Iron deficiency (ID) at both birth and 9 months predicts right frontal EEG asymmetry in infancy.

This study considered effects of timing and duration of iron deficiency (ID) on frontal EEG asymmetry in infancy. In healthy term Chinese infants, EEG was recorded at 9 months in three experimental conditions: baseline, peek-a-boo, and stranger approach. Eighty infants provided data for all conditions. Prenatal ID was defined as low cord ferritin or high ZPP/H. Postnatal ID was defined as ≥ two abnormal iron measures at 9 months. Study groups were pre- and postnatal ID, prenatal ID only, postnatal ID only, and not ID. GLM repeated measure analysis showed a main effect for iron group. The pre- and postnatal ID group had negative asymmetry scores, reflecting right frontal EEG asymmetry (mean ± SE: -.18 ± .07) versus prenatal ID only (.00 ± .04), postnatal ID only (.03 ± .04), and not ID (.02 ± .04). Thus, ID at both birth and 9 months was associated with right frontal EEG asymmetry, a neural correlate of behavioral withdrawal and negative emotions.

Estimating the prevalence of iron deficiency in the first two years of life: technical and measurement issues.

National-level data on iron deficiency is not available for most countries and many rely on the prevalence of anemia as a proxy estimate, assuming that approximately 50% of anemia cases are caused by iron deficiency. Anemia, however, has multiple causal factors and the risk attributable to any one cause will depend on its relative importance in a population in relation to other causes. The present review summarizes current estimates on the prevalence of iron deficiency and anemia in children younger than 2 years and evaluates the strengths and weaknesses of currently available indicators of iron deficiency in children. Anemia prevalence is insufficient to estimate the prevalence of iron deficiency in children younger than 2 years. The methods widely used to assess iron deficiency at the population level rely on venous blood samples and are complicated and costly to implement.

Deferoxamine iron chelation increases delta-aminolevulinic acid induced protoporphyrin IX in xenograft glioma model.

Exogenous administration of delta-aminolevulinic acid (delta-ALA) leads to selective accumulation of protoporphyrin IX (PpIX) in brain tumors, and has shown promising results in increasing extent of resection in fluorescence-guided resection (FGR) of brain tumors. However, this approach still suffers from heterogeneous staining and so some tumor margins may go undetected because of this variation in PpIX production. The aim of this study was to test the hypothesis that iron chelation therapy could increase the level of fluorescence in malignant glioma tumors. Mice implanted with xenograft U251-GFP glioma tumor cells were given a 200 mg kg(-1) dose of deferoxamine (DFO), once a day for 3 days prior to delta-ALA administration. The PpIX fluorescence observed in the tumor regions was 1.9 times the background in animal group without DFO, and 2.9 times the background on average, in the DFO pre-treated group. A 50% increase in PpIX fluorescence contrast in the DFO group was observed relative to the control group (t-test P-value = 0.0020). These results indicate that iron chelation therapy could significantly increase delta-ALA-induced PpIX fluorescence in malignant gliomas, pointing to a potential role of iron chelation therapy for more effective FGR of brain tumors.

Synergistic increase in osteosarcoma cell sensitivity to photodynamic therapy with aminolevulinic acid hexyl ester in the presence of hyperthermia.

OBJECTIVE: We observed that two osteosarcoma cell lines from the same tumor displayed marked differences in their sensitivities to photodynamic therapy (PDT) with aminolevulinic acid hexyl ester (hALA-PDT). We investigated why these two closely related lines had different hALA-PDT sensitivities and whether the PDT phototoxicity of the less sensitive cell line could be increased by a simultaneous application of hyperthermia (HT). METHODS: Flow cytometry was used to evaluate the intracellular accumulation of protoporphyrin IX (PpIX), a metabolic product of aminolevulinic acid, in two human mandibular osteosarcoma cell lines (HOSM-1 and HOSM-2) treated with HT, hALA-PDT, or hALA-PDT combined with HT (PDT + HT). With hALA-PDT, cells treated with 0.2 mM hALA were irradiated with a light dose of 10-80 J/cm(2) from a near-infrared irradiator. With PDT + HT, the cells were treated as for hALA-PDT except that the temperature was raised to 43.5 degrees C during irradiation. RESULTS: At 6 h after hALA treatment, HOSM-2 cells carried about 1.53-fold more PpIX than HOSM-1 cells. With hALA-PDT, the survival rate for HOSM-1 cells treated with 80 J/cm(2) irradiation was 35.7%, while that for HOSM-2 cells treated with 40-80 J/cm(2) was below 12%. With PDT + HT, the survival rate for HOSM-1 and HOSM-2 cells treated with 80 J/cm(2) irradiation was 14.1% and 10.7%, respectively. CONCLUSION: A combination therapy comprising hALA-PDT + HT treatment may be very useful for the treatment of tumors containing cells that are insensitive to hALA-PDT, such as the HOSM-1 cell line described in this study.

Determinants of anemia among preschool children in the Philippines.

OBJECTIVE: Our objective was to identify the determinants of anemia among rural Filipino children aged 12-71 months. METHODS: A cross-sectional survey was conducted among 2090 preschool children from 8 rural villages in Cebu, an area non-endemic for malaria and schistosomiasis. Hemoglobin (Hb) concentration was determined using a HemoCue hemoglobinometer and zinc protoporphyrin (ZPP) concentration was measured with a hematofluorometer. A 3-day non-consecutive 24-hour food recall interview with the child's primary caregiver was done to estimate the child's dietary intake. Stool analysis for presence of soil-transmitted helminths was performed through a concentration technique. A separate interview on household socio-economic status with the child's primary caregiver was conducted. RESULTS: Mean Hb concentration was 12.0 g/dL (SD 1.3). 16.1% were anemic. Age and sex had a significant interaction in their effect on Hb concentration. Females had higher Hb concentration between 12 to 23 months of age. Hb levels equalize between the 2 genders at around 24 months and increase with similar increments until 71 months of age. All dietary parameters improved Hb concentration with increasing intake. In the multiple regression, however, only the index for bioavailable iron and vitamin C intakes remained independent factors. None of the helminths or combination of helminths had significant effects on Hb concentration. Among the socio-economic variables, maternal educational attainment and water supply were significant independent factors. Mean ZPP concentration was 72.07 (SD 46.45) and 30.8% were iron deficient. As with Hb concentration, age and sex had a significant interaction in their effect on ZPP concentration, with females having lower ZPP levels before 24 months of age. Bioavailable iron (animal iron + 0.3*plant iron) had a significant effect on ZPP concentration at levels of at least 15% of the iron requirement. This was seen even after controlling for multivitamin supplementation. CONCLUSION: The control of anemia among preschoolers can be achieved through a combination of various nutritional interventions such as micronutrient supplementation, food fortification and nutrition education. Our findings emphasize the importance of a multi-sectoral approach to nutritional problems--the importance of empowering women (through engagement and education) and of maintaining a healthy physical environment (water and sanitation) are often peripheral concerns of nutritionists. Our study highlights the importance of supporting initiatives that address these issues not only for their core benefit, but also for the potential benefit to nutrition.

Adiposity in women and children from transition countries predicts decreased iron absorption, iron deficiency and a reduced response to iron fortification.

BACKGROUND: Overweight is increasing in transition countries, while iron deficiency remains common. In industrialized countries, greater adiposity increases risk of iron deficiency. Higher hepcidin levels in obesity may reduce dietary iron absorption. Therefore, we investigated the association between body mass index (BMI) and iron absorption, iron status and the response to iron fortification in populations from three transition countries (Thailand, Morocco and India). METHODS: In Thai women (n=92), we examined the relationship between BMI and iron absorption from a reference meal containing approximately 4 mg of isotopically labeled fortification iron. We analyzed data from baseline (n=1688) and intervention (n=727) studies in children in Morocco and India to look for associations between BMI Z-scores and baseline hemoglobin, serum ferritin and transferrin receptor, whole blood zinc protoporphyrin and body iron stores, and changes in these measures after provision of iron. RESULTS: In the Thai women, 20% were iron deficient and 22% were overweight. Independent of iron status, a higher BMI Z-score was associated with decreased iron absorption (P=0.030). In the Indian and Moroccan children, 42% were iron deficient and 6.3% were overweight. A higher BMI Z-score predicted poorer iron status at baseline (P<0.001) and less improvement in iron status during the interventions (P<0.001). CONCLUSIONS: Adiposity in young women predicts lower iron absorption, and pediatric adiposity predicts iron deficiency and a reduced response to iron fortification. These data suggest the current surge in overweight in transition countries may impair efforts to control iron deficiency in these target groups. Interactions of the 'double burden' of malnutrition during the nutrition transition may have adverse consequences.

A cohort study of haemoglobin and zinc protoporphyrin levels in term Zambian infants: effects of iron stores at birth, complementary food and placental malaria.

OBJECTIVE: To examine zinc-protoporphyrin (ZPP) and haemoglobin levels, and to determine predictors of iron deficiency anaemia (IDA) in Zambian infants. SUBJECTS AND METHODS: Ninety-one women and their normal birth weight (NBW) infants were followed bi-monthly during the first 6 months of life, and iron status, food intake, malaria parasitaemia and growth were monitored. At 4 months, the infants were divided into two groups, and the data were analysed according to whether or not they were exclusively breastfed. RESULTS: Almost two-third of infants were born with low iron stores as defined by ZPP levels, and this proportion increased with age. Over 50% had developed IDA by 6 months. Exclusive breastfeeding at 4 months could be a protective factor for IDA (odds ratio (OR): 0.2; 95% confidence interval (CI): 0.0-1.1). Exclusively breastfed infants had higher haemoglobin values at 4 and 6 months (mean difference 0.6; 95% CI: 0.1-1.2 g/dl and mean difference 0.9; 95% CI: 0.2-1.7 g/dl, respectively), compared with infants with early complementary feeding. In univariate analysis, past or chronic placental malaria appeared to be a predictor of IDA at 4 and 6 months, but the significance was lost in multivariate analysis. CONCLUSIONS: Zambian NBW infants are born with low iron stores and have a high risk to develop IDA in the first 6 months of life. Continuation of exclusive breastfeeding after 4 months is associated with a reduction of anaemia. The effect of placental malaria infection on increased risk of infant IDA could not be proven.

Lead poisoning as a result of infertility treatment using herbal remedies.

INTRODUCTION: Lead poisoning remains a major problem in our society due to the lack of awareness of its ill effects among the clinical community. The non-availability of blood lead levels at primary health centers results in many lead-poisoned cases being treated symptomatically, without the diagnosis being sought. CASE REPORT: We report a case here, presenting with colicky abdominal pain receiving symptomatic treatment initially; lead poisoning was confirmed following the estimation of blood lead levels. DISCUSSION: The consumption of unbranded herbal medicines as a causative factor for lead poisoning was confirmed by high lead content in the medicines. Surely, this demands public health programs to disseminate strategies to the medical personnel about the clinical implication of lead poisoning, as well as the general public for awareness and to prevent lead poisoning.

Drug-induced protoporphyria in beagle dogs.

As part of regulatory safety testing program, a 13-week oral toxicity study with a new antipsychotic drug candidate was performed in beagle dogs. During this study, dark red/brown feces were recorded in treated dogs and increases in liver parameters (alanine aminotransferase, alkaline phosphatase, bilirubin) were measured biochemically. At the end of the study, livers of high-dose (50 mg/kg) animals were (mottled) dark brown, sometimes with pale foci. Histopathological examination of these livers showed dark globular pigment deposits in the hepatocellular cytoplasm and within the bile canaliculi. Varying numbers of inflammatory cell infiltrates were additionally present in association with the deposits. These pigment deposits showed birefringency with characteristic "Maltese Cross"-like structures under polarized light. Electronmicroscopy revealed the typical, so-called "sunburst" pattern with radiating double-lined crystalline structures. These morphologic characteristics strongly indicated at the presence of porphyrins, which was definitely confirmed biochemically. Published reports of drug-induced hepatic porphyria in dogs are rare. The possible underlying mechanism in the dog and man is discussed.

Hepatic porphyria induced by the herbicide tralkoxydim in small mammals is species-specific.

Tralkoxydim is the active ingredient in a postemergent herbicide used in cereal crops. During preregistration trials, tralkoxydim was observed to cause hepatic porphyria and cholestasis in laboratory mice. Porphyria was not seen in similarly exposed rats or hamsters, but data were not collected regarding the susceptibility of any wild small mammal species to the tralkoxydim-induced porphyria. To address this data gap, we exposed small mammals to tralkoxydim, to 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC; a known porphyrinogenic chemical), or to sunflower oil alone. We studied small mammal species that might be exposed following agricultural applications of the herbicide, including the white-footed mouse (Peromyscus leucopus), the deer mouse (P. maniculatus), and the meadow vole (Microtus pennsylvanicus). Because of their known susceptibility to both tralkoxydim- and DDC-induced porphyria, commercially supplied Mus musculus (CD-1 Swiss mice) were exposed as positive-control animals. We also exposed offspring of wild-caught M. musculus to compare their responses to those of the commercially supplied animals. Potential hepatotoxicity was determined by assessing the accumulation of liver protoporphyrin. Of the species tested, only M. musculus was susceptible to the porphyrinogenic action of tralkoxydim, and no significant accumulation of protoporphyrin was observed in any of the other species exposed to the herbicide.

Influence of CaNa2 EDTA on topical 5-aminolaevulinic acid photodynamic therapy.

BACKGROUND: We assessed whether the CaNa2 EDTA could improve the accumulation of protoporphyrin IX (PpIX) and photosensitisation in HEp-2 cells as well as the depth of treatment of skin cancers on the topical 5-Aminolaevulinic acid (5-ALA) PDT. METHODS: HEp-2 cells were incubated with 5-ALA (0-1 mmol/L) and CaNa2EDTA (0-1 mmol/L) for 4 hours, intracellular protoporphyrin IX content was quantified by extraction, and cell viability was assessed by use of the methyl-tetrazolium (MTT) assay four hours after exposure to light. In comparison with the pictures before and after treatment, depth of treatment could be determined using a Acuson Sequioa 512 phase-array system in paired experiments. RESULTS: PpIX accumulation increased with increasing extracellular concentrations of ALA (0-1 mmol/L). Adding 1 mmol/L of CaNa2EDTA increased 30% PpIX accumulation over the same period of incubation in the concentration of 1 mmol/L ALA. Significant difference was observed between the 5-ALA alone group and 5-ALA combined CaNa2 EDTA group in the PpIX accumulation (P < 0.01). Cell viability after exposure to light decreased with adding CaNa2 EDTA, a statistical difference in a same fluence above 1.2 J/cm2 between two groups was demonstrated (P < 0.05, P < 0.01 respectively). Depth of treatment of skin cancers were increased in CaNa2 EDTA-treated group. CONCLUSION: CaNa2 EDTA could improve the PpIX accumulation and photosensitisation in HEp-2 cells. Clinically, CaNa2 EDTA could increase the depth of treatment in the cutaneous cancers.

Detection of protoporphyrin IX in envelope membranes of pea chloroplasts.

Envelope membranes were prepared from mature pea chloroplasts. The tetrapyrrole contents of envelope membranes were analysed. The envelope membranes of pea chloroplasts contained substantial amounts of protoporphyrin IX and trace amounts of Mg-protoporphyrin IX and its monoester in addition to protochlorophyllide. The protoporphyrin IX content of envelope membranes was 89.25 pmol (mg protein)(-1). Its content in pea envelope membrane was higher than that of protochlorophyllide. The proportion of monovinyl and divinyl forms of protochlorophyllide present in pea chloroplast envelope membrane was 3:7. The significance of the presence of protoporphyrin IX in the envelope membrane is discussed in relation to plastidic Chl biosynthesis.

Erythropoietic protoporphyria treated with narrow-band (TL-01) UVB phototherapy.

Erythropoietic protoporphyria is a rare photodermatosis for which treatment options are limited. The present report describes the clinical features of a patient with erythropoietic protoporphyria and liver function test abnormalities associated with treatment with beta-carotene. Subsequent treatment with narrow-band UVB phototherapy resulted in marked subjective improvement in photosensitivity, which was confirmed by abolition of demonstrated abnormalities on monochromator phototesting. The therapeutic options for photosensitivity in erythropoietic protoporphyria are reviewed and discussed.

Iron and zinc status of women and men who followed cholesterol-lowering diets.

OBJECTIVE: To determine if men and women taught the National Cholesterol Education Program step 2 diet experienced diminished iron or zinc status after 2 years of consuming the diet. DESIGN: Subjects attended 8 weeks of diet classes and followup counseling. Four-day food records, zinc protoporphyrin/ heme (ZPP/H, a measure of iron stores), hematocrit, and plasma zinc values were collected at baseline, 1 year, and 2 years. SUBJECTS: Participants in a study of the efficacy of the step 2 diet (213 men and 151 women) who had low-density lipoprotein cholesterol levels above the 75th percentile, and who were not taking lipid-altering medication. STATISTICAL ANALYSES: Paired t tests to compare preintervention and postintervention test results. Two-sample t tests to compare gender, menopause status, and hormone-use groups. Analysis of variance was performed to compare plasma zinc levels among diet-adherence subgroups. Individual evaluation of outlying values and backward stepwise regression to determine the independent effects of variables were also completed. RESULTS: Mean dietary intake approached or exceeded step 2 guidelines. Density of iron intake increased. Density of zinc intake was unchanged, but mean intake was low. Mean ZPP/H and hematocrit did not change for premenopausal or postmenopausal women with or without supplemental hormones; men's values changed only slightly. Abnormal ZPP/H and hematocrit occurred sporadically. Mean plasma zinc levels did not differ from baseline for women, increased slightly for men, and did not differ by level of dietary adherence. No plasma zinc concentrations were below normal. APPLICATION: In this large-scale, 2-year prospective study of women and men, no adverse effects on intake or plasma indicators of iron and zinc were detected as a result of subjects being taught the NCEP step 2 diet.

Changes in biochemical indicators of iron status during iron repletion and depletion in monkeys.

Different modes of iron depletion and repletion were studied in monkeys to understand the sequential changes in and the relative importance of different biochemical indicators of iron status. Six control monkeys were divided into two groups, one was fed an iron-deficient diet (group 1) and the other underwent phlebotomy in addition to receiving an iron-deficient diet (group 2). Previously iron-depleted monkeys were subdivided into 4 groups of 3 animals each. While one group was continued on the iron-deficient diet (group 3), the second group received parenteral iron (group 4), the third group (group 5) received a sufficient-iron-containing diet, and the fourth group was fed 50% of the iron requirement. All indicators of iron status like hemoglobin (Hb), erythrocyte protoporphyrin (EPP), serum transferrin saturation and serum ferritin were monitored periodically, in addition to liver and bone marrow iron. all the indicators except serum ferritin and liver iron showed a decrease in group 2. On the other hand, animals receiving parenteral iron (group 4) showed an increase in all the parameters except serum ferritin. The dietary supplementation produced an increase in Hb and a decrease in EPP only (groups 5 and 6). There was a significant positive correlation between changes in bone marrow iron and Hb concentration depending on the severity of depletion and repletion. Both serum ferritin and liver iron did not respond to changes in dietary iron. Another parameter which responded to repletion was EPP. Serum ferritin and liver iron did not respond to changes in dietary iron or was not sensitive to subclinical iron deficiency. The results indicate that change in Hb is more sensitive to detect the deficiency of iron. It was also observed that different parameters respond variably under different modes of depletion and repletion.

Lead poisoning associated with the use of Ayurvedic metal-mineral tonics.

OBJECTIVE: The aim of this study was to confirm the connection between lead poisoning and the use of traditional Ayurvedic metal mineral tonics. METHODS: The study group comprised 29 subjects (26 adults and three children) who had previously taken Ayurvedic metal mineral tonics. All subjects were tested for lead absorption by blood lead, erythrocyte delta-aminolevulinic acid dehydratase activity and erythrocyte protoporphyrin. RESULTS: Eighteen samples of Ayurvedic preparations were obtained from 15 subjects and analyzed for lead content. Five adult subjects with established lead poisoning received chelation therapy. In Ayurvedic preparations a wide range of lead content was found (0.9-72,990 micrograms Pb/g; 0.35-29,900 micrograms Pb/capsule or tablet). The blood lead, erythrocyte delta-aminolevulinic acid dehydratase and erythrocyte protoporphyrin of the subjects, grouped according to the remedies used, correlated with the lead content in the preparations (p < 0.001). Chelation therapy was successful in normalization of laboratory findings and clinical recovery. CONCLUSION: Ayurvedic metal-mineral tonics are again identified as a potential source of high lead. The import of such tonics should be strictly controlled.

A longitudinal study of iron status in healthy Danish infants: effects of early iron status, growth velocity and dietary factors.

In a cohort of term infants (n=91), followed from birth to 12 months, iron intake was examined by 24-h food records, and iron status by blood samples (haemoglobin (Hb)), mean corpuscular volume (MCV), serum values for iron, ferritin and transferrin, and erythrocyte protoporphyrin) at 2, 6 and 9 months. At 9 months of age, 5% had anaemia (Hb<105 g/l), but none had developed iron deficiency according to strict definitions used in this study (serum ferritin < 13 micrograms/l and transferrin saturation < 10%). Infants with high serum ferritin, serum transferrin and erythrocyte protoporphyrin values at one blood sampling also had high values at the following sample (tracking, r=0.45-0.80), suggesting that iron stores at delivery are an important determinant of iron stores during late infancy. Factors related to changes in serum ferritin were investigated by multiple linear regression. From 2 to 6 months, serum ferritin was negatively associated with knee-heel growth velocity (p=0.006) and positively with intake of infant formula (p=0.04). From 6 to 9 months it was negatively associated with intake of bread (p=0.001), and there was a trend for a positive association with intake of meat (p=0.07) and fish (p=0.08) and for a negative association with intake of cow's milk (p=0.07). In conclusion, those with a high growth velocity and a dietary pattern with a high intake of bread and a low intake of meat and fish had lower ferritin values and thereby an increased risk of depleting their iron stores later during infancy.

Protoporphyrin biosynthesis in melanoma B16 cells stimulated by 5-aminolevulinic acid and chemical inducers: characterization of photodynamic inactivation.

The stimulation of protoporphyrin (PP) biosynthesis in B16 melanoma cells in order to facilitate photodynamic cell killing was studied. Biosynthesis and accumulation of PP in the melanoma cells was increased from 8 to 15 pmol/mg protein by the use of dimethyl-sulfoxide (DMSO), a differentiation-inducer. Treatment of the cells with the porphyrogenic agent allylisopropyl-acetamide (AIA) stimulated an additional PP increase. The most remarkable enhancement of intracellular PP was achieved by the supplementation of 5-aminolevulinic acid (5-ALA) to the growth medium following the addition of DMSO and AIA during the induction phase. The intracellular concentration of PP exceeded 21,950 pmol/mg protein following combined stimulation by DMSO/AIA and 5-ALA. The porphyrins produced in the incubated cells, in serum-depleted medium, consisted of 95% PP; 88% of it was recovered from the cells and only 7% was excreted into the medium. Photosensitization of the B16 melanoma cells containing high PP concentrations was effective even at low light doses. Potassium (K) efflux was the first measurable sign of cell damage determined by X-ray microanalysis (XRMA) following fast liquid-nitrogen fixation. During a 1 min interval, 70% of cellular K was lost. After 5 min illumination, complete cell destruction was detected by scanning electron microscopy (SEM) and XRMA. The photodamaged cells showed influx of Na, Cl and Ca ions accompanying the immediate K losses. Ultrastructural cell damage was manifested by disintegration of the outer membrane. Total cell death of B16 melanoma cells was achieved by chemical induction of endogenous PP and photosensitization.

Identification and characterization of the iron compounds in bone marrow by means of Mössbauer spectrometry.

In order to determine and to demonstrate the cellular iron molecular states in hematopoietic bone marrow, direct investigations were performed by means of different and complementary spectroscopic techniques: optical absorption, electron spin resonance and Mössbauer spectrometry. In fact, the latter appears to have been the most informative. In addition to the hemoglobin forms, five- and six-coordination ligand protoporphyrins IX (monomeric and polymeric stacking, respectively) were observed. A small amount of non-hemic high-spin iron III storage component (ferritin) was measured. No diferric transferrin was detected. A ferrous compound was also observed and attributed to the mitochondrial iron pool.

Observation on bovine congenital erythrocytic protoporphyria in the blonde d'Aquitaine breed.

Three blonde d'Aquitaine calves (one male and two females) about four months old, exhibited skin lesions just after birth, the site and nature of which suggested photosensitisation. Their porphyrin metabolism indicated a marked decrease in the activity of lymphocytic ferrochelatase, leading to a diagnosis of congenital erythrocytic protoporphyria. The associated nervous disorders of the 'recurrent epileptiform seizure' type are discussed in the light of complementary histological and biochemical tests.