Cobalamin status is negatively correlated with vascular endothelial-cadherin in vegetarian and vegan women with vitamin B12 deficiency.

Optimal vitamin B12 status is important for vascular health. Vascular endothelial (VE) cadherin is an adherent junction protein involved in the maintenance of a functional endothelium. We hypothesized that vitamin B12 deficiency can negatively affect markers of vascular function, such as VE-cadherin. Within a human intervention study, we explored the possible association between cobalamin status (i.e., vitamin B12, holotranscobalamin, and homocysteine) and VE-cadherin (as marker of vascular health) in vegetarians/vegans (VEG) with B12 deficiency. The associations were evaluated at baseline and after 90-day supplementation with 2000 µg/wk of vitamin B12. On the whole, an inverse association between VE-cadherin and holotranscobalamin (P = .014) and a positive association between VE-cadherin and homocysteine (P = .041) was documented at baseline. VEG women showed higher levels of VE-cadherin compared with VEG men (P = .044), suggesting an increase in endothelial permeability. The intervention with vitamin B12 restored serum vitamin levels and improved the overall cobalamin status, whereas it did not affect VE-cadherin levels. The inverse association between holotranscobalamin and VE-cadherin was also maintained after intervention in women, corroborating the strong correlation between these 2 parameters. The results obtained seem to suggest a possible association between cobalamin status and VE-cadherin even if the intervention with B12 failed to positively affect VE-cadherin levels. Thus, further studies are needed to corroborate these findings and clarify the contribution of a vitamin B12 intervention on VE-cadherin levels in this target population. This trial was registered at ISRCTN registry (ISRCTN75099618).

Digestive and gastroprotective effects of Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (syn. A. moschata Wulfen) (Asteraceae): From traditional uses to preclinical studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (syn. A. moschata Wulfen) (Asteraceae) is an alpine endemic plant whose aerial parts are harvested by the locals mainly for the digestive properties. Despite its widespread use, few studies have been conducted to date to verify its bioactivity. AIM OF THE STUDY: The purpose of the work was to meet the tradition confirming with experimental data the popular belief that the consumption of this species offers beneficial effects to the gastrointestinal system. MATERIALS AND METHODS: Using Soxhlet apparatus, the dried aerial parts of A. erba-rotta subsp. moschata were successively extracted with petroleum ether (PET), dichloromethane (DCM) and methanol (MeOH). The essential oil (EO) was obtained by hydrodistillation using a Clevenger apparatus while infusion (AE) was prepared following the traditional local recipe. Their chemical characterization was performed by various techniques including SPME-GC/MS, GC/MS and HPLC/MS-MS. An in vitro biological screening was carried out. The influence of AE on lipid digestion was monitored by titration of free fatty acids (FFA) during pancreatic lipase activity with the pH-stat method. For all extracts and EO, the anti-Helicobacter pylori activity was assessed by the broth microdilution method, the influence on cell viability was evaluated against NCI-N87, OE21 and Caco-2 cell lines and a preliminary toxicity evaluation was done using Brine Shrimp lethality (BSL) assay. The anti-inflammatory potential was evidenced by interleukin IL-1- induced IL8 expression on Caco-2 cells. RESULTS: AE increased by 15% the FFA releasing compared to the pancreatic lipase alone. PET, DCM and MeOH extracts as well as AE and EO were considered active against the growth of both antimicrobial susceptible and resistant strains of H. pylori with MIC values starting from 16 µg/mL. PET and DCM (IC50 = 89 µg/mL and 96 µg/mL, respectively, against Caco-2 cell line) extracts showed the high effect on cell viability while the EO reduced in 50% of cell viability at 1.48 µL/mL (NCI-N87 cells), 1.42 µL/mL (OE21 cells), and 3.44 µL/mL (Caco-2 cells) corroborating the BSL results. In different degrees, all extracts and EO inhibited the IL-1ß-stimulated IL-8 production in Caco-2 cells. CONCLUSIONS: The obtained data are encouraging and provide a scientific basis for the traditional use of A. erba-rotta subsp. moschata as a digestive agent although they need to be further corroborated by studies involving the investigation of both the in vivo activities and the role of the compounds detected in the extracts.

Overview of Human Intervention Studies Evaluating the Impact of the Mediterranean Diet on Markers of DNA Damage.

The Mediterranean diet (MD) is characterized by high consumption of fruits, vegetables, cereals, potatoes, poultry, beans, nuts, lean fish, dairy products, small quantities of red meat, moderate alcohol consumption, and olive oil. Most of these foods are rich sources of bioactive compounds which may play a role in the protection of oxidative stress including DNA damage. The present review provides a summary of the evidence deriving from human intervention studies aimed at evaluating the impact of Mediterranean diet on markers of DNA damage, DNA repair, and telomere length. The few results available show a general protective effect of MD alone, or in combination with bioactive-rich foods, on DNA damage. In particular, the studies reported a reduction in the levels of 8-hydroxy-2'⁻deoxyguanosine and a modulation of DNA repair gene expression and telomere length. In conclusion, despite the limited literature available, the results obtained seem to support the beneficial effects of MD dietary pattern in the protection against DNA damage susceptibility. However, further well-controlled interventions are desirable in order to confirm the results obtained and provide evidence-based conclusions.

Effect of two different sublingual dosages of vitamin B12 on cobalamin nutritional status in vegans and vegetarians with a marginal deficiency: A randomized controlled trial.

BACKGROUND & AIMS: Vegetarians and vegans are more vulnerable to vitamin B12 deficiency with severe risks of megaloblastic anemia, cognitive decline, neuropathy, and depression. An easy and simple method of supplementation consists of taking one weekly dosage of 2000 µg. However, single large oral doses of vitamin B12 are poorly absorbed. The present research evaluates the ability of two different sublingual dosages of vitamin B12 (350 µg/week vs 2000 µg/week) in improving cyanocobalamin (vitamin B12) nutritional status in vegans and vegetarians with a marginal deficiency. METHODS: A 12-week randomized, double-blind, controlled, parallel intervention trial was performed. Forty subjects with marginal vitamin B12 deficiency were enrolled and randomly divided into two groups: test group Ld (low dose, 350 µg/week) and control group Hd (high dose, 2000 µg/week) vitamin B12 supplementation. Blood samples were collected at baseline and after 15, 30, 60, and 90 days from the intervention for the determination of vitamin B12, related metabolic markers, and blood cell counts. RESULTS: Two-way analysis of variance showed a significant effect of time (P < 0.0001) and of time × treatment interaction (P = 0.012) on serum concentration of vitamin B12 that increased after 90-day supplementation (Ld and Hd) compared to baseline. Both the supplements increased (P < 0.0001, time effect) the levels of holotranscobalamin, succinic acid, methionine and wellness parameter, while decreased (P < 0.0001, time effect) the levels of methylmalonic acid, homocysteine and folate compared to baseline. No difference was observed between groups (Ld vs Hd). No effect was detected for vitamin B6 and blood cell count. CONCLUSIONS: In our experimental conditions, both supplements were able to restore adequate serum concentrations of vitamin B12 and to improve the levels of related metabolic blood markers in subjects with a marginal deficiency. The results support the use of a sublingual dosage of 50 µg/day (350 µg/week) of cobalamin, instead of 2000 µg/week (provided as a single dose), to reach a state of nutritional adequacy of vitamin B12 in this target population. This study was registered at www.isrctn.org as ISRCTN75099618.

Role of polyphenols and polyphenol-rich foods in the modulation of PON1 activity and expression.

Paraoxonase 1 (PON1) is a high-density lipoprotein (HDL)-associated enzyme involved in the protection of low-density lipoprotein and HDLs against lipid peroxidation. Several studies documented the capacity of polyphenols to stimulate PON1 transcription activation. The objective of the present review is to provide the main evidence about the role and the potential mechanism of action of polyphenols and polyphenol-rich foods in the modulation of PON1 gene expression and activity. A total of 76 in vitro and in vivo studies were included in the review. Overall, while evidence obtained in vitro is limited to quercetin and resveratrol, those deriving from animal models seem more convincing for a wide range of polyphenols but only at pharmacological doses. Evidence from human studies is promising but deserves more substantiation about the role of polyphenol-rich foods in the regulation of PON1 activity and expression. Research focused on the understanding of the structure-activity relationship of polyphenols with PON1 and on the mechanisms at the base of PON1 modulation is warranted. Well-designed human intervention studies are encouraged to corroborate the findings of polyphenols also at physiological doses.

Berry Fruit Consumption and Metabolic Syndrome.

Metabolic Syndrome is a cluster of risk factors which often includes central obesity, dyslipidemia, insulin resistance, glucose intolerance, hypertension, endothelial dysfunction, as well as a pro-inflammatory, pro-oxidant, and pro-thrombotic environment. This leads to a dramatically increased risk of developing type II diabetes mellitus and cardiovascular disease, which is the leading cause of death both in the United States and worldwide. Increasing evidence suggests that berry fruit consumption has a significant potential in the prevention and treatment of most risk factors associated with Metabolic Syndrome and its cardiovascular complications in the human population. This is likely due to the presence of polyphenols with known antioxidant and anti-inflammatory effects, such as anthocyanins and/or phenolic acids. The present review summarizes the findings of recent dietary interventions with berry fruits on human subjects with or at risk of Metabolic Syndrome. It also discusses the potential role of berries as part of a dietary strategy which could greatly reduce the need for pharmacotherapy, associated with potentially deleterious side effects and constituting a considerable financial burden.

Bilberry adulteration: identification and chemical profiling of anthocyanins by different analytical methods.

Extracts of the bilberry fruit have protective effects against retinopathy and vascular complications; therefore, they are important ingredients in food supplements. Recently, there have been several reported cases of adulteration. Thus, to characterize the anthocyanin profile, and the relative percentages of these pigments, this study analyzed bilberry fruits from different countries by liquid chromatography coupled to a diode array detector and a mass spectrometer detector. A total of 15 anthocyanins were identified, and a fingerprint profile was used for the quality control of the target material. Fourteen bilberry extracts and 12 finished products labeled as bilberry from different marketing manufacturers were analyzed. Approximately 50% of these extracts differed significantly from the reference bilberry, suggesting possible adulteration. Approximately 60% of the extracts and 33% of the food supplements presented a lower anthocyanin content than declared. The adulterations were observed mainly with extracts of mulberry and chokeberry.

Effect of Time and storage temperature on anthocyanin decay and antioxidant activity in wild blueberry ( Vaccinium angustifolium ) powder.

This study evaluated the effects of storage on total and single anthocyanin (ACN) content, and total antioxidant activity (TAA) of freeze-dried wild blueberry (WB) powder maintained at 25, 42, 60, and 80 °C for 49 days. Storage reduced single and total ACN content at all of the temperatures; it was slower at 25 °C (-3% after 2 weeks), whereas it was faster at 60 °C (-60%) and at 80 °C (-85%) after 3 days. The values of half-life time (t1/2) were found to be 139, 39, and 12 days at 25, 42, and 60 °C, respectively, utilizing the Arrhenius equation. No significant effects were detected on TAA by temperature increase. In conclusion, this study provides important information on the stability of WB powder at 25 °C; this is interesting scientific research for the food industry.

Horse meat consumption affects iron status, lipid profile and fatty acid composition of red blood cells in healthy volunteers.

This study investigated the effect of moderate consumption of horse meat on iron status, lipid profile and fatty acid composition of red blood cells in healthy male volunteers. Fifty-two subjects were randomly assigned to two groups of 26 subjects each: a test group consuming two portions of 175 g/week of horse meat, and a control group that abstained from eating horse meat during the 90 days trial. Before and after 90 days, blood samples were collected for analysis. Horse meat consumption significantly (p ≤ 0.05) reduced serum levels of total and low-density lipoprotein cholesterol ( - 6.2% and - 9.1%, respectively) and transferrin ( - 4.6%). Total n - 3, long chain polyunsaturated fatty acids n - 3 and docosahexeanoic acid content in erythrocytes increased (p ≤ 0.05) by about 7.8%, 8% and 11%, respectively. In conclusion, the regular consumption of horse meat may contribute to the dietary intake of n - 3 polyunsaturated fatty acids and may improve lipid profile and iron status in healthy subjects.

Improvement of lymphocyte resistance against H(2)O(2)-induced DNA damage in Sprague-Dawley rats after eight weeks of a wild blueberry (Vaccinium angustifolium)-enriched diet.

The effect of wild-blueberry consumption on the resistance of H(2)O(2)-induced DNA damage was evaluated in the Sprague-Dawley (SD) rat. Thirty-two, four week-old SD rats were randomly assigned to two groups of sixteen rats each and fed a control diet (C) or a wild-blueberry (WB) diet for four or eight weeks. Following the dietary treatment, plasma antioxidant capacity was assessed by the TRAP assay. Ex vivo protection from H(2)O(2)-induced DNA damage was evaluated in lymphocytes by means of the comet assay. No significant effect was detected in plasma antioxidant capacity at four and eight weeks in both dietary groups. In contrast, the level of DNA damage was significantly lower in rats fed the WB diet compared with those on the C diet after eight weeks (11.± 1.0% vs 17.2 ± 2.4% DNA in tail, p ≤ 0.05) but not after four weeks. In conclusion, while WB consumption did not affect plasma antioxidant activity, it improved lymphocyte protection against oxidative damage, but only after eight weeks.

Anthocyanin absorption, metabolism, and distribution from a wild blueberry-enriched diet (Vaccinium angustifolium) is affected by diet duration in the Sprague-Dawley rat.

The effect of wild blueberry consumption on anthocyanin (ACNs) distribution and metabolism in Sprague-Dawley (SD) rats was investigated. Thirty-two rats were fed for 4 or 8 weeks with a control (C) or a wild blueberry-enriched diet (8%) (WB). Anthocyanin profile in plasma, urine, feces, brain, and liver was evaluated by LC-MS/MS, and significantly increased in urine and not in feces after 8 weeks on the WB diet compared to that in 4 weeks, but no anthocyanins were detected in plasma, liver, and brain samples either in the C or WB groups. Metabolites of ACNs were detected in the plasma, urine, feces, and tissues of both the C and WB groups, but the urinary excretion of hippuric acid increased significantly after 4 and 8 weeks of WB consumption. Thus, it seems that ACNs are metabolized by the intestinal microflora to respective phenyl-alkyl acids, which can be further metabolized to benzoic acid. In conclusion, ACNs are bioavailable in rats, and the extent of their metabolism and excretion is based on diet duration. Additionally, urinary hippuric acid content could represent a potential biomarker of ACNs absorption and metabolism in the SD rat under the present experimental conditions.

In vivo prion protein intestinal uptake in fish.

Intestinal uptake of abnormal prion protein (PrPSc), the pathological agent involved in transmissible spongiform encephalopathies (TSEs), has been investigated in rainbow trout (Oncorhynchus mykiss). Experimental procedures were conducted in vivo by immunohistological PrPSc localization in intestine and pyloric caeca after forced feeding of infected material. Results indicate that PrPSc was absorbed by the intestinal mucosa and that it persisted in the fish gastrointestinal tract for up to 3 days in pyloric caeca and for up to 7 days in the distal intestine. It did not remain longer than 15 days in the fish intestine; furthermore, it did not cross the intestinal barrier.

Scrapie infectivity is quickly cleared in tissues of orally-infected farmed fish.

BACKGROUND: Scrapie and bovine spongiform encephalopathy (BSE) belongs to the group of animal transmissible spongiform encephalopathy (TSE). BSE epidemic in the UK and elsewhere in Europe has been linked to the use of bovine meat and bone meals (MBM) in the feeding of cattle. There is concern that pigs, poultry and fish bred for human consumption and fed with infected MBM would eventually develop BSE or carry residual infectivity without disease. Although there has been no evidence of infection in these species, experimental data on the susceptibility to the BSE agent of farm animals other than sheep and cow are limited only to pigs and domestic chicken. In the framework of a EU-granted project we have challenged two species of fish largely used in human food consumption, rainbow trout (Oncorhynchus mykiss) and turbot (Scophthalmus maximus), with a mouse-adapted TSE strain (scrapie 139A), to assess the risk related to oral consumption of TSE contaminated food. In trout, we also checked the "in vitro" ability of the pathological isoform of the mouse prion protein (PrPSc) to cross the intestinal epithelium when added to the mucosal side of everted intestine. RESULTS: Fish challenged with a large amount of scrapie mouse brain homogenate by either oral or parenteral routes, showed the ability to clear the majority of infectivity load. None of the fish tissues taken at different time points after oral or parenteral inoculation was able to provoke scrapie disease after intracerebral inoculation in recipient mice. However, a few recipient mice were positive for PrPSc and spongiform lesions in the brain. We also showed a specific binding of PrPSc to the mucosal side of fish intestine in the absence of an active uptake of the prion protein through the intestinal wall. CONCLUSION: These results indicate that scrapie 139A, and possibly BSE, is quickly removed from fish tissues despite evidence of a prion like protein in fish and of a specific binding of PrPSc to the mucosal side of fish intestine.

Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC and its confirmation by MS.

During the period of October-July 2000, 240 samples of dairy ewes milk, obtained from farms of Enna (Sicily, Italy), were checked for Aflatoxin M(1) (AFM(1)) by HPLC using a fluorimetric detector. The limit of detection and the limit of quantification were 250 ng/L for AFM(1). All the positive milk samples for AFM(1) were confirmed by LC-MS. AFM(1) was detected in 81% of milk samples, ranging from 2 to 108 ng/L. Three samples were over the legal limits (50 ng/L). Mean contamination of samples obtained from stabulated ewes was higher than that from grazing ewes (35.27 vs. 12.47 ng/L). Furthermore, samples collected in the period September-October showed higher contamination than samples collected during the other months (42.68 vs. 10.55 ng/L). Both differences are related to the administration of compound feed. Based on current toxicological knowledge we concluded that the AFM(1) contamination levels recorded in ewe milk did not present a serious human health hazard. However, as ewe milk is exclusively used to produce cheese due to its higher protein content, and also considering the preferential binding of AFM(1) to casein during coagulation of milk, a potentially high concentration effect could occur, thus the surveillance of contamination levels should be more continuous and widespread.

Cyanidins: metabolism and biological properties.

Cyanidin and its glycosides belong to the anthocyanins, a widespread class of water-soluble plant compounds that are responsible for the brilliant color (red, orange, blue) of fruits and flowers. They are widely ingested by humans as it has been estimated a daily intake around 180 mg, mainly deriving from fruits and red wines. This paper reviews the literature on the biological activities, absorption and metabolism of cyanidins, with emphasis to the antioxidant, antimutagenic and other protective activities ascribed to these compounds. Their role in contrasting development of cancer and other pathologies is also reviewed. It is concluded that a great deal of work is still necessary to i) definitively clarify the metabolism of cyanidins in human beings; ii) assess the dietary burden and variations within and between populations; iii) evaluate the relationship between cyanidin glycosides-rich food consumption and incidence of given pathologies. The amount of work to be performed is even more significant when considering a possible therapeutic use of cyanidin glycosides-based drugs. With this aim, information on absorption, distribution, metabolism and excretion of cyanidin-glycosides administered by main possible routes are largely insufficient. However, consisting findings allow looking at cyanidins as dietary compounds with a potential beneficial role for human health.

Comparison of lutein bioavailability from vegetables and supplement.

Lutein is a carotenoid present in dark green leafy vegetables and it may be involved in the prevention of several diseases related to oxidative stress. The aim of this study was to evaluate bioavailability of lutein from different food sources (150 g spinach and 200 g broccoli) and a supplement in oil (300 mg VEGEX), all providing about 9 mg lutein. Eight healthy females were instructed to eat a low-carotenoid diet for the period of experimentation. On three different occasions, three weeks apart, volunteers ate the lutein sources together with 10 g olive oil and 40 g bread. Blood samples were collected just before eating, every two hours for 12 hours, and at 24, 32, 56, 80 and 104 hours. Lutein concentration increased significantly after six to eight hours and peaked after 10-12 hours, with the highest concentration reached after VEGEX intake. Lutein concentration remained significantly elevated for up to 80 hours (VEGEX and spinach). On the whole, our results suggest that the intake of one single dose of lutein from different sources is able to bring about a significant plasma response in the short term.

Procyanidins from Vitis vinifera seeds: in vivo effects on oxidative stress.

The purpose of this study was to evaluate the effect of supplementation with procyanidins from Vitis vinifera on markers of oxidative stress. Ten healthy volunteers received a daily dose of 110 mg of procyanidins for 30 days. Fasting venous blood samples were taken before and at the end of the supplementation period and after 7 days of wash-out. The total antioxidant activity and the plasma concentrations of alpha-tocopherol were not modified. Conversely, the levels of alpha-tocopherol in red blood cell membranes increased significantly from 1.8 +/- 0.1 to 2.8 +/- 0.2 mg/g. Similarly, the lymphocyte oxidized DNA [8-oxo-7,8-dihydro-2'-deoxyguanosine/2'-deoxyguanosine ratio] was reduced from 7.23 +/- 2.47 to 2.34 +/- 0.51, and the red blood cell membrane fatty acid composition shifted to a higher level of polyunsaturated fatty acids. On the basis of these results, it may be suggested that dietary procyanidins exert their antioxidant protection in vivo by sparing liposoluble vitamin E and reducing DNA oxidative damage.

Effect of the ratio of casein phosphopeptides to calcium (w/w) on passive calcium transport in the distal small intestine of rats.

OBJECTIVE: We investigated the influence of different ratios between casein phosphopeptides and calcium (CPP/Ca) in intestinal lumen on passive Ca transport across the distal small intestine of rats in vitro. METHODS: We measured the amounts of passive Ca absorbed across the everted distal small intestine of rats with the use of radioactive 45Ca as tracer. Four CPP/Ca ratios (5, 10, 15, and 20 w/w) and three mineral concentrations (4, 8, and 12 mmol/L) were tested on groups of six rats. RESULTS: We found a significant effect of the CPP/Ca ratio on mineral absorption (P < 0.001). The amounts of Ca absorbed increased from a ratio of 5 to a ratio of 15 and decreased with a ratio of 20, with a similar trend for every concentration tested. The more evident effect was found with the concentration of 12 mmol/L of Ca (P < 0.001 to 0.05). CONCLUSION: The positive effect of CPPs on passive Ca absorption seems to depend on the relative amounts of both species in intestinal lumen. In this study, the ratio 15 was identified as the most efficient at increasing mineral transport. This evidence might be considered before suggesting CPP as a Ca dietary integrator, although more studies in vitro and in vivo are needed.