SELENIUM Present Status and Perspectives in Biology and Medicine G. N. Schrauzer Editor includes Proceedings of the International Symposium on Present Status and Perspectives of Selenium in Biology and Medicine May 2-3, 1987, Nonnweiler, West Germany Humana Press· Clifton, New Jersey ISBN-13: 978-0-89603-154-8 e-ISBN-13: 978-1-4612-4606-0 DOl: 10.1007/978-1-4612-4606-0 (for copyright page) Copyright © 1988 The Humana Press Inc. All Rights Reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the copyright owner. Preface The papers and reviews in the present volume are in part regular contributions submitted for publication in Biological Trace Element Research, and in part invited papers from participants of the International Symposium "Present Status and Perspectives of Sele nium in Biology and Medicine," held on May 2 and 3, 1987, at the Eu ropean Academy, Nonnweiler /Trier, FRG. The Symposium focused on ecological and biomedical aspects of selenium with emphasis on the manifestations of selenium deficiency in different countries, the distribution of selenium in human organs and body fliuds, the roles of selenium in health and disease, its anti carcinogenic action, effects of supplementation, and therapeutic applications. Organized jointly by the International Association of Bioinor ganic Scientists and BioSynopsia, a Nonprofit Subsidiary of the Tech nology Center, Stuttgart, the scientific board consisted of Professors W. Hartfiel, Bonn, FRG, G. N. Schrauzer, La Jolla, CA, USA, and H. Zumkley, Muenster, FRG. G. N. Schrauzer Editor CONTENTS v Editor's Preface 1 W. Hartfiel* and N. Bahners Selenium Deficiency in the Federal Republic of Germany 13 Af.L.Jackson Selenium: Geochemical Distribution and Association with Human Heart and Cancer Death Rates and Longevity in China and the United States 23 O. Oster, * G. Schmiedel, and W. Prellwitz The Organ Distribution of Selenium in German Adults 47 O. Oster, * G. Schmiedel, and W. Prellwitz Correlations of Blood Selenium with Hematological Parameters in West German Adults 83 Josef Kiem Selenium in Platelets 89 Yong Xian Wang and Josef Kiem* Effect of Selenium Supplementation on Platelet Selenium, Glutathione Peroxidase, and Aggregation 97 Alberto Afas, Jiuyu Jiang, and Bibudhendra Sarkar* Selenite Metabolism in Rat and Human Blood 111 P. Schramel, * S. Hasse, and J. Ovcar-Pavlu Selenium, Cadmium, Lead, and Mercury Concentrations in Human Breast Milk, in Placenta, Maternal Blood, and the Blood of the Newborn 125 J. Clausen and S. A. Nielsen Comparison of Whole Blood Selenium Values and Erythrocyte Glutathione Peroxidase Activities of Normal Individuals on Supplementation with Selenate, Selenite, L-Selenomethionine, and High Selenium Yeast 139 H. Zumkley Clinical Aspects of Selenium Metabolism 147 H. B. von Stockhausen Selenium in Total Parenteral Nutrition 157 H. Koehler, * H.-J. Peters, H. Pankau, and H.-J. Duck Selenium in Cardiology and Angiology 167 Brad M. Dworkin, * William S. Rosenthal, Gary P. Wormser, Lisa ·Weiss, Miguel Nunez, Carol Joline, and Anthony Herp Abnormalities of Blood Selenium and Glutathione Peroxidase Activity in Patients with Acquired Immunodeficiency Syndrome and Aids-Related Complex 179 Jorgen Clausen, * Gunde Egeskov Jensen, and Soren Achim Nielsen Selenium in Chronic Neurologic Diseases: Multiple Sclerosis and Batten's Disease 205 S. Chaitchik, * C. Shenberg, Y. Nir-EI, and M. Mantel The Distribution of Selenium in Human Blood Samples of Israeli Population Comparison between Normal and Breast Cancer Patients 213 S. M. Lin and M. H. Yang Arsenic, Selenium, and Zinc in Patients with Blackfoot Disease 223 Gerald Batist Selenium: Preclinical Studies of Anticancer Therapeutic Potential 231 S. Y. Yu, * Y. J. Chu, and W. G. Li Selenium Chemoprevention of Liver Cancer in Animals and Possible Human Applications 243 S. Y. Yu, * P. Ao, L. M. Wang, S. L. Huang, H. C. Chen, X. P. Lu, and Q. Y. Liu Biochemical and Cellular Aspects of the Anticancer Activity of Selenium 257 Dorthe Arenholt-Bindslev, * Borge Larsen, Mohamed Abdulla, and Arne Jepsen Antagonistic Effect of Selenite on Tumor Promoter Induced Cell Proliferation in Cultures of Rat Tongue Epithelium 267 J. Chmielnicka, * G. Zareba, M. Witasik, and E. Brzeznicka Zinc-Selenium Interaction in the Rat 277 O. Thorlacius-Ussing* and F. Taagehoj Jensen Selenium in the Anterior Pituitary of the Rat after a Single Injection of75Se Sodium Selenite 289 Momoko Chiba, * Nobuko Kamiya, and Masakazu Kikuchi Experimental Study on Interactions between Selenium and Tin in Mice 303 Author Index 305 Subject Index ©Copyright 1988 by the Humana Press Inc. All rights of any nature whatsoever reserved. 0163-4984/88/1501-0001$02.40 Selenium Deficiency in the Federal Republic of Germany W. HARTFIEL * AND N. BAHNERS Institute of Animal Nutrition, Department of Feedstock Research, University of Bonn, West Germany Received May 1, 1987; Accepted August 4, 1987 ABSTRACT A mean selenium of .123 mg/kg dry wt was observed in 195 samples of agricultural soils, and a mean of .158 mg Se/kg dry wt in 304 samples of grassland soils collected at 354 sites in various regions of the Federal Republic of Germany. For grassland soil, a north/south gradient of Se concentrations was observed. In the industrialized re gions of the North, higher Se levels were generally observed, the highest value of .652 mg Se/kg dry wt at a site in Northrhine Westphalia. The mean selenium content of grass from the respective collection sites was .045 mglkg dry wt in all regions of the FRG, a level insufficient for the maintenance of health of farm animals. The absence of a correlation between the soil- and grass-Se con tents indicates that Se uptake by plants is not solely dependent on the presence of Se. Grass may be deficient in Se even if grown on Se-rich soils. Fixation of Se by acidic soils appears to be a major factor; the high Se levels in the soils of industrialized areas is not bioavailable. Based on these findings, it is concluded that locally produced feedstock must be supplemented with Se to prevent the outbreak of deficiency diseases in farm animals. Index Entries: Selenium in grasses and soils; selenium in animal feedstock; selenium fixation by acidic soils; selenium deficiency in the Federal Republic of Germany. *Author to whom all correspondence and reprint requests should be addressed. Biological Trace Element Research 1 Vol. 15, 1988 2 Hartfiel and Bahnefs INTRODUCTION The present study was initiated after symptoms of selenium defi ciency were noted among cows in our care who had been feeding on pas tures in the vicinity of our Institute. Whereas cows maintained on com mercial feed in stables remained normal and healthy, a high incidence of placental retention and subsequent low fertility occurred in grass-fed ani mals. After the afflicted animals remained unresponsive to conventional corrective measures by local veterinarians, we suspected selenium deficiency as a possible cause, even though the territory of the Federal Republic of Germany (FRG) was still generally regarded to be "Se adequate." Because of this widely held view, selenium was not given high priority. The element in fact is not even mentioned in the most re cent official reports of the German Nutritional Society (1). However, our survey of the literature revealed that very little is actu ally known on the selenium status of the FRG. In virtually all discussions of the subject, reference is made to papers by Oelschlaeger and Menke (2,3), who in 1969 reported on the selenium contents of animal feedstock and foods. In feed such as straw, tubers, and roots collected in the vicin ity of Stuttgart, these authors reported Se concentrations of only .09 mg/kg dry wt, which are clearly in the deficiency range. High or ade quate selenium levels were only found in feed additives such as fish meal or raw mineral phosphate (2). In foods taken from local markets, mean Se contents ranged from .1 to .2 mg/kg dry wt (3). Although these amounts are sufficient to protect animals against acute selenium deficiency diseases, they are not neces sarily adequate to prevent chronic or latent deficiency conditions. It should also be noted that in this study the origin of the foods was not specified. As some could have been imported, it thus would be impossi ble to draw any firm conclusions. In view of this state of affairs, we thus decided to determine the selenium status in the FRG, focusing on the en tire human food chain. Our first studies dealt with the Se contents of ara ble soil, grassland soil, and grass. Since cereals and forage crops are im portant sources of selenium for animals and man, the selenium concentrations of soils and of grass, and knowledge of the factors that determine Se assimilation by plants, are of fundamental importance. In order to obtain reliable data on these subjects, special emphasis was placed on analytical methodology and collection of samples. Thus, apart from selecting the most accurate and sensitive method of analysis avail able, we also carefully avoided errors during sample collection. In par ticular, all samples were taken from precisely defined locations, and, to permit estimates of Se bioavailability, for each sample of grass, a corre sponding sample of soil from underneath was taken as well. In addition, in vitro fertilization experiments will be reported that were conducted to determine the factors that control Se assimilation. Biological Trace Element Research Vol. 15, 1988 Selenium Deficiency in the P.R. G. 3 MATERIALS AND METHODS Methods of Analysis Among the available methods of selenium analysis compiled by Gissel-Nielsen et aI. (4) (see Table 1), we selected atomic absorption spectrometry with hydride generation with sodium borohydride in view of its high sensitivity. The standard conditions for Se determination are given in Table 2. All samples were wet-ashed using a mixture of the concentrated acids (suprapure) of HN03, H2S04, and HCl04 in the ratios of 4:1:1, as recommended by Raptis et al. (5). Sample Digestion Air-dry samples, usually .2-1.0 g, were weighed into a digestion flask ("Duran 50") of 75 mL capacity to which 3-5 (HN03-prepurified) glass beads of 2 mm diameter were added. Subsequently, the mixture of acids (10 mL) was carefully added to the samples, and each digestion flask was placed on a heating block whose temperature was programmed to reach the temperature of 95-100° within 45 min, and to maintain this temperature for another 30 min. Thereafter, each sample was checked for residual organic matter. In most cases, the digestion solution was homo geneous and yellow; occasionally, however, careful manual agitation of the vessel was necessary to wash down particles that had migrated to upper parts of the digestion vessels, thus escaping digestion. In such cases, heating at 100° was continued until a homogeneous solution was Table 1 Methods of Selenium Analysis as Compiled by Gissel-Nielsen et al. (4) Approximate sensitivity, Method ng Neutron activation analysis, NAA 10 Fluorescence spectroscopy, FS 2-5 Atomic absorption spectrometry, AAS Flame 500a Flameless, graphite .5a Hydride: flame 2 quartz-cell .02a Inductive coupled plasma, ICP 50 Gas chromatography, GC 1 X-ray fluorescence spectrometry, XRFS 2500 XRFS after preconcentration 10 Differential pulse cathode stripping voltanetry, 5 DPCSV aApproximate sensitivity/mL of sample solution. Biological Trace Element Research Vol. 15, 1988 4 Hartfiel and Bahners Table 2 Standard Conditions for Selenium Determinations Employed Instrument Perkin Elmer 403 Adjustments Wavelength: 196.0 nm Slit width: 2.0 nm Light source Electrode-free discharge lamp Hydride generator MHS-20 Conditions Reducing agent: NaBH4 Cell temperature: 9000 Reaction time: 8 sec Reaction solution: 3% NaBH4 in 1% NaOH Volume of solution: 2.5 mL Carrier gas: Nitrogen Flushing time 1" 35 sec 21> 30 sec 'Prior measurement. After measurement. b obtained. The block temperature was subsequently raised to 1500 to com plete the digestion. After cooling, hexavalent selenium was reduced to the tetravalent state by the addition of 2 mL of suprapure cone. HCl. The digestion vessels were again heated to 1500 for 6 h to remove all re maining HN0 to prevent signal-depressing effects of this acid (6). After 3 the reduction, the clear digests were transferred into volumetric flasks of 25 mL capacity and filled to the mark by the addition of doubly distilled water. Prior to the assay, each sample was placed into two reaction ves sels. Into each vessel, 1 mL of suprapure HCl and 2-3 drops of antifoam ing agent were added. Standardization Calibration curves were obtained by subjecting the solutions of known Se content to the same digestion conditions as employed for the samples. Reproducibility, Accuracy, and Precision of Results Using rice flour with a Se content of .4 ± .1 mg Se/kg dry wt as a standard reference material in amounts from .3 to .8 g, with and without added additional known amounts of Se, the reproducibility, precision, and accuracy of the results was generally very good (see Table 3). During the three years of our study we also participated in international interlaboratory control studies, the results of which are summarized in Table 4. It may be seen that our results were generally higher than the reported means. We attribute this to the efficiency of our digestion proc ess and the avoidance of losses of Se during digestion. Biological Trace Element Research Vol. 15, 1988 en ~~ ~. o ~Q lb' ::::J -Q S· S-Ib :-rJ ;::0 P U1 e S mg 012 0tO 010 014 014 012 021 019 014 013 .......... d, ± ± ± ± ± ± ± ± ± ± ve 9 3 0 7 0 5 5 4 0 1 r 9908393473 Obse 5 .34 .36 .410 .35 .44 .44.55 .43 .45 .5 : n e S g n d ar g, 0 0 0 d n 482 n i 1 a h ble 3 ons with Rice Flour St Additions to Se Standard hing, ng Se post as 40 80 120 Taati as n r i o m i r r p e t e ium D d. Se nts of ples, Se 20 00 85 10 00 00 00 00 00 00 n cemg 1223222222 le alntam .......... e Cos S c Trial le , g mpht 3 5 7 8 5 5 5 5 5 5 g .......... ai Se w 1 . ± S T g k / e r S u og lm f e-4 c. i R OJ 6' l o· !!l ~ @ !!l ~ ~ @' lfl ~ 9- ~ :-.\n i