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Suppressing toxic aldehydic lipid oxidation product (LOP) generation in culinary oils is now considered vital, since the deleterious effects arising from their ingestion are implicated in a wide range of disease conditions. Partial substitution involves the replenishment of thermally-stressed culinary oils with corresponding unheated ones. This technique was tested by employing 10%, 25%, 50%, and 75% (v/v) partial substitutions of coconut, olive, rapeseed, and sunflower oils at 180℃ for a 300 min continuous thermo-oxidation duration. Oil samples were analysed by proton nuclear magnetic resonance (

Glucuronidation is a Phase 2 metabolic pathway responsible for the metabolism and excretion of testosterone to a conjugate testosterone glucuronide. Bioavailability and the rate of anabolic steroid testosterone metabolism can be affected upon UGT glucuronidation enzyme alteration. However, there is a lack of information about the in vitro potential assessment of UGT2B17 inhibition by salicylic acid. The purpose of this study is to investigate if UGT2B17 enzyme activity is inhibited by salicylic acid. A UGT2B17 assay was developed and validated by HPLC using a C18 reversed phase column (SUPELCO 25 cm × 4.6 mm, 5 μm) at 246 nm using a gradient elution mobile phase system: (A) phosphate buffer (0.01 M) at pH = 3.8, (B) HPLC grade acetonitrile and (C) HPLC grade methanol. The UGT2B17 metabolite (testosterone glucuronide) was quantified using human UGT2B17 supersomes by a validated HPLC method. The type of inhibition was determined by Lineweaver–Burk plots. These were constructed from the in vitro inhibition of salicylic acid at different concentration levels. The UGT2B17 assay showed good linearity (R2 &gt, 0.99), acceptable recovery and accuracy (80–120%), good reproducibility and acceptable inter and intra-assay precision (&lt, 15%), low detection (6.42 and 2.76 μM) and quantitation limit values (19.46 and 8.38 μM) for testosterone and testosterone glucuronide respectively, according to ICH guidelines. Testosterone and testosterone glucuronide were found to be stable up to 72 h in normal laboratory conditions. Our investigational study showed that salicylic acid uncompetitively inhibited UGT2B17 enzyme activity. Thus, drugs that are substrates for the UGT2B17 enzyme have negligible potential effect of causing interaction with salicylic acid in humans.

Cannabis products (marijuana, weed, hashish) are among the most widely abused psychoactive drugs in the world, due to their euphorigenic and anxiolytic properties. Recently, hair analysis is of great interest in analytical, clinical, and forensic sciences due to its non-invasiveness, negligible risk of infection and tampering, facile storage, and a wider window of detection. Hair analysis is now widely accepted as evidence in courts around the world. Hair analysis is very feasible to complement saliva, blood tests, and urinalysis. In this review, we have focused on state of the art in hair analysis of cannabis with particular attention to hair sample preparation for cannabis analysis involving pulverization, extraction and screening techniques followed by confirmatory tests (e.g., GC–MS and LC–MS/MS). We have reviewed the literature for the past 10 years’ period with special emphasis on cannabis quantification using mass spectrometry. The pros and cons of all the published methods have also been discussed along with the prospective future of cannabis analysis.

OBJECTIVES: Christian Orthodox fasting (COF), a periodical vegetarian subset of the Mediterranean diet, has been proven to exert beneficial effects on human health. Athonian fasting is a pescetarian COF variation, where red meat is strictly restricted throughout the year. Previous studies have examined the COF nutritional synthesis and health effects in general population fasters (GF) and Athonian monks (AM), separately. The aim of this study is to comparatively evaluate the characteristics and effects of this nutritional advocacy between the two populations. METHODS: The study included 43 male GFs (20-45 y of age) and 57 age-matched male AMs following COF. Dietary intake data were collected in both groups during a restrictive (RD) and a nonrestrictive (NRD) day. Nutritional, cardiometabolic, and anthropometric parameters were compared between the two cohorts. RESULTS: AM presented lower daily total caloric intake for both RD (1362.42 ± 84.52 versus 1575.47 ± 285.96 kcal, P < 0.001) and NRD (1571.55 ± 81.07 versus 2137.80 ± 470.84 kcal, P < 0.001) than GF.They also demonstrated lower body mass index (23.77 ± 3.91 versus 28.92 ± 4.50 kg/m2, P

Infection caused by high-risk human papillomaviruses (HPVs) are implicated in the aetiology of cervical cancer. Although current methods of treatment for cervical cancer can ablate lesions, preventing metastatic disseminations and excessive tissue injuries still remains a major concern. Hence, development of a safer and more efficient treatment modality is of vital importance. Natural products from plants are one of the principal sources of precursors to lead compounds with direct pharmaceutical application across all disease classes. One of these plants is Ficus carica, whose fruit latex, when applied on HPV-induced skin warts, has shown potential as a possible cure for this virus related lesions. This study explores the in vitro biological activities of fig latex and elucidates its possible mechanisms of action on cervical cancer cell lines CaSki and HeLa positive for HPV type 16 and 18, respectively. Our data shows that fig latex inhibits properties that are associated with HPV-positive cervical cancer transformed cells such as rapid growth and invasion and substantially downregulated the expression of p16 and HPV onco-proteins E6, E7. These findings suggest Ficus carica latex has the potential to be used in the development of therapeutic modalities for the possible treatment, cure and prevention of HPV related cervical cancer.

Vitamin D deficiency is being recognized as a global issue and has been implicated in many health issues. Hence, there is an increased interest in developing sensitive, reproducible, and non-invasive assays to measure Vitamin D levels. This study aimed to apply a sensitive liquid chromatography-mass spectrometric assay to hair samples to develop and validate a clinical assay to provide a quarterly average level of vitamin D in one test. Hair samples were collected from 70 male university students/young adults and pulverized/sonicated in methanol/water for 2 h to extract Vitamin D metabolites. A sensitive liquid chromatographic-mass spectrometric assay was employed to quantitate vitamin D and metabolites. Of the eight Vitamin D and metabolites screened, only the primary, clinically significant form of vitamin D (25OHD3) was detected and quantified in hair samples in the range of 17–1541 pg/mg. One-third of the hair samples (21 out of 70) had Vitamin D levels below the LLOD of the assay (10 pg/mg). The mean and standard deviation values for hair (25OHD3) were 276.7 ± 329.9, respectively. This pilot study reveals the potential of the vitamin D hair test in clinical assays as a complementary test to a vitamin D blood test, which would provide a quarterly average.

Objective\ud To explore the potential of combining non-contact profilometry (NCP) and confocal laser scanning microscopy (CLSM) data to measure the entire erosive process non-destructively and to validate findings using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), scanning electron microscopy (SEM) and surface microhardness (SMH) using the same samples throughout.\ud \ud Methods\ud Polished bovine enamel samples (n = 35) were divided into groups (7/group) with similar SMH values. Samples underwent individual erosive challenges (1 % citric acid, pH3.8) for 1, 5, 10, 15 or 30 min under stirring and aliquot extracts were analysed for Ca and P by ICP-AES. SMH was used to measure erosive softening. Profilometry was used to assess bulk volume loss (BVL). Images were captured by SEM. Samples were stained with rhodamine-B (0.1 mM, 24 h) and images captured by CLSM. Image processing was used to determine changes in fluorescent volume for the first 10 μm (ΔFV10) for each enamel sample which were combined with BVL to calculate total lesion volume (TLV). ANOVA, linear regression and Pearson correlation analysis were used where applicable.\ud \ud Results\ud Surface softening, [Ca], [P], BVL and ΔFV10μm increased with acid erosion duration which were significant by 10 min (P

Differences in lipid oxidation products (LOPs) and trace metal concentrations of French fry samples found between two global chain fast-food restaurants in the UK were investigated using high-resolution proton nuclear magnetic resonance ( H NMR) and inductively coupled plasma-optical emission spectrometry (ICP-OES) analyses, respectively, of extracts derived therefrom. Over the course of 3 days and 3 different diurnal time periods, samples of French fries (FFs) were analyzed, and comparisons of two different oil extraction methods were undertaken for the two restaurants involved. The magnitude of concentrations of LOPs extracted from FFs is discussed. Significant differences between 6/7 aldehyde classifications, and aluminum, manganese, vanadium, lead, iron, copper and nickel levels between samples from the two restaurants are also reported. Redox-active transition and further trace metal concentrations inversely correlated with FF oil sample LOP contents; this suggested an antioxidant rather than a pro-oxidant role for them. [Abstract copyright: Copyright © 2021 Le Gresley, Ampem, De Mars, Grootveld and Naughton.]

High-resolution 1H nuclear magnetic resonance (NMR) analysis was employed to molecularly screen the lipid, lipid oxidation product (LOP), and antioxidant compositions of four natural (unrefined) cod liver oil (CLO) products. Products 1&ndash, 3 were non-fermented CLOs, whilst Product 4 was isolated from pre-fermented cod livers. Supporting analytical data that were acquired included biogenic amine, flavanone, tannin, phenolic antioxidant, &alpha, tocopherol, and oxygen radical absorbance capacity (ORAC) determinations by recommended HPLC, LC/MS/MS, or spectrophotometric methods. SDS-PAGE, HPLC, and 1H NMR analyses investigated and determined collagenous antioxidants and their molecular mass ranges. 1H NMR analysis of aldehydic LOPs was employed to explore the susceptibilities/resistivities of each CLO product to peroxidation that is induced by thermal stressing episodes (TSEs) at 180&deg, C, or following prolonged (42 day) storage episodes at 4 and 23 &deg, C. Product 4 displayed extremely high ORAC values, which were much greater than those of Products 1&ndash, 3, and that were predominantly ascribable to significant levels of peroxidation-blocking and/or aldehyde-consuming collagenous polypeptides/peptides and ammoniacal agents therein. Significantly lower levels of toxic aldehydes were generated in the pre-fermented Product 4 during exposure to TSEs, or the above long-term storage episodes. These results confirmed the enhanced peroxidative resistivity of a fermented, antioxidant-fortified natural CLO product over those of non-fermented unrefined products. Product 4: Green Pasture Blue Ice&trade, Fermented Cod Liver Oil.

Absence of adequate maternal vitamin D supplementation and decreased maternal ultraviolet exposure during pregnancy are key determinants for the manifestation of neonatal hypovitaminosis D at birth. These parameters may vary, according to country-specific dietary patterns, health policies and sunshine exposure. We aimed to investigate differences in calcium metabolism and anthropometric profiles according to neonatal vitamin D status at birth, in a sunny region of Northern Greece. A secondary aim was to identify maternal parameters as risk factors for developing neonatal vitamin D deficiency at birth. A total of 129 mother-neonate pairs were included in the study and classified into three groups, according to neonatal 25-hydroxy-D [25(OH)D)] concentrations at birth [deficiency (30 nmol/l), insufficiency (30-50 nmol/l) and sufficiency (50 nmol/l)]. Neonatal biochemical and anthropometric profiles and maternal demographic, social, dietary and biochemical profiles were comparatively evaluated between the three groups. Univariate and multivariate logistic regression was performed to identify independent associations of maternal factors with neonatal vitamin D status. Vitamin D deficient-neonates manifested higher parathyroid hormone (7.20 ± 2.60 vs 5.50 ± 1.50 pg/ml, p = 0.01) and lower corrected calcium (10.70 ± 0.70 vs 11.30 ± 1.30 mg/dl, p = 0.02) concentrations compared with vitamin d-insufficient neonates. Mothers of vitamin D deficient and insufficient neonates had a lower total of 25(OH)D (31.7 ± 19.2 and 36.5 ± 22.3 vs 53.3 ± 39.0 nmol/l, p 0.01) and 25(OH)D

High-resolution NMR analysis has been used, for the first time, to identify, putatively, two new secondary aldehydic lipid oxidation products in culinary oils. The impact of heating and cooling times on the thermal stability, fatty acid composition and lipid oxidation product (LOP) concentrations have been analysed for continuous and discontinuous heating periods (180 °C). The susceptibility of the selected oils to thermal oxidation for the different heating episodes has been evaluated via the detection and determination of LOPs, particularly cytotoxic and genotoxic aldehydes. The identities and quantities of these LOPs evolved throughout a 2.0 hour period. Results acquired indicated that sunflower oil was more resistant to discontinuous oxidation than rapeseed and olive oils, however overall discontinuous heating resulted in more LOPs.

Compensatory changes in appetite and energy intake do not appear to occur in the short-term after acute exercise; however, responses have not been compared in South Asians, a group at high risk of central obesity and type 2 diabetes, with white Europeans. This study examined appetite perceptions, energy intake and appetite-related hormones after moderate-to-vigorous intensity cycling in South Asian versus white European men. Fifteen South Asians (mean(SD) 29(8) years; 25.4(4.5) kg m ) and fifteen white Europeans (33(10) years; 26.1(3.8) kg m ) matched for age and body mass index completed two 7 h trials (control and exercise). Participants rested throughout both trials apart from completing 60 min cycling at 2-3 h in the exercise trial. A standardised breakfast was consumed at 0 h and an ad libitum buffet meal at 4 h. Appetite perceptions and appetite-related hormones were measured at predetermined intervals. Exercise suppressed acylated ghrelin (d = 0.19, P < 0.001) and increased total peptide YY (PYY) (d = 0.14, P = 0.004), insulin (d = 0.09, P = 0.046) and glucose concentrations (d = 0.31, P < 0.001) (main effect of trial), without stimulating compensatory increases in energy intakes in either group (group-by-trial interactions). South Asians exhibited lower absolute energy intake and higher insulin concentrations than white Europeans (main effect group d ≥ 0.63, P ≤ 0.003), whereas group-by-time interactions revealed lower acylated ghrelin concentrations at 3 and 4 h (d ≥ 0.75, P ≤ 0.038) and higher glucose concentrations at 0.75 and 2 h (d ≥ 0.67, P ≤ 0.008) in South Asian than white European men. These findings demonstrate that acute exercise induces a short-term energy deficit and similar appetite responses in South Asian and white European men. [Abstract copyright: Copyright © 2021 Elsevier Ltd. All rights reserved.]

Maternal vitamin D deficiency is considered to be the key determinant of the development of neonatal vitamin D deficiency at birth and during early infancy. Specific vitamin D receptor (VDR) gene polymorphisms have been associated with adverse pregnancy and offspring outcomes. The aim of this study was to evaluate the effect of maternal and neonatal VDR polymorphisms (ApaI, TaqI, BsmI, FokI, Tru9I) on maternal and neonatal vitamin D status. VDR polymorphisms were genotyped in 70 mother-neonate pairs of Greek origin, and classified according to international thresholds for Vitamin D status. Mean neonatal and maternal 25-hydroxy-vitamin D [25(OH)D] concentrations were 35 ± 20 and 47 ± 26 nmol/l, respectively. Neonatal VDR polymorphisms were not associated with neonatal 25(OH)D concentrations. In contrast, mothers with the Fokl FF polymorphism had a 70 % lower risk of vitamin D deficiency [25(OH)D

The inhibitory effect of new chemical entities on rat liver P450 marker activities was investigated in a functional approach towards drug development. Treatment of colorectal cancer (CRC) and chemoprevention using salicylic acid has gained a lot of attention, mainly in the prevention of the onset of colon cancer. Thus, an in vitro inhibitory effect of salicylic acid on rat CYP2C11 activity was examined by using high performance liquid chromatography (HPLC). High performance liquid chromatography analysis of a CYP2C11 assay was developed on a reversed phase C18 column (SUPELCO 25 cm &times, 4.6 mm &times, 5 &micro, m) at 243 nm using 32% phosphate buffer (pH 3.36) and 68% methanol as a mobile phase. The CYP2C11 assay showed good linearity for all components (R2 &gt, 0.999). Substrates and metabolites were found to be stable for up to 72 hours. Additionally, the method demonstrated good reproducibility, intra- and inter-day precision (&lt, 15%), acceptable recovery and accuracy (80%&ndash, 120%), and low detection (1.3501 &micro, M and 3.2757 &micro, M) and quantitation limit values (4.914 &micro, M and 9.927 &micro, M) for 16&alpha, hydroxytestosterone and testosterone, respectively. Salicylic acid acts reversibly as a noncompetitive (weak) inhibitor with Ki = 84.582 &plusmn, 2.67 &micro, M (concentration of inhibitor to cause 50% inhibition of original enzyme activity (IC50) = 82.70 &plusmn, M) for CYP2C11 enzyme activity. This indicates a low potential to cause toxicity and drug&ndash, drug interactions.

South Asians have a greater cardiovascular disease (CVD) and type 2 diabetes (T2D) risk than white Europeans, but the mechanisms are poorly understood. This study examined ethnic differences in free fatty acids (FFAs) metabolic profile (assessed using liquid chromatography-mass spectrometry), appetite-related hormones and traditional CVD and T2D risk markers in blood samples collected from 16 South Asian and 16 white European men and explored associations with body composition, objectively-measured physical activity and cardiorespiratory fitness. South Asians exhibited higher concentrations of five FFAs (laurate, myristate, palmitate, linolenic, linoleate, p &le, 0.040), lower acylated ghrelin (ES = 1.00, p = 0.008) and higher leptin (ES = 1.11, p = 0.004) than white Europeans, total peptide YY was similar between groups (p = 0.381). South Asians exhibited elevated fasting insulin, C-reactive protein, interleukin-6, triacylglycerol and ratio of total cholesterol to high-density lipoprotein cholesterol (HDL-C) and lower fasting HDL-C (all ES &ge, 0.74, p &le, 0.053). Controlling for body fat percentage (assessed using air displacement plethysmography) attenuated these differences. Despite similar habitual moderate-to-vigorous physical activity (ES = 0.18, p = 0.675), V ˙ O2max was lower in South Asians (ES = 1.36, p = 0.001). Circulating FFAs in South Asians were positively correlated with body fat percentage (r2 = 0.92), body mass (r2 = 0.86) and AUC glucose (r2 = 0.89) whereas in white Europeans FFAs were negatively correlated with total step counts (r2 = 0.96). In conclusion, South Asians exhibited a different FFA profile, lower ghrelin, higher leptin, impaired CVD and T2D risk markers and lower cardiorespiratory fitness than white Europeans.

Inhibition of cytochrome P450 (CYP) alters the pharmacokinetic parameters of the drug and causes drug&ndash, drug interactions. Salicylic acid been used for the treatment of colorectal cancer (CRC) and chemoprevention in recent decades. Thus, the aim of this study was to examine the in vitro inhibitory effect of salicylic acid on CYP2E1 activity in rat liver microsomes (RLMs) using high-performance liquid chromatography (HPLC). High-performance liquid chromatography analysis of a CYP2E1 assay was developed on a reversed phase C18 column (SUPELCO 25 cm &times, 4.6 mm &times, 5 &micro, m) at 282 nm using 60% H2O, 25% acetonitrile, and 15% methanol as mobile phase. The CYP2E1 assay showed a good linearity (R2 &gt, 0.999), good reproducibility, intra- and inter-day precision (&lt, 15%), acceptable recovery and accuracy (80&ndash, 120%), and low detection (4.972 &micro, M and 1.997 &micro, M) and quantitation limit values (15.068 &micro, M and 6.052 &micro, M), for chlorzoxazone and 6-hydroxychlorzoxazone, respectively. Salicylic acid acts as a mixed inhibitor (competitive and non-competitive inhibition), with Ki (inhibition constant) = 83.56 &plusmn, 2.730 &micro, M and concentration of inhibitor causing 50% inhibition of original enzyme activity (IC50) exceeding 100 &micro, M (IC50 = 167.12 &plusmn, 5.460 &micro, M) for CYP2E1 enzyme activity. Salicylic acid in rats would have both low and high potential to cause toxicity and drug interactions with other drugs that are substrates for CYP2E1.

Understanding the potential for adverse drug reactions (ADRs), from herb-drug interactions, is a key aspect of medicinal plant safety, with particular relevance for public health in countries where medicinal plant use is highly prevalent. We undertook an in-depth assessment of extracts of Hyptis verticillata Jacq., via its impact on activities of key cytochrome P450 (CYP) enzymes (CYPs 1A1, 1A2, 1B1, 3A4 and 2D6), its antioxidant properties (determined by DPPH assays) and chemical characterisation (using LC-MS). The dried plant aqueous extract demonstrated potent inhibition of the activities of CYPs 1A1 (7.6 µg/mL), 1A2 (1.9 µg/mL), 1B1 (9.4 µg/mL) and 3A4 (6.8 µg/mL). Further analysis of other crude extracts demonstrated potent inhibition of CYP1A2 activity for a dried plant ethanol extract (1.5 µg/mL), fresh plant ethanol extract (3.9 µg/mL), and moderate activity for a fresh plant aqueous extract (27.8 µg/mL). All four extracts demonstrated strong antioxidant activity, compared to the positive control (ascorbic acid, 1.3 µg/mL), with the dried plant ethanol extract being the most potent (1.6 µg/mL). Analysis of the dried plant aqueous extract confirmed the identity of seven phytochemicals, five lignans and two triterpenes. Individual screening of these phytochemicals against the activity of CYP1A2 identified yatein as a moderate inhibitor (71.9 μM), likely to contribute to the plant extract’s potent bioactivity. Further analysis on the impact of this plant on key drug metabolizing enzymes in vivo appears warranted for likely ADRs, as well as furthering development as a potential chemopreventive agent.

International audience; CONTEXT: Adult body fat is associated with birth anthropometry, suggesting a role for metabolic regulators including vitamin D and the adipokines-adiponectin and irisin-which have been reported to interact but, as yet, data remain controversial.OBJECTIVE: To study (i) the relationship between vitamin D, its binding protein (VDBP) and the adipokines, adiponectin, and irisin in mothers and neonates at birth and (ii) their effects on neonate anthropometric outcomes.DESIGN: Cross-sectional study for healthy mothers with full-term and uncomplicated births.SETTING: Primary care.SUBJECTS: Seventy pairs of newly delivered neonates and their mothers.MAIN OUTCOMES FEATURES: Biochemical markers from maternal and cord: VDBP, adiponectin, irisin, calcium, albumin, parathyroid hormone, 25OHD, 1,25(OH)D. Maternal demographic and social characteristics and neonate anthropometric parameters were recorded.RESULTS: Maternal VDBP levels (364.1 ± 11.9 μg/ml) demonstrated a strong positive correlation with maternal adiponectin (4.4 ± 0.4 μg/ml) and irisin (308.8 ± 50.8 ng/ml) concentrations, which remained significant (p CONCLUSIONS: The main findings of this study are (i) the perspective of a potential independent interaction of VDBP and adiponectin in both mothers and neonates and (ii) the lack of a causative model effect of both maternal/neonatal vitamin D status and adipokine profile on neonatal anthropometry at birth, as a surrogate marker of future metabolic health of the offspring.

In clinical settings, nitrous oxide gas is a safe anesthetic used during childbirth, in dentistry, and to relieve anxiety in emergencies. Colloquially known as “hippy crack”’ or “laughing gas,” it is increasingly taken recreationally for its euphoric and relaxing effects and hallucinogenic properties. Using a self-reported survey, we gathered quantitative and qualitative information on users and non-users of hippy crack among a young population regarding: consumption patterns, knowledge, risk awareness and intentions toward future abuse. Quantitative responses from a total of 140 participants were analyzed for frequencies and relationships, whereas qualitative data were evaluated via identifying the reoccurring themes. Overall, 77.1% (n = 108) had heard of hippy crack and 27.9% (n = 39) admitted to past-year use. Prior users mostly indicated intended future use, had an average low number of past-year uses but some with > 20 occasions, had a varied number of inhalations per occasion (often 1–10) with an effect lasting up to 5 min, and a majority preferred social rather than lone use. For non-users, 79.2% said they would take hippy crack with the vast majority (94%) preferring a social setting. The results show a concerning gap between available evidence and awareness of side effects. Despite serious reported side effects, including psychosis and myeloneuropathy—especially on the young developing brain—only a minority (29.3%) was aware of any side effects. In contrast, in a hypothetical scenario depicting a first social encounter with hippy crack, the qualitative responses were in contrast to qualitative outcomes revealing that participants would try (n = 30)/not try (n = 25) it, would feel under pressure to try it (n = 6) with only 11 opting to exit the situation. In summary, this first report of trends and perceptions of the use of hippy crack among young adults in the England highlights a lack of concern with side effects, coupled to a willingness to partake. Because typical users are young with risks to the still developing brain, education about the nitrous oxide abuse is warranted to prevent impaired brain development. Further studies to investigate the possible effects of nitrous oxide on the developing brain in young adults would advance meaningful prevention.

The purported health benefits of red wine owing to its antioxidant capacity attract persistent attention. The aims of this study were (1) to explore the contribution of individual phenolic constituents to the antioxidant capacity of red wine and (2) to investigate if non-phenolic components have oxidant or antioxidant effects. Red wine and a faux wine model systems of 11 constituent phenolic compounds were challenged with increasing concentrations of selected oxidants, namely hydrogen peroxide, redox-active transition metal ions and Fenton systems. Antioxidant activity was assessed through loss of phenolic components upon oxidant challenge measured using a reversed-phase high-performance liquid chromatography (HPLC) method. Elemental analysis of the red wine showed that the concentration of metal ions was in the range of 0.01–1.11 mM, whereas the concentration of antioxidant phenolics in the wine was in the range of 0.10–4.40 mM. As expected, addition of all five oxidant systems to both red wine and faux wine resulted in considerable loss of phenolics as quantified by HPLC. However, the pattern of phenolic loss was largely similar for both red wine and the faux wine across the five oxidant systems (frequently ca. 50–60% with the initial addition of oxidant), with only marginal effects on antioxidant activity after further additions of oxidants. The data revealed that the non-phenolic wine constituents had little influence over oxidative processes under the conditions studied, suggesting that the antioxidant properties observed in the tested red wine could be attributable to the phenolics.

Introduction The autobiographical Implicit Association Test (aIAT) is a variant of the Implicit Association Test reportedly capable of detecting an individual's concealed autobiographical event with very high accuracy. A previous attempt to utilize this measurement technique for the identification of cocaine users rendered an alarming rate of false positives. In this study, we aimed to explore the potential reasons behind the measurement's inaccuracy. Methods Two versions of the cocaine aIAT were devised with different category labels (descriptive ‘guilty/innocent’ and self-referenced ‘as if you were/were not’). Forty-one cocaine abstinent participants (43.9% male; mean age = 28.17 ± 7.36) were randomly assigned to one of the two conditions. Self-declared cocaine abstinence was confirmed for the 12-month period preceding data collection through hair analysis. Participants were also administered bespoke implicit and explicit cocaine user attitude measures, the self-esteem IAT and the Rosenberg self-esteem scale. Results The category labels which elicited self-referenced knowledge showed low accuracy (19%) compared to the 65% of the ‘guilty/innocent’ labels proposed by original authors. The self-referenced aIAT version significantly correlated with the self-concept measures. The aIAT outcomes were independent from attitudes toward cocaine users. Conclusions Category labels play an influential role in determining the test's accuracy, demonstrating that participants’ propositional knowledge and self-concept are involved during test performance. The aIAT does not appear to tap directly into an individual's implicit memory when relevant memory is not available. Although the test cannot be recommended for detecting drug use, further research should investigate underlying mechanisms and other potentials of the technique.

Acetobacter aceti was successfully used to induce transcriptional analysis, plant biochemical profile and nutritional contents in barley. Association of barley and microbial strain AC8 was reported best combination among other eight microbial strains of Bacillus in this study. AC8 microbial strain was identified as A. aceti from Fungal Culture Bank, Institute of Agricultural Science, University of the Punjab, Lahore, Pakistan. Different strains of A. aceti i.e., AC1, AC2, AC3… AC8 were analyzed as plant inducers and AC8 was screened out as best inducer in barley. It induced highest quantities of plant biochemical (i.e., phytosterols, phenolics, alkaloids and terpenoids) in barley. Current study revealed that among eight microbial strains AC8 had maximum potential to increase ascorbic acid, panthothenic acid, pyridoxine, thiamine and riboflavin in barley than other microbial strains. AC8 screened out among other microbial strains on the basis of its high vitamins induction potential. AC3 plus AC6 were reported second in the recorded list although other strains had a chronological reduction in vitamins as AC2>AC7>AC4>AC5 and AC1. Further evaluation of AC8 was done to check its efficiency for biochemicals, nutritional and isozyme contents induction in barley. Statistical analysis was performed using ANOVA and DMRT through DSAASTAT.

Greek Orthodox fasting (OF), which involves 180-200 days of fasting per year, is dictated by the Christian Orthodox religion. For the first time, this cross-sectional study examines the characteristics and the effects of OF on anthropometry, cardiometabolic markers and calcium homeostasis in Athonian monks (AMs).Daily intakes of energy, macro- and micronutrients of a day during a weekend of Nativity Fast, defined as non-restrictive day (NRD), and a weekday during Great Lent, labeled as restrictive day (RD) were recorded.The daily energy intake of 70 AM (age=38.8±9.7 years) was low during both RD and NRD (1265.9±84.5 vs 1660±81 kcal, respectively, P0.001). Paired samples t-test showed statistically significant difference between daily intakes in RD and NRD: carbohydrates (159.6±21.8 vs 294.3±23.4 g, P0.0001) and saturated fat (12.7±0.0 vs 16.4±0.0 g, P0.0001) were lower, whereas protein (89.2±1.3 vs 72.35±1.3 g, P0.001) was higher during RD. A subsample of 50 monks (age=38.7±10.6 years) formed a study cohort for cardiometabolic and calcium homeostasis assessment. Body weight (74.3±12.9 kg) and body mass index (BMI; 23.8±4.1 kg/mUnaffected by variation in lifestyle factors, the results of this unique study offers clear evidence for the health benefits of the strict Athonian OF through optimal lipid and glucose homeostasis.

Following the experiment that have been designed for immunological screening test followed by a GC-MS confirmation method for the simultaneous analysis of delta-9-tetrahydrocannabinol (9-THC) , 11-nor-9-Carboxy-THC (THC-COOH), 11-Hydroxy-THC (OH-THC), cocaine, benzoylecgonine (BZ) in human hair and avoiding the significant factors responsible for drug degradation by acid and alkali hydrolysis and replace it with enzymatic hair digestion to obtain optimal recovery conditions, that resulted a false positive results after the radioimmunoassay pre-analysis associated with enzymatic hair digestion. Enzymatic hair digestion was used to hydrolyse 18 Turkish samples using proteinase K, Dithiothreitol and Tris HCl buffer. At the beginning 18 out of 18 samples showed to be screened positive on ELISA, towards the end using a threshold of 0.1ng/mg in 50mg hair, 2 positive results were found in 18 cases. It was revealed that the 2 samples were positive for 9-THC and THC-COOH. Cross reaction lead to false positive results in the pre screening step as a result of the degradation of the antibodies in the precoated ELISA micro plate. Sixteen out of 18 positive detections could not be confirmed by GC-MS. In this study we investigated cause behind the drug screening malfunction.

In vitro studies show that diclofenac inhibits enzymatic steroid glucuronidation. This study was designed to investigate the influence of diclofenac on the excretion of stanozolol and 3'-hydroxystanozolol via analyses in hair, blood and urine in vivo in a rat study. Brown Norway rats were administered with stanozolol (weeks 1-3) and diclofenac (weeks 1-6). Weekly assessment of steroid levels in hair was complemented with spot urine and serum tests. Levels of both stanozolol and 3'-hydroxystanozolol steadily increased in hair during stanozolol treatment and decreased post-treatment, but remained readily detectable for 6 weeks. In contrast, compared to control rats, diclofenac significantly reduced urinary excretion of 3’-hydroxystanozolol which was undetectable in most samples. This is the first report of diclofenac altering steroid metabolism in vivo, detrimentally affecting detection in urine, but not in hair which holds considerable advantages over urinalysis for anti-doping tests.

Summary Pomegranates (Punica granatum) have been known for centuries for their healing properties. The phenolic components of pomegranate are believed to be responsible for their antioxidant activities, hence playing a major role in reducing oxidative stress-related disease, such as cardiovascular diseases, cancer, and neurodegenerative diseases. In this study, thin-layer chromatography (TLC) was used to, first, separate and identify the phenolic constituents of pomegranate juice and, second, to assess the antioxidant efficacies of the identified compounds. Different oxidant and hydroxyl radical generating systems (Fe3+, Cu2+, H2O2, Fe2+-H2O2, and Cu2+-H2O2) were used in assessing the efficacies of phenolic compounds found in pomegranate juice. A 10 × 10 cm and 20 × 20 cm sized silica gel 60 F254 TLC plates with toluene-ethyl acetate-formic acid (60:40:10 v/v/v) as a mobile phase were used for the chromatographic separation. Two compounds, ellagic acid and gallic acid, were separated and identified. When ...

Vitamin D status during pregnancy is linked to bone mineralization of developing fetus, which justifies targeting sufficient levels of vitamin D in pregnant women. Despite high level of sunshine in the Mediterranean regions, maternal hypovitaminosis D remain common in these countries. The aim of this narrative review was to provide potential explanations for this phenomenon in an effort to guide future public health policies and vitamin D intakes during pregnancy. We searched Medline for publications regarding hypovitaminosis D during pregnancy in the Mediterranean region. Available studies confirmed the high prevalence of hypovitaminosis D among pregnant women in the Mediterranean regions (50-65% in most studies), resulting in severe skeletal and nonskeletal health events among the offspring. Reasons for this may rely on maternal darker skin pigmentation, poor dietary vitamin D intake, veiled clothing and reduced sunshine exposure, health policies and increased prevalence of obesity. Public health organizations should be aware of this phenomenon and develop specific policies to prevent hypovitaminosis D and its adverse outcomes in maternal and neonatal health.

Solobacterium moorei has recently been implicated as a causative agent of halitosis. In vitro experiments to evaluate the role of S. moorei in halitosis have, however, been complicated by a paucity of information on the ideal conditions for culturing this organism. This work aimed to optimize a liquid culture medium for S. moorei, and to determine the growth-curve of the organism. Further, the ability of S. moorei to generate volatile sulfur compounds was investigated and compared quantitatively to other oral anaerobes by an optimized head-space gas chromatography method. Serum-supplementation of standard liquid growth media gave greater growth of S. moorei than non-supplemented broths, with the best medium found to be serum-supplemented tryptone soya broth. S. moorei was able to metabolize cysteine directly to hydrogen sulfide, but was unable to produce methanethiol from methionine. S. moorei produced 2-3 times more hydrogen sulfide (normalized for colony forming units) than Porphyromonas gingivalis and Veillonella dispar, but considerably less than Fusobacterium nucleatum. The study has identified reliable growth conditions for culture of S. moorei, which were employed to show that S. moorei has the requisite biochemistry consistent with a potential role in halitosis.

Prevalence estimation models, using randomized or fuzzy responses, provide protection against exposure to respondents beyond anonymity and represent a useful research tool in socially sensitive situations. However, both guilty and innocent noncompliance can have a profound impact on prevalence estimations derived from these models. In this article, we introduce the maximum-likelihood extension of the single sample count (SSC-MLE) estimation model to detect and attribute noncompliance through testing 5 competing hypotheses on possible ways of noncompliance. We demonstrate the ability of the SSC-MLE to estimate and attribute noncompliance with a single sample using the observed distribution of affirmative answers on recent recreational drug use from a sample of university students (N = 1,441). Based on the survey answers, the drug use prevalence was estimated at 17.62% (± 6.75%), which is in line with relevant drug use statistics. Only 2.51% (± 1.54%) were noncompliant, of which 0.55% (± 0.44%) was attributed to guilty noncompliance (i.e., have used drugs but did not admit) and 2.17% (± 1.44%) to innocent noncompliers with no drug use in the past 3 months to hide. The SSC-MLE indirect estimation method represents an important tool for estimating the prevalence of a broad range of socially sensitive behaviors. Subsequent applications of the SSC-MLE to a range of transgressive behaviors with varying sensitivity will contribute to establishing the SSC-MLE's performance properties, along with obtaining empirical evidence to test the underlying assumption of independence of noncompliance from involvement. Freely downloadable, user-friendly software to facilitate applications of the SSC-MLE model is provided.

Glomerular filtration rate (GFR) is usually determined by estimation of iothalamate (IOT) clearance. We have developed and validated an accurate and robust method for the analysis of IOT in human plasma and urine. The mobile phase consisted of methanol and 50 mM sodium phosphate (10:90; v/v). Flow rate was 1.2 mL/min on a C18 reverse phase column, Synergi-hydro (250 × 4.6 mm) 4 µm 80 Å, with an ultraviolet detector set to 254 nm. Acetonitrile was used for the deproteination and extraction of IOT from human plasma and urine. Precision and accuracy were within 15% for IOT in both plasma and urine. The recoveries of IOT in urine and plasma ranged between 93.14% and 114.74 and 96.04–118.38%, respectively. The linear range for urine and plasma assays were 25–1500 and 1–150 µg/mL respectively. The lower limits of detection were 0.5 µg/mL for both urine and plasma, with no interference from plasma and urine matices. This method has been fully validated according to FDA guidelines and the new HPLC assay has been applied to a new formulation of IOT (Conray™ 43), to calculate GFR in healthy volunteers. The new method is simple, less expensive and it would be instrumental in future clinical and pharmacokinetic studies of iothalamate in kidney patients.

UDP glucuronosyltransferases (UGTs) are a family of enzymes that glucuronidate a variety of exogenous and endogenous compounds, including androgens; which makes them more hydrophilic, changes biological activity, and aids biological excretion. UGT2B17 has been identified as the major enzyme for testosterone glucuronidation, where UGT2B15 has been suggested to play a minor role. A deletion polymorphism in the UGT2B17 gene and an amino acid change in the UGT2B15 gene at codon 85 (aspartate>tyrosine) has been described where the resulting reduced rate of glucuronidation was implicated in the increased risk of prostate cancer. This review aims to analyse the impact that UGT2B15 and UGT2B17 genetic polymorphisms have on the risk of prostate cancer, through the use of studies previously conducted assessing the level of expression of each independent UGT2B enzyme, whereby ethnicity was taken into account. Positive and negative outcomes of clinical studies have been outlined for both UGT2B15 and UGT2B17, where conflicting evidence has led to inconclusive results for determining the consequence of UGT2B enzymes in prostate cancer risk. Additionally, inhibitors of testosterone glucuronidation such as non-steroidal, antiinflammatory drugs (diclofenac and ibuprofen), tea and red wine extracts have been identified to potentially affect the level of circulating testosterone by inhibiting UGT2B enzymes, thus potentially exacerbating prostate cancer risk. Future trials would involve a wider examination of other food substances and pharmaceuticals that can be attributed to the inhibition of UGT2B enzymes.

A sensitive and reproducible method has been developed and validated for the simultaneous quantification of the key antiretroviral drugs abacavir and tenofovir in hair using LC-MS/MS. The method was validated according to the US Food and Drug Administration (FDA) guidelines for the parameters: specificity, stability, limits of detection (LOD), limits of quantification (LOQ), linearity, accuracy, precision and recovery. Hair samples (50mg) were decontaminated and subjected to methanolic extraction, where 1 ml methanol was added along with the internal standard abacavir-d4 at a final concentration of 0.15 ng/mg hair. After 16 h, the drugs were recovered by liquid-liquid extraction using ammonium acetate buffer and a mixture of methyl tert-butyl ether:ethyl acetate (1:1). The samples were reconstituted with 200 μl acetonitrile:water (1:1) prior to injection for LC-MS/MS. The LOD and LOQ values were 0.06 and 0.12 ng/mg (drug/hair) for both drugs. Calibration curves were linear in the concentration range of 0.12-4.0 ng/mg of drug/hair with regression coefficient (r(2)) value of 0.999 for both drugs. The data for accuracy, precision and recovery were within the FDA limits. The concentrations of the drugs in the hair samples ranged from 0.12 ng/mg to 4.48 ng/mg and 0.32 ng/mg to 1.67 ng/mg for tenofovir and abacavir, respectively. This is the first full report of a method for the simultaneous determination of these two key antiretroviral drugs in hair. The newly developed method is useful for future routine analysis of tenofovir and abacavir in human hair and could be used in therapeutic drug monitoring and adherence to medicines studies, which would be helpful in decision making regarding treatment change in combination anti-retroviral therapies.

Objectives This paper compares two indirect prevalence estimation methods that offer protection beyond anonymity and are suitable for self-administration, for investigating the epidemiology of transgressive or socially sensitive behaviours. Design In this self-report study, 513 participants (58.7% male) from sports clubs across the UK and southern Ireland were asked to complete an anonymous survey containing the recently developed Single Sample Count (SSC), along with a comparative method Unrelated Question Model (UQM), using prohibited performance-enhancing drugs/substances (PED) as sensitive and hormone-boosting herbal supplements (HS) as non-sensitive control questions. Method The survey comprised of sections of SSC, UQM, social projection and simple network scale up methods. Respondents were asked to indicate whether they preferred the SSC or UQM for more protection and ease of completion. Results A large discrepancy was observed in prevalence estimates for PED using the UQM (58.4%) and SSC (19.8%), but not for HS (54.9% and 54.0%, respectively). The SSC prevalence estimate for PED was in keeping with the results from social projection (13.8% in own sport; 26.1% in all sports) and network scale up (19.3% for known and suspected doping combined). A clear preference was logged for SSC. Conclusion SSC, but not UQM, showed good concurrent validity with social projection and personal networks for PED; and good discriminant validity with HS. The observed discrepancy could be explained by strategic responding which can inflate the proportion of ‘yes’ answers in the UQM. Adaptation of the UQM for self-administration may lead to an unwanted upward response distortion via strategic responding.

Dear Editor, We read with interest the recent article by Katrinaki et al. (1). The authors report on cross-sectional values of serum 25-hydroxyvitamin D [25(OH)D] concentration from a large Mediterranean cohort in Crete, Greece. One of the major findings of the study was the sharp decline of parathyroid hormone (PTH) concentrations below the upper reference range of 65 pg/ml, with a non-significant decrease above ~20 ng/ml 25(OH)D. This led the authors to conclude that a cut-off level of 20 ng/ml is optimal to avoid secondary hyperparathyroidism in this specific population. The study provides useful novel, large scale results, from a Mediterranean region where vitamin D deficiency is highly prevalent (2). These findings however, contradict a previous large scale study in the U.S. which found that PTH decreased monotonically to 25(OH)D concentrations above 70 ng/mL (3). Although of potential interest, a continuous report of the PTH curve with regard to higher 25(OH)D concentrations is not provided in the paper by Katrinaki et al. (1) and the authors report metabolic parameters inversely correlated with 25OHD in Figure 3, with no plateau or saturation (1) . In the same previous large scale study (3), a significant part of the population with normal PTH levels ( 30 ng/mL (4). Further specific concerns regarding the interpretation of study findings, lie in the accuracy of the analyses and in particular the 25(OH)D results determined by chemiluminescence especially the claim that ‘the majority or protein-bound 25-(OH)D was released’ .The evidence for this statement is absent as is a reference to qualify the comment contradicted in the literature. In conclusion, we believe that available evidence (3-5), does not support that 25(OH) D concentrations of 20 ng/mL could be considered as the normal cut-off value for estimating vitamin D status. Additional studies of vitamin D replacement and treatment showing changes in specific patient-important outcomes (4) at certain levels of 25(OH)D are needed and would provide higher quality evidence that would lead to stronger recommendations.

Background\ud A gas chromatography mass spectrometry (GCMS) method for the determination of diclofenac in human plasma has been developed and validated.\ud \ud Results\ud This method utilizes hexane which is a relatively less toxic extraction solvent compared to heptane and benzene. In addition, phosphoric acid and acetone were added to the samples as deproteination agents, which increased the recovery of diclofenac. These revised processes allow clean extraction and near-quantitative recovery of analyte (approx. 89–95 %). Separation was achieved on a BP-1 column with helium as carrier gas. The molecular ion peaks of the indolinone derivatives of diclofenac ion (m/z 277) and the internal standard, 4-hydroxydiclofenac ion (m/z 439) were monitored by a mass-selective detector using selected ion monitoring (SIM) mode. The linear range for the newly developed and highly sensitive assay was between 0.25–50 ng/mL. The detection and lower quantifiable limits were 0.125 and 0.25 ng/mL, respectively. The inter-day and intra-day coefficients of variation for high, medium and low quality control concentrations were less than 9 %. The robustness and efficacy of this sensitive GCMS method was further demonstrated by using it for a pharmacokinetic study of an oral dosage form of diclofenac, 100 mg of modified-release capsules (Rhumalgan XL), in human plasma.\ud \ud Conclusions\ud This method is rapid, sensitive, specific, reproducible and robust, and offers improved sensitivity over previous methods. This method has considerable potential to be used for detailed pharmacokinetics, pharmacodynamics and bioequivalence studies of diclofenac in humans.

Research into the Female Athlete Triad (FAT) often posits that the condition is one of the unwanted consequences of increased physical activity and the prevailing preference for a lean body among female athletes; as well as the result of mounting pressure for constant performance improvement, which is often coupled to a misconception that low body weight would help to achieve this goal. This paper challenges the prevailing concept of the FAT for being inexact and over-specific, giving the impression that only athletes are affected by this condition, whilst the narrow focus on the co-occurrence of disordered eating–amenorrhea–osteoporosis can potentially lead to incorrect diagnoses of females suffering from, or at risk of developing, the condition. As the common underlying factor in athletes and non-athlete females suffering from FAT conditions is chronically low energy availability (via increased physical activity and/or disordered or restricted eating), we propose a unified framework that focuses on this common characteristic. Under the umbrella term ‘Female Energy Deficiency’ (FED), the expanded FAT and related concepts such as Anorexia Athletica and atypical eating disorder may be reconciled. The suggested framework can facilitate the understanding of this convoluted field within and outside the athletic community and offers flexibility for future developments. To support our proposition, we discuss the: i) expansion of the components to capture the extent and depth of this health condition, ii) expansion of the ‘at risk’ population, and iii) effective prevention, along with the need for early diagnosis and treatment.