A study of relationships between bone-related vitamins and minerals, related risk markers, and subsequent mortality in older British people: the National Diet and Nutrition Survey of People Aged 65 Years and Over.

SUMMARY: Follow-up of a British national survey of older people found that in men, all-cause mortality was predicted by baseline plasma concentrations of phosphorus, albumin, creatinine and α1-antichymotrypsin, and food energy intake and in women by plasma alkaline phosphatase, creatinine, α1-antichymotrypsin, 25-hydroxy-vitamin D (marginally), and phosphorus intake. INTRODUCTION: Predictive power, for all-cause mortality, of bone-related vitamin and mineral indices and intakes, measured at baseline (primary objective), was studied in the British National Diet and Nutrition Survey (community-living subset) of People Aged 65 Years and Over. A secondary objective was to identify cross-sectional relationships between indices at baseline to help explain mortality predictions. METHODS: Mortality status was recorded for 1,054 (mean age 76.6 ± 7.4 years, 49.0% female) participants from baseline survey in 1994/1995 until September 2008. Seventy-four per cent of male and 62% of female participants died. Cox proportional hazards models were used to relate baseline nutrient and risk marker estimates to subsequent survival. Results below 1.0 signified lower risk at greater nutrient (status or intake) values and vice versa. RESULTS: In both sexes, all-cause mortality was significantly predicted by body weight and mid-upper arm circumference. In men, it was predicted by baseline plasma concentrations (per SD) of: phosphorus (hazard ratio 1.18, 95% confidence interval (CI) = 1.06-1.30), albumin (hazard ratio 0.84, 95% CI = 0.74-0.94), creatinine (hazard ratio 1.20, 95% CI = 1.08-1.33) and α(1)-antichymotrypsin (hazard ratio 1.21, 95% CI = 1.11-1.33). In women, it was predicted by plasma albumin (hazard ratio 0.83, 95% CI = 0.72-0.96), alkaline phosphatase (hazard ratio 1.08, 95% CI = 1.01-1.16), creatinine (hazard ratio 1.37, 95% CI = 1.13-1.66), α(1)-antichymotrypsin (hazard ratio 1.27, 95% CI = 1.11-1.45) and marginally by 25-hydroxy-vitamin D (hazard ratio 0.87, 95% CI = 0.75-1.00). In men, it was predicted by dietary intake (per SD) of food energy; in women, by intake of phosphorus. Adjustment for plasma α(1)-antichymotrypsin or plasma creatinine reduced the significance of plasma phosphorus in men. CONCLUSION: Mortality prediction by higher plasma phosphorus in older British men may imply impaired renal function and/or acute phase status. Further studies are needed on which associations are causal and modifiable.

5-Diazo-4-oxo-L-norvaline: reactive asparagine analog with biological specificity.

The L-asparagine analog, 5-diazo-4-oxo-L-norvaline, specifically inactivates L-asparaginase and inhibits the growth of L-asparagine-dependent or L-asparaginase-sensitive tumor cells in culture. With 5-(14)C-labeled compound, a biphasic incorporation into sensitive cells occurs, but the inhibition of cell multiplication is manifest much later than the rapid phase of incorporation of the analog.

Patients' and sleeping partners' experience of treatment for sleep-related breathing disorders with a mandibular repositioning splint.

AIM: To determine in detail the complications associated with the use of mandibular repositioning splints (MRS) to treat sleep-related breathing disorders. METHOD: This prospective cross-sectional cohort study audits the management with mandibular repositioning splints of 121 patients suffering from sleep-related breathing disorders. Investigation of patients' and sleeping partners' perspectives on treatment was undertaken with the use of a questionnaire based study. RESULTS: Sixty-eight per cent of respondents reported that they were compliant with treatment; various side effects were reported of which excess salivation was the most common. Investigation of sleeping partners' perspectives revealed that 70% felt that their partners' snoring was improved and 47% felt that their partner's breathing pauses during sleep were reduced. Sixty-four per cent of the sleeping partners also reported that their own sleep pattern had improved since their partner's treatment. CONCLUSION: Mandibular repositioning splints used in the manner described by this paper are demonstrated to have a good compliance rate, provide successful treatment and exhibit only minor, reversible side effects.

The effect of hypoxia on collagen synthesis in cultured 3T6 fibroblasts and its relationship to the mode of action of ascorbate.

When exposed to low oxygen tension, in the absence of added ascorbic acid 3T6 mouse fibroblast cultures in late log phase respond by increased lactate production and increased hydroxylation of proline in nascent collagen, which is paralleled by an increase in prolyl hydroxylase activity. After 6 h recovery from the anoxic stimulus, however, cultures still yield more prolyl hydroxylase than controls, but the effect on hydroxylation of nascent collagen has disappeared. These observations help to dissect the dual role of ascorbate which can stimulate hydroxylation both by increasing the amount of active enzyme and by a cofactor-like role; in addition, these observations may be relevant to wound healing.

Breast milk as a source of vitamins, essential minerals and trace elements.

Human breast milk provides all of the vitamins and essential minerals and trace elements (micronutrients) that are required by the normal term infant, until weaning. With a few exceptions, excessive micronutrient supplies to the mother, or a moderate deficiency in her diet, do not greatly alter the supply to the infant. Thus, the infant is well-protected by maternal homeostatic processes, although the mechanisms of these are not yet well understood. Considerable progressive changes in concentration occur for some of the micronutrients during the course of lactation. Because the concentration of these nutrients, and of other substances that modify their absorption by the infant, such as binding proteins, differs considerably between human milk, animal milk and, hence, commercial milk formulae, there is great interest in the quantitative significance of micronutrient supplies, and their variability in breast milk, in the quest for improvement of commercial formulations. The aim of this review is to summarize the available information about the factors that determine breast milk contents of micronutrients.

Prevalences of hyperhomocysteinemia, unfavorable cholesterol profile and hypertension in European populations.

BACKGROUND: Hyperhomocysteinemia (HHCY) is a risk factor for cardiovascular diseases (CVD). HHCY may interact with hypertension (HTEN) and an unfavorable cholesterol profile (UNFAVCHOL) to alter the risk of CVD. OBJECTIVES: To estimate the prevalences of HHCY (1) isolated and (2) in combination with UNFAVCHOL and/or HTEN in different age categories. To provide information that may improve the screening and treatment of subjects at risk of CVD. DESIGN: Cross-sectional data on 12,541 men and 12,948 women aged 20 + y were used from nine European studies. RESULTS: The prevalence of isolated HHCY was 8.5% in subjects aged 20-40 y, 4.7% in subjects aged 40-60 y and 5.9% in subjects aged over 60 y. When combining all age groups, 5.3% had isolated HHCY and an additional 5.6% had HHCY in combination with HTEN and/or UNFAVCHOL. The combinations of risk factors increased with age and, except for HHCY&UNFAVCHOL, were more prevalent than predicted by chance. Of the young subjects (20-40 y), 24% suffered from one or more of the investigated CVD risk factors. This figure was 75.1% in the old subjects (60+ years). CONCLUSIONS: A substantial number of subjects in selected European populations have HHCY (10.9%). In half of these cases, subjects suffer also from other CVD risk factors like UNFAVCHOL and HTEN. Older people in particular tend to have more than one risk factor. Healthcare professionals should be aware of this when screening and treating older people not only for the conventional CVD risk factors like UNFAVCHOL and HTEN but also HHCY, as this can easily be reduced through increased intake of folic acid via supplement or foods fortified with folic acid.

The relationship between severity of obstructive sleep apnoea/hypopnoea syndrome (OSAHS) and lateral cephalometric radiograph values: a clinical diagnostic tool.

BACKGROUND: This prospective cross-sectional cohort study examined the relationship between radiographic anatomy and the severity of OSAHS. The severity of OSAHS can be measured subjectively in terms of the Epworth scale and objectively in terms of the apnoea/hypopnoea Index (AHI). METHODS: 121 lateral cephalometric radiographs were traced under uniform conditions and a series of 56 landmarks identified, from which 48 angular and linear measurements were made. Significant changes occurred when comparison of these measurements with the severity of OSAHS were made. RESULTS: Body mass index (BMI), the maxillary-mandibular planes angle (MMPA) and the linear measurement between points 7 and 12, (the pharyngeal dimension measured from the tip of the soft palate to the corresponding horizontal point on the posterior pharynx), increased significantly with increasing severity of OSAHS as measured by the Epworth score. Overjet, lower lip length, and the distance from the hyoid bone to a point B on the mandible all increased significantly with increasing severity of OSAHS, as measured by the AHI. The hyoid bone was found to rotate counter clockwise as the severity of OSAHS increased, as a result the distance between the most anterior superior point on the hyoid bone and the maxillary plane was seen to decrease as severity of OSAHS increased in terms of AHI. CONCLUSIONS: Some radiographic anatomical features show significant change as the severity of OSAHS increases and these features could be used in the identification of patients who have severe OSAHS. The Logit equation derived from the findings of this study may also be a useful clinical tool in predicting the likelihood of a subject suffering from severe OSAHS.

Nutritional efficacy of a fortified weaning rusk in a rural area near Beijing.

A micronutrient-fortified rusk for weanling children was tested in a rural area near Beijing. Two hundred twenty-six children aged 6-13 mo were randomly assigned by village to either a micronutrient-fortified or an unfortified rusk, daily for 3 mo. The fortified rusk contained extra zinc; iron; calcium; vitamins A, D, and B-12; thiamin, riboflavin, niacin, and folic acid. Infants receiving the fortified rusk exhibited no decline in hemoglobin concentrations during the study whereas those receiving the unfortified rusk exhibited a significant decline. Improvements were also seen in erythrocyte porphyrin, plasma vitamin A, and riboflavin status, but these improvements were seen in both of the groups. Micronutrient fortification was probably beneficial for iron status and the fortified rusk promises to be an effective vehicle for supplementation.

PIP: In February-June 1990, in China, researchers assigned, by village, 226 6-13 month old, full-term, healthy infants from 33 villages of the Mi-yun rural area near Beijing to receive daily either a micronutrient-fortified or an unfortified rusk for 3 months. 15% of all infants were initially anemic, but not severely so. Extra zinc; iron; calcium; vitamins A, D, and B-12; thiamin; riboflavin; niacin; and folic acid fortified the rusk. The study aimed to determine the efficacy of the micronutrient fortification. Mean hemoglobin levels decreased considerably in the infants in the unfortified rusk group (12.85 g/L vs. 12.95 g/L; p .01), but remained the same for the fortified rusk group. The change in the unfortified rusk group's hemoglobin levels was much greater than that of the fortified rusk group (p .01). The significance of the supplement-group effect fell when the researchers included initial ferritin and free erythrocyte porphyrin concentrations (p = .04-.08). There was a considerable reduction in free erythrocyte porphyrin in both groups of infants (p .001) and the response was basically the same for both groups. Even though the fortified rusk group experienced a significant increase in plasma vitamin A (.093 mcmol/L; p .01), the difference in response between the 2 groups was not significant. Infants in the fortified rusk group experienced a considerable fall in vitamin E levels (p .001) which was a significantly greater decline than that observed in the unfortified rusk group (2.6 mcmol/L vs. .79 mcmol/L; p = .012). The erythrocyte glutathione reductase index of riboflavin status improved significantly in the fortified rusk group (.07; p = .05), but it was not significantly different from that of the unfortified group. These results suggested that fortifying a commercial weaning risk with micronutrients benefited the iron status of these children.

Biochemical evidence of thiamine depletion during the Cuban neuropathy epidemic, 1992-1993.

During an epidemic outbreak of neuropathy in Cuba during 1992-1993, blood and urine samples were collected from 107 persons with confirmed neuropathy, from 106 control subjects without clinical abnormality who were broadly matched with the affected persons by age and domicile, and from 537 unmatched subjects, also free from clinical abnormality. The unmatched subjects lived in two locations in Cuba; at each location they were drawn from two age ranges: 11-15-y-old secondary school students and 16-64-y-old adults. Measurements of urinary thiamine and blood transketolase and its activation with thiamine pyrophosphate were made. For the neuropathy subjects, these measurements were repeated after 3 wk of rehabilitation. All groups showed biochemical evidence of thiamine depletion affecting 30-70% of their members, which is a high prevalence. Severity of biochemical depletion was, however, no greater in the neuropathy subjects than in the control subjects (P > 0.05). However, it was greater in Pinar del Rio, where the incidence of disease was higher, than in the city of Havana, where less disease was seen. Although the majority of the affected subjects responded biochemically to a daily oral multivitamin supplement containing thiamine (P < 0.001), in some cases normal biochemical status was not achieved even after 3 wk of intensive treatment. In the affected group, thiamine status was inversely correlated with the amount of alcohol consumed (P = 0.007). Thiamine status at the outset was correlated with clinical outcome after treatment. Although neither thiamine depletion nor alcohol abuse were likely to have been the sole cause of the neuropathy epidemic, they may have been contributory factors. Thiamine supplementation or food fortification may therefore be necessary in Cuba.

Riboflavin status of adolescent vs elderly Gambian subjects before and during supplementation.

Riboflavin status was measured by activation coefficient of erythrocyte glutathione reductase (EGRAC) in elderly and adolescent rural Gambians whose intakes were low. Fifty-one adolescents and 52 elderly subjects were each subdivided into six supplementation groups to receive 0.25, 0.5, 0.75, 1.0, 1.5, or 2.5 mg riboflavin/d on 5 consecutive days weekly for 5 wk. Before supplementation, EGRAC values were grossly abnormal and differed (p less than 0.005) between the two female groups [elderly subjects, 1.94 +/- 0.33 (means +/- SD); adolescents. 1.72 +/- 0.27] but not between the two male groups. With supplementation greater than 0.25 mg/d the values became nearly equal in the two age groups. Only at total intakes between 1.8 and 2.5 mg riboflavin/d did the subjects attain acceptable status, with a mean EGRAC of 1.3-1.4. Clearly there is a need for further study of the discrepancy between observed index values during supplementation and the accepted ranges of normality at intakes close to the recommended amounts.

Riboflavin status in Gambian pregnant and lactating women and its implications for Recommended Dietary Allowances.

Riboflavin status by the erythrocyte glutathione reductase (NAD(P)H2:glutathione oxidoreductase EC1.6.4.2) test, and dietary riboflavin intake, were measured in 156 pregnant and lactating women in Keneba, a rural Gambian village, and in 59 pregnant and lactating women in Cambridge, England. The Keneba women were studied longitudinally, for up to 19 months. In Keneba, where the mean daily riboflavin intake was 0.5 mg, the mean erythrocyte glutathione reductase activation coefficient was 1.78 and there was a marked deterioration of biochemical status near parturition. In Cambridge, where the mean intake was 2.3 mg/day, the mean activation coefficient was 1.19. A vitamin fortified diet supplement, given to lactating women in Keneba for 8 months, raised their mean riboflavin intake for 0.5 to 1.5 mg/day and reduced the mean activation coefficient to 1.42. It is therefore likely that an intake even greater than the current United Kingdom or United States recommended daily amounts would be needed to achieve biochemical 'normality' in these women.

PIP: Riboflavin status by the erythrocyte glutathione reductase (NAD(P)H2 glutathione oxidoreductase ECI.6.4.2) test, and dietary riboflavin intake, were measured in 156 pregnant and lactating women in Keneba, a rural Gambian village, and in 59 pregnant and lactating women in Cambridge, England. The Keneba women were studied longitudinally, for up to 19 months. In Keneba, where the mean daily riboflavin intake was 0.5 mg, the mean erythrocyte glutathione reductase activation coefficient was 1.78 and there was a marked deterioration of biochemical status near parturition. In Cambridge, where the mean intake was 2.3 mg/day, the mean activation coefficient was 1.19. A vitamin fortified diet supplement, given to lactating women in Keneba for 8 months, raised their mean riboflavin intake for 0.5 to 1.5 mg/day and reduced the mean activation coefficient to 1.42. It is therefore likely that an intake even greater than the current United Kingdom or U.S. recommended daily amounts would be needed to achieve biochemical 'normality' in these women.

Riboflavin requirements of lactating Gambian women: a controlled supplementation trial.

As part of a study to determine the minimum allowance of riboflavin which is adequate for lactating women in a rural African environment, 60 subjects living in two Gambian villages were given either 2 mg riboflavin or a placebo daily on a double-blind basis for 12 wk. Their riboflavin intake from dietary sources was about 0.5 mg/day. In the supplemented group, the mean activation coefficient (AC) of erythrocyte glutathione reductase fell from 1.62 to 1.19 within 3 wk, and 90% had mean AC's below 1.3 throughout supplementation, whereas the placebo group maintained mean AC's between 1.6 and 1.9. Clinical signs associated with riboflavin deficiency improved more rapidly in the supplemented group; their breast milk riboflavin levels increased, and their infants' AC's were reduced, compared with those of the placebo group. After withdrawal of the supplement, the maternal and infants' AC's rose toward those of the placebo group. Thus a total riboflavin intake of about 2.5 mg/day during lactation is sufficient to maintain normal biochemical status in most Gambian women.

Does vitamin C reduce blood pressure? Results of a large study of people aged 65 or older.

OBJECTIVE: To characterize relationships among blood pressure, pulse rate, vitamin C status and other protective and risk factors for older British people, from a national survey. DESIGN: A cross-sectional analysis of survey data. SETTING: A population study, representative of mainland Britain. SUBJECTS: Among 914 people of both sexes living in the community, 373 were taking blood-pressure-lowering drugs and were therefore excluded from the analyses. INTERVENTIONS: Completion of an interview on health, lifestyle and dietary habits, recording of a 4-day dietary record, anthropometry and taking of a blood sample to determine haematological and biochemical status. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressures, pulse rate, indices of micronutrient status including plasma ascorbate concentration, nutrient intake and haematology. RESULTS: Plasma ascorbate concentration was inversely correlated to systolic and diastolic blood pressures and pulse rate. Other covariates of blood pressure included age, sex, domicile, plasma retinol, fibrinogen and gamma-tocopherol concentrations, erythrocyte count, prothrombin time and urine sodium: creatinine ratio. Covariates of pulse rate included sex, domicile, plasma fibrinogen and platelet count. Blood pressure was also correlated to intake of vitamin C. CONCLUSIONS: Plasma ascorbate concentration and intake of vitamin C are covariates of blood pressure in older people living in Britain. New intervention studies are now needed, to test for possible causalities.

Documentation of blood culture results.

AIMS: To evaluate the adequacy of documentation of blood culture results in patients' medical notes. METHODS: A pro-forma was completed following review of medical notes at 24 and 48 hours after a blood culture had been reported as positive. The study was performed on blood cultures received at the Department of Microbiology, Royal Hallamshire Hospital, Sheffield, from two local hospitals. Two periods were studied: (A) May to June 1993 and (B) September to October 1993. RESULTS: There were 43 results studied in period A and 79 in period B, giving a total of 122 results studied. Overall, 72 (59%) of 122 results were recorded in the medical notes at 24 hours. Of those results deemed highly significant, 40 (63%) of 63 were recorded. There was no significant difference in the documentation of results if the result was given personally or via the telephone. Nor was there any difference in documentation between different medical grades. Throughout the study there were six inaccurate records. The cumulative documentation over 48 hours of positive results was 54 (86%) of 63 of highly significant, 27 (69%) of 39 of uncertain significance, and 11 (55%) of 20 probable contaminant results. CONCLUSIONS: Documentation of blood culture results is currently suboptimal.

Measurement of ascorbic acid kinetics in man using stable isotopes and gas chromatography/mass spectrometry.

The feasibility of studying ascorbic acid kinetics in man using stable isotope-labelled tracers and gas chromatographic (GC) separation followed by mass spectrometric (MS) quantitation was assessed. Preliminary studies with 13C-labelled material showed that although better precision at low levels could be achieved using the GC/combustion/MS technique, consideration of likely enrichments in a human study made the simpler GC/MS method just as suitable. On this basis, a small pilot study of the kinetics in man was carried out. The enrichment of the ascorbic acid in plasma was measured for a 24 h period after oral administration of 13C-labelled material. The results were fitted to a simple three-compartment model and rate constants and pool sizes were deduced. The results obtained are comparable to those obtained in other published studies, from which we conclude that the technique may be useful as a non-invasive method for the assessment of nutritional status in a variety of populations.

Endothelial cell adhesion molecules in healthy adults during acute hyperhomocysteinemia and mild hypertriglyceridemia.

OBJECTIVE: Hyperhomocysteinemia and hypertriglyceridemia are independently associated with atherosclerotic disease. The process of atherogenesis involves inflammation and endothelial dysfunction. We tested whether concurrent acute hyperhomocysteinemia and mild hypertriglyceridemia increase the concentrations of circulatory cellular adhesion molecules in healthy subjects. STUDY DESIGN AND METHODS: Twelve healthy volunteers aged 37.5 years (range, 25-51) participated in the present study. The concentrations of plasma total homocysteine (p-tHcy), serum triglycerides, circulatory cellular adhesion molecules (CAMs), and concentrations of nitrate were measured at 0 (fasting), 2, 4, and 6 h after loading with (1) methionine, (2) fat, (3) methionine + fat, and (4) water (control). Wash out period between each loading was >or=1 week. RESULTS: Percent relative changes from baseline in the concentrations of p-tHcy, 2, 4, and 6 h after methionine and methionine + fat were significantly different from after water and fat loading. Changes in the concentrations of serum triglycerides 2 h after fat loading were significantly different from water loading, whereas methionine + fat loading caused a significant difference after 2, 4, and 6 h. We detected a synergistic increase in the triglyceride area response to methionine + fat loading. We detected also a significant difference in percent relative changes in the concentrations of P-selectin (PSEL) (P = 0.02), E-selectin (ESEL) (P = 0.003), and vascular cell adhesion molecule-1 (VCAM-1) (P = 0.005) 6 h after methionine + fat loading as compared to water loading. There was an additive increase in the PSEL area response to methionine + fat loading. Furthermore, area response to VCAM was greater to methionine loading than water loading (P = 0.01). A decrease in the concentration of NO(3) was more pronounced after methionine + fat loading and a significantly decreased area response of nitrate to methionine + fat loading was detected than to area response to water loading (P = 0.002). CONCLUSION: Inflammatory activation of the endothelium takes place during concurrent transient hyperhomocysteinemia and mild hypertriglyceridemia.

Survival of Salmonella species in eggs poached using a microwave oven.

The use of microwave ovens is becoming increasingly popular, but there is little data on the bactericidal effect of this mode of cooking. Following a family outbreak of gastroenteritis due to Salmonella enteritidis PT4, where eggs poached in a microwave oven were the suspected source, we investigated the survival of Salmonella spp. in artificially contaminated eggs cooked in a microwave oven. The survival of six serotypes of Salmonella at various inocula were studied, after cooking the eggs using two recognized poaching methods. Salmonellae were readily recovered after cooking if the yolk was still soft, whichever serotype, inoculum or cooking method was used. The survival of organisms was related to the number of organisms present in the raw eggs. The advice regarding eating lightly cooked eggs should be the same whether microwave or more conventional cooking techniques are used.

The efficacy of a hospital dry cleaning cycle in disinfecting material contaminated with bacteria and viruses.

It is not known how efficiently a standard dry cleaning cycle can kill bacteria or viruses. In-situ experiments were carried out to determine the cidal activity of such a cycle using perchloroethylene solvent against five clinical bacterial isolates and a DNA bacteriophage. Viable counts of bacteria recovered from material after dry cleaning were reduced by 3-8 logs, with up to 10(3) colony forming units (cfu) surviving per strip. Numbers of bacteriophage were only reduced by 10-100 fold. The resistance of the bacteriophage to solvent and heat (60 degrees C) was compared with that of polio and herpes simplex viruses in vitro. Polio virus and bacteriophage, but not herpes simplex virus, survived exposure to perchloroethylene at room temperature for 40 min. Dry cleaning with perchloroethylene is not bactericidal and is particularly poorly virucidal for non-enveloped viruses.

The relationship among dental status, nutrient intake, and nutritional status in older people.

Dental health status may influence nutrition. The objective of this part of the National Diet and Nutrition Survey was to assess if there is a relationship between dental status in people 65 years and older and intake of certain nutrients and any link between dental status and blood-derived values of key nutrients. Random national samples of independently living subjects and those living in institutions had dental examinations, interviews, four-day food diaries, and blood and urine analyzed. In the sample living independently, intakes of most nutrients were lower in edentate than dentate subjects. Intake of non-starch polysaccharides, protein, calcium, non-heme iron, niacin, and vitamin C was significantly lower in edentate subjects. People with 21 or more teeth consumed more of most nutrients, particularly of non-starch polysaccharide. This relationship in intake was not apparent in the hematological analysis. Plasma ascorbate and plasma retinol were the only analytes significantly associated with dental status.

Micronutrient deficiencies in the aetiology of anaemia in a rural area in The Gambia.

Early studies in a rural community in The Gambia led to reports of anaemia in certain sections of the population. More recent investigations concerned with identifying micronutrient deficiencies have elaborated these earlier findings. The major determinant of anaemia is an iron deficiency, leading to a microcytic hypochromic haematological picture. However, riboflavin deficiency, which is very widespread, and which leads to clinical signs in some subjects, appears to impose some limitations on absorption and utilization of iron. A subclinical deficiency of ascorbic acid is also observed and this may further compromise iron status. In addition very poor folate status leads to a macrocytosis in some individuals and may limit the haematological response to iron supplementation.