Purification and characterization of a corrinoid compound from Chlorella tablets as an algal health food

Vitamin B(12) content of an algal health food, Chlorella tablets (Chlorella sp.), was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor-chemiluminescence methods. The values of 200.9-211.6 microg/100 g dry weight determined by the chemiluminescence method were similar to the values (201.3-285.7 microg/100 g dry weight) determined by the microbiological method. A corrinoid compound was purified to homogeneity from the Chlorella tablets and characterized. The purified corrinoid compound was identified as vitamin B12, on the basis of silica gel 60 TLC, C18 reversed-phase HPLC, 1H NMR spectroscopy, and UV-Vis spectroscopy.     

Certification of B-group vitamins (B1, B2, B6, and B12) in four food reference materials

In 1989, the Community Bureau of Reference started a research program to improve the quality of vitamin analysis in food. To achieve this task, vitamin methodology was evaluated and tested by interlaboratory studies and the preparation of certified reference materials, which will be used for quality control of vitamin measurements. The main improvements in methodology were achieved by testing and standardizing the extraction condition and enzymatic hydrolysis procedures. Results for each individual material are derived from five replicate determinations using at least two independent methods: liquid chromatography (HPLC) and microbiological assay for vitamins B1, B2, and B6; and radioprotein binding and microbiological assays for vitamin B12. The certificate of analysis for four reference materials gives mass fraction values for water-soluble vitamins. These certified values were based on the acceptable statistical agreement of results from collaborating laboratories. Certified values with uncertainties (mg/kg dry matter) for each CRM are as follows: 4.63 (0.20) and 4.10 (0.51) for vitamins B1 and B6, respectively, in CRM 121 (wholemeal flour); 6.51 (0.24), 14.54 (0.3), 6.66 (0.43), and 0.034 (0.003) for vitamins B1, B2, B6, and B12, respectively, in CRM 421 (milk powder); 3.07 (0.17) and 4.80 (0.40) for vitamins B1 and B6, respectively, in CRM 485 (lyophilized mixed vegetables), and 8.58 (0.55), 106.8 (2.8), 19.3 1.5), and 1.12 (0.044) for vitamins B1. B2, B6, and B12, respectively, in CRM 487 (lyophilized pig liver).     

Broth from canned clams is suitable for use as an excellent source of free vitamin B12

Vitamin B(12) was assayed and characterized in the broth of canned clams (boiled plain). The broth contained considerable amounts of vitamin B(12) (2.7-14.1 μg/100 g, 1.3-6.7 μg/can). HPLC and LC/ESI-MS/MS chromatograms demonstrated that the clam broth contained true vitamin B(12). Gel filtration experiments indicated that most (72%) of the vitamin B(12) found in the broth was recovered in free vitamin B(12) fractions. These results indicate that the clam broth would be suitable for use an excellent source of free vitamin B(12) for elderly persons with food-bound vitamin B(12) malabsorption.     

Excretion of (14)C-fumonisin B(1), (14)C-hydrolyzed fumonisin B(1), and (14)C-fumonisin B(1)-fructose in rats.

14C-Fumonisin B(1) (FB(1)) was produced by Fusarium proliferatum M-5991 in modified Myro liquid medium and purified to >95% purity with a specific activity of 1.7 mCi/mmol. Nine male and nine female F344/N rats were each dosed by gavage with 0.69 micromol of (14)C-FB(1), (14)C-hydrolyzed FB(1), or (14)C-FB(1)-fructose/kg body weight. Urinary excretion of (14)C-FB(1) and (14)C-FB(1)-fructose was 0.5% and 4.4% of the total dose, respectively, and was similar between male and female rats. Urinary excretion of (14)C-hydrolyzed HFB(1) was significantly greater (P > 0.05) in female rats as compared with male rats (17.3% vs 12.8% of the total dose, respectively). There were no significant (P > 0.05) differences in biliary excretion of the three fumonisin compounds with a mean of 1. 4% of the dose excreted at 4 h after dosing. Lesser amounts continued to be excreted up to 9.25 h after dosing. Although biliary excretion of the (14)C-FB(1), (14)C-hydrolyzed FB(1), and (14)C-FB(1)-fructose was similar, increased urinary excretion of the (14)C-hydrolyzed FB(1) as compared to (14)C-FB(1) and (14)C-FB(1)-fructose indicated a greater absorption of the hydrolyzed form.     

Mass spectrometric determination of the predominant adrenergic protoalkaloids in bitter orange (Citrus aurantium)

The predominant adrenergic protoalkaloid found in the peel and fruit of bitter orange, Citrus aurantium, is synephrine. Synephrine is reputed to have thermogenic properties and is used as a dietary supplement to enhance energy and promote weight loss. However, there exists some concern that the consumption of dietary supplements containing synephrine or similar protoalkaloids may contribute to adverse cardiovascular events. This study developed and validated a positive-ion mode liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the quantitative determination of the major (synephrine) and minor (tyramine, N-methyltyramine, octopamine, and hordenine) adrenergic protoalkaloids in a suite of National Institute of Standards and Technology (NIST) bitter orange Standard Reference Materials (SRMs): SRM 3258 Bitter Orange Fruit, SRM 3259 Bitter Orange Extract, and SRM 3260 Bitter Orange Solid Oral Dosage Form. The limit of quantitation (LOQ) for all protoalkaloids is approximately 1 pg on-column, except for octopamine (20 pg on-column). Additionally, the method has a linear dynamic range of > or =3 orders of magnitude for all of the protoalkaloids. Individual, as well as "total", protoalkaloid levels (milligrams per kilogram) in the NIST SRMs were determined and compared to the levels measured by an independent liquid chromatography/fluorescence detection (LC/FD) method. Satisfactory concordance between the LC/MS/MS and LC/FD protoalkaloid measurements was demonstrated. LC/MS/MS analysis of the protoalkaloids in the SRMs resulted in mean measurement imprecision levels of < or =10% coefficient of variation (% CV).     

Total and inorganic arsenic in marketed food and associated health risks for the Catalan (Spain) population

Inorganic arsenic (iAs) is considered to be a human carcinogen. In this paper, total (As) and iAs contents of 215 food products and drinks (i.e., seafood, fruits and vegetables, meat products, oils and fats, rice and rice products, seasonings, and alcoholic drinks) marketed in Catalonia (Spain) were quantified by inductively coupled plasma-mass spectrometry. The analytical method described was used for different food products, obtaining feasible results without the need to couple LC-ICP-MS for iAs. Daily As and iAs intakes for the average adult Catalan consumer were estimated at 354 and 6.1 μg/day/person, respectively, using consumption data from the Catalan Nutrition Survey (ENCAT). The highest As content was found in seafood, contributing 96% of dietary As intake, whereas rice presented the highest iAs values, corresponding to 67% of dietary iAs intake. As cooking process may affect iAs content, boiled rice was evaluated, showing an iAs reduction (up to 86%) when using higher water volumes (30:1 water/rice ratio) than those used in previous studies. This iAs exposure was slightly below the exposure risk range stated by the European Food Safety Authority (0.3-8 μg/kg of body weight/day), although the possibility of a risk to the population with high rice consumption cannot be excluded.     

Natural occurrence of fumonisins B(1) and B(2) in domestic maize of Taiwan.

Samples of maize grown in various districts of Taiwan were collected and analyzed for the presence of fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) using high-performance liquid chromatography. Forty-nine (44.5%) and 2 (1.8%) of 110 samples were found to contain FB(1) (109-1148 ng/g) and FB(2) (222-255 ng/g), respectively. The frequency of detection and also the maximum FB(1) concentration were found in samples from Penton (2/2, 262 ng/g), followed by Chiayi (18/26, 264 ng/g), Tainan (8/16, 160 ng/g), Hualinen (5/14, 1148 ng/g), Taitung (7/20, 109 ng/g), and Yunlin (9/26, 361 ng/g). Of the 110 samples examined, only 2 samples from Hualinen had been detected containing FB(2). During an analysis of the distribution pattern of FB(1), it became apparent that >79% of tested samples had FB(1) concentrations <100 ng/g, whereas 2.7% (or 3 samples) contained FB(1) >300 ng/g. These results clearly illustrated that domestically produced maize for human consumption is frequently contaminated with FB(1).     

反相高效液相色谱法测定饲料级维生素B_（12）

本实验采用反相高效液相色谱法测定饲料级维生素Ｂ（１２）。该方法用Ｗａｔｅｒｓ紫外检测器，波长３６０μｍ，工作曲线相关系数０．９９９８，方法回收率９８．２１％，相对标准差０．３２。     

Jug r 4, a legumin group food allergen from walnut (Juglans regia Cv. Chandler)

Allergy to walnut is the most frequently reported tree nut allergy in the United States. Walnut 2S albumin, a vicilin-like protein, and a lipid transfer protein allergen have previously been described. Our objective was to clone and express a cDNA encoding a legumin group protein, assess IgE-binding with sera from walnut allergic patients, and investigate cross-reactivity with selected nuts. Primers were used to obtain the cDNA by 5' and 3' rapid amplification of cDNA ends from walnut mRNA. The cDNA was subcloned into the pMAL-c2X vector and the recombinant fusion protein, named rJug r 4, was expressed in Escherichia coli. The obtained cDNA encoded a precursor protein with a predicted molecular weight of 58.1 kD, which showed significant sequence homology to hazelnut and cashew legumin allergens. Serum IgE from 21 of 37 (57%) patients bound the rJug r 4 fusion protein. In vitro cross-reactivity was demonstrated with hazelnut, cashew, and peanut protein extracts.     

坡缕石对匙叶翼首草生长、产量和品质的影响

试验采用田间单因素随机区组设计,研究了加施破缕石0、750、1500、2250和3000 kg hm-2五个处理对匙叶翼首草产量及品质的影响。结果表明:缕石在750~3000 kg hm-2施用量下,均能提高匙叶翼首草的产量和品质,且产量随着坡缕石的增加而增加;加坡缕石为3000 kg hm-2时的产量最高较对照极显著增产26.56%(P<0.01),品质也最优;在匙叶翼首草栽培中科学合理的将N、P和K肥料与破缕石配施是提高其产量和品质的一个重要的农艺措施之一。     

Marsh woundwort, <Emphasis Type="Italic">Stachys palustris</Emphasis> L. (Lamiaceae): an overlooked food plant

The aim of this article is to study the geographical distribution and historical patterns of use of a little known root crop native to Europe—marsh woundwort Stachys palustris L. The species grows in wet grasslands and arable fields. It produces edible tubers. Both ethnographic literature and archival sources were searched. Seventeen reliable references concerning the consumption of S. palustris in southern and south–eastern Poland were found. The tubers were usually dried and powdered, and then added to soups or to bread dough. They were also eaten as raw snacks. The plant was used mainly during food shortages in spring, until the turn of the nineteenth and twentieth century, and later only as an occasional raw snack until the 1970s. Marsh woundwort was also eaten in western Ukraine (one reference). There are many references to the edibility of marsh woundwort in the eighteenth and nineteenth century European economic botany literature, particularly in Sweden and Great Britain. These publications tried to popularize the use of S. palustris as food. However, there is no firm evidence from these countries that marsh woundwort was used as food there. Marsh woundwort was also used, throughout northern and central Europe, as pig fodder and as a medicinal plant, particularly for healing wounds. Further studies on the nutritive value of this forgotten crop should be undertaken, particularly that there is little knowledge of the chemical composition of both marsh woundwort and its Asian relative Stachys affinis widely cultivated in China as a vegetable.     

Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate

Impatiens noli-tangere is scarce in the UK and probably only native to the Lake District and Wales. It is the sole food plant for the endangered moth Eustroma reticulatum. Significant annual fluctuations in the size of I. noli-tangere populations endanger the continued presence of E. reticulatum in the UK. In this study, variation in population size was monitored across native populations of I. noli-tangere in the English Lake District and Wales. In 1998, there was a crash in the population size of all metapopulations in the Lake District but not of those found in Wales. A molecular survey of the genetic affinities of samples in 1999 from both regions and a reference population from Switzerland was performed using AFLP and ISSR analyses. The consensus UPGMA dendrogram and a PCO scatter plot revealed clear differentiation between the populations of I. noli-tangere in Wales and those in the Lake District. Most of the genetic variation in the UK (H_T=0.064) was partitioned between (G_ST=0.455) rather than within (H_S=0.034) regions, inferring little gene flow occurs between regions. There was similar bias towards differentiation between metapopulations in Wales, again consistent with low levels of interpopulation gene flow. This contrasts with far lower levels of differentiation in the Lake District which suggests modest rates of gene flow may occur between populations. It is concluded that in the event of local extinction of sites or populations, reintroductions should be restricted to samples collected from the same region. We then surveyed climatic variables to identify those most likely to cause local extinctions. Climatic correlates of population size were sought from two Lake District metapopulations situated close to a meteorological station. A combination of three climatic variables common to both sites explained 81-84% of the variation in plant number between 1990 and 2001. Projected trends for these climatic variables were used in a Monte Carlo simulation which suggested an increased risk of I. noli-tangere population crashes by 2050 at Coniston Water, but not at Derwentwater. Implications of these findings for practical conservation strategies are explored.     

Development of a simple method for the determination of genistein, daidzein, biochanin A, and formononetin (biochanin B) in human urine.

A simple method was developed for the determination of free and/or total isoflavones daidzein, genistein, and their respective 4'-methoxy derivatives biochanin A and formononetin (biochanin B) at low levels in human urine. A solid-phase extraction on octadecyl silica (C(18)) columns was used for the isolation of the phytoestrogens from the matrix. An extraction on a ChemElut 1010 column connected on-line to a Florisil cartridge by a Teflon stopcock was used for effective eluate purification. A mixture of dichloromethane and ethyl acetate was used for elution of the isoflavones from the columns in tandem. The isoflavones were determined as trimethylsilyl (TMS) ethers using GC/MS-SIM after separation on an HP-5MS fused silica column. TMS ethers were obtained by using BSTFA containing 1% of TMCS. For the determination of free isoflavones 6-hydroxyflavone was used as internal standard, whereas robigenin was used in the case of total isoflavone determination. Recoveries for free isoflavones under study varied from 63.5 to 89.6% at the 25 ng mL(-)(1) level and from 63.5 to 89. 2% at the 5 ng mL(-)(1) level in urine. Analytical curves were linear between 5 and 25 ng mL(-)(1). Detection limits varied from 1 ng mL(-)(1) for formononetin to 2.3 ng mL(-)(1) for daidzein. Recoveries for total isoflavone determination after enzymatic hydrolysis with glucuronidase from Helix pomatia ranged from 56.5 to 77.1% at the 25 ng mL(-1) level.     

松材线虫及其近缘种热激蛋白90基因(hsp90)克隆与序列分析

热激蛋白90家族(heat shock protein 90(HSP)family)是一种进化保守的蛋白质,在分子伴侣功能、细胞循环调控、抗原传递方面发挥着重要作用。本研究用RT-PCR和RACE技术,克隆得到了松材线虫(Bursaphelenchus xylophilus)、拟松材线虫(Bursaphelenchus mucronatus)和豆伞滑刃线虫(Bursaphelenchus doui)热激蛋白90基因的cDNA全长序列,分别命名为Bx-hsp90、Bm-hsp90和Bd-hsp90,(GenBank登录号分别为:GU013561、HMO34943和HM347331),其cDNA全长分别为2255、2193和2232bp,开放阅读框均为2127bp,编码708个氨基酸,5'端非编码区的长度分别为33、13和13bp,3'端非编码区的长度分别为95、53和92bp。其DNA全长分别为2440、2388和2407bp,包含3个内含子。三者氨基酸序列的一致性为98.78%。进化分析表明,松材线虫、拟松材线虫、豆伞滑刃线虫与秀丽小杆线虫(Caenorhabditis elegans)的亲缘关系较近,推测三...     

Evaluation of health, nutrition and food security programmes in a complex emergency: the case of Congo as an example of a chronic post-conflict situation

OBJECTIVE: To describe the case of Congo as an example of the assessment and appropriateness of donor operational and sectoral strategies in a complex emergency. DESIGN AND SETTING: The paper reports the findings of an external evaluation of operations financed by the European Commission Humanitarian Office in the Democratic Republic of Congo (DRC). RESULTS: The Congolese health system is suffering from severe deterioration. What is functioning in the public health context is donor-dependent with high costs and limited coverage. Despite a relatively favourable agro-climatic situation, food shortage and famine severely affect the nutritional status of large population groups. In this context, humanitarian programmes have generally improved access to health care and the nutritional status of beneficiaries. The reduction of malnutrition in project areas is often demonstrated even though the context did not permit consolidation of these results. Malnutrition continues to claim a massive cost of lives owing to the effect of widespread food insecurity that follows a circular cause-and-effect pattern of very low food production and extreme poverty. CONCLUSIONS: The current context in DRC does not correspond yet to 'post-crisis': neither at population level with regard to indicators of poverty, malnutrition, disease and death, nor at institutional level, with regard to state support to institutions. In these situations, the international community is often called upon to replace the state as service provider. Integrated humanitarian actions should be the future of relief projects in DRC. Health, nutrition and food security components should be considered a standard public health intervention strategy representing the most sensible approach to address the needs of the affected population.