Demand-oriented riboflavin supply of organic broiler using a feed material from fermentation of Ashbya gossypii.

Alternatives to riboflavin (vitamin B2) production by recombinant microorganisms are needed in organic poultry production, but are cost-intensive, so that a demand-oriented riboflavin supply is necessary. Details on the riboflavin requirements of organic poultry are not available. A feed material with high native riboflavin content from fermentation of the filamentous fungus Ashbya gossypii was studied. Two runs with 800 Ranger Gold™ broilers each (40 pens with 20 animals) were conducted. The fattening period was divided into starter (S), grower (G) and finisher (F) stage. In the first run, a basal diet without riboflavin supplementation (NATIVE; 3.27, 3.50 and 3.16 mg riboflavin/kg DM in S, G and F) was compared to diets with supplementation at low (LOW; 5.30, 4.85 and 5.19 mg/kg in S, G and F), medium (MEDIUM; 7.56, 6.88 and 7.56 mg/kg in S, G and F) and high (HIGH; 10.38, 9.14 and 9.93 mg/kg in S, G and F) dosage. In the second run, different combinations of low and medium riboflavin supplementation were used in S, G and F diets: S-LOW (4.50 mg riboflavin/kg DM), G-MEDIUM (6.66 mg/kg), F-MEDIUM (5.71 mg/kg) (Treatment A), S-LOW (4.50 mg riboflavin/kg DM); G-LOW (4.92 mg/kg), F-LOW (4.01 mg/kg) (Treatment B); S-MEDIUM (6.37 mg/kg), G-MEDIUM (7.37 mg/kg), F-MEDIUM (5.07 mg/kg) (Treatment C); S-MEDIUM (6.37 mg/kg), G-LOW (5.28 mg/kg), F-LOW (4.22 mg/kg) (Treatment D). Body weight, feed and water consumption were recorded weekly, health and welfare indicators were scored bi-weekly. Slaughter traits were assessed for five males and females per pen. In the first run, NATIVE animals showed symptoms of riboflavin deficiency and lower live weights in the second week of age. Riboflavin contents of this group were increased to avoid further deficiency and recovery was observed. Feed conversion was better in HIGH (2.07) compared with NATIVE and LOW (2.11). At slaughter, treatments differed neither for foot pad dermatitis nor plumage cleanliness. In the second run, daily weight gains did not differ between treatments in any of the weeks. Feed conversion ranged between 1.99 and 2.04. Riboflavin deficiency was not observed in the second run, while treatment D showed superior economic efficiency. In conclusion, native contents of feed components (3.27 mg/kg DM) were not sufficient to meet the riboflavin demand and a total content of 4.50 mg/kg DM was identified as safe lower threshold. The levels rather according to commercial recommendations were not additionally beneficial to performance and health.

[Prevention of neural tube defects: regional policies in folic acid enrichment and supplementation]. / Prävention von Neuralrohrdefekten: Länderpolitiken zur Folsäureanreicherung und -supplementierung.

BACKGROUND: The USA and Canada had already started an obligatory food fortification with folic acid in 1998; In 2009, Australia and New Zealand also started to do so. METHODS: A survey was carried out among members of The European Consumers' Organisation (BEUC). RESULTS: Most of the European countries go along with the recommendation of 200 µg folic acid (or 400 µg folic acid equivalents) for adults and 300 µg (600 µg) for pregnant women. To prevent neural tube defects, an additional supplementation of 0,4 mg folic acid is recommended for women before conception. So far, none of the European countries has implemented an obligatory folic acid enrichment of grain or other food, but this step is under discussion. CONCLUSION: In a European market with free trading of goods it is of utmost importance that especially those (socially deprived) women in most need of folic acid, are reached. A common European decision for/against fortification should be considered. Public Health ethics demand not only good evidence for the benefit, but also a good estimation of the (potential) risks. Due to a paucity of good risk estimation, no European country has plans to decide in favour of an obligatory fortification on its own.

[Neural tube defects in Austria: Assumption and calculations on the prevention potential through folic acid enrichment and supplementation]. / Neuralrohrdefekte in Österreich: Annahmen und Berechnungen zum Verhinderungspotenzial durch Folsäureanreicherung und-supplemente.

BACKGROUND: Countries with obligatory fortification of food (USA, Canada) document a significant decrease of neural tube defects in newborns. METHODS: In this study the Daly or, respectively, the Wald method was employed for calculating the potential of fortification/and supplementation for prevention in Austria. RESULTS: According to the EUROCAT study, in Austria the overall prevalence of neural tube defects (live birth, still births and induced abortions due to neural defect) is assumed to be 7.95 per 10,000 live and still births, and the prevalence of 3.9 per 10,000 live births - that is 62 or, respectively, 30 in absolute numbers per 78,000 births per year. In 2006, 26 live-born children with neural tube defects were actually reported in Austria by Statistik Austria. CONCLUSION: Different folic acid fortification and supplementation strategies can avoid 1.2-1.4 per 10,000 (9-11 cases) of neural tube defects (live and still births). Folic acid supplements are effective to decrease the amount of neural tube defects, however, only when pregnancies are planned. Thus, evidence-based neural tube defects are more common among lower social groups. An obligatory fortification of food could therefore reach unplanned pregnancies and women facing social problems. A reason to justify this population-based intervention where people need not decide for themselves could be the reduction of social imbalances. There are, of course, advantages and disadvantages of obligatory fortification of food and, therefore, all circumstances have to be taken into consideration.

A recombinant allergen chimer as novel mucosal vaccine candidate for prevention of multi-sensitivities.

BACKGROUND: As conventional immunotherapy is less efficacious in patients with allergic multi-sensitivities compared with mono-sensitized subjects, new intervention strategies are needed. Therefore, an allergen chimer was genetically engineered for treatment of multi-sensitization with birch and grass pollen on the basis of mucosal tolerance induction. METHODS: The major birch pollen allergen Bet v 1 served as a scaffold for N- and C-terminal linkage of the immunodominant peptides of the grass pollen allergens Phl p 1 and Phl p 5 and this new construct was cloned and expressed in Escherichia coli. After purification, physicochemical and immunological characterization the chimer was used for intranasal tolerance induction prior to poly-sensitization with Bet v 1, Phl p 1 and Phl p 5. RESULTS: The immunological characterization revealed that the conformation of Bet v 1 within the chimer was comparable to that of natural as well as recombinant Bet v 1. The chimer was immunogenic in mice for T and B cell responses to the three allergens. Intranasal application of the chimer prior to poly-sensitization significantly suppressed humoral and cellular allergen-specific Th2 responses and prevented development of airway inflammation upon allergen challenge. Moreover, local allergen-specific IgA antibodies were induced by the chimer. The mechanisms of poly-tolerance induction seemed to be mediated by regulatory cytokines, since TGF-beta and IL-10 mRNA in splenocytes were upregulated and tolerance was transferable with these cells. CONCLUSION: The data indicate that such allergen chimers harboring several unrelated allergens or allergen peptides could serve as mucosal polyvalent vaccines for prevention of multi-sensitivities.

Modulation of allergic immune responses by mucosal application of recombinant lactic acid bacteria producing the major birch pollen allergen Bet v 1.

BACKGROUND: Probiotic lactic acid bacteria (LAB) are able to modulate the host immune system and clinical trials have demonstrated that specific strains have the capacity to reduce allergic symptoms. Therefore, we aimed to evaluate the potential of recombinant LAB producing the major birch pollen allergen Bet v 1 for mucosal vaccination against birch pollen allergy. METHODS: Recombinant Bet v 1-producing Lactobacillus plantarum and Lactococcus lactis strains were constructed. Their immunogenicity was compared with purified Bet v 1 by subcutaneous immunization of mice. Intranasal application of the live recombinant strains was performed to test their immunomodulatory potency in a mouse model of birch pollen allergy. RESULTS: Bet v 1 produced by the LAB was recognized by monoclonal anti-Bet v 1 and IgE antibodies from birch pollen-allergic patients. Systemic immunization with the recombinant strains induced significantly lower IgG1/IgG2a ratios compared with purified Bet v 1. Intranasal pretreatment led to reduced allergen-specific IgE vs enhanced IgG2a levels and reduced interleukin (IL)-5 production of splenocytes in vitro, indicating a shift towards non-allergic T-helper-1 (Th1) responses. Airway inflammation, i.e. eosinophils and IL-5 in lung lavages, was reduced using either Bet v 1-producing or control strains. Allergen-specific secretory IgA responses were enhanced in lungs and intestines after pretreatment with only the Bet v 1-producing strains. CONCLUSIONS: Mucosal vaccination with live recombinant LAB, leading to a shift towards non-allergic immune responses along with enhanced allergen-specific mucosal IgA levels offers a promising approach to prevent systemic and local allergic immune responses.

Maternal nutritional risk factors for small for gestational age babies in a developed country: a case-control study.

AIMS: To assess the effect of maternal diet during pregnancy on the risk of delivering a baby who is small for gestational age (SGA). METHODS: Case-control study of 844 cases (SGA) and 870 controls (appropriate size for gestational age (AGA)). Only term (37+ completed weeks of gestation) infants were included. Retrospective food frequency questionnaires were completed at birth on the diet at the time of conception and in the last month of pregnancy. RESULTS: At the time of conception, mothers of AGA infants ate significantly more servings of carbohydrate rich food and fruit, and were more likely to have taken folate and vitamin supplements than mothers of SGA infants. There was some evidence that mothers of AGA infants also ate more servings of dairy products, meat, and fish (0.05 < p < 0.1). However, after adjustment for maternal ethnicity, smoking, height, weight, hypertension, and occupation, fish intake (p = 0.04), carbohydrate-rich foods (p = 0.04), and folate supplementation (p = 0.02) were associated with a reduced risk of SGA. In the last month of pregnancy, only iron supplementation was associated with a reduced risk of SGA (p = 0.05) after adjustment for potential confounders. CONCLUSIONS: This study suggests that small variations in maternal diets within the normal range during pregnancy in developed countries are associated with differences in birth weight.

Comparison of plasma biomarkers with dietary assessment methods for fruit and vegetable intake.

OBJECTIVE: To assess the variability and validity of plasma-based biomarkers of antioxidant vitamin, and fruit and vegetable intake. SETTING: Leeds, Wakefield, Huddersfield and Bradford, England. SUBJECTS: A total of 54 free-living, nonsmoking women recruited from participants of the UK Women's Cohort Study (UKWCS). METHODS: Two fasting blood samples were taken at two time points, 18 months apart. A 4-day food diary was completed prior to the first blood sample and a 24-h recall was conducted at the time of the second blood collection. All blood samples were analysed for ascorbic acid and four carotenoids. Associations between antioxidant vitamin intake from all food sources and supplements, as well as fruit and vegetable intake, and plasma levels of the antioxidant vitamins were assessed. RESULTS: Using the 4-day diary, positive associations were found between micronutrient intake from all food sources and plasma concentrations of ascorbic acid (P<0.01) and beta-carotene (P<0.01). No associations were seen between plasma micronutrient levels and specifically fruit and vegetable intakes. In general, associations between plasma levels and intakes assessed by the 24-h recall were less marked than those based on the 4-day diary. CONCLUSIONS: Plasma ascorbic acid and beta-carotene are good indicators of previous vitamin C and beta-carotene intake, from all food sources. However, caution is required in extrapolating these results to include individual food groups, rich in these vitamins. The results imply that the practice of using plasma biomarkers simply as a proxy measure of dietary intake is not valid and emphasise that plasma biomarkers are not simply a reflection of dietary intake, but also of a number of physiological processes. Biomarkers in nutrition epidemiological studies are however useful to measure nutrient status at the tissue level.

Anti-AIDS agents. Part 47: Synthesis and anti-HIV activity of 3-substituted 3',4'-Di-O-(S)-camphanoyl-(3'R,4'R)-(+)-cis-khellactone derivatives.

Six 3-substituted 3',4'-di-O-(S)-camphanoyl-(+)-cis-khellactone derivatives (3-8) were synthesized from 3-methyl DCK (2). 3-Hydroxymethyl DCK (6) exhibited potent anti-HIV activity in H9 lymphocytes with EC(50) and TI values of 1.87 x 10(-4) microM and 1.89 x 10(5), respectively. These values are similar to those of DCK and better than those of AZT in the same assay.

A critical evaluation of the application of biomarkers in epidemiological studies on diet and health.

One of the problems which may beset epidemiological studies is the difficulty of accurately measuring the dietary intakes of participants. Biomarkers of diet promise to provide a more accurate measure of dietary intake and a more objective one in that they are not reliant on the subject's memory. This review considers some issues of importance in epidemiology when information is obtained from biomarkers. The approach taken is to use examples both of normal dietary constituents and of contaminants in relation to a range of diet and health questions to illustrate these points. A brief overview of the role of sample collection, processing and storage is given including some generic recommendations for maximising the reliability of subsequent analytical data. Using the examples of phytoestrogens and iodine the question of whether biomarkers can accurately reflect the intake of the dietary constituents of interest at the population level or at the individual level is considered. The relationship of the biomarker to the natural history of the disease is exemplified using the role of folate in neural tube defects. Finally, intakes of vitamin D and heterocyclic amines are used to illustrate the integration of biomarkers into epidemiological studies of prostate and colorectal cancer, respectively. It is concluded that biomarkers may provide a more accurate and objective measure of diet than estimates of current or usual intake but that this approach also has limitations. A combination of methods will probably prove to be most valuable and this approach is being taken in current large prospective studies.

Health technology assessment in Austria.

The Austrian healthcare system relies mainly on physicians in private practice and on various services provided by hospitals. The social health insurance scheme is compulsory, covering 99% of the population. The system is very decentralized. While the federal state provides the framework, the nine autonomous provinces are responsible for administering health and social services. There is ongoing public discussion about centralizing the healthcare system to make it more efficient and to enforce structural reforms. Because of concerns about healthcare expenditures, in 1997 the Performance-Related Hospital Financing System (LKF), a system similar to the diagnosis-related group system, was introduced for hospitals, including a plan for large medical devices. It is too early to evaluate the success of this new system, although some effects of the LKF system that could have been anticipated, such as shortened lengths of stay and more hospitalizations, have been seen. Previously, health technologies have been almost uncontrolled in Austria. The evaluation of health technologies as an instrument to support or to control their dissemination and use or to help define policies is not institutionalized or systematically used. It seems clear that structural reforms of the Austrian healthcare system are needed. Health technology assessment should be part of such reforms.

Aflatoxin-albumin adduct formation after single and multiple doses of aflatoxin B(1) in rats treated with Thai medicinal plants.

The objective was to conduct an assessment of the ability of two Thai medicinal plants, Cymbopogon citratus Stapf and Murdannia loriformis, to modulate levels of serum aflatoxin-albumin (AF-albumin) adducts following aflatoxin B(1) (AFB(1)) exposure in rats. The influence of the plant extracts on AF-albumin adduct formation after a single exposure to 250 microg/kg body weight (bw) AFB(1) was measured over a 48-h period. Rats received M. loriformis extract (3 g/kg bw) or C. citratus Stapf extract (5 g/kg bw) daily for the week prior to the AFB(1) administration. In control rats, maximum adduct levels were observed 12 h post-AFB(1) treatment but in the animals receiving Murdannia extract, maximum levels occurred earlier, at 4 h post-treatment. No such effect was observed with the Cymbopogon extract. Daily treatment of rats with AFB(1) at 250 microg/kg bw for 3 weeks caused serum AF-albumin adduct levels to accumulate over a 10-14 day period and reach plateau levels 4.4-fold higher than observed after a single dose. Treatment with Murdannia extract for 1 week before and then throughout the AFB(1) exposure period resulted in a slight decrease in the AF-albumin adduct levels in the first week of the intervention. After that time, however, the reduction in adduct levels in the Murdannia extract group did not differ significantly from controls. No significant alteration in the biomarker levels was seen with the Cymbopogon extract treatments compared to control rats.

Prospective, controlled, double-blind study of i.v. tenoxicam for analgesia after thoracotomy.

We have shown that a single i.v. dose of tenoxicam 20 mg, after thoracotomy, when compared with placebo in 20 patients (with one exclusion), was associated with a reduction in consumption of papaveretum, assessed by patient-controlled analgesia, of 2.2 mg h-1 (22%) to 4 h and 1.4 mg h-1 (23%) to 12 h after operation (repeated measures analysis of variance: P less than 0.01). There was no reduction from 12 to 24 h. There was no significant difference between groups in pain scores or in side effects.

Lack of effects of selenium on N-nitrosomethylbenzylamine-induced tumorigenesis, DNA methylation, and oncogene expression in rats and mice.

The effects of dietary selenium deficiency and excess on N-nitrosomethylbenzylamine-(NMBA) induced esophageal neoplasia in rats and forestomach tumors in mice and the effects of dietary selenium on DNA adduct formation and on the activities of DNA adduct-repairing enzyme and oncogene expression in rat esophagus were investigated. The esophageal and forestomach tumors were induced by administration of NMBA by gavage with a total dose of 39 mg/kg body wt in rats and 12 mg/kg body wt in mice. Neither selenium dietary deficiency (Se < 0.02 ppm) nor selenium excess (2.0 ppm) showed any significant effect on the incidence of tumors or number of tumors per tumor-bearing animal. For the DNA adduct formation studies, rats were given a dose of NMBA intraperitoneally after six weeks on the different selenium-containing diets. No significant difference in the amount of the DNA adduct O6-methyldeoxyguanosine was found among the different selenium-treated groups. In a parallel group of rats that did not receive NMBA, the levels of esophageal O6-methyldeoxyguanosine DNA methyltransferase were not significantly altered by dietary selenium levels. The c-myc oncogene expression in rat esophagus was induced by the administration of NMBA (3 mg/kg body wt) by gavage once a week for eight weeks. Dietary selenium did not show any effects on its expression. On the basis of the results of these studies, dietary selenium has no effects in the NMBA-induced tumor model.

Liver cancer in Thailand. II. A case-control study of hepatocellular carcinoma.

Potential risk factors for hepatocellular carcinoma were investigated in a case-control study among inhabitants of north east Thailand. Sixty-five cases from 3 hospitals, with matched controls, were included. Infection with hepatitis-B virus was the major risk factor-chronic carriers of HB surface antigen had an estimated relative risk of 15.2. Infection with hepatitis-C virus appeared to be rare. No increase in risk was found with recent aflatoxin intake, as estimated by consumption of possibly contaminated foods, or by measuring aflatoxin-albumin adducts in serum. Regular use of alcohol (2 or more glasses of spirits per week) was associated with a non-significant elevation in risk (o.r. = 3.4, 95% c.i. 0.8-14.6), but the number of regular drinkers in the population was small. The meaning of an apparent protection conferred by certain food items is uncertain, but a possible role of betel nut in the aetiology deserves further investigation.

Evaluation of methods for quantitation of aflatoxin-albumin adducts and their application to human exposure assessment.

Aflatoxin (AF) albumin adducts are found in peripheral blood after exposure to aflatoxin B1 (AFB1) and the measurement of these adducts is potentially a useful tool in the epidemiological study of the role of AFB1 in the etiology of liver cancer. Three complementary approaches to the quantitation of AF-albumin adducts are described: (a) enzyme-linked immunosorbent assay (ELISA) performed directly on intact albumin (direct ELISA); (b) ELISA performed on an albumin hydrolysate (hydrolysis ELISA); (c) high-performance liquid chromatographic fluorescence detection of AF-lysine adduct after albumin hydrolysis and immunoaffinity purification. These techniques have been validated by direct comparison with rat albumin samples modified to a known extent. Detection limits of approximately 100, 5.0, and 5.0 pg AF/mg human albumin were determined for the three methods, respectively. Samples obtained from individuals from Thailand, The Gambia, Kenya, and France have been used to validate the measurement of AF-albumin adducts by these three methods. Levels of 7 to 338 pg AF/mg albumin were observed in the former two countries while no adducts were detected in samples from France. The relative properties of the three assays, with special regard to their application in epidemiological studies, are considered. A combination of the hydrolysis ELISA for large scale screening followed by confirmatory analyses in positive samples by high-performance liquid chromatographic fluorescence is suggested as an optimum methodology.