Crocetin Exerts Its Anti-inflammatory Property in LPS-Induced RAW264.7 Cells Potentially via Modulation on the Crosstalk between MEK1/JNK/NF-κB/iNOS Pathway and Nrf2/HO-1 Pathway.

Crocetin is a main bioactive component with a carotenoid skeleton in Gardenia jasminoides, a typical traditional Chinese medicine with a long history in Southeast Asia. Crocetin is being commonly consumed as spices, dyes, and food colorants. Recent pharmacological studies had implied that crocetin may possess potent anti-inflammatory properties; however, the underlying molecular mechanism is not fully elucidated. In the present study, the regulatory effect of crocetin on redox balance was systematically investigated in lipopolysaccharide- (LPS-) stimulated RAW264.7 cells. The results showed that crocetin dose-dependently inhibited LPS-induced nitric oxide production and inducible nitric oxide synthase (iNOS) expression in RAW264.7 cells. Molecular data revealed that crocetin exerted its anti-inflammatory property by inhibiting the MEK1/JNK/NF-κB/iNOS pathway and activating the Nrf2/HO-1 pathway. The shRNA-knockdown (KD) of MEK1 and ERK1 confirmed that the activation of MEK1 and inhibition of JNK mediated the anti-inflammatory effect of crocetin. Moreover, the pull-down assay and computational molecule docking showed that crocetin could directly bind to MEK1 and JNK1/2. It is noticed that both KD and knockout (KO) of HO-1 gene blocked this action. More detailed data have shown that HO-1-KO blocked the inhibition of p-IκB-α by crocetin. These data indicated that crocetin exerted its anti-inflammatory property via modulating the crosstalk between the MEK1/JNK/NF-κB/iNOS pathway and the Nrf2/HO-1 pathway, highlighting HO-1 as a major player. Therefore, the present study reveals that crocetin can act as a potential candidate for redox-balancing modulation in charge of its anti-inflammatory and chemopreventive effect, which strengthens its potency in the subsequent clinic application in the near future.

Improved quantification method of crocins in saffron extract using HPLC-DAD after qualification by HPLC-DAD-MS.

Crocins in commercial liquid saffron extracts (Saffr'activ®) were identified using high-performance liquid chromatography (HPLC) with a diode array detector (DAD) and mass spectrometry (MS). The impact of storage on the qualities of the saffron extract were studied with HPLC-DAD-MS by exposing trans-4-GG crocin to environmental factors. Light and temperature induced degradation after only one week. Trans-4-GG crocin was totally hydrolyzed when stored at 60 °C and exposed to light. A quick and reliable method using HPLC-DAD was then developed to improve quantification of crocins in commercial liquid saffron extracts. An internal standard quantification method that uses a response factor, corrected with the molecular weight of each crocin, improved results for old saffron extracts.

Biofortified and fortified maize consumption reduces prevalence of low milk retinol, but does not increase vitamin A stores of breastfeeding Zambian infants with adequate reserves: a randomized controlled trial.

BACKGROUND: Replacement of conventional staples with biofortified or industrially fortified staples in household diets may increase maternal breast milk retinol content and vitamin A intakes from complementary foods, improving infant total body stores (TBS) of vitamin A. OBJECTIVES: To determine whether biofortified or industrially fortified maize consumption by Zambian women and their breastfeeding infants could improve milk retinol concentration and infant TBS. METHODS: We randomly assigned 255 lactating women and their 9-mo-old infants to a 90-d intervention providing 0 µg retinol equivalents (RE)/d as conventional maize or ∼315 µg RE/d to mothers and ∼55 µg RE/d to infants as provitamin A carotenoid-biofortified maize or retinyl palmitate-fortified maize. Outcomes were TBS, measured by retinol isotope dilution in infants (primary), and breast milk retinol, measured by HPLC in women (secondary). RESULTS: The intervention groups were comparable at baseline. Loss to follow-up was 10% (n = 230 mother-infant pairs). Women consumed 92% of the intended 287 g/d and infants consumed 82% of the intended 50 g/d maize. The baseline geometric mean (GM) milk retinol concentration was 1.57 µmol/L (95% CI: 1.45, 1.69 µmol/L), and 24% of women had milk retinol <1.05 µmol/L. While mean milk retinol did not change in the biofortified arm (ß: 0.11; 95% CI: -0.02, 0.24), the intervention reduced low milk retinol (RR: 0.42; 95% CI: 0.21, 0.85). Fortified maize increased mean milk retinol (ß: 0.17; 95% CI: 0.04, 0.30) and reduced the prevalence of low milk retinol (RR: 0.46; 95% CI: 0.25, 0.82). The baseline GM TBS was 178 µmol (95% CI: 166, 191 µmol). This increased by 24 µmol (± 136) over the 90-d intervention period, irrespective of treatment group. CONCLUSIONS: Both biofortified and fortified maize consumption improved milk retinol concentration. This did not translate into greater infant TBS, most likely due to adequate TBS at baseline. This trial was registered at clinicaltrials.gov as NCT02804490.

Enhanced retinoid response by a combination of the vitamin A ester retinyl propionate with niacinamide and a flavonoid containing Ceratonia siliqua extract in retinoid responsive in vitro models.

OBJECTIVES: Retinoids have been used for decades as efficacious topical agents to treat photoaged skin. The purpose of our present research is to evaluate whether the activity of the vitamin A ester retinyl propionate (RP) can be enhanced by niacinamide (Nam) and a flavonoid containing Ceratonia siliqua (CS) fruit extract in retinoid responsive in vitro models. METHODS: Retinyl propionate was tested alone and in combination with Nam and CS in an RARα reporter cell line for promoter activation and compared to trans-retinoic acid (tRA) activation. These treatments were also tested in keratinocytes for gene expression profiling by qPCR using a panel of 40 retinoid responsive genes. RESULTS: tRA or RP elicited RARα reporter activation in a dose-dependent manner. The combination of 0.5 µM or 2 µM RP with 10 mM Nam had a 56% and 95% signal increase compared to RP, respectively. The addition of 1% CS to 0.5 µM or 2 µM RP with 10 mM Nam elicited a further increase of 114% and 156%, respectively, over RP and Nam combinations. All retinoids elicited an increase in expression of 40 retinoid sensitive genes over control levels. Of the 40 genes, 27 were enhanced by either 0.1 µM RP or 0.5 µM RP with 10 mM Nam and 1% CS. Nam or CS had very modest activity in both models. CONCLUSION: The combination of RP with Nam and CS showed a higher retinoid response than RP in two separate retinoid responsive in vitro models. We hypothesize Nam and CS enhances RP activity by modulating metabolism to tRA via increasing NAD+ pools and inhibiting reduction of retinal (RAL) back to retinol, respectively. The findings provide evidence that this combination may have enhanced efficacy for treating the appearance of photoaged skin.

OBJECTIFS: Les rétinoïdes sont utilisés depuis des décennies comme agents topiques efficaces pour traiter la peau photo-âgée. Le but de notre recherche actuelle est d'évaluer si l'activité du propionate rétinyl ester de vitamine A (RP) peut être augmentée par le niacinamide (Nam) et un flavonoïde contenant un extrait de fruit de Ceratonia Siliqua (CS) dans les modèles in vitro sensibles aux rétinoïdes. MÉTHODES: RP a été testé seul et en combinaison avec Nam et CS dans une ligne de cellule rapporteur de RARα pour l'activation du promoteur et par rapport à l'activation de l'acide transrétinoïque(tRA). Ces traitements ont également été testés dans les kératinocytes pour le profilage d'expression génique par qPCR à l'aide d'un panel de 40 gènes rétinoïdes sensibles. RÉSULTATS: tRA ou RP ont provoqué l'activation du promoteur RARα d'une manière dépendante de la dose. La combinaison de 0,5 µM ou 2 µM de RP avec 10 mM de Nam a permis une augmentation respectivement de 56% et 95% du signal par rapport à RP. L'ajout de 1 % de CS à 0,5 µM ou 2 µM de RP avec 10 mM de Nam a permis une nouvelle augmentation de 114 % et 156 %, respectivement, qu'avec la combinaison RP et Nam. Tous les rétinoïdes ont provoqué une augmentation de l'expression de 40 gènes sensibles aux rétinoïdes sur les niveaux de contrôle. Sur les 40 gènes, 27 ont été améliorés soit par 0,1 µM de RP ou 0.5 µM de RP avec 10 mM de Nam et 1% de CS. Nam ou CS avaient une activité très modeste dans les deux modèles. CONCLUSION: La combinaison de RP avec Nam et CS a montré une réponse rétinoïde plus élevée que RP dans deux modèles in vitro séparés sensibles aux rétinoïdes. Nous émettons l'hypothèse que Nam et CS améliorent l'activité RP en modulant le métabolisme de tRA par l'augmentation des groupement NAD+ et en inhibant la réduction du rétinal (RAL) en rétinol, respectivement. Les résultats fournissent la preuve que cette combinaison peut améliorer l'efficacité du traitement de l'aspect de la peau photo-âgée.

Cereal bran protects vitamin A from degradation during simmering and storage.

Food supplementation with vitamin A is an efficient strategy to combat vitamin A deficiency. The stability of vitamin A during cooking and storage is, however, low. We here show that cereal bran protects retinyl palmitate (RP) during simmering and storage. Native wheat bran stabilized RP the most during simmering. About 75% RP was recovered after 120 min of cooking, while all RP was lost after 80 min in the absence of bran. Heat-treated rice bran protected RP the best during forced storage, with a 35% recovery after 8 weeks. RP was degraded entirely in the absence of bran in less than one week. Results suggested that the physical entrapment of oil within the large wheat bran particles protects RP from the action of water and pro-oxidants during simmering. During storage, the high amount and diversity of lipid components present in rice bran are presumably responsible for its protective effect.

Cultivars of biofortified cowpea and sweet potato: Bioavailability of iron and interaction with vitamin A in vivo and in vitro.

The objective of this study was to evaluate the interaction of pro-vitamin A-rich sweet potato on iron bioavailability of biofortified cowpeas, using in vitro Caco-2 cells and in vivo depletion-repletion rat model. Mixtures of conventional rice with cultivars of iron-biofortified (Aracê, Xiquexique, and Tumucumaque) or conventional (Guariba) cowpeas with or without sweet potato biofortified with pro-vitamin A carotenoids were evaluated. The ratio of ferritin/total protein in Caco-2 cells was used as the index of cellular Fe uptake in the in vitro assay. The animal study evaluated the hemoglobin gain, the relative biological value, and the gene expression of transferrin and ferritin proteins by reverse transcription polymerase chain reaction. In the in vitro study, Xiquexique cowpea presented higher bioavailability of iron in the absence of sweet potato, and no difference was observed between the other cultivars of cowpea with and without sweet potato. The in vivo bioavailability (relative biological value of hemoglobin regeneration efficiency) differed statistically only between Guariba groups added to sweet potato and Tumucumaque. Ferritin mRNA expression did not differ between the test and control (ferrous sulfate) groups. Regarding the transferrin mRNA expression, there was a difference between the test and control groups except for the Xiquexique group. The association of rice and beans with sweet potato rich in carotenoids favored the gene expression of proteins involved in the iron metabolism, as well as its bioavailability, corroborating beneficial effects of this mixture. Xiquexique cowpea was shown to be the most promising compared to the other cultivars, exhibiting higher iron content in the digestible fraction, better in vitro bioavailability of iron, and transferrin gene expression. PRACTICAL APPLICATION: Data from the study indicated greater in vitro bioavailability of iron for Xiquexique cowpea and sweet potato mixtures, in addition to the greater regeneration efficiency of hemoglobin in vivo as the bioavailability of iron among biofortified beans, highlighting the promising benefits of biofortification.

Changes in fatty acids composition, antioxidant potential and induction period of UHT-treated tea whitener, milk and dairy drink.

BACKGROUND: In developing and developed countries, several versions of safe and shelf-stable Ultra High Temperature, UHT-treated products are manufactured. Terminologies and formulations of UHT-treated tea whitener, milk and dairy drink considerably vary. Comprehensive studies have been performed on UHT-treated milk; however, fatty acids compositional changes and oxidation status of UHT-treated tea whitener and dairy drink at different storage intervals have not been reported in literature. METHODS: UHT-treated tea whitener, milk and dairy drink samples (450 each) of the same manufacturing date were purchased from the market and stored at ambient temperature (25-30 °C) for 90 days. At the time of collection, all the samples were only one week old. Samples of UHT-treated tea whitener, milk and dairy drink were regarded as treatments and every treatment was replicated five times. Chemical composition, fatty acid profile, 2, 2-Diphenyl-1-picrylhydrazyle (DPPH) radical scavenging activity, total antioxidant activity, reducing power, antioxidant activity in linoleic acid system and induction period were determined at 0, 45 and 90 days of storage. RESULTS: Fat content in freshly collected samples of UHT treated-tea whitener, milk and dairy drink were 6 and 3.5%. UHT treated milk had highest total antioxidant capacity, antioxidant activity in linoleic acid and 2, 2-Diphenyl-1-picrylhydrazyle (DPPH) free radical scavenging activity followed by UHT tea whitener and dairy drink. In freshly collected samples of UHT-treated milk, concentrations vitamin A and E were 0.46 µg/100 g and 0.63 mg/100 g, respectively. UHT-treated tea whitener had the lowest concentrations of vitamin A and E. With the progression of storage period, amount of vitamin A and E decreased. In freshly collected samples, amount of short, medium and unsaturated fatty acids in UHT-treated milk were 10.54, 59.71 and 27.44%, respectively. After 45 days of storage of UHT-treated milk, the loss of short, medium and unsaturated fatty acid was 7%, 7.1 and 5.8%, respectively. After 90 days of storage of UHT-treated milk, the loss of short, medium and unsaturated fatty acid was 8.53, 13.51 and 11.88%, accordingly. After 45 days of storage of UHT-treated tea whitener, the loss of medium and unsaturated fatty acid was 1.6 and 0.99%, respectively. After 90 days of storage, the loss of medium and unsaturated fatty acids were 8.2 and 6.6%, respectively. The induction period of fresh UHT-treated tea whitener, milk and dairy drink was 15.67, .74 and 7.27 h. Strong correlations were recorded between induction period and peroxide value of UHT-treated products. CONCLUSION: This investigation disclosed that UHT-treated tea whitener had 6% fat content with no short-chain fatty acids. Antioxidant capacity of UHT-treated milk was higher than dairy drink and tea whitener. Due to the presence of partially hydrogenated fat, oxidative stability of UHT-treated tea whitener was better than UHT-treated milk and dairy drink. Vitamin A and E was not found in UHT-treated tea whitener. For the anticipation of oxidative stability of UHT-treated milk, dairy drink and tea whitener, induction period/ Rancimat method can be used.

Consumer Perceptions and Acceptability of Traditional Dishes Prepared with Provitamin A-Biofortified Maize and Sweet Potato.

Vitamin A deficiency (VAD) is prevalent in South Africa, particularly among predominantly poor rural communities. Provitamin A (PVA)-biofortified crops could be used to address VAD; however, there are challenges of poor consumer acceptability. This study investigated the effect of replacing white maize and cream-fleshed sweet potato (CFSP) with PVA-biofortified maize and orange-fleshed sweet potato (OFSP), respectively, on consumer acceptability and perceptions of traditional dishes of rural communities in South Africa. Consumer acceptability of PVA-biofortified phutu (a crumbly maize porridge) served with either curried cabbage, chicken or bambara groundnut, separately, and boiled OFSP was evaluated by black South African adults (n = 120) using a five-point facial hedonic scale. Focus group discussions (FGDs) were conducted with 56 subjects recruited from the consumer panel to assess consumer perceptions of the food samples. The majority of the participants rated the composite dishes containing PVA-biofortified phutu as "4 = good" and the acceptability of the composite dishes varied significantly (p < 0.05). Compared to other age groups, the 50-59-year age group showed higher preference for white phutu and chicken curry, whereas the 30-39-year age group showed higher preference for PVA-biofortified phutu and chicken curry. The acceptability of OFSP and CFSP was similar. The study participants showed positive perceptions of the OFSP, as well as PVA-biofortified phutu if served with either curried chicken or cabbage. The findings suggest that PVA-biofortified maize and OFSP can replace white maize and CFSP, respectively, in selected traditional dishes of the rural communities studied to alleviate VAD.

Does vitamin fortification affect light oxidation in fluid skim milk?

Off-flavors in milk related to light oxidation form due to photoxidation of native riboflavin and tetrapyrroles, resulting in an array of lipid oxidation compounds. Recent work has established that fortification with water-dispersible vitamin A can result in off-flavors in fluid skim milk caused by vitamin A degradation products in the vitamin premix. The objective of this study was to determine the role of vitamin fortification on light oxidation of high temperature, short time pasteurized fluid skim milk. First, the aroma profiles and aroma-active volatile compounds in light-exposed vitamin premixes were determined by exposing the premixes to fluorescent (FL) or light-emitting diode (LED) light at 2,000 lx at 4°C for 0, 2, 4, 8, or 24 h. A trained panel (n = 6) documented aroma profiles of each vitamin premix at each time point. Headspace solid-phase microextraction followed by gas chromatography-olfactometry and gas chromatography-mass spectrometry (GC-MS) were performed to characterize aroma-active compounds in light-exposed vitamin premixes. In the second experiment, commercial vitamin premixes (vitamin A and vitamin D in oil and water matrices) were used to fortify skim milk (vitamin A: 3,000 IU/946 mL; vitamin D: 600 IU/946 mL). Skim milk was pasteurized, homogenized, and packaged in 946-mL high-density polyethylene jugs. Milks were exposed to FL or LED light at 2,000 lx at 4°C for 4, 12, 24, or 48 h. Controls with and without vitamins and light shielding were included. Riboflavin and vitamin A and D degradation were quantified via ultra-high-performance liquid chromatography. A trained panel (n = 8) documented sensory profiles of milks at each time point. Lipid oxidation volatile compounds were quantified via solid-phase microextraction with GC-MS. Vitamin degradation volatile compounds were quantified via solvent-assisted sorptive stir bar extraction with GC-MS. Riboflavin, vitamin A, and vitamin D degradation were consistent with that reported in previous studies. We found no effect of vitamin fortification on development of typical light oxidation-related off-flavors (cardboard and mushroom) or lipid oxidation-related volatiles (hexanal and heptanal). A perfumey/floral flavor was documented in the oil-based vitamin A-fortified milk, suggesting that light exposure affected the off-flavors contributed by water- versus oil-based vitamin fortification. These results show no evidence that vitamin fortification at current levels provides any protection against light oxidation-related off-flavors in fluid milk.

Potentiality of front-face fluorescence and mid-infrared spectroscopies coupled with partial least square regression to predict lipid oxidation in pound cakes during storage.

The potentialities of front-face fluorescence (FFF) and mid-infrared (MIR) spectroscopies coupled with partial least square regression (PLSR) were compared to predict the lipid oxidation of pound cakes. The level of lipid oxidation in pound cakes determined using classical methods showed some changes. Similarly, the fluorescence emission (305-490 nm) and excitation (252-390 nm) spectra and MIR spectra scanned in the 4000-700 cm-1 region showed some changes in pound cakes as a function of both storage time and the type of oil used in the formulation. The application of PLSR to the MIR spectra, provided excellent predictive results for free fatty acid (R2 = 0.97) and peroxide values (R2 = 0.87). Similar results were obtained from both tryptophan and MIR spectra for the prediction of TOTOX (R2 > 0.86) demonstrating the efficiency of the MIR and FFF spectroscopies to qualify and quantify the level of lipid oxidation in pound cakes.

Prevention of hepatic stellate cell activation using JQ1- and atorvastatin-loaded chitosan nanoparticles as a promising approach in therapy of liver fibrosis.

Preventing hepatic stellate cell (HSC) activation represents a promising approach to resolve liver fibrosis. Several drugs have been reported to delay/prevent HSCs activation, however with limited clinical benefits. The latter may be in part attributed to the limited ability of such drugs in targeting more than one pathway of HSC activation. Added to that, is their inability of reaching their target cell in sufficient amounts to induce a therapeutic effect. In this work, chitosan NPs were loaded with JQ1 and atorvastatin, two drugs that have been reported to prevent HSCs activation, however via different mechanisms. NPs were then modified with different densities of retinol (Rt) for active targeting of HSCs. The NP HSCs targeting ability as a function of Rt density was assessed in vitro on primary HSCs and in vivo in carbon tetrachloride (CCl4) induced fibrotic mouse models. In vitro NPs modified with a low Rt density (LRt-NPs) showed ≈2 folds enhanced HSCs uptake in comparison to unmodified NPs, whereas NPs modified with a high Rt density (HRt-NPs) showed ≈0.8 folds change in uptake relative to unmodified NPs. Similarly, in vivo LRt-NPs showed higher accumulation in fibrotic livers in comparison to healthy livers whereas HRt-NPs and unmodified NPs showed lower accumulation in fibrotic livers relative to healthy controls respectively. Finally, the ability of drug-loaded NPs in preventing HSCs activation was assessed by monitoring the reduction in α-smooth muscle actin (α-SMA) expression by Western blot. NPs loaded with both JQ1 and atorvastatin showed reduction in α-SMA expression. In addition, a synergistic reduction in α-SMA was observed when cells were co-treated with JQ1 and atorvastatin loaded NPs.

Hepatic Vitamin A Concentrations in Vervets (Chlorocebus aethiops) Supplemented with Carotenoids Derived from Oil Palm.

Commonly used in biomedical research, vervets (Chlorocebus aethiops) are omnivorous but primarily meet their vitamin A requirements from provitamin A carotenoids. Hypervitaminosis A has occurred in vervets that consume feed high in preformed vitamin A. We investigated the vitamin A status of vervets supplemented daily with various antioxidants derived from palm oil. Male vervets (n = 40) were placed for 23 wk on a high-fat diet (34.9% energy) containing 645 µ g retinol activity equivalents (RAE), with 515 µ g RAE from preformed vitamin A. Vervets were randomized to 5 treatments (duration, 20 mo): control; 100 mg d-α-tocopheryl acetate; 100 mg oil palm (Elaeis guineensis)-derived vitamin E; 50 mg oil palm-derived vitamin E + 50 mg carotenoid complex + unrestricted palm-derived water-soluble antioxidants; and 5) unrestricted water-soluble antioxidants. Livers (n = 38) were analyzed for vitamin A, α-retinol (α-vitamin A), and carotenoids. Median hepatic vitamin A and total carotenoid concentrations were 6.49 µ mol/g and 4.30 nmol/g, respectively. Compared with controls, vervets fed the carotenoid complex had higher hepatic vitamin A (11.9 ± 5.1 µ mol/g), α -vitamin A (1.3 ± 0.7 µ mol/g), α -carotene (11.5 ± 5.3 nmol/g), ß-carotene (15.6 ± 8.6 nmol/g), and total carotenoids (28.1 ± 13.9 nmol/g) but lower lutein (0.66 ± 0.28 nmol/g) and zeaxanthin (0.24 ± 0.06 nmol). NHP may benefit from replacement of preformed vitamin A with carotenoids in feeds; however, bioconversion efficiency in these models should be investigated to determine optimal levels.

Vitamin A nutrition among East Timor refugee children.

BACKGROUND AND OBJECTIVES: Children age 6 to 72 months, living in refugee camps are at increased risk of developing vitamin A deficiency (VAD), resulting in increased morbidity and mortality. Due to poverty, often this population group has limited access to foods containing vitamin A from animal-based food sources and do not commonly consume available foods containing beta-carotene. To date, there is a paucity of data on vitamin A intake in young refugee children. To determine vitamin A intake in children ages 6 to 72 months at refugee camps in East Timor. METHODS AND STUDY DESIGN: A cross sectional study was carried out among children ages 6 to 72 months at refugee camps near Dili, East Timor. A detailed vitamin A intake questionnaire was ascertained from the primary caretaker, and the criteria and indicator cut off values suggested by World Health Organization (WHO) were used to classify the populations' vitamin A risk. RESULTS: Although animal sources of vitamin A were limited due to costs, all 89 children commonly consumed fruit containing vitamin A sources more than 3 times a week. Most children (69.7%) had been breast fed regularly, while 30.3% combined with bottle milk. 80.9% of children received vitamin A supplementation. CONCLUSION: Children in East Timor refugee camps have adequate vitamin A intake.

Usual nutrient intake adequacy among young, rural Zambian children.

Inadequate nutrient intakes put children at risk for impaired growth and development. We described diet, usual intakes of energy and macro- and micronutrients and prevalence of nutrient intake adequacies among 4-8-year-old Zambian children. Children not yet in school and living in Mkushi District, Central Province, Zambia were enrolled into an efficacy trial of pro-vitamin A biofortified maize. Children in the non-intervened arm were included in this analysis (n 202). Dietary intake data were collected by tablet-based 24-h recall on a monthly basis over the 6-month trial. Observed nutrient intakes were derived from reported food quantities, standard recipes and food composition tables. Usual nutrient intake distributions were modelled based on observed intakes. Prevalence of inadequacy was estimated by comparing the usual nutrient intake distribution to the nutrient requirement distribution. Frequency and quantity of consumption of commonly reported foods were described and key sources of energy and nutrients were identified. Median usual energy intake was 6422 kJ/d (1535 kcal/d). Most childrens' macronutrient intakes fell within recommended ranges (74-98 %). Estimated prevalences of inadequate intakes of Fe, folate, vitamin B12 and Ca were 25, 57, 76 and >99 %, respectively. Estimated prevalences of inadequacy for other micronutrients were low (0·1-2·2 %). Commonly consumed foods included maize, vegetable oil, tomatoes, rape leaves and small fish (>0·6 servings/d), whereas meat, eggs or dairy were rarely eaten (<0·2 servings/d). These findings suggest that the heavily plant-based diet of rural Zambian children provides inadequate Ca, folate, vitamin B12 and Fe to meet recommended nutrient intakes.

Indigenous leafy vegetables of Eastern Africa - A source of extraordinary secondary plant metabolites.

Indigenous African leafy vegetables vary enormously in their secondary plant metabolites whereat genus and the species have a great impact. In African nightshade (Solanum scabrum), spiderplant (Cleome gynandra), amaranth (Amaranthus cruentus), cowpea (Vigna unguiculata), Ethiopian kale (Brassica carinata) and common kale (Brassica oleracea) the specific secondary metabolite profile was elucidated and gained detailed data about carotenoids, chlorophylls, glucosinolates and phenolic compounds all having an appropriate contribution to health beneficial properties of indigenous African leafy vegetables. Exemplarily, various quercetin glycosides such as quercetin-3-rutinoside occur in high concentrations in African nightshade, spiderplant, and amaranth between ~1400-3300µg/g DW. Additionally the extraordinary hydroxycinnamic acid derivatives such as glucaric isomers and isocitric acid isomers are found especially in amaranth (up to ~1250µg/g DW) and spiderplant (up to 120µg/g DW). Carotenoids concentrations are high in amaranth (up to101.7µg/g DW) and spiderplants (up to 64.7µg/g DW) showing high concentrations of ß-carotene, the pro-vitamin A. In contrast to the ubiquitous occurring phenolics and carotenoids, glucosinolates are only present in the Brassicales species Ethiopian kale, common kale and spiderplant characterized by diverse glucosinolate profiles. Generally, the consumption of a variety of these indigenous African leafy vegetables can be recommended to contribute to different benefits such as antioxidant activity, increase pro-vitamin A and anticancerogenic compounds in a healthy diet.

Antioxidant capacity and fatty acids characterization of heat treated cow and buffalo milk.

BACKGROUND: Antioxidant capacity of milk is largely due to vitamins A, E, carotenoids, zinc, selenium, superoxide dismutase, catalase, glutathione peroxidase and enzyme systems. Cow milk has antioxidant capacity while the antioxidant capacity of buffalo milk has been studied in a limited way. The information regarding the effect of pasteurization and boiling on antioxidant capacity of cow and buffalo milk is also scared. METHODS: Cow and buffalo milk was exposed to two different heat treatments i.e. 65 °C for 30 min and boiling for 1 min. After heat treatments, milk samples were cooled down to 4 °C packaged in transparent 250 ml polyethylene PET bottles and stored at 4 °C for 6 days. Milk composition, total flavonoid content, total antioxidant capacity, reducing power, DPPH free radical scavenging activity, antioxidant activity in linoleic acid, vitamin C, A, E, selenium, Zinc, fatty acid profile, peroxide value and sensory characteristics were studied in raw, pasteurized and boiled cow and buffalo milk at 0, 3 and 6 days of storage period. RESULTS: Total antioxidant capacity (TAC) of raw, pasteurized and boiled milk for cow (42.1, 41.3 and 40.7%) and buffalo (58.4, 57.6 and 56.5%) samples was found, respectively. Reducing power (RP) of raw cow and buffalo milk was 6.74 and 13.7 while pasteurization and boiling did not showed significant effect on RP of both cow and buffalo milk. DPPH activity of raw, pasteurized and boiled milk for cow (24.3, 23.8 and 23.6%) and buffalo (31.8, 31.5 and 30.4%) samples was noted, respectively. Storage period up to 3 days was non-significant while DPPH assay after 6 days of storage period indicated significant decline in antioxidant activity of milk samples. Antioxidant activity in linoleic acid (AALA) of buffalo and cow milk were recorded 11.7 and 17.4%, respectively. Pasteurization and boiling did not showed any impact on antioxidant capacity of cow and buffalo milk. The Loss of vitamin C in pasteurization (40 and 42%) and boiling (82 and 61%) of cow and buffalo milk was recorded, respectively. Concentration of vitamin A and E in pasteurized cow and buffalo milk was not significantly different from raw milk samples of cow and buffalo. Concentration of selenium and zinc was not influenced by the heat treatment in both cow and buffalo milk samples. After 3 days of refrigerated storage, antioxidant capacity of both cow and buffalo milk decreased. Concentrations of short-chain and medium-chain fatty acids increased in pasteurized and boiled cow and buffalo milk, while long-chain fatty acids decreased in pasteurized and boiled cow and buffalo milk, with no effect on colour and flavor score. Peroxide value of pasteurized and boiled cow and buffalo milk was not influenced by the storage up to 3 days. CONCLUSIONS: These results suggest that buffalo milk had a higher antioxidant capacity than cow milk and pasteurized milk should be consumed within 3 days of refrigerated storage for better antioxidant perspectives.

Nonenzymatic ß-Carotene Degradation in Provitamin A-Biofortified Crop Plants.

Provitamin A biofortification, the provision of provitamin A carotenoids through agriculture, is regarded as an effective and sustainable intervention to defeat vitamin A deficiency, representing a global health problem. This food-based intervention has been questioned in conjunction with negative outcomes for smokers and asbestos-exposed populations of the CARET and ATBC trials in which very high doses of ß-carotene were supplemented. The current notion that ß-carotene cleavage products (apocarotenoids) represented the harmful agents is the basis of the here-presented research. We quantitatively analyzed numerous plant food items and concluded that neither the amounts of apocarotenoids nor ß-carotene provided by plant tissues, be they conventional or provitamin A-biofortified, pose an increased risk. We also investigated ß-carotene degradation pathways over time. This reveals a substantial nonenzymatic proportion of carotene decay and corroborates the quantitative relevance of highly oxidized ß-carotene polymers that form in all plant tissues investigated.

Comparative Safety and Efficacy Profile of a Novel Oil in Water Vaccine Adjuvant Comprising Vitamins A and E and a Catechin in Protective Anti-Influenza Immunity.

Non-replicating vaccines, such as those based on recombinant proteins, require adjuvants and delivery systems, which have thus far depended on mimicking pathogen danger signals and strong pro-inflammatory responses. In search of a safer and more efficacious alternative, we tested whether vaccinations with influenza recombinant hemagglutinin (HA) mixed with a novel vegetable oil in water emulsion adjuvant (Natural Immune-enhancing Delivery System, NIDS), based on the immune-enhancing synergy of vitamins A and E and a catechin, could protect against intra-nasal challenge with live influenza virus. Vaccinations of inbred Brag Albino strain c (BALB/c) mice, with HA mixed with NIDS compared to other adjuvants, i.e., a squalene oil in water emulsion (Sq. oil), and the Toll Like Receptor 3 (TLR3) agonist Poly (I:C), induced significantly lower select innate pro-inflammatory responses in serum, but induced significantly higher adaptive antibody and splenic T Helper 1 (TH1) or TH2, but not TH17, responses. Vaccinations with NIDS protected against infection, as measured by clinical scores, lung viral loads, and serum hemagglutination inhibition titers. The NIDS exhibited a strong dose sparing effect and the adjuvant action of NIDS was intact in the outbred CD1 mice. Importantly, vaccinations with the Sq. oil, but not NIDS, induced a significantly higher Serum Amyloid P component, an acute phase reactant secreted by hepatocytes, and total serum IgE. Thus, the NIDS may be used as a clinically safer and more efficacious vaccine adjuvant against influenza, and potentially other infectious diseases.

The research and implementation continuum of biofortified sweet potato and maize in Africa.

The enhancement of sweet potato and maize with provitamin A carotenoids has been part of HarvestPlus's research continuum since the formation of the biofortification project. This review includes case studies of biofortification strategies used for sweet potato in Uganda and orange maize in Zambia. The current status of the science and release of biofortified varieties was reviewed by three scientists who were part of the HarvestPlus program for more than a decade with input from a scientist who experienced orange maize dissemination in Zambia. High ß-carotene varieties of sweet potato were introduced into South Africa and Mozambique, and efficacy and effectiveness studies, respectively, showed promise to improve vitamin A status, followed by dissemination efforts in Uganda. A randomized, controlled effectiveness trial tested extension models to promote sweet potato and assessed vitamin A intake among Ugandans. Orange maize breeding was initially a challenge, but considering that the carotenoid biosynthetic pathway was present in maize germplasm, breeders quickly bred higher amounts of provitamin A into the maize that was ultimately released in Zambia. Initial resistance occurred because orange maize was associated with yellow maize, which had negative connotations associated with food aid and animal feed, and consumers preferred white maize. Currently, both orange crops are available on the market.

Short-Term Daily Consumption of Provitamin A Carotenoid-Biofortified Maize Has Limited Impact on Breast Milk Retinol Concentrations in Zambian Women Enrolled in a Randomized Controlled Feeding Trial.

BACKGROUND: Provitamin A carotenoid-biofortified maize is a conventionally bred staple crop designed to help prevent vitamin A deficiency. Lactating women are a potential target group, because regularly eating biofortified maize may increase vitamin A in breast milk-a critical source of vitamin A for breastfeeding infants. OBJECTIVE: We assessed whether daily consumption of biofortified orange maize would increase the retinol concentration in the breast milk of Zambian women. METHODS: Lactating women (n = 149) were randomly assigned to receive orange maize delivering 600 µg retinol equivalents (REs)/d as carotenoid plus placebo (OM), low-carotenoid white maize plus 600 µg REs/d as retinyl palmitate (VA), or white maize plus placebo (WM). Boiled maize (287 g dry weight/d) was served as 2 meals/d, 6 d/wk for 3 wk. We measured initial and final breast milk plasma retinol and ß-carotene concentrations, and plasma inflammatory protein concentrations. RESULTS: Groups were comparable at enrollment, with an overall geometric mean milk retinol concentration of 0.95 µmol/L (95% CI: 0.86, 1.05 µmol/L); 56% of samples had milk retinol <1.05 µmol/L. Median capsule and maize intake was 97% and 258 g dry weight/d, respectively. Final milk ß-carotene did not vary across groups (P = 0.76). Geometric mean (95% CI) milk retinol concentration tended to be higher in the OM [1.15 µmol/L (0.96, 1.39 µmol/L)] and VA [1.17 µmol/L (0.99, 1.38 µmol/L)] groups than in the WM group [0.91 µmol/L (0.72, 1.14 µmol/L); P = 0.13], and the proportion of women with milk retinol <1.05 µmol/L was 52.1%, 42.9%, and 36.7% in the WM, OM, and VA groups, respectively (P-trend = 0.16). CONCLUSIONS: Daily biofortified maize consumption did not increase mean milk retinol concentration in lactating Zambian women; however, there was a plausible downward trend in the risk of low milk retinol across intervention groups. This trial was registered at clinicaltrials.gov as NCT01922713.