records were obtained from each participant.. Food records wereanalyzed by using NUTRITIONIST IV software (version 2.011993; N-Squared Computing,Salem,OR). Vitamin and mineralsupplement use was documented and evaluated as appropriate.
Clinical and biochemical measures
Fasting,peripheral venous blood samples were collected inthe morning between 0700 and 0900 by venipuncture. Completeblood counts,a chemistry panel,a serum immunoglobulin analy-sis,and a complement fraction analysis were performed by theLoma Linda University Medical Center Clinical Laboratoryaccording to standardized procedures.Serum ferritin was analyzed with an enzyme immunoassay kit(Milenia NKFE1; Diagnostic Products Corporation,Los Angeles)by using a microplate reader (model 2380; Bio-Tek,Winooski,VT). Serum folic acid and vitamin B-12 concentrations were deter-mined by simultaneous radioassays (Quantaphase-II,1911040;Bio-Rad Laboratories,Richmond,CA) using a gamma counter(model LB1213; EGNG Berthold,Wildbad,Germany). Traceelement–free tubes (Becton Dickinson,Rutherfold,NJ) wereused to collect blood for plasma zinc analysis by atomic absorp-tion spectrophotometry (model AA-475; Varian,Sunneyvale,CA)(13). Standard reference material (bovine serum standard ref-erence material no. 1598,National Institute of Standards andTechnology,Gaithersburg,MD) was used to check the accuracyand precision of the determinations. Serum methylmalonic acid,2-methylcitrate homocysteine,and cystathionine were measuredby gas chromatography–mass spectrometry at Metabolite Labo-ratories,Inc,at the University of Colorado Health Sciences Cen-ter,Denver (14,15).
Mitogen assay and natural killer cell activity
Lymphocytes for blastogenic response tests and killer cellassays were isolated from blood with heparin by using Ficoll-Paque (Pharmacia Fine Chemicals,Piscataway,NJ) and resus-pended in RPMI-1640 culture medium (Gibco,Grand Island,NY). Lymphocyte proliferation was measured by [
H]thymidineincorporation after stimulation with phytohemagglutinin,con-canavalin A and pokeweed mitogens (16). The stimulation index(SI) was calculated as follows:SI=[cpm (mitogen stimu-lated)/cpm (control)].Natural killer cytolytic activity was determined in peripheralblood mononuclear cells by using K562 target cells in a
Crrelease assay (17). The percentage of
Cr release or percentagelysis at multiple effector-to-target ratios was determined by usingthe following equation:[(sample
100. Cytotoxicity was expressed aslytic units (LU) and these were defined as the number of cellsrequired to cause 20% target cell lysis calculated by an exponen-tially fit equation and expressed as LU/10
peripheral bloodmononuclear leukocytes.
Statistical analyses were done by using SPSS for WINDOWS(Statistical Package for the Social Sciences,version 6.0 1996;SPSS,Inc,Chicago). Group means and SDs were calculated.Independent-sample
tests were conducted to evaluate differ-ences between the vegan and nonvegetarian groups. Multipleregression was used to evaluate the influence of diet,age,orbody mass index (BMI; in kg/m
) on selected immune measures.
Twenty-five vegans (10 men,15 women) and 20 nonvegetari-ans (10 men,10 women) who met the eligibility criteria wereincluded in the study. Subject characteristics are summarized in
. Vegans were defined as those who excluded meat,fish,poultry,dairy products,and eggs from their diets whereas non-vegetarians regularly included all food categories. There were nosignificant differences between the 2 groups in age,physicalactivity level,or blood lipid concentrations. The vegan group,however,had a significantly lower BMI than the nonvegetariangroup. The average number of years of vegan diet was 4.2 witha range of 1–25 y. Most of the vegans had followed a vegetariandiet before becoming vegans. The average number of years of following vegetarian dietary practices was 12.1 y with a range of 1–37 y. Of the 20 nonvegetarians,7 regularly used multivitamin-mineral supplements,compared with 4 of 25 in the vegan group.One nonvegetarian and 6 vegans took single-nutrient supple-ments of iron,calcium,or vitamin C. None of the nonvegetarianstook a separate vitamin B-12 supplement,whereas 9 of the 25vegans reported that they did so.
Dietary pattern and nutrient intake
The food pattern of the vegans compared with that of the non-vegetarians is shown in
. Vegans consumed no ﬂesh foodsand practically no dairy foods or eggs. Vegans,however,consumedmore servings per day of grains and breads,vegetables,fruit,legumes,and nuts and seeds. Only vegans consumed soymilk,tofu,and meat analogs. Meat analogs are commercially available foodsprepared from soy protein,wheat gluten,legumes,and other ingre-dients and designed to substitute for meat in the diet.Results from 4 d of food records were averaged for each per-son and used for group comparisons as shown in
. Theintake of female vegans compared with that of female nonvege-tarians was significantly lower in protein,total fat,saturated fat,and monounsaturated fat both in quantity and as a percentage of energy. Female vegans consumed more dietary fiber and lessNUTRITIONAL STATUS OF VEGANS AND NONVEGETARIANS587S
Characteristics of vegan and nonvegetarian subjectsNonvegetariansVegans(
941Blood lipids (mmol/L)Total cholesterol4.80
0.40Duration of vegetarian diet (y)012.1 (1–25)
Duration of vegan diet (y)04.2 (1–37)Multivitamin-mineral supplement users74Single-nutrient (calcium,iron,or 16vitamin C) supplement usersVitamin B-12 supplement users09
Signiﬁcantly different from nonvegetarians,
< 0.001 (
–; range in parentheses.
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