore-forming (MAS) populations of cocoa bean fermentation were lower than 1 log spore/g in Farm 1 but higher than 4 log spore/g in Farm 2. B. cereus isolated from cocoa powder was also recovered from cocoa beans, nibs, and samples after roasting, refining, and pressing, which indicated that B. cereus spores persist throughout cocoa processing. Phylogenetic group IV was the most frequent (73%), along with processing. Strains from phylogenetic group III (14 %) did not show the ces gene's presence.

Investigating luxS gene expression in lactobacilli along lab-scale cocoa fermentations.

Previous metagenomic analyses have suggested that lactobacilli present potential for Quorum Sensing (QS) in cocoa fermentation, and in the present research, laboratory scale fermentations were carried out to monitor the expression of luxS, a universal marker of QS. For that, 96 h-fermentations were studied, as follows: F0 (non inoculated control), F1 (inoculated with yeasts, lactic acid bacteria, and acetic acid bacteria), F2 (inoculated with yeasts and acetic acid bacteria), F3 (inoculated with yeasts only). The parameters evaluated were: plate counting, quantification of key enzymes and analysis of volatile organic compounds associated with key sensory descriptors, using headspace gas chromatography-mass spectrometry (GC-MS). Furthermore, QS was estimated by the quantification of the expression of luxS genes by Reverse Transcriptase Real-Time PCR. The results demonstrated that microbial succession occurred in pilot scale fermentations, but no statistical differences for microbial enumeration and α-diversity index were observed among experiments and control. Moreover, it was not possible to make conclusive correlations of enzymatic profile and fermenting microbiota, likely due to the intrinsic activity of plant hydrolases. Regarding to the expression of luxS genes, in Lactiplantibacillus plantarum they were active along the fermentation, but for Limosilactobacillus fermentum, luxS was expressed only at early and middle phases. Correlation analysis of luxS expression and production of volatile metabolites evidenced a possible negative association of Lp. Plantarum with fermentation quality. In conclusion, these data corroborate former shotgun metagenomic analysis by demonstrating the expression of luxS by lactobacilli in pilot scale cocoa fermentation and evidence Lp. Plantarum is the main lactic acid bacteria related to its expression.

Genomic structural variants constrain and facilitate adaptation in natural populations of Theobroma cacao, the chocolate tree.

Genomic structural variants (SVs) can play important roles in adaptation and speciation. Yet the overall fitness effects of SVs are poorly understood, partly because accurate population-level identification of SVs requires multiple high-quality genome assemblies. Here, we use 31 chromosome-scale, haplotype-resolved genome assemblies of Theobroma cacao-an outcrossing, long-lived tree species that is the source of chocolate-to investigate the fitness consequences of SVs in natural populations. Among the 31 accessions, we find over 160,000 SVs, which together cover eight times more of the genome than single-nucleotide polymorphisms and short indels (125 versus 15 Mb). Our results indicate that a vast majority of these SVs are deleterious: they segregate at low frequencies and are depleted from functional regions of the genome. We show that SVs influence gene expression, which likely impairs gene function and contributes to the detrimental effects of SVs. We also provide empirical support for a theoretical prediction that SVs, particularly inversions, increase genetic load through the accumulation of deleterious nucleotide variants as a result of suppressed recombination. Despite the overall detrimental effects, we identify individual SVs bearing signatures of local adaptation, several of which are associated with genes differentially expressed between populations. Genes involved in pathogen resistance are strongly enriched among these candidates, highlighting the contribution of SVs to this important local adaptation trait. Beyond revealing empirical evidence for the evolutionary importance of SVs, these 31 de novo assemblies provide a valuable resource for genetic and breeding studies in Tcacao.

Steaming Maintains Fatty Acids, Antioxidants, and Proximate Content in Snack Bar Products from Cocoa Beans.

Chocolate products on the market are generally in the form of chocolate bars as snacks made from cocoa powder. Fat and powder are separated first through a pressing process to obtain the cocoa powder. Cocoa powder loses most of its fat content during processing. Therefore, the study aimed to determine the effect of steaming time on the cocoa bean content of fatty acids, free fatty acids, proximate levels, and antioxidant activity of snack bar products made from steamed cocoa beans. Seven steaming time intervals for cocoa beans were studied. The results showed that a longer steaming time affects the fatty acids, saturated fatty acids, antioxidants, and proximate in cocoa beans. Steaming time treatment at 45 minutes increased oleic acid, palmitic acid, and antioxidant activity. In addition, reducing free fatty acids represents a quality improvement that meets international Codex Alimentarius standards, offering a competitive advantage in the market. The food industry can adopt this steaming technique to develop snack bars and new products that are healthier and more sustainable by using steaming as an effective processing method in maintaining and increasing the nutritional value of products.

Chemical Composition of Fat Bloom on Chocolate Products Determined by Combining NMR and HPLC-MS.

To reduce unwanted fat bloom in the manufacturing and storage of chocolates, detailed knowledge of the chemical composition and molecular mobility of the oils and fats contained is required. Although the formation of fat bloom on chocolate products has been studied for many decades with regard to its prevention and reduction, questions on the molecular level still remain to be answered. Chocolate products with nut-based fillings are especially prone to undesirable fat bloom. The chemical composition of fat bloom is thought to be dominated by the triacylglycerides of the chocolate matrix, which migrate to the chocolate's surface and recrystallize there. Migration of oils from the fillings into the chocolate as driving force for fat bloom formation is an additional factor in the discussion. In this work, the migration was studied and confirmed by MRI, while the chemical composition of the fat bloom was measured by NMR spectroscopy and HPLC-MS, revealing the most important triacylglycerides in the fat bloom. The combination of HPLC-MS with NMR spectroscopy at 800 MHz allows for detailed chemical structure determination. A rapid routine was developed combining the two modalities, which was then applied to investigate the aging, the impact of chocolate composition, and the influence of hazelnut fillings processing parameters, such as the degree of roasting and grinding of the nuts or the mixing time, on fat bloom formation.

Integrating new variables into a framework to support cacao denomination of origin: a case study in Southwest Colombia.

BACKGROUND: Cocoa quality plays a pivotal role in establishing denominations of origin, with genotypes, geography, climate and soil conditions being key variables. However, these factors have not been comprehensively explored in defining cacao denominations of origin. The present study addresses this gap by laying the foundation for cacao denomination of origin, focusing on the Buenaventura region on Colombia's Pacific coast. Our goal is to provide a holistic understanding of the elements underpinning cacao denomination of origin, emphasizing Buenaventura's unique cocoa quality and geographical significance. RESULTS: Through the Buenaventura case, we propose a robust framework applicable to other cacao-producing regions, elevating the recognition and value of cacao denomination of origin. Our framework encompasses geography, agronomy, genetics, microbial diversity, pests and diseases and cocoa quality. In a pioneering move, we propose a cacao denomination of origin in Colombia, specifically examining Bajo Calima, Sabaletas and Cisneros within Buenaventura region. Buenaventura stands out for its cocoa quality, characterized by fruity flavors attributed to the rich biodiversity of the lowland rainforest. CONCLUSION: Our analysis indicates specific geographical indicators for each of the study zones, with Buenaventura identified as a region with natural characteristics to produce fine flavour cocoa products. Each zone exhibited a high differentiation and diversity of cacao cultivars. Buenaventura has the potential to be designated as a future denomination of origin for cacao from the Pacific region of Colombia, characterized by its unique fruity-aroma chocolates. Our framework is adaptable to other cacao-producing regions, facilitating the establishment of denominations of origin within the cocoa industry and agriculture. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The severity and yield effects of the chocolate spot disease in faba bean affected by intercropping and nitrogen input.

BACKGROUND: The aim is to study the disease suppression efficiency, yield loss rate, and yield benefits of intercropped faba bean against chocolate spot under nitrogen (N) input, to clarify the effectiveness of intercropping faba beans in controlling chocolate spot and its contribution to yield increase. RESULTS: Four N input levels and disease suppression treatments were discovered when faba bean-wheat intercropped was used. Adding N enhanced the chocolate spot's area under disease progression curve (AUDPC) by 27.1-69.9%. In contrast to monoculture, intercropping reduced the AUDPC of the chocolate spot by 32.4-51.0% (P < 0.05). Interestingly, the relative control efficacy (RCE) of intercropping at the EShan site was better. With disease suppression or non-suppression, N input increased grain yield loss and its components in faba bean. The total yield advantage effect (TE) and disease-suppression effect (DSE) of faba bean intercropped at the two experimental sites were significantly increased under N input (N1 and N2 level). The proportion of recovery yield due to intercropping suppressed disease (DSE/TE ratio) in EShan (52.1%) was higher than that in Xundian (40.9%), and the DSE of intercropping played an indispensable role in the two sites. Regression analysis of AUDPC and grain yield loss amount showed that one unit increase in chocolate spot AUDPC could cause a grain yield loss of 0.38-0.86 kg ha-1 . The partial land equivalent ratio (pLER) of intercropping faba beans at the EShan site was > 0.33. CONCLUSION: In conclusion, intercropped faba bean with N treatment (45-90 kg ha-1 ) was the best choice for maximizing the intercropping disease control effect. © 2023 Society of Chemical Industry.

Challenges and potential for detecting and quantifying titanium dioxide in food.

BACKGROUND: Titanium dioxide (TiO2) is banned in some countries but its use is still permitted in others. The global food supply chain is therefore challenged with the need to use rapid and reliable testing methods to either detect the presence of TiO2 or to quantify its concentration. The goal of this study was to determine the feasibility of using color, texture profile analysis, Raman microscopy, and X-ray fluorescence (XRF) spectroscopy to detect and quantify TiO2 in fillings used in the pastry and confectionery industry. In this study, two types of fillings were investigated: vanilla based and chocolate based. All fillings were prepared in four different variations - without TiO2 and with three concentrations as follows: 0.25 g*kg-1, 0.5 g*kg-1, or 0.75 g*kg-1 TiO2 per sample. The methods were selected for their ability to analyze the samples in a short period of time. RESULTS: All of the methods showed moderate to high potential for detecting TiO2 in the samples. The results reveal how TiO2 affects the food matrix color and texture. Use of Raman microscopy confirms its detectability, although concentrations of TiO2 do not follow a pattern. X-ray fluorescence spectroscopy showed the greatest potential as it can not only detect TiO2 but can also quantify its concentration in the samples. CONCLUSIONS: The highest potential for quantifying the concentration of this food additive was achieved with XRF. © 2024 Society of Chemical Industry.

One-bite-sized 3D printed finger foods, oriented to malnutrition, sarcopenia and frailty prevention in the older people.

BACKGROUND: In this study, innovative chocolate, citrus and mixture flavoured tofu-based nutritionally customised, dysphagia-oriented, comfortably consumed, appetising, one-bite-sized finger foods, oriented to malnutrition, sarcopenia and frailty prevention in older people were created by using 3D printing technology. Developed products were characterised by evaluating chemical composition and physical properties and performing sensory evaluation among geriatric clinic residents (≥60 years). RESULTS: The dietary composition of the developed foods was: 19-21 g (100 g)-1 protein, 6-8 g (100 g)-1 fibre, 8-9 g (100 g)-1 fat, 11 mg (100 g)-1 iron, 14 mg (100 g)-1 zinc, 70 µg (100 g)-1 selenium. Foods were also enriched with branched-chain amino acids, such as leucine, isoleucine and valine. All formulated foods were classified as level 6 by International Dysphagia Diet Standardisation Initiative classification. Chocolate-flavoured food was much harder (4914 g) with lower adhesiveness value (-33.6 g s), compared to the citrus- or mixture-flavoured foods. Older people evaluated all finger foods as very easy handled by hand, soft, easy to swallow, having a moderate flavour intensity and a weak afterfeel. Despite the fact that the chocolate food was evaluated as having the highest hardness and gumminess values by the instrumental method, this difference was not noticeable to the evaluators. However 7% of the participants said that 3D printed foods were sticky to dentures. CONCLUSION: The results suggest that it is possible to create nutrient-dense comfortably consumed 3D printed foods, oriented to malnutrition, sarcopenia and frailty prevention in older people. © 2024 Society of Chemical Industry.

A bite of dark chocolate? Black humour in mental health services.

OBJECTIVE: Black humour permits expression of what may otherwise be unspeakable and is observed and used by staff working in mental health services. The aim of this study was to identify how humour, particularly black humour, was perceived by different health professionals in psychiatric practice. METHODS: Participants were invited to complete a survey. Data was collated and statistically analysed by a biostatistician. Chi square and univariate tests were performed to identify associations between categories. RESULTS: The sub-question relating to the benefits of black humour was analysed. Main findings were that the majority of staff perceived black humour to be beneficial in psychiatric practice (n = 564 of 710 total; 79.4%), particularly if they used general and black humour with patients, families and colleagues. Those who observed black humour being used collegially about patients and families were more likely to find black humour beneficial; and even those uncomfortable with black humour being used by colleagues were more likely to see the benefits of black humour. CONCLUSION: Black humour was perceived to be beneficial in mental health settings when used mindfully, sensitively and in context. Further study with patients and relatives may shed light on how widely the perception of benefit is shared.

Cell surface ATP synthase-released H+ and ATP play key roles in cocoa butter intake-mediated regulation of gut immunity through releases of cytokines in rat.

Proper food intake is important for maintaining good health in humans. Chocolate is known to exert anti-inflammatory effects; however, the mechanisms remain unclear. In this study, we aimed to investigate the effects of cocoa butter intake on gut immunity in rats and rabbits. Cocoa butter intake increased the lymph flow, cell density, and IL-1ß, IL-6 and IL-10 levels in mesenteric lymph. Clodronate, a macrophage depletion compound, significantly enhanced the release of all cytokines. The immunoreactivities of macrophage markers CD68 and F4/80 in the jejunal villi were significantly decreased with clodronate. Piceatannol, a selective cell surface ATP synthase inhibitor significantly reduced the cocoa butter intake-mediated releases of IL-1ß, IL-6 and IL-10. The immunoreactivities of cell surface ATP synthase were observed in rat jejunal villi. Shear stress stimulation on the myofibroblast cells isolated from rat jejunum released ATP and carbon dioxide depended with H+ release. In rabbit in vivo experiments, cocoa butter intake increased the concentrations of ATP and H+ in the portal vein. The in vitro experiments with isolated cells of rat jejunal lamina propria the pH of 3.0 and 5.0 in the medium released significantly IL-1ß and IL-6. ATP selectively released IL-10. These findings suggest that cocoa butter intake regulates the gut immunity through the release and transport of IL-1ß, IL-6, and IL-10 into mesenteric lymph vessels in a negative feedback system. In addition, the H+ and ATP released from cell surface ATP synthase in jejunal villi play key roles in the cocoa butter intake-mediated regulation of gut immunity.

Nano solid phase micro membrane tip and electrochemical methods for vanillin analysis in chocolate samples.

A polymer-based nanosensor and electrochemical methods were developed for the quantitative analysis of vanillin. The sample preparation was done using nano solid phase micro membrane tip extraction (NSPMMTE). A novel poly(phenylalanine)/TiO2/CPE sensor was built as the working electrode for the first time for the analysis of the vanillin substance. The electrochemical behavior and analytical performance of vanillin were examined in detail by cyclic voltammetry (CV) and differential pulse stripping voltammetry (DPSV) techniques via the oxidation process. The optimized modules of the DPSV technique that affected the vanillin peak current and peak potential were pH, pulse amplitude, step potential, and deposition time. The electroactive surface areas of bare CPE, TiO2/CPE, and poly(phenylalanine)/TiO2/CPE electrodes were found to be 0.135 cm2, 0.155 cm2, and 0.221 cm2, respectively. The limit of detection (LOD) was 32.6 µg/L in the 0.25-15.0 mg/L working range at pH 7.0. The selectivity of the proposed DPSV method for the determination of vanillin on the modified electrode was investigated in the presence of various organic and inorganic substances, and the determination of vanillin with high recovery was achieved with less than 5% relative error. The analytical application was applied in chocolate samples and the DPSV method was found highly efficient, reproducible, and selective.