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Concept: Black pepper

365

The evidence base for the health effects of spice consumption is insufficient, with only one large population-based study and no reports from Europe or North America. Our objective was to analyze the association between consumption of hot red chili peppers and mortality, using a population-based prospective cohort from the National Health and Nutritional Examination Survey (NHANES) III, a representative sample of US noninstitutionalized adults, in which participants were surveyed from 1988 to 1994. The frequency of hot red chili pepper consumption was measured in 16,179 participants at least 18 years of age. Total and cause-specific mortality were the main outcome measures. During 273,877 person-years of follow-up (median 18.9 years), a total of 4,946 deaths were observed. Total mortality for participants who consumed hot red chili peppers was 21.6% compared to 33.6% for those who did not (absolute risk reduction of 12%; relative risk of 0.64). Adjusted for demographic, lifestyle, and clinical characteristics, the hazard ratio was 0.87 (P = 0.01; 95% Confidence Interval 0.77, 0.97). Consumption of hot red chili peppers was associated with a 13% reduction in the instantaneous hazard of death. Similar, but statistically nonsignificant trends were seen for deaths from vascular disease, but not from other causes. In this large population-based prospective study, the consumption of hot red chili pepper was associated with reduced mortality. Hot red chili peppers may be a beneficial component of the diet.

Concepts: Cohort study, Epidemiology, Medical statistics, Relative risk, Chili pepper, Black pepper, Cayenne pepper, Red Hot Chili Peppers

213

The genus Capsicum is New World in origin and represents a complex of a wide variety of both wild and domesticated taxa. Peppers or fruits of Capsicum species rarely have been identified in the paleoethnobotanical record in either Meso- or South America. We report here confirmation of Capsicum sp. residues from pottery samples excavated at Chiapa de Corzo in southern Mexico dated from Middle to Late Preclassic periods (400 BCE to 300 CE). Residues from 13 different pottery types were collected and extracted using standard techniques. Presence of Capsicum was confirmed by ultra-performance liquid chromatography (UPLC)/MS-MS Analysis. Five pottery types exhibited chemical peaks for Capsicum when compared to the standard (dihydrocapsaicin). No peaks were observed in the remaining eight samples. Results of the chemical extractions provide conclusive evidence for Capsicum use at Chiapas de Corzo during a 700 year period (400 BCE-300 CE). Presence of Capsicum in different types of culinary-associated pottery raises questions how chili pepper could have been used during this early time period. As Pre-Columbian cacao products sometimes were flavored using Capsicum, the same pottery sample set was tested for evidence of cacao using a theobromine marker: these results were negative. As each vessel that tested positive for Capsicum had a culinary use we suggest here the possibility that chili residues from the Chiapas de Corzo pottery samples reflect either paste or beverage preparations for religious, festival, or every day culinary use. Alternatively, some vessels that tested positive merely could have been used to store peppers. Most interesting from an archaeological context was the presence of Capsicum residue obtained from a spouted jar, a pottery type previously thought only to be used for pouring liquids.

Concepts: Mexico, Capsicum, Capsaicin, Chili pepper, Black pepper, Indigenous peoples of the Americas, Mexican cuisine, Jalapeño

168

Next generation sequencing has an advantageon transformational development of species with limited available sequence data as it helps to decode the genome and transcriptome. We carried out the sequencing using illuminaHiSeq™ 2000 to generate the first leaf transcriptome of black pepper ( L.), an important spice variety native to South India and also grown in other tropical regions. Despite the economic and biochemical importance of pepper, a scientifically rigorous study at the molecular level is far from complete due to lack of sufficient sequence information and cytological complexity of its genome. The 55 million raw reads obtained, when assembled using Trinity program generated 2,23,386 contigs and 1,28,157 unigenes. Reports suggest that the repeat-rich genomic regions give rise to small non-coding functional RNAs. MicroRNAs (miRNAs) are the most abundant type of non-coding regulatory RNAs. In spite of the widespread research on miRNAs, little is known about the hair-pin precursors of miRNAs bearing Simple Sequence Repeats (SSRs). We used the array of transcripts generated, for the prediction and detection of ‘43 pre-miRNA candidates bearing different types of SSR motifs’. The analysis identified 3913 different types of SSR motifs with an average of one SSR per 3.04 MB of thetranscriptome. About 0.033% of the transcriptome constituted ‘pre-miRNA candidates bearing SSRs’. The abundance, type and distribution of SSR motifs studied across the hair-pin miRNA precursors, showed a significant bias in the position of SSRs towards the downstream of predicted ‘pre-miRNA candidates’. The catalogue of transcripts identified, together with the demonstration of reliable existence of SSRs in the miRNA precursors, permits future opportunities for understanding the genetic mechanism of black pepper and likely functions of ‘tandem repeats’ in miRNAs.

Concepts: Scientific method, Gene, Genetics, Molecular biology, Genome, RNA, MicroRNA, Black pepper

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BACKGROUND AND PURPOSE: Capsaicin, the active ingredient of chilli, and medium-chain triglycerides (MCT) have been shown to increase diet-induced thermogenesis (DIT), improve satiety and decrease energy intake. Combinations of thermogenic ingredients have previously been investigated such as mustard and chilli, or capsaicin and green tea with positive effects. The aim of this study was to investigate the combined effects of chilli and MCT feeding on DIT and satiety in healthy volunteers. METHODS: Seven healthy volunteers were tested on four occasions following an overnight fast. Volunteers were fed a breakfast containing chilli and MCT oil, chilli and sunflower oil, bell pepper and sunflower oil or bell pepper and MCT oil. Satiety and gastrointestinal comfort were measured using visual analogue scales (VAS) and category scales. Baseline energy expenditure, and DIT and fat oxidation were measured for 6 h using indirect calorimetry. RESULTS: There were significant differences in DIT between the meals (P = 0.003) which increased from 7.0 % for pepper-sunflower oil to 10.7 % for chilli-MCT oil. The predominant differences existed between the chilli-MCT oil and chilli-sunflower oil (P = 0.013), between chilli-MCT oil and pepper-sunflower oil (P = 0.007) and between pepper-sunflower oil and pepper-MCT oil (P = 0.004). There was a significant difference in fat oxidation between the pepper-sunflower oil and pepper-MCT oil (P = 0.032). There were no differences in any VAS satiety parameters or gastrointestinal comfort ratings. CONCLUSION: Adding chilli and MCT to meals increases DIT by over 50 % which over time may cumulate to help induce weight loss and prevent weight gain or regain.

Concepts: Nutrition, Fatty acid, Triglyceride, Carbon, Thermogenesis, Capsicum, Scoville scale, Black pepper

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A number of factors likely affect the liking of capsaicin-containing foods such as social influences, repeated exposure to capsaicin, physiological differences in chemosensation, and personality. For example, it is well known that repeated exposure to capsaicin and chilies can result in chronic desensitization. Here, we explore the relationship between multiple personality variables - body awareness/consciousness, sensation seeking, and sensitivity to punishment, and sensitivity to reward - and the liking and consumption of capsaicin-containing foods. As expected, a strong relationship was found between liking of spicy foods and frequency of chili consumption. However, no association was observed between frequency of chili consumption and the perceived burn/sting of sampled capsaicin. Nor was there any association between perceived burn/sting of capsaicin and any of the personality measures. Private Body Consciousness did not relate to any of the measures used in the current study. Sensation Seeking showed positive correlations with the liking of spicy foods, but not non-spicy control foods. Sensitivity to Punishment showed no relation with frequency of chili consumption, and nonsignificant negative trends with liking of spicy foods. Conversely, Sensitivity to Reward was weakly though significantly correlated with the liking of a spicy meal, and similar nonsignificant trends were seen for other spicy foods. Frequency of chili consumption was positively associated with Sensation Seeking and Sensitivity to Reward. Present data indicate individuals who enjoy spicy foods exhibit higher Sensation Seeking and Sensitivity to Reward traits. Rather than merely showing reduced response to the irritating qualities of capsaicin as might be expected under the chronic desensitization hypothesis, these findings support the hypothesis that personality differences may drive differences in spicy food liking and intake.

Concepts: Scientific method, Food, Sense, Capsaicin, Chili pepper, Dissociative identity disorder, Black pepper, Spice

28

A series of compounds related to piperine and antiepilepsirine was screened in a monoamine oxidase A and B assay. Piperine is an alkaloid from the source plant of both black and white pepper grains, Piper nigrum. Piperine has been shown to have a wide range of activity, including MAO inhibitory activity. The z-factor for the screening assay was found to be greater than 0.8 for both assays. Notably, the compounds tested were selective towards MAO-B, with the most potent compound having an IC(50) of 498nM. To estimate blood-brain barrier (BBB) permeability, we used a PAMPA assay, which suggested that the compounds are likely to penetrate the BBB. A fluorescent bovine serum albumin (BSA) high-throughput screening (HTS) binding assay showed an affinity of 8μM for piperine, with more modest binding for other test compounds. Taken together, the data described here may be useful in gaining insight towards the design of selective MAO-B inhibitory compounds devoid of MAO-A activity.

Concepts: ELISA, Serum albumin, Neurotransmitter, Bovine serum albumin, Dopamine, Monoamine oxidase, Black pepper, Monoamine oxidase A

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This work examines the synthesis of 3,4-methylenedioxy-N-methylamphetamine (MDMA) from common starting materials that may be utilised by clandestine laboratory operators. Piperonal was prepared from two common starting materials, piperine (from pepper) and vanillin (a common flavouring). Piperine was converted to piperonal by ozonolysis and oxidative cleavage with potassium permanganate and tetrahydrofuran. Vanillin was converted to piperonal by demethylation with pyridine and aluminium chloride followed by methylenation with dichloromethane. The resulting piperonal samples were converted via a commonly encountered route to MDMA. The impurities that indicate a particular route were identified and the feasibility of each method was also assessed.

Concepts: Redox, Aluminium, Laboratory, Methamphetamine, MDMA, Potassium permanganate, Manganese, Black pepper

27

Activation of the pregnane X receptor (PXR) and subsequently its target genes, including those encoding drug transporters and metabolizing enzymes, while playing substantial roles in xenobiotics detoxification, might cause undesired drug-drug interactions. Recently, an increased awareness has been given to dietary components for potential induction of diet-drug interactions through activation of PXR. Here, we studied, whether piperine (PIP), a major component extracted from the widely-used daily spice black pepper, could induce PXR-mediated expression of cytochrome P450 3A4 (CYP3A4) and multidrug resistance protein 1 (MDR1). Our results showed that PIP activated human PXR (hPXR)-mediated CYP3A4 and MDR1 expression in human hepatocytes, intestine cells, and a mouse model; PIP activated hPXR by recruiting its coactivator SRC-1 in both cellular and cell-free systems; PIP bound to the hPXR ligand binding domain in a competitive ligand binding assay in vitro. The dichotomous effects of PIP on induction of CYP3A4 and MDR1 expression observed here and inhibition of their activity reported elsewhere challenges the potential use of PIP as a bioavailability enhancer and suggests that cautions should be taken for PIP consumption during drug treatment in patients, particularly those who favor daily pepper spice or rely on certain pepper remedies.

Concepts: DNA, Protein, Gene, Metabolism, Cytochrome P450, Steroid, CYP3A4, Black pepper

27

A reliable in vitro regeneration system for the economical and medicinally important Piper nigrum L. has been established. Callus and shoot regeneration was encouraged from leaf portions on Murashige and Skoog (MS) medium augmented with varied concentrations of plant growth regulators. A higher callus production (90 %) was observed in explants incubated on MS medium incorporated with 1.0 mg L(-1) 6-benzyladenine (BA) along with 0.5 mg L(-1) gibberellic acid after 4 weeks of culture. Moreover, a callogenic response of 85 % was also recorded for 1.0 mg L(-1) BA in combination with 0.25 mg L(-1) α-naphthalene acetic acid (NAA) and 0.25 mg L(-1) 2,4-dichlorophenoxyacetic acid or 0.5 mg L(-1) indole butyric acid (IBA) along with 0.25 mg L(-1) NAA and indole acetic acid. Subsequent sub-culturing of callus after 4 weeks of culture onto MS medium supplemented with 1.5 mg L(-1) thiodiazoran or 1.5 mg L(-1) IBA induced 100 % shoot response. Rooted plantlets were achieved on medium containing varied concentrations of auxins. The antioxidative enzyme activities [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)] revealed that significantly higher SOD was observed in regenerated plantlets than in other tissues. However, POD, CAT, and APX were higher in callus than in other tissues. A high-performance liquid chromatography (HPLC) fingerprint analysis protocol was established for quality control in different in vitro-regenerated tissues of P. nigrum L. During analysis, most of the common peaks represent the active principle “piperine.” The chemical contents, especially piperine, showed variation from callus culture to whole plantlet regeneration. Based on the deviation in chromatographic peaks, the in vitro-regenerated plantlets exhibit a nearly similar piperine profile to acclimated plantlets. The in vitro regeneration system and HPLC fingerprint analysis established here brought a novel approach to the quality control of in vitro plantlets, producing metabolites of interest with substantial applications for the conservation of germplasm.

Concepts: Superoxide dismutase, Chromatography, High performance liquid chromatography, Carboxylic acid, Butyric acid, Acids in wine, Black pepper, Piperine

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The aim of this study was to optimize the antioxidant activity of Piper nigrum L. essential oil extracted using the supercritical carbon dioxide (SC-CO2) technique. Response surface methodology was applied using a three-factor central composite design to evaluate the effects of three independent extraction variables: pressure of 15-30MPa, temperature of 40-50°C and dynamic extraction time of 40-80min. The DPPH radical scavenging method was used to evaluate the antioxidant activity of the extracts. The results showed that the best antioxidant activity was achieved at 30MPa, 40°C and 40min. The extracts were analyzed by GC-FID and GC-MS. The main components extracted using SC-CO2 extraction in optimum conditions were β-caryophyllene (25.38±0.62%), limonene (15.64±0.15%), sabinene (13.63±0.21%), 3-carene (9.34±0.04%), β-pinene (7.27±0.05%), and α-pinene (4.25±0.06%). The essential oil obtained through this technique was compared with the essential oil obtained using hydro-distillation. For the essential oil obtained by hydro-distillation, the most abundant compounds were β-caryophyllene (18.64±0.84%), limonene (14.95±0.13%), sabinene (13.19±0.17%), 3-carene (8.56±0.11%), β-pinene (9.71±0.12%), and α-pinene (7.96±0.14%). Radical scavenging activity of the extracts obtained by SC-CO2 and hydro-distillation showed an EC50 of 103.28 and 316.27µgmL(-1) respectively.

Concepts: Carbon dioxide, Carbon, Response surface methodology, Supercritical fluid, Supercritical carbon dioxide, Inorganic solvents, Black pepper, Central composite design