Obesity is associated with altered gene expression in human tastebuds.

BACKGROUND: The role of taste perception in the development and persistence of obesity is currently unclear due to conflicting results from psychophysical and other studies. No study to date has assessed whether there is an underlying fundamental difference in the physiology of taste tissue between lean and obese individuals. METHOD/SUBJECTS: We analysed the transcriptomic profile (RNA-seq) of human fungiform taste papillae biopsied from lean (n = 23) and obese (n = 13) Caucasian females (age range 18-55) to identify differences in gene expression. RESULTS: Obesity status was the major contributor to variance in global gene expression between individuals. A total of 62 genes had significantly different gene expression levels between lean and obese (P < 0.0002), with the specific taste associated genes phospholipase C beta 2 (PLCß2) and sonic hedge-hog (SHH) having significantly reduced expression in obese group. Genes associated with inflammation and immune response were the top enriched biological pathways differing between the lean and the obese groups. Analysis of a broader gene set having a twofold change in expression (2619 genes) identified three enriched theme groups (sensory perception, cell and synaptic signalling, and immune response). Further, analysis of taste associated genes identified a consistent reduction in the expression of taste-related genes (in particular reduced type II taste cell genes) in the obese compared to the lean group. CONCLUSION: The findings show obesity is associated with altered gene expression in tastebuds. Furthermore, the results suggest the tastebud microenvironment is distinctly different between lean and obese persons and, that changes in sensory gene expression contribute to this altered microenvironment. This research provides new evidence of a link between obesity and altered taste and in the future may help design strategies to combat obesity.

A low-fat diet up-regulates expression of fatty acid taste receptor gene FFAR4 in fungiform papillae in humans: a co-twin randomised controlled trial.

Fatty acid taste (FAT) perception is involved in the regulation of dietary fat intake, where impaired FAT is associated with increased fatty food intake. There are a number of FAT receptors identified on human taste cells that are potentially responsible for FAT perception. Manipulating dietary fat intake, and in turn FAT perception, would elucidate the receptors that are associated with long-term regulation of FAT perception. The present study aimed to assess associations between diet-mediated changes to FAT receptors and FAT perception in humans. A co-twin randomised controlled trial was conducted, where each matching twin within a pair were randomly allocated to either an 8-week low-fat (LF; <20 % energy fat) or an 8-week high-fat (HF; >35 % energy fat) diet. At baseline and week 8, fungiform papillae were biopsied in the fasted state and FAT receptor gene expressions (cluster of differentiation 36 (CD36), free fatty acid receptor 2 (FFAR2), FFAR4, G protein-coupled receptor 84 (GPR84) and a delayed rectifying K+ channel (K+ voltage-gated channel subfamily A member 2; KCNA2)) were measured using RT-PCR; and FAT threshold (FATT) was assessed using three-alternate forced choice methodology. Linear mixed models were fitted, adjusting for correlation between co-twins. Intake was compliant with the study design, with the LF and HF groups consuming 14·8 and 39·9 % energy from fat, respectively. Expression of FFAR4 increased by 38 % in the LF group (P = 0·023; time-diet interaction P = 0·063). ΔFFAR4 (Δ, week 8-baseline) was associated with Δfat intake (g) ( = -159·4; P < 0·001) and ΔFATT ( = -8·8; P = 0·016). In summary, FFAR4 is involved in long-term diet-mediated changes to FAT perception. Manipulating dietary fat intake, and therefore FFAR4 expression, might aid in reducing taste-mediated passive overconsumption of fatty foods.

Expression of the candidate fat taste receptors in human fungiform papillae and the association with fat taste function.

Significant experimental evidence supports fat as a taste modality; however, the associated peripheral mechanisms are not well established. Several candidate taste receptors have been identified, but their expression pattern and potential functions in human fungiform papillae remain unknown. The aim of this study is to identify the fat taste candidate receptors and ion channels that were expressed in human fungiform taste buds and their association with oral sensory of fatty acids. For the expression analysis, quantitative RT-PCR (qRT-PCR) from RNA extracted from human fungiform papillae samples was used to determine the expression of candidate fatty acid receptors and ion channels. Western blotting analysis was used to confirm the presence of the proteins in fungiform papillae. Immunohistochemistry analysis was used to localise the expressed receptors or ion channels in the taste buds of fungiform papillae. The correlation study was analysed between the expression level of the expressed fat taste receptors or ion channels indicated by qRT-PCR and fat taste threshold, liking of fatty food and fat intake. As a result, qRT-PCR and western blotting indicated that mRNA and protein of CD36, FFAR4, FFAR2, GPR84 and delayed rectifying K+ channels are expressed in human fungiform taste buds. The expression level of CD36 was associated with the liking difference score (R -0·567, ß=-0·04, P=0·04) between high-fat and low-fat food and FFAR2 was associated with total fat intake (ρ=-0·535, ß=-0·01, P=0·003) and saturated fat intake (ρ=-0·641, ß=-0·02, P=0·008).

Gaining Wings to FLY: Using Drosophila Oogenesis as an Entry Point for Citizen Scientists in Laboratory Research.

Citizen science is a productive approach to include non-scientists in research efforts that impact particular issues or communities. In most cases, scientists at advanced career stages design high-quality, exciting projects that enable citizen contribution, a crowdsourcing process that drives discovery forward and engages communities. The challenges of having citizens design their own research with no or limited training and providing access to laboratory tools, reagents, and supplies have limited citizen science efforts. This leaves the incredible life experiences and immersion of citizens in communities that experience health disparities out of the research equation, thus hampering efforts to address community health needs with a full picture of the challenges that must be addressed. Here, we present a robust and reproducible approach that engages participants from Grade 5 through adult in research focused on defining how diet impacts disease signaling. We leverage the powerful genetics, cell biology, and biochemistry of Drosophila oogenesis to define how nutrients impact phenotypes associated with genetic mutants that are implicated in cancer and diabetes. Participants lead the project design and execution, flipping the top-down hierarchy of the prevailing scientific culture to co-create research projects and infuse the research with cultural and community relevance.

The Role of Multiply-Fortified Table Salt and Bouillon in Food Systems Transformation.

Our global food system lacks the critically needed micronutrients to meet the daily requirements of the most at-risk populations. Diets also continue to shift toward unhealthy foods, including the increased intake of salt. While most countries exceed the WHO's recommended levels, sodium does play an essential physiological role. Table salt and other salt-containing condiments, such as bouillon, also have cultural importance, as they are used to enhance the flavor of foods cooked at home. Given their universal consumption across income classes and both urban and rural populations, these condiments are an integral part of the food system and should, therefore, be part of its transformation. Fortification of salt and salt-containing condiments can play a catalytic role in the delivery of population-wide nutritional and health benefits. With relatively consistent levels of intake across the population, these condiments hold high potential for delivering micronutrients beyond iodine while also reducing concerns related to high micronutrient intake, particularly so in countries where the industries are relatively consolidated. As a flexible and complementary strategy to an evolving food system, fortification levels can also be adjusted over time to ensure micronutrient delivery targets continue to be achieved as the system improves, whether through lower intakes of sodium in line with WHO recommendations, enhanced consumption of nutrient-dense foods, and/or broader adoption of biofortified crops. Future areas of innovation are required to realize this vision, including developing affordable salt substitutes to meet cost requirements of consumers in low-and middle-income countries, improving the stability and bioavailability of the micronutrients in condiments so that delivery targets can be reached without affecting sensory attributes, and the development of efficient systems for monitoring population intake and micronutrient status to inform fortification program design and management. Rather than being considered antithetical to the transformation, multiply-fortified salt and bouillon can strengthen our ability to meet the cultural, sensory, nutritional, and health needs of an evolving food system.

Odour enhances the sense of presence in a virtual reality environment.

Virtual reality (VR) headsets provide immersive audio-visual experiences for users, but usually neglect to provide olfactory cues that can provide additional information about our environment in the real world. This paper examines whether the introduction of smells into the VR environment enhances users' experience, including their sense of presence through collection of both psychological and physiological measures. Using precise odour administration with an olfactometer, study participants were exposed to smells while they were immersed in the popular PlayStation VR game "Resident Evil 7". A within-subject study design was undertaken where participants (n = 22) walked-through the same VR environment twice, with or without the introduction of associated congruent odour stimuli. Directly after each gameplay, participants completed a questionnaire to determine their sense of presence from the overall gameplay and their sense of immersion in each of the virtual scenes. Additionally, physiological measurements (heart rate, body temperature and skin electrodermal activity) were collected from participants (n = 11) for each gameplay. The results showed the addition of odours significantly increased participants' sense of spatial presence in the VR environment compared to VR with no odour. Participants also rated the realism of VR experience with odour higher compared to no odour, however odour addition did not result in change in emotional state of participants (arousal, pleasure, dominance). Further, the participants' physiological responses were impacted by the addition of odour. Odour mediated physiological changes were dependent on whether the VR environment was novel, as the effect of odour on physiological response was lost when participants experienced the aroma on the second gameplay. Overall, the results indicate the addition of odours to a VR environment had a significant effect on both the psychological and physiological experience showing the addition of smell enhanced the VR environment. The incorporation of odours to VR environments presents an opportunity to create a more immersive experience to increase a person's presence within a VR environment. In addition to gaming, the results have broader applications for virtual training environments and virtual reality exposure therapy.

Sodium Reduction in Bouillon: Targeting a Food Staple to Reduce Hypertension in Sub-saharan Africa.

Bouillon cubes are a staple ingredient used in Sub-saharan African countries providing flavor enhancement to savory foods. Bouillon has been identified as a vehicle for fortification to overcome micronutrient deficiencies in Sub-saharan Africa. However, bouillon has a high sodium content (and in addition with other foods) contributes to dietary sodium intake above recommended guidelines. High dietary sodium intake is a key risk factor for hypertension and cardiovascular disease (CVD). Africa has the highest rates of hypertension and CVD globally with nearly half the adult population above 25 years affected. This review presents current state of research on sodium reduction strategies in bouillon. The key challenge is to reduce sodium levels while maintaining optimal flavor at the lowest possible production cost to ensure bouillon continues to be affordable in Sub-saharan Africa. To produce lower sodium bouillon with acceptable flavor at low cost will likely involve multiple sodium reduction strategies; direct reduction in sodium, sodium replacement and saltiness boosting flavor technologies. Efforts to reduce the sodium content of bouillon in Sub-saharan Africa is a worthwhile strategy to: (i) lower the overall sodium consumption across the population, and (ii) deliver population-wide health benefits in a region with high rates of hypertension and CVD.

In-Mouth Volatile Production from Brassica Vegetables (Cauliflower) and Associations with Liking in an Adult/Child Cohort.

Interactions between Brassica vegetables and human saliva can affect in-mouth odor development, which in turn may be linked to individual perception and liking. S-Methyl-l-cysteine sulfoxide is a unique substrate present in Brassicas that produces odor-active sulfur volatiles due to the activity of enzymes present in plant tissue and due to bacteria, which may be present to varying extents in an individual's oral microbiome. Proton transfer reaction mass spectrometry was applied to measure individual differences in sulfur volatile production in real time when fresh human saliva was incubated ex vivo with raw cauliflower for a cohort of child-adult pairs. Large differences in the rate of sulfur volatile production were measured between individuals, but not between age groups. Significant positive relationships were found for volatile production between the adult-child pairs, suggesting a degree of commonality in saliva composition and oral microbiome activity. Furthermore, significant negative relationships were measured between the amount of in-mouth sulfur volatile production and liking for raw cauliflower in children.

Isocotoin suppresses hepatitis E virus replication through inhibition of heat shock protein 90.

Hepatitis E virus (HEV) causes 14 million infections and 60,000 deaths per year globally, with immunocompromised persons and pregnant women experiencing severe symptoms. Although ribavirin can be used to treat chronic hepatitis E, toxicity in pregnant patients and the emergence of resistant strains are major concerns. Therefore there is an imminent need for effective HEV antiviral agents. The aims of this study were to develop a drug screening platform and to discover novel approaches to targeting steps within the viral life cycle. We developed a screening platform for molecules inhibiting HEV replication and selected a candidate, isocotoin. Isocotoin inhibits HEV replication through interference with heat shock protein 90 (HSP90), a host factor not previously known to be involved in HEV replication. Additional work is required to understand the compound's translational potential, however this suggests that HSP90-modulating molecules, which are in clinical development as anti-cancer agents, may be promising therapies against HEV.

Corrigendum to "Influence of saliva on individual in-mouth aroma release from raw cabbage (Brassica oleracea var. capitata f. rubra L.) and links to perception" [Heliyon 4 (12) (December 2018) e01045].

[This corrects the article DOI: 10.1016/j.heliyon.2018.e01045.].

Influence of saliva on individual in-mouth aroma release from raw cabbage (Brassica oleracea var. capitata f. rubra L.) and links to perception.

Raw or minimally processed vegetables are popular for health reasons and for their unique textural and flavor attributes. While many aroma volatiles are produced in situ when plant tissues are mechanically disrupted, enzymes expressed in bacteria in oral microbiota such as cysteine-ß-lyase (EC 4.4.1.13) may also contribute to aroma formation in-mouth during consumption. Interactions between raw cabbage and fresh human saliva (n = 21) were measured ex vivo by real-time monitoring of sulfur volatile production by proton transfer reaction mass spectrometry (PTR-MS). Inter-individual differences in the concentration of sulfur volatiles from the breakdown of S-methyl-L-cysteine sulfoxide (SMCSO) in fresh cabbage by saliva were characterized and a 10-fold difference in the extent of sulfur volatile production was measured across individuals. The overall intensity and garlic odor of raw cabbage was positively correlated with the concentration of sulfur volatiles after incubation with fresh human saliva. A buildup of SMSCO-derived sulfur volatiles in vivo, over twenty repeated mouthfuls was demonstrated, indicating that these reactions can affect sensory perception within the timescale of eating. These findings show the perceived odor experienced when eating cabbage differs, thus resulting in a unique flavor experience between individuals.

Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription.

Approximately 20 million hepatitis E virus (HEV) infections occur annually in both developing and industrialized countries. Most infections are self-limiting, but they can lead to chronic infections and cirrhosis in immunocompromised patients, and death in pregnant women. The mechanisms of HEV replication remain incompletely understood due to scarcity of adequate experimental platforms. HEV undergoes asymmetric genome replication, but it produces an additional subgenomic (SG) RNA encoding the viral capsid and a viroporin in partially overlapping open reading frames. Using a novel transcomplementation system, we mapped the intragenomic subgenomic promoter regulating SG RNA synthesis. This cis-acting element is highly conserved across all eight HEV genotypes, and when the element is mutated, it abrogates particle assembly and release. Our work defines previously unappreciated viral regulatory elements and provides the first in-depth view of the intracellular genome dynamics of this emerging human pathogen.IMPORTANCE HEV is an emerging pathogen causing severe liver disease. The genetic information of HEV is encoded in RNA. The genomic RNA is initially copied into a complementary, antigenomic RNA that is a template for synthesis of more genomic RNA and for so-called subgenomic RNA. In this study, we identified the precise region within the HEV genome at which the synthesis of the subgenomic RNA is initiated. The nucleotides within this region are conserved across genetically distinct variants of HEV, highlighting the general importance of this segment for the virus. To identify this regulatory element, we developed a new experimental system that is a powerful tool with broad utility to mechanistically dissect many other poorly understood functional elements of HEV.

Mechanism of fat taste perception: Association with diet and obesity.

Energy homeostasis plays a significant role in food consumption and body weight regulation with fat intake being an area of particular interest due to its palatability and high energy density. Increasing evidence from humans and animal studies indicate the existence of a taste modality responsive to fat via its breakdown product fatty acids. These studies implicate multiple candidate receptors and ion channels for fatty acid taste detection, indicating a complex peripheral physiology that is currently not well understood. Additionally, a limited number of studies suggest a reduced ability to detect fatty acids is associated with obesity and a diet high in fat reduces an individual's ability to detect fatty acids. To support this, genetic variants within candidate fatty acid receptors are also associated with obesity reduced ability to detect fatty acids. Understanding oral peripheral fatty acid transduction mechanisms and the association with fat consumption may provide the basis of novel approaches to control development of obesity.

A Comparison of Collection Techniques for Gene Expression Analysis of Human Oral Taste Tissue.

Variability in human taste perception is associated with both genetic and environmental factors. The influence of taste receptor expression on this variability is unknown, in part, due to the difficulty in obtaining human oral tissue that enables quantitative expression measures of taste genes. In a comparison of six current techniques (Oragene RNeasy Kit, Isohelix swab, Livibrush cytobrush, tongue saliva, cheek saliva collection, and fungiform papillae biopsy), we identify the fungiform papillae biopsy is the optimal sampling technique to analyse human taste gene expression. The fungiform papillae biopsy resulted in the highest RNA integrity, enabling amplification of all the assessed taste receptor genes (TAS1R1, TAS1R2, TAS1R3, SCNN1A and CD36) and taste tissue marker genes (NCAM1, GNAT3 and PLCß2). Furthermore, quantitative expression was observed in a subset of taste genes assessed from the saliva collection techniques (cheek saliva, tongue saliva and Oragene RNA kit). These saliva collection techniques may be useful as a non-invasive alternative sampling technique to the fungiform papillae biopsy. Identification of the fungiform papillae biopsy as the optimal collection method will facilitate further research into understanding the effect of gene expression on variability in human taste perception.

Association of SLC11A1 promoter polymorphisms with the incidence of autoimmune and inflammatory diseases: a meta-analysis.

Solute carrier family 11 member a1 (SLC11A1) exerts pleiotropic effects on macrophage function. Expression of SLC11A1 is regulated by a (GT)(n) microsatellite promoter repeat polymorphism of which nine alleles have been described. Enhanced activation of macrophages, associated with increased expression from allele 3, may be functionally linked to the development of autoimmune and inflammatory diseases. Conversely, low expression, driven by allele 2, may afford resistance. We have performed a meta-analysis to determine the association of SLC11A1 promoter alleles 2 and 3 with autoimmunity and inflammation. A random effects pooled odds ratio (OR) of 1.04 (95% confidence interval [CI]=0.20) for allele 3 suggested a weak association of this allele with an increased risk of disease. Calculation of the OR in the absence of asymmetry yielded a random effects pooled OR of 0.88 (95% CI=0.66), effectively reversing the above association. A fixed effects pooled OR of 0.90 (95% CI=0.24) was obtained for allele 2, suggesting a weak predominance of disease in the absence of this allele. Application of the trim-and-fill method resulted in a fixed effects OR of 0.80 (95% CI=0.22), thus strengthening this association. Associations of allele 3 with autoimmune and inflammatory diseases reported in several association studies may be attributable to some form of bias amongst published results.

Comparison of hospital episode statistics and central cardiac audit database in public reporting of congenital heart surgery mortality.

OBJECTIVE: To verify or refute the value of hospital episode statistics (HES) in determining 30 day mortality after open congenital cardiac surgery in infants nationally in comparison with central cardiac audit database (CCAD) information. DESIGN: External review of paediatric cardiac surgical outcomes in England (HES) and all UK units (CCAD), as derived from each database. SETTING: Congenital heart surgery centres in the United Kingdom. DATA SOURCES: HES for congenital heart surgery and corresponding information from CCAD for the period 1 April 2000 to 31 March 2002. HES was restricted to the 11 English centres; CCAD covered all 13 UK centres. MAIN OUTCOME MEASURE: Mortality within 30 days of open heart surgery in infants aged under 12 months. RESULTS: In a direct comparison for the years when data from the 11 English centres were available from both databases, HES omitted between 5% and 38% of infants operated on in each centre. A median 40% (range 0-73%) shortfall occurred in identification of deaths by HES. As a result, mean 30 day mortality was underestimated at 4% by HES as compared with 8% for CCAD. In CCAD, between 1% and 23% of outcomes were missing in nine of 11 English centres used in the comparison (predominantly those for overseas patients). Accordingly, CCAD mortality could also be underestimated. Oxford provided the most complete dataset to HES, including all deaths recorded by CCAD. From three years of CCAD, Oxford's infant mortality from open cardiac surgery (10%) was not statistically different from the mean for all 13 UK centres (8%), in marked contrast to the conclusions drawn from HES for two of those years. CONCLUSIONS: Hospital episode statistics are unsatisfactory for the assessment of activity and outcomes in congenital heart surgery. The central cardiac audit database is more accurate and complete, but further work is needed to achieve fully comprehensive risk stratified mortality data. Given unresolved limitations in data quality, commercial organisations should reconsider placing centre specific or surgeon specific mortality data in the public domain.