High Concentrations Of Vitamin Research Articles

DETERMINATION OF HEAVY METALS AND C VITAMIN IN ALLIUM PLANTS (GARLIC AND ONION)

Analyses of plants render important information in toxicological and environmental investigations since plants are a very significant link in the trophic chain. The aim of this study was to analyze quantitatively the distribution of heavy metals and ascorbic acid in Allium plants: Allium sativum L. (garlic) and Allium cepa L. (onion). Heavy metals accumulation in garlic and onion was quantified by flame atomic absorption spectrometry (FAAS) after the chemical mineralization of the samples with nitric acid and hydrogen peroxide in a Digesthal device. To determinate of vitamin C we used a titrimetric method with potassium brommat-bromide solution in acid medium, when vitamin C is oxidized to dehidroascorbic acid by bromine. Our results indicate a high concentration of vitamin C in leaves of the studied Allium plants.

Vitamin C Is an Important Cofactor for Both Adrenal Cortex and Adrenal Medulla

The adrenal gland is among the organs with the highest concentration of vitamin C in the body. Interestingly, both the adrenal cortex and the medulla accumulate such high levels of ascorbate. Ascorbic acid is a cofactor required both in catecholamine biosynthesis and in adrenal steroidogenesis. Here we provide an overview on the role of vitamin C in the adrenal cortex and medulla derived from in vitro and in vivo studies. In addition, recent insights gained from an animal model with a deletion in the gene for the ascorbic acid transporter will be summarized. Mutant mice lacking the plasma membrane ascorbic acid transporter (SVCT2) have severely reduced tissue levels of ascorbic acid and die soon after birth. There is a significant decrease of tissue catecholamine levels in the adrenals. On the ultrastructural level, adrenal chromaffin cells in SVCT2 null mice show depletion of catecholamine storage vesicles, signs of apoptosis, and increased glycogen storage. Decreased plasma levels of corticosterone and altered morphology of mitochondrial membranes indicate additional effects of the deficiency on adrenal cortical function. The data derived from these animal models and various cell culture studies confirm a crucial role for vitamin C for both the adrenal cortex as well as the adrenal medulla further underlining the interdependence of the two endocrine systems united in one gland.

Amlodipine and carvedilol prevent cytotoxicity in cortical neurons isolated from stroke-prone spontaneously hypertensive rats.

We previously reported that vitamin E prevents apoptosis in neurons during cerebral ischemia and reperfusion in stroke-prone spontaneously hypertensive rats (SHRSP). In this paper, we analyzed the effects of antihypertensives as well as vitamin E, which were added to neuron cultures after reoxygenation (20% O2) following hypoxia (1% O2). When added after hypoxia before reoxygenation, vitamin E conferred significant protection to neuronal cells. It was also shown that vitamin E conferred complete protection from neural cell death when added hypoxia and again before reoxygenation. At higher concentrations of vitamin E, strong neuroprotection was observed. Moreover, we verified that pretreatment with either amlodipine, carvedilol or dipyridamole consistently prevented cell death during hypoxia and reoxygenation (H/R). On the other hand, nilvadipine, a dihydropyridine-type calcium entry blocker, had no apparent effect on neuroprotection during H/R. The order of neuroprotective potency was vitamin E > dipyridamole > carvedilol > or = amlodipine > nilvadipine. In parallel experiments, we examined whether these antihypertensive agents were more effective when combined with vitamin E and dipyridamole. The results suggested that in our in vitro model system, antioxidants were the most important agents for the reduction of oxygen-free radical damage in cortical neurons. These findings suggest that amlodipine and carvedilol, with their antioxidant properties and antihypertensive activity, would be useful to inhibit neuronal cell death in the treatment of cerebrovascular stroke and neurodegenerative diseases in hypertensive patients.

Caspase‐3 mediates retinoid‐induced apoptosis in the organogenesis‐stage mouse limb

Caspases are key mediators in the regulation and execution of apoptosis, a crucial part of the morphogenetic process during limb development. Caspase-8 and -9 are upstream caspases. Caspase-8 mediates the extrinsic pathway of apoptosis triggered by signaling through TNF-R1 family receptors. Caspase-9 is activated during the intrinsic pathway downstream of mitochondria. Caspase-3 is an effector caspase that initiates degradation of the cell in the final stages of apoptosis. Vitamin A is a potent teratogen that causes limb reduction defects in embryos exposed during organogenesis. Previous in vitro studies have shown that exposure of the organogenesis-stage murine limb to vitamin A results in excessive levels of apoptosis. The goal of this work was to characterize the involvement of caspase-3, -8, and -9, as well as cytochrome-c release from the mitochondria, in the apoptotic cascade induced by vitamin A. Limb buds from gestational day 12 CD-1 mice were cultured in a chemically defined medium in the absence or presence of vitamin A. Cultures were terminated after 6 days to examine the effect of the drug on gross morphology. Apoptosis was detected by TUNEL staining after culture for 24 hr. Caspase activation was determined by Western blotting and localized by immunohistochemistry of control and treated limbs. The release of cytochrome-c into the cytoplasm was assessed by Western blotting after cell-fractionation. Limbs cultured in the presence of vitamin A showed a dose-dependent growth reduction and dysmorphogenesis of the cartilaginous anlagen. Apoptosis was increased in the interdigital, anterior, and posterior marginal zones and in the apical ectodermal ridge. Western-blotting confirmed the presence of activated caspase-3 that increased with time in culture and vitamin A concentration. Cleaved caspase-3 immunoreactivity colocalized with TUNEL stained limb regions and increased dramatically with increasing drug concentrations. In contrast, procaspase-8 and -9 were not activated. Exposure to high concentrations of vitamin A did, however, increase cytoplasmic cytochrome-c, suggesting mitochondrial involvement. Caspase-3 is a key effector caspase in the apoptotic pathway induced by Vitamin A. While caspases-8 and -9 are not responsible for the activation of caspase-3 in response to the drug, cytochrome-c release from mitochondria may play an upstream role.

Downregulation of Superoxide Dismutase Activity and Gene Expression in Cultured Rat Brain Astrocytes after Incubation with Vitamin C

Reactive oxygen species have been linked with neuropathological changes in the central nervous system. Epidemiological studies supported the beneficial effect of supplementation of antioxidants. Superoxide dismutase (SOD) is an endogenous enzyme which can scavenge reactive oxygen species. This study investigated the effect of supplementation with ascorbic acid (vitamin C) on the changes of SOD in cultured neurological cells. Rat brain astrocytes (RBA-1 cells) were incubated with vitamin C and divided into four groups: a control group (without vitamin C) and three treatment groups with vitamin C at 40, 80, and 160 µmol/l. After short-term (2 days) and long-term (7 days) incubation, SOD activity, SOD mRNA level by Northern blotting, and SOD protein amounts by Western blotting were measured. After 2 days of incubation, vitamin C resulted in a decrease in the activity of SOD in a concentration-dependent manner (Mn-SOD from 14.8 ± 1.2 to 13.2 ± 0.5 U/mg protein and Cu/Zn-SOD from 64.8 ± 1.2 to 51.7 ± 0.9 U/mg protein; p < 0.05), and vitamin C also attenuated the Cu/Zn-SOD mRNA level from 100 to 86.3 ± 6.7%; p < 0.01), whereas the protein amounts of these two SODs remained unchanged. After 7 days of incubation with vitamin C, the SOD activity of RBA-1 cells decreased significantly (Mn-SOD from 14.9 ± 0.3 to 11.8 ± 0.3 U/mg protein and Cu/Zn SOD from 61.8 ± 1.8 to 54.6 ± 0.9 U/mg protein; p < 0.01), and the mRNA level was also attenuated (Mn-SOD from 100 to 86.8 ± 8.7% and Cu/Zn-SOD from 100 to 84.7 ± 4.8%; p < 0.01). These results suggest that 2 and 7 days of incubation with relatively high concentrations of vitamin C may downregulate activity and gene expression of SOD in cultured RBA-1 cells.

Vitamin D 3 and 25-hydroxyvitamin D 3 in raw and cooked pork cuts

The contents of vitamin D 3 and its metabolically active metabolite 25-hydroxyvitamin D 3 (25OHD 3) were examined by HPLC in different parts of four common raw pork cuts (loin boneless, leg inside, thin belly, neck) and in cooked meat (loin boneless). In whole raw pork cuts, varying in fat content from 2.2 to 26.5 g/100 g, concentrations of vitamin D 3 from 0.05 to 0.21 μg/100 g were measured. Pork cuts also contained significant amounts of 25OHD 3, from 0.07 to 0.14 μg/100 g. Further, the study demonstrated that most of the vitamin D 3 and 25OHD 3 is located in the fatty tissues, and that rind, despite its limited fat content, has a high concentration of vitamin D 3 and 25OHD 3. Cooking increased vitamin D 3 and 25OHD 3 calculated per 100 g of tissue in all parts and in the whole cut (in whole cuts in raw and cooked meat, respectively: vitamin D 3: 0.15 (0.08–0.24) μg/100 g and 0.18 (0.11–0.28) μg/100 g; P=0.33; 25OHD 3: 0.09 (0.06–0.18) μg/100 g and 0.13 (0.10–0.18) μg/100 g; P=0.02); however, correcting for differences in dry matter content, ameliorated all significant differences. 25OHD 3 has a higher (from 1.5 to 5 times) biological activity than vitamin D 3. Meat 25OHD 3 contributes significantly to vitamin D activity. Food databases should include concentrations of both vitamin D and 25OHD.

Vitamins reverse endothelial dysfunction through regulation of eNOS and NAD(P)H oxidase activities.

Antioxidant vitamins C and E have protective properties in genetic hypertension associated with enhanced oxidative stress. This study investigated whether vitamins C and/or E modulate vascular function by regulating enzymatic activities of endothelial nitric oxide synthase (eNOS) and NAD(P)H oxidase using thoracic aortas of 20- to 22-week-old male spontaneously hypertensive rats (SHR) and their matched normotensive counterparts, Wistar-Kyoto rats (WKY). SHR aortas had impaired relaxant responses to acetylcholine but not to sodium nitroprusside, despite an approximately 2-fold increase in eNOS activity and NO release. The levels of superoxide anion (O2-), a potent NO scavenger, and NAD(P)H oxidase activity were also 2-fold higher in SHR aortas. Mechanical but not pharmacological inactivation of endothelium (by rubbing and 100 micromol/L L-NAME, respectively) significantly abrogated O2- in both strains. Treatments of SHR aortas with NAD(P)H oxidase inhibitors, namely diphenyleneiodinium and apocynin, significantly diminished O2- production. The incubation of SHR aortas with different concentrations of vitamin C (10 to 100 micromol/L) and specifically with high concentrations of vitamin E (100 micromol/L) improved endothelial function, reduced superoxide production as well as NAD(P)H oxidase activity, and increased eNOS activity and NO generation in SHR aortas to the levels observed in vitamin C- and E-treated WKY aortas. Our results reveal endothelial NAD(P)H oxidase as the major source of vascular O2- in SHR and also show that vitamins C and E are critical in normalizing genetic endothelial dysfunction through regulation of eNOS and NAD(P)H oxidase activities.

Efferent loop small intestinal vitamin D receptor concentration and bone mineral density after Billroth II (Polya) gastrectomy in humans.

Animal studies have demonstrated that the highest concentration of vitamin D receptors (and greatest capacity for active calcium absorption) occurs in the proximal duodenum. By passing the duodenum following Polya/Billroth II gastrectomy could result in the development of a metabolic bone disease and low bone mineral density (BMD). We thus compared the vitamin D receptor (VDR) concentration in mucosal biopsies taken at endoscopy from two functionally corresponding areas of the small intestine: the jejunum (or efferent loop) in 21 patients with a history of Polya/Billroth II gastrectomy and the second part of the duodenum in age/sex-matched control subjects. We also measured the BMD by dual energy X-ray absorptiometry. The mean VDR concentration was not significantly different between the two groups (patients vs controls, fmol/mg protein, mean +/- SE: 34.99 +/- 2.57 vs 34.67 +/- 3.71; P = 0.22), even when subgrouped as males (36.22 +/- 3.16 vs 31.2 +/- 4.24; P = 0.351) or females (31.93 +/- 4.7 vs 43 +/- 6.76; P = 0.193). In Polya/Billroth II gastrectomy patients, the VDR concentration in the efferent loop declined with age (r = -0.78, P = 0.02). In the same group, BMD, as compared with matched controls, was significantly reduced at the lumbar spine (Z-score: patients vs controls: -1.138 vs 0.099, P = 0.01), but not at the femoral neck (Z-score: -0.69 vs 0.7, P = 0.084). There was no correlation between VDR and time since operation or BMD. These results suggest that following Polya/Billroth II gastrectomy, the functional capacity of the jejunal efferent loop in reference to VDR concentration is similar to that of the second part of the duodenum in normal subjects. Therefore, the reduced BMD in our patients, also a common finding in other studies, may not be secondary to the reduced capacity of the VDR system that facilitates the active calcium transport pathway in the proximal small intestine.

Mutagenicity, antioxidant potential, and antimutagenic activity against hydrogen peroxide of cashew (Anacardium occidentale) apple juice and cajuina.

Fresh and processed cashew (Anacardium occidentale) apple juice (CAJ) are among the most popular drinks in Brazil. Besides their nutritional benefits, these juices have antibacterial and antitumor potential. The chemical constituents of both the fresh juice and the processed juice (cajuina) were analyzed and characterized as complex mixtures containing high concentrations of vitamin C, various carotenoids, phenolic compounds, and metals. In the present study, these beverages exhibited direct and rat liver S9-mediated mutagenicity in the Salmonella/microsome assay with strains TA97a, TA98, and TA100, which detect frameshifts and base pair substitution. No mutagenicity was observed with strain TA102, which detects oxidative and alkylating mutagens and active forms of oxygen. Both CAJ and cajuina showed antioxidant activity as determined by a total radical-trapping potential assay. To test whether this antioxidant potential might result in antimutagenesis, we used a variation of the Salmonella/microsome assay that included pre-, co-, and posttreatment of hydrogen peroxide-exposed Salmonella typhimurium strain TA102 with the juices. CAJ and cajuina protected strain TA102 against mutation by oxidative damage in co- and posttreatments. The antimutagenic effects during cotreatment with hydrogen peroxide may be due to scavenging free radicals and complexing extracellular mutagenic compounds. The protective effects in posttreatment may be due to stimulation of repair and/or reversion of DNA damage. The results indicate that CAJ and cajuina have mutagenic, radical-trapping, antimutagenic, and comutagenic activity and that these properties can be related to the chemical constituents of the juices.

Vitamin E and MDA Concentrations in Plasma of Healthy Young Adult, Elderly and Pregnancy

Vitamin E is one of essential micronutrients. It is an antioxidant which potentially protects bioinolecules against oxidative damage. One ofparameters ofoxidafive damage is MDA or lipid peroxide. In this study, concentrations of plasma vitamin E as an antioxidant and MDA as one of oxidative damage markers were measured. The subjects consisted offive groups: (I) male medical students (young adult), (II) female medical students (young adult), (III) nursery inhabitants in Yogyakarta province (Abiyoso), (IV) nursery inhabitants in Manado, North Sulawesi Province (Senja Cerah), as the old ages, and (V) pregnant women within third trimester (of Prenatal Care Unit in Sardjito Hospital). The concentrations of Vitamin E (mg/dl) and MDA (nmol/ml) were as follows: Young Adult Male (n= 28) 20,8 + 3,8 and 0,190 + 0,057 ; Young adult Female (n-28) 21,9 + 4,5 and 0,123 + 0,057 ; Old age Yogyakarta (n=41) 30,6 + 10,4 and 0,291 + 0,062 ; Old age Manado (n=40) 26,5 + 4,5 and 0,391 + 0,081 ; Pregnant Women (=28) 39,9 + 11,3 and 0,165 -f 0,067. Groups of people who were hyperlipemic tend to have high concentration of Vitamin E, while group IV who eat less vegetable protein and more PUFA tend to have higher MDA concentration.

Vitamin A physiology in the precocious harbour seal (Phoca vitulina): a tissue-based biomarker approach

Vitamin A is a nutrient essential to all mammals for growth and development, as well as for the maintenance of reproductive, endocrine, and immune systems. Environmental contaminant-related disruption of vitamin A has been observed in many wildlife species and can therefore be used as a biomarker of toxic effects. However, the natural processes regulating vitamin A uptake, storage, and distribution among compartments are poorly understood in marine mammals. In this study, 20 young healthy harbour seals (Phoca vitulina) were captured to establish a compartment-based model providing a foundation for a mechanistic understanding of vitamin A physiology and disruption. Vitamin A (retinol, retinyl palmitate, and (or) retinoic acid) was quantified in blood plasma and in biopsy samples of liver, blubber, and skin. Although the highest concentrations of vitamin A were found in liver, blubber represents a more important storage depot, with an estimated 66% of the total retinoid content of the compartments measured. We suggest that vitamin A physiology in the precocious harbour seal has evolved to deal with high vitamin A availability during a short nursing period and to sustain growth during the postweaning fast. Positive correlations in vitamin A concentrations among liver, blubber, and skin support the use of less invasive biopsy sampling of just blubber or skin, which can provide physiologically relevant information in biomarker studies of free-ranging marine mammals.

Vitamin A metabolism is altered in brown Norway and long-Evans rats infused with naftidrofuryl or erythromycin intravenously.

Enzymatic retinyl ester hydrolysis is a key reaction for maintaining cellular retinol homeostasis. The ability of naftidrofuryl and erythromycin to inhibit retinol liberation by retinyl ester hydrolase (REH) in vitro suggests an ability to interfere with vitamin A metabolism in vivo, particularly during hepatic processing. To address this question, systemic and local response to these agents were studied in Brown Norway (BN) and Long-Evans (LE) rats. The study was conducted in two parts: a drug-loading phase and a washout phase. Analysis of variance of the time course changes in plasma retinol during the post-treatment period (Days 10-18) showed rat strain (p < 0.04) and time (p < 0.001; strain-by-time interactive effect, p < 0.001) to be significant factors, but drug exposure (p = 0.19) was not significant. Endpoints included hepatic REH activity, size and composition of the liver vitamin A stores, and retinoid content of the kidneys. Rats recovering from naftidrofuryl dosing demonstrated a lower REH activity than did animals recovering from erythromycin treatment (p < 0.009). The major side effect of erythromycin is vitamin A accumulation in the liver (p < 0.001) and reductions in retinol reserves (p < 0.02) were among the consequences of naftidrofuryl treatment. In the kidney of LE rats, there were higher concentrations of vitamin A (p < 0.003) secondary to naftidrofuryl exposure. Together our data suggest that clinically achievable concentrations of the drugs, given as a continuous infusion, produce aberrations in vitamin A metabolism.

Poster Sessions CP10: Blood–Brain Barrier. Expression of vitamin C transporter SVCT2 in hypothalamic glial cells. An immunohistochemical and kinetic analysis

Kinetic analysis of vitamin C uptake has demonstrated that specialized cells take up ascorbic acid (AA), the reduced form of vitamin C, through sodium‐AA cotransporters. Recently, two different isoforms of sodium‐vitamin C cotransporters (SVCT 1, 2) that mediate high affinity Na+‐dependent l‐ascorbic acid have been cloned. SVCT2 was detected mainly in choroid plexus cells and neurons, however, there are no evidences of SVCT2 expression in glial cells. High concentrations of vitamin C has been demonstrated in brain hypothalamic area. The hypothalamic glial cells, known as alpha and beta tanycytes, are specialized ependymal cells that bridge the cerebrospinal fluid and the portal blood of the median eminence. Our hypothesis postulates that tanycytes take up reduced vitamin C from the portal blood and cerebrospinal fluid generating an high concentration of this vitamin in brain hypothalamic area. In situ immunohistochemical analyses demonstrated that SVCT2 transporter is selectively expressed in apical region of tanycytes. A newly developed primary culture of mouse hypothalamic tanycytes was used to confirm the expression and function of SVCT2 isoform in these cells. Reduced vitamin C uptake was temperature and sodium dependent. Kinetic analysis showed an apparent Km of 20 μm and a Vmax of 45 pmol/min per million cells for the transport of ascorbic acid. The expression of SVCT2 was confirmed by immunoblots and RT–PCR. Tanycytes may perform a neuroprotective role concentrating the vitamin C in the hypothalamic area.Acknowledgements: Supported by Grands FONDECYT 1010843 and DIUC‐GIA 201.034.006‐1.4 from Concepción University.

Resveratrol and vitamin C as antioxidants in blood platelets

We investigated the effects of two different compounds with potential antioxidative action (found in many fruits, which can be used in therapy of cardiovascular disorders): trans-resveratrol (3,4′,5-trihydroxystilbene) and vitamin C on oxidative stress in blood platelets. Oxidative stress in blood platelets was estimated by the measurement of: (1) the generation of superoxide radicals (O 2 − ) (reduction of cytochrome c) and other reactive oxygen species—ROS: H 2O 2, singlet oxygen and organic radicals (chemiluminescence), (2) the production of thiobarbituric acid reactive substances (TBARS) and the level of conjugate dienes as markers of lipid peroxidation. We have shown that vitamin C at the concentrations of 750–3000 μM, but not at the concentration of 100 μM (prooxidative action) significantly inhibited peroxidation of lipids (measured by TBARS and conjugate dienes) and the production of ROS in blood platelets. After the incubation of blood platelets for 30 min at 37 °C with vitamin C at the concentration of 3000 μM, inhibition of ROS generation (above 90%) was achieved. The inhibition of ROS production caused by resveratrol at physiological plasma concentrations 0.05–2 μM was lower than by vitamin C. At the highest concentration of vitamin C used (3000 μM), the inhibition of O 2 − generation was about 40%. Moreover, we did not observe any synergistic action of resveratrol and vitamin C at antioxidative dose (3000 μM) on the inhibition of lipid peroxidation and the production of O 2 − or ROS in blood platelets. Resveratrol as an antioxidant reduced oxidative stress in blood platelets caused by vitamin C at prooxidative dose (100 μM).

Sensory quality of marinated frozen stored chicken thighs as affected by dietary fish fat and vitamin E

The effects of diets containing fish meal (0 or 4%), fish silage (0 or 4%) and vitamin E (60 or 200 mg kg–1) and the processing effect of marinating with sodium citrate (0.24 or 0.48%) or ascorbate (0.31 or 0.62%) have been studied to test the hypothesis that oxidative stability of frozen stored chicken thighs can be improved by such treatments. A trained sensory panel assessed the samples after storage at –25 °C for 1 week, 3 and 6 months. Feed with 4% fish meal resulted in increased fish flavour and odour of the thighs while 4% fish silage had a smaller effect on these attributes. Fish meal and fish silage added together into the feed by an amount of 4% each, caused strong fish flavour and odour in the product and accelerated the rancidity process. High concentration of vitamin E (200 mg kg–1) in the feed reduced rancidity when 4% fish products had been added to the feed, but no effect was noted when 4% fish meal plus 4% fish silage had been added together. High concentration of ascorbate in the brine (0.62%) decreased sensory score for rancidity attributes (hay, grass, soap and paint), while high concentration of citrate (0.48%) increased these parameters in frozen stored chicken thighs.

Effects of piscivory on the fatty acid profiles and antioxidants of avian yolk: studies on eggs of the gannet, skua, pelican and cormorant

AbstractPiscivorous birds consume diets which are rich in highly‐polyunsaturated n‐3 fatty acids; these play vital roles in embryonic development but are very susceptible to oxidative damage. The effects of such diets on the fatty acid composition and antioxidant content of the yolk were investigated in the northern gannet Morus bassanus, the great skua Catharacta skua, the American white pelican Pelecanus erythrorhynchos and the double‐crested cormorant Phalacrocorax auritus. The phospholipid fraction of the eggs of these four species contained high proportions of the n‐3 fatty acid, docosahexaenoic acid, which formed 7.5–11.3% (w/w) of the fatty acids of this fraction. The presence of eicosapentaenoic and docosapentaenoic acids also contributed to the total n‐3 content of the phospholipid. The n‐6 polyunsaturate, arachidonic acid, formed between 8% and 19% (w/w) of the phospholipid fatty acids. For the pelican and cormorant, this is consistent with the consumption of freshwater fishes in which arachidonic acid may be a significant acyl constituent. This finding is, however, more difficult to explain for the gannet and skua which largely consume marine fish with a low arachidonic acid content. The yolks of all four species contained relatively high concentrations of vitamin E (90.2–302.3 μg g−1 wet yolk) which was mainly present as α‐tocopherol. The eggs of the pelican and cormorant were especially enriched in carotenoids (150.9 and 115.7 μg g−1 wet yolk, respectively).

High pressure extraction of vitamin E-rich oil from Silybum marianum

Abstract The efficiency of high pressure extraction in the production of oil with a high concentration of vitamin E, from seeds of Silybum marianum , was investigated. The solvents used in extraction were liquid or supercritical carbon dioxide and liquid propane. Operating parameters were 100, 200, 300 bar and 25, 40, 60 and 80°C for CO 2 and 60, 150, 200 bar and 40, 60 and 80°C for propane, respectively. The influence of process parameters on the total yield of extraction, vitamin E composition and acid value of the extracts was investigated. Results were compared with those obtained for conventional extraction with organic solvents (n-hexane, petroleum ether, dichloromethane, chloroform). The mathematical model based on the adsorption–desorption equilibrium of oil from solid tissue, the diffusion of oil to the surface and mass transfer through external film into the bulk, described the experimental extraction results well.

Use of the brine shrimp, Artemia spp., in marine fish larviculture

Since no artificial feed formulation is yet available to completely substitute for Artemia, feeding live prey to young fish larvae still remains essential in commercial hatchery operations. The nutritional quality of commercially available Artemia strains being relatively poor in eicosapentaenoic acid (EPA, 20:5 n-3) and especially docosahexaenoic acid (DHA, 22:6 n-3), it is essential and common practice to enrich these live prey with emulsions of marine oils. In Artemia, the most commonly applied boosting technique is a 24-h enrichment period after hatching. However, the variability of enrichment studied in one Artemia strain (Great Salt Lake, Utah, USA) by the ICES Working Group on Mass Rearing of Juvenile Fish, showed a high variability in fatty acid bioaccumulation under laboratory or commercial conditions. To avoid the variation originating from differences in commercial preparations, standardized ICES emulsions with different HUFA and DHA/EPA ratios have been formulated and are available for research purposes. It should be emphasized, however, that the enrichment technique has limitations as Artemia are selectively catabolizing some of the nutrients such as DHA and phospholipids. Research on the kinetics of DHA catabolism in various Artemia strains has shown that DHA catabolism is strain-dependent and could partially be overcome by the use of strains of different geographical origin. Nowadays, various enrichment emulsions have been formulated differing in the fatty acid composition of their triglycerides. In this respect, the traditional formulations rich in EPA have been replaced by new products rich in DHA and arachidonic acid. To reduce the risks for oxidation of these fatty acids, higher concentrations of vitamin E are incorporated into the emulsions. Also, vitamin C has been incorporated in booster formulations that increase the level of ascorbic acid in Artemia to 2000 ppm. All these changes in the formulation of the enrichment diets offer more possibilities to cover the needs of different species and help to reduce problems related to diseases, stress resistance, malformation, and pigmentation in numerous fish species. Although continuous disinfection of Artemia during hatching and enrichment is becoming a routine operation in many hatcheries, the interference of bacteria in hatching and enrichment remains an important study object for which probionts might also give some solutions. As more attention is given to the use of on-grown Artemia as a cheaper alternative to the use of nauplii, simple cost-effective production techniques have been developed. The use of the right size of on-grown Artemia for feeding ensures a better energy balance in food intake and assimilation, thereby improving the performance of the fish. Furthermore, its palatability induces a good and fast feeding response. These characteristics, coupled with the use of bioencapsulation techniques to enhance the quality of the on-grown Artemia, make this organism an optimum diet for nursery of the fish.

Dietary vitamin E, IgE concentrations, and atopy

Vitamin E inhibits IgE responses to allergic stimuli in animals. We investigated the relation between dietary vitamin E intake and serum IgE concentrations and atopy, measured as allergen skin sensitisation, in a random sample of 2633 adults. Higher concentrations of vitamin E intake were associated with lower serum IgE concentrations and a lower frequency of allergen sensitisation. These findings may explain the beneficial effect of dietary vitamin E on the incidence of asthma.

Vitamin D 2 modulates melittin–membrane interactions

In the present work, mutual interaction of melittin, a pore forming hemolytic toxin from bee venom, and vitamin D 2, an antioxidant steroid, with dipalmitoyl phosphatidylcholine (DPPC) multilamellar liposomes has been investigated. Turbidity and Fourier transform infrared (FTIR) spectroscopic measurements, in combination with thermodynamic calculations, were used to monitor the modulating effect of vitamin D 2 on a melittin–DPPC membrane system. The results indicate that melittin on its own decreases the main phase transition to lower temperatures and also dramatically decreases the stability of the membrane. It has an overall disordering effect on the phospholipid membrane structures. Inclusion of vitamin D 2 at low concentrations (3, 6 mol%) into melittin containing DPPC liposomes slightly shifts the main phase transition to lower temperatures. High concentration of vitamin D 2 (9 mol%) has a more dramatic effect in shifting the main phase transition to lower temperature. It also causes a significant broadening in the phase transition curve. The present study also demonstrates that, with the addition of vitamin D 2 into melittin–DPPC system, absorbance value in turbidity study and the frequency of the CH 2 stretching band in FTIR study changes in a manner that are consistent with a reduction in the membrane perturbing effect of melittin on DPPC liposomes. Vitamin D 2 diminishes the disordering effect of melittin on DPPC lipids and produces a more ordered membrane system. These results were confirmed with thermodynamic calculations.