Study of the Selectivity and Bioactivity of Polyphenols Using Infrared Assisted Extraction from Apricot Pomace Compared to Conventional Methods.

The valorization of industrial food byproducts by means of environment-friendly extraction methods is becoming a major interest because of its environmental and economic values. In this study, the efficiency of many technologies, such as ultrasounds (US), microwaves (MW), and infrared (IR), was compared, in terms of polyphenol yield and bioactivity from apricot pomace. IR was the most effective method with the highest polyphenol (10 mg GAE/g DM), flavonoid (6 mg CE/g DM), and tannin (3.6 mg/L) yields. In terms of efficacy, IR was followed by MW, US, then solid-liquid (S/L) extraction. IR extract from apricot pomace exhibited the highest inhibitory activity against all the studied gram-positive strains (Methicillin Resistant Staphylococcus aureus, Staphylococcus aureus, Methicillin-resistant Staphylococcus epidermidis, and Staphylococcus epidermidis) and a one gram-negative strain (Escherichia coli). Moreover, IR extracts had by far the highest antiradical activity (AC) (40%) followed by MW (31%), US (28%), and then S/L (15%). High-performance liquid chromatography (HPLC) permitted the identification and quantification of rutin in all extracts; whereas catechin was detected in those of IR (3.1 µg/g DM), MW (2.1 µg/g DM), and US (1.5 µg/g DM). Epicatechin was exclusively found in IR extract (4 µg/g DM), suggesting the selectivity of IR towards this compound. Scanning electron microscopy (SEM) revealed that the IR technique induced the highest cellular and structural damage in apricot pomace, which could explain the effectiveness of this technology.

Worsening of the Cardiovascular Profile in a Developing Country: The Greater Beirut Area Cardiovascular Cohort.

BACKGROUND: Lebanon has no established governmental noncommunicable diseases surveillance and monitoring system to permit reporting on noncommunicable diseases rates. The last World Health Organization-supported surveillance report showed worrying trends in cardiovascular disease (CVD) risk factors. OBJECTIVES: A cardiovascular cohort was established to permit CVD outcomes studies in an urban sample in the Lebanese capital and the study in hand presents the baseline CVD risk factors of this cohort. METHODS: A cross-sectional study was carried out including 501 Lebanese adults (64.3% women) from the Greater Beirut area using random multistage probability sampling. Interviews, physical exams, and blood withdrawal were conducted to collect information on demographic and lifestyle factors, body mass index, blood pressure, fasting blood glucose, blood lipids, as well as history of coronary artery diseases, hypertension, diabetes mellitus type 2, dyslipidemia, and stroke. Means with SD for continuous variables and frequencies and percentages for categorical variables are reported. RESULTS: The prevalence CVD risk factors including obesity, smoking, diabetes mellitus type 2, hypertension, and dyslipidemia prevalence in the Greater Beirut area was higher than that reported for the general population. Important sex and age differences were also observed, whereby older participants and women had higher rates of obesity, diabetes mellitus type 2, and dyslipidemia and younger participants and men were engaged more in cigarette smoking and alcohol consumption. Interestingly, water pipe smoking was similarly prevalent among genders. CONCLUSIONS: The overall prevalence of CVD risk factors in this urban population is higher than reported in the 2010 World Health Organization Stepwise Approach to Surveillance report on the Lebanese population, indicating that the urban population in the capital carries a higher burden of CVD risk. In addition, sex and age difference rates of CVD risk factors highlight the need for tailored public health measures to tackle the sex- and age-based CVD risk factors.

Bisphenol A urinary level, its correlates, and association with cardiometabolic risks in Lebanese urban adults.

Bisphenol A (BPA) is an endocrine disruptor with multiple purported metabolic effects. This study aimed to measure BPA among Lebanese population, to identify its predictors, and to explore any link to metabolic disorders. A representative sample of 501 adults from Lebanon was recruited in a cross-sectional study. Urinary BPA was measured, and data were collected for anthropometric measurements, medical history, food intake, and laboratory markers of metabolic conditions. BPA data was divided into tertiles. A total of 89% of the subjects had detectable urinary BPA levels, with an overall mean of 3.67 ± 4.75 µg/L and a mean creatinine-adjusted BPA of 2.90 ± 4.79 µg/g. There was a significant positive association with female gender and older age for being in the highest BPA tertile. BPA level was linked to metabolic syndrome (MetS), obesity, type-2 diabetes (T2D), hypertension, and dyslipidemia. After adjustment, the trend remained for BPA in association with MetS and T2D. Though urinary BPA in the Lebanese population was higher in older women, the levels were similar to world-reported figures. Our results suggest a link with metabolic disorders but not at a significant level. These findings call for longitudinal and broader sample measurements.

PREVALENCE OF DIABETES IN GREATER BEIRUT AREA: WORSENING OVER TIME.

OBJECTIVE: The Middle East North Africa region has one of the highest rates of diabetes, both in prevalence and in rate of increase. The aim of this study was to examine the prevalence and associated risk factors of type 2 diabetes mellitus (T2D) in the adult population of Beirut. METHODS: A random sample of 501 men and women aged 18 to 79 years was examined in a cross-sectional manner. The sample was then divided into 3 groups based on T2D self-report, glycosylated hemoglobin (HbA1c), and fasting glucose (no diabetes [ND], at risk for diabetes [RD], and probable diabetes [PD]). These were compared to determine the various associated risks. RESULTS: The sample consisted of 64.3% women, with an average age of 45.4 ± 15 years, and a mean body mass index (BMI) of 29.4 ± 5.9 kg/m2. The subjects were predominantly from a low socio-economic status, and more than half smoked either cigarettes or a waterpipe. The percentages of the 3 groups were as follows: 41.7%, 40.3%, and 18.0% for ND, RD, and PD, respectively. Out of 90 subjects diagnosed with PD, 26 did not know they had diabetes. Independent, positively associated risk factors were age, BMI, heart rate, hypertension, triglyceride, and high-density lipoprotein cholesterol. CONCLUSION: The prevalence of T2D was high in this study and seems to be increasing compared to prior diabetes reports. Overall, the whole sample had a high prevalence of cardiovascular risk factors such as smoking, obesity, and physical inactivity. However, subjects with diabetes had significantly more components of the metabolic syndrome. ABBREVIATIONS: ADA = American Diabetes Association CRP = C-reactive protein DD = definite diabetes FPG = fasting plasma glucose HbA1c = glycosylated hemoglobin MENA = Middle-East North Africa ND = no diabetes PD = probable diabetes RD = at risk of diabetes T2D = type 2 diabetes mellitus TSH = thyroid-stimulating hormone.

Effects of fatty acids on benzo[a]pyrene uptake and metabolism in human lung adenocarcinoma A549 cells.

Dietary supplementation with natural chemoprotective agents is receiving considerable attention because of health benefits and lack of toxicity. In recent in vivo and in vitro experimental studies, diets rich in n-3 polyunsaturated fatty acids have been shown to provide significant anti-tumor action. In this investigation, the effects of control fatty acids (oleic acid (OA), linoleic acid (LA)) and n-3 PUFA, e.g., docosahexaenoic acid (DHA) on the uptake and metabolism of the carcinogenic polycyclic aromatic hydrocarbon, benzo[a]pyrene (BaP) was investigated in A549 cells, a human adenocarcinoma alveolar basal epithelial cell line. A549 cells activate BaP through the cytochrome P450 enzyme system to form reactive metabolites, a few of which covalently bind to DNA and proteins. Therefore, multiphoton microscopy spectral analysis combined with linear unmixing was used to identify the parent compound and BaP metabolites formed in cells, in the presence and absence of fatty acids. The relative abundance of select metabolites was associated with altered P450 activity as determined using ethoxyresorufin-O-deethylase activity in cells cultured in the presence of BSA-conjugated fatty acids. In addition, the parent compound within cellular membranes increases significantly in the presence of each of the fatty acids, with the greatest accumulation observed following DHA treatment. DHA treated cells exhibit significantly lower pyrene-like metabolites indicative of lower adducts including DNA adducts compared to control BSA, OA or LA treated cells. Further, DHA reduced the abundance of the proximate carcinogen BaP 7,8-dihydrodiol and the 3-hydroxybenzo[a]pyrene metabolites compared to other treatments. The significant changes in BaP metabolites in DHA treated cells may be mediated by the effects on the physicochemical properties of the membrane known to affect enzyme activity related to phase I and phase II metabolism. In summary, DHA is a highly bioactive chemo-protective agent capable of modulating BaP-induced DNA adducts.

Optimization of Saanen sperm genes amplification: evaluation of standardized protocols in genetically uncharacterized rural goats reared under a subtropical environment.

The purpose of this research is to optimize quantitatively the amplification of specific sperm genes in reference genomically characterized Saanen goat and to evaluate the standardized protocols applicability on sperms of uncharacterized genome of rural goats reared under subtropical environment for inclusion in future selection programs. The optimization of the protocols in Saanen sperms included three production genes (growth hormone (GH) exons 2, 3, and 4, αS1-casein (CSN1S1), and α-lactalbumin) and two health genes (MHC class II DRB and prion (PrP)). The optimization was based on varying the primers concentrations and the inclusion of a PCR cosolvent (Triton X). The impact of the studied variables on statistically significant increase in the yield of amplicons was noticed in four out of five (80%) optimized protocols, namely in those related to GH, CSN1S1, α-lactalbumin, and PrP genes (P < 0.05). There was no significant difference in the yield of amplicons related to MHC class II DRB gene, regardless of the variables used (P > 0.05). The applicability of the optimized protocols of Saanen sperm genes on amplification of uncharacterized rural goat sperms revealed a 100% success in tested individuals for amplification of GH, CSN1S1, α-lactalbumin, and MHC class II DRB genes and a 75% success for the PrP gene. The significant success in applicability of the Saanen quantitatively optimized protocols to other uncharacterized genome of rural goats allows for their inclusion in future selection, targeting the sustainability of this farming system in a subtropical environment and the improvement of the farmers livelihood.

Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line.

Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an "advanced unmixing process", identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 h. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to the analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.

Stimulation of postprandial in vivo glycogenesis and lipogenesis of rats fed high fructose diet with varied phosphate content.

It has been reported that increased fructose intake is associated with the development of the metabolic syndrome. The phosphate (P) sequestering capacity of fructose is likely to affect the phosphorylation capacity of different metabolites, and this, in turn, may be the basis for several metabolic derangements, especially in the P requiring reactions, for example, glycogenesis and lipogenesis. We hypothesized that P enrichment of the diet can balance P status and, consequently, affect glycogenesis and lipogenesis. An animal experiment was executed in which adult male Sprague-Dawley rats were maintained for 4 days on high-fructose diets with different P content (0.15%, 0.165%, 0.30%, and 1.65%). At the end of the feeding period, overnight fasted rats were tube fed a test meal, injected with (3)H(2)O and euthanized 1 hour later. Final plasma glucose, insulin, uric acid, and triacylglycerol concentrations, as well as in vivo rates of glycogen and lipid synthesis and hepatic glycogen content, were measured. Results showed that increased P content of the diet was associated with an increase in postprandial epididymal fat pad (P = .007) and hepatic lipogenesis (P = .029), as well as glycogenesis (P = .024). In conclusion, P content of the diet was found to stimulate both glycogenesis and lipogenesis. These alterations in carbohydrate and fat metabolism point to the potential of P in influencing nutritional status.

Plasma copper, zinc, and selenium levels and correlates with metabolic syndrome components of lebanese adults.

Zinc, copper, and selenium statuses were reported to be linked to the development of chronic diseases, especially coronary heart disease (CHD). Metabolic syndrome, a known CHD risk factor, was found to be highly prevalent in Lebanon. Nevertheless, no data are available on the statuses of plasma zinc, copper, and selenium, especially in terms of their relation to the components of the metabolic syndrome. A sample of 398 men and women aged 18-65 years was drawn from 23 health centers across Lebanon; anthropometric measurements and biochemical analyses of fasting plasma samples were performed. Subjects were found to have normal plasma statuses of copper and selenium but were at elevated risk of zinc deficiency. Plasma selenium levels correlated positively with all the components of the metabolic syndromes, while that of copper correlated only with total, high-density lipoprotein and low-density lipoprotein cholesterol. Plasma zinc did not correlate with any of the metabolic syndrome components.

High-performance liquid chromatography method for quantifying sphingomyelin in rat brain.

A rapid, reproducible and accurate high-performance liquid chromatographic (HPLC) method for the quantitative determination of sphingomyelin in rat brain was developed and validated using normal-phase silica gel column, acetonitrile-methanol-water (65:18:17 (v/v)) at a flow rate of 1 ml/min, isocratic elution, UV detection at 207 nm and 1,2-dimyristoyl-sn-glycero-3-phosphocholine as an internal standard. Total run time was 10.0 min. The calibration curve was linear over the range of 0.025-0.4 mg/ml sphingomyelin (R2>0.99). The intra-day coefficient of variation ranged from 1.4% to 2.2%. The average inter-day coefficient of variation over a period of 4 days was 3.1%. The practical limit of detection was 0.005 mg/ml with a quantification limit of 0.01 mg/ml.

Effects of benzo-a-pyrene on oxytocin-induced Ca2+ oscillations in myometrial cells.

Benzo-a-pyrene (BaP) is a polycyclic aromatic hydrocarbon that exists as a major environmental pollutant. The effect of this carcinogen/mutagen upon myometrial Ca(2+) signaling in a human myometrial cell line (PHM1) was examined. Exposure of cells to BaP did not alter basal Ca(2+) levels or the inositol(1,4,5) trisphosphate-releasable Ca(2+) pool. However, BaP significantly decreased the initial oxytocin-induced Ca(2+) transient and the frequency of oxytocin-induced Ca(2+)oscillations as well as delayed their onset. To determine the specific effects of BaP, pharmacologic agents that target intracellular Ca(2+) homeostasis mechanisms were used. Genistein (a non-specific tyrosine kinase inhibitor) and AG1478 (an epidermal growth factor receptor blocker) markedly reduced the oxytocin-induced Ca(2+) oscillations in control, but had no effect in BaP treated cells. Addition of epidermal growth factor or serum before or after oxytocin restored the Ca(2+) oscillations in BaP treated cells to a level similar to control cells, while the K(+) channel blocker tetraethylammonium chloride, partially restored the Ca(2+) response. These data suggest that the tyrosine kinase pathway, which is part of the G-protein coupled receptor pathway response to oxytocin in PHM1 cells, is a target of BaP action and that EGF or serum can restore the oxytocin-induced Ca(2+) oscillations.

Lysine fortification reduces anxiety and lessens stress in family members in economically weak communities in Northwest Syria.

Lysine is a limiting amino acid in diets based on wheat as the staple. In experimental animals, prolonged dietary lysine inadequacy increases stress-induced anxiety. If observed in humans, such a result would have a strong implication for the relationship between nutrition and communal quality of life and mental health. As part of a 3-month randomized double-blind study, we tested whether lysine fortification of wheat reduces anxiety and stress response in family members in poor Syrian communities consuming wheat as a staple food. In the lysine-fortified group, the plasma cortisol response to the blood drawing as a cause of stress was reduced in females, as was sympathetic arousal in males as measured by skin conductance. Lysine fortification also significantly reduced chronic anxiety as measured by the trait anxiety inventory in males. These results suggest that some stress responses in economically weak populations consuming cereal-based diets can be improved with lysine fortification.

Circulating antibodies directed against nitrosylated antigens in trypanosome-infected mice.

Nitric oxide has been implicated as an effector cytotoxic molecule in trypanosomiasis. In this work, we investigated the presence of circulating antibodies directed against nitrosylated epitopes as biological indicators for nitric oxide (NO) production in the sera of trypanosome-infected mice. We tested these sera with synthetic antigens, such as S-nitrosated protein or nitrosylated conjugates of amino acids that possess a high affinity to NO, by an immunoenzymatic assay. We detected antibodies directed against nitroso epitopes in the sera of infected mice, as compared to non-infected control mice. The antibody response was linked to the IgM isotype. Our results indicate the production of NO and its derivatives in trypanosomiasis. This production may potentially induce the synthesis of nitroso epitopes in vivo and favor the development of a humoral immune response.

Characterization of calcium oscillations in normal and benzo[a]pyrene-treated clone 9 cells.

Intracellular Ca2+ oscillations induced by oxytocin and vasopressin were analyzed in a rat liver cell line (Clone 9) in order to identify mechanisms by which benzo[a]pyrene (BaP) alters Ca2+ signaling patterns in these cells. Clone 9 cells exhibit an initial Ca2+ spike, followed by Ca2+ oscillations upon oxytocin or vasopressin treatment. The range of frequencies (maximum 110 mHz) was dependent on agonist concentration with a constant amplitude less than or equal to the amount of Ca2+ generated from the inositol trisphosphate (InsP(3))-sensitive pool. This study examined contributions of extracellular and intracellular pools to the frequency of Ca2+ oscillations and the role of membrane channels, second messengers, and different pharmacological reagents on the regulation of oscillation frequency in both control and BaP-treated cells. Results indicated that the Ca2+ oscillations are mainly due to inositol 1,4,5-triphosphate (InsP(3))-sensitive stores and that extracellular Ca2+ contributes to refilling of this intracellular Ca2+ pool. The frequency of Ca2+ oscillations is also sharply affected by protein kinase C activated by phospholipase C. In BaP-treated Clone 9 cells, basal Ca2+ levels were elevated and the frequency of Ca2+ oscillations was suppressed in a dose-dependent fashion. Suppression of Ca2+ oscillations is due, at least in part, to an effect of BaP on enhanced opening of K+ channels. This was confirmed by showing that inhibition of the K+ channel opening by tetraethylammonium chloride can reverse the effect of BaP on oxytocin-induced Ca2+ oscillations, and potentially decrease the toxicity of BaP.