Serum Soluble Transferrin Receptor Concentrations Are Elevated in Congolese Children with Glucose-6-Phosphate Dehydrogenase Variants, but Not Sickle Cell Variants or α-Thalassemia.

Background: Anemia is common in Congolese children, and inherited blood disorders may be a contributing cause. The presence of sickle cell variants, X-linked glucose-6-phosphate dehydrogenase (G6PD) deficiency and α-thalassemia, has been previously reported. G6PD A- deficiency is characterized by the co-inheritance of G6PD 376 and 202 variants and is common in sub-Saharan Africa.Objective: We aimed to measure the associations between inherited blood disorders and hemoglobin, ferritin, and soluble transferrin receptor (sTfR) concentrations in Congolese children.Methods: Venous blood was collected from 744 children aged 6-59 mo from 2 provinces. We measured biomarkers of nutritional and inflammation status and malaria. Pyrosequencing was used to detect sickle cell variants. Polymerase chain reaction was used to detect G6PD variants and α-thalassemia deletions.Results: Overall, 11% of children had a sickle cell variant, 19% of boys were G6PD A- hemizygotes, 12% and 10% of girls were G6PD A- hetero- or homozygotes, respectively, and 12% of children had α-thalassemia. Multivariable linear regression models (adjusted for age, province, altitude, malaria, and biomarkers of nutritional and inflammation status) showed that G6PD A- hemizygous boys and G6PD 376 homozygous girls had higher sTfR concentrations [geometric mean ratios (95% CIs): 1.20 (1.03, 1.39) and 1.25 (1.02, 1.53), respectively] than children with no G6PD variants. Hemoglobin and ferritin concentrations were not independently associated with any of the inherited blood disorder genotypes.Conclusions: We found that 2 G6PD variant genotypes were associated with elevated sTfR concentrations, which limits the accuracy of sTfR as a biomarker of iron status in this population.

In Rwandese Women with Low Iron Status, Iron Absorption from Low-Phytic Acid Beans and Biofortified Beans Is Comparable, but Low-Phytic Acid Beans Cause Adverse Gastrointestinal Symptoms.

BACKGROUND: Phytic acid (PA) is a major inhibitor of iron bioavailability from beans, and high PA concentrations might limit the positive effect of biofortified beans (BBs) on iron status. Low-phytic acid (lpa) bean varieties could increase iron bioavailability. OBJECTIVE: We set out to test whether lpa beans provide more bioavailable iron than a BB variety when served as part of a composite meal in a bean-consuming population with low iron status. METHODS: Dietary iron absorption from lpa, iron-biofortified, and control beans (CBs) (regular iron and PA concentrations) was compared in 25 nonpregnant young women with low iron status with the use of a multiple-meal crossover design. Iron absorption was measured with stable iron isotopes. RESULTS: PA concentration in lpa beans was ∼10% of BBs and CBs, and iron concentration in BBs was ∼2- and 1.5-fold compared with CBs and lpa beans, respectively. Fractional iron absorption from lpa beans [8.6% (95% CI: 4.8%, 15.5%)], BBs [7.3% (95% CI: 4.0%, 13.4%)], and CBs [8.0% (95% CI: 4.4%, 14.6%)] did not significantly differ. The total amount of iron absorbed from lpa beans and BBs was 421 µg (95% CI: 234, 756 µg) and 431 µg (95% CI: 237, 786 µg), respectively, and did not significantly differ, but was >50% higher (P < 0.005) than from CBs (278 µg; 95% CI: 150, 499 µg). In our trial, the lpa beans were hard to cook, and their consumption caused transient adverse digestive side effects in ∼95% of participants. Gel electrophoresis analysis showed phytohemagglutinin L (PHA-L) residues in cooked lpa beans. CONCLUSION: BBs and lpa beans provided more bioavailable iron than control beans and could reduce dietary iron deficiency. Digestive side effects of lpa beans were likely caused by PHA-L, but it is unclear to what extent the associated digestive problems reduced iron bioavailability. This trial was registered at clinicaltrials.gov as NCT02215278.

The challenge of managing headache disorders in a tertiary referral neurology clinic in Rwanda.

OBJECTIVE: To determine the pattern, management, and outcome of headaches among patients treated at Outpatient Neurology Clinic. METHODS: A retrospective study was conducted at the Out-Patient Neurology Clinic of the Butare University Teaching Hospital, University of Rwanda, Butare, Rwanda between February and May 2015. We extracted the demographic data, headache characteristics, and associated conditions, prior pain-relieving medication use, waiting time before consultation, the results of paraclinical investigations, final diagnosis according to the International Classification of Headache Disorders, management, and 3-month clinical outcome from the medical records of all patients who consulted for headache over 36-month period. Epi Data and Statistical package for Social sciences software version 21.0 (SPSS Inc, Chicago, IL, USA) software were used for data processing. RESULTS: Headache disorders represent a quarter of all neurological consultations. Patients were predominantly female (67%) and young (78% <45 years old). One-third (34%) presented with chronic tension-type headache. Neuroimaging demonstrated an abnormality in a significant minority (14%). Amitriptyline was the most commonly used drug (60%) in management. Forty percent of those patients followed for 3 months did not experience any clinical improvement. CONCLUSION: Headache is among the most common medical complaints in the Outpatient Neurology Clinic, with a wide array of underlying diagnoses, and a significant yield on neuroimaging. A significant proportion of those suffering from headache disorders have poor short-term outcomes. Novel approaches, such as headache support groups and alternative pharmacological agents, should be investigated for these patients.

Phytic acid concentration influences iron bioavailability from biofortified beans in Rwandese women with low iron status.

BACKGROUND: The common bean is a staple crop in many African and Latin American countries and is the focus of biofortification initiatives. Bean iron concentration has been doubled by selective plant breeding, but the additional iron is reported to be of low bioavailability, most likely due to high phytic acid (PA) concentrations. OBJECTIVE: The present study evaluated the impact of PA on iron bioavailability from iron-biofortified beans. METHODS: Iron absorption, based on erythrocyte incorporation of stable iron isotopes, was measured in 22 Rwandese women who consumed multiple, composite bean meals with potatoes or rice in a crossover design. Iron absorption from meals containing biofortified beans (8.8 mg Fe, 1320 mg PA/100 g) and control beans (5.4 mg Fe, 980 mg PA/100 g) was measured with beans containing either their native PA concentration or with beans that were ∼50% dephytinized or >95% dephytinized. RESULTS: The iron concentration of the cooked composite meals with biofortified beans was 54% higher than in the same meals with control beans. With native PA concentrations, fractional iron absorption from the control bean meals was 9.2%, 30% higher than that from the biofortified bean meals (P < 0.001). The quantity of iron absorbed from the biofortified bean meals (406 µg) was 19% higher (P < 0.05) than that from the control bean meals. With ∼50% and >95% dephytinization, the quantity of iron absorbed from the biofortified bean meals increased to 599 and 746 µg, respectively, which was 37% (P < 0.005) and 51% (P < 0.0001) higher than from the control bean meals. CONCLUSIONS: PA strongly decreases iron bioavailability from iron-biofortified beans, and a high PA concentration is an important impediment to the optimal effectiveness of bean iron biofortification. Plant breeders should focus on lowering the PA concentration of high-iron beans. This trial was registered at clinicaltrials.gov as NCT01521273.

A new high-sensitive nephelometric method for assaying serum C-reactive protein based on phosphocholine interaction.

BACKGROUND: The measurement of C-reactive protein (CRP) concentrations has been of interest as a classical marker of acute phase response; in addition, it has been of particular interest in cardiovascular risk stratification where high-sensitive measurements are necessary. Since CRP is able to bind phospholipids (mainly phosphocholine) in the presence of calcium ions, we explored the possibilities of developing a high-sensitive affordable nephelometric CRP assay based on diluted soy oil emulsions. METHODS: Serum (or heparinized plasma) was mixed with Intralipid 20% in Tris-calcium buffer (pH 7.5). After 12 min of incubation at 37°C, the CRP-phospholipid complexes were measured by nephelometry (840 nm) using a BN II nephelometer (Siemens). Results (n=97) were compared with those obtained using a typical immunoturbidimetric method (Roche). RESULTS: Imprecision of the functional nephelometric assay was evaluated using three human serum pools. Within-run coefficients of variation (CVs) for level 1, 2 and 3 were 6.1%, 4.7% and 4.5%, respectively, and between-run CVs were 17.6%, 18.8% and 11.3%, respectively. Good agreement was obtained between the functional nephelometric and the immunoturbidimetric CRP assay in a concentration range from 0.1 mg/L to 50 mg/L (r=0.884). A logit-log calibration curve was made between 0.056 mg/L and 1.785 mg/L. The limit of detection was 0.5 mg/L. CONCLUSIONS: The functional nephelometric CRP assay allowed high-sensitive CRP determinations in serum and plasma. Since the assay is species independent, the described functional CRP assay could be used for veterinary purposes as well.

Stable iron isotope studies in Rwandese women indicate that the common bean has limited potential as a vehicle for iron biofortification.

Biofortification of plants is a new approach to combat iron deficiency. Common beans (Phaseolus vulgaris) can be bred with a higher iron concentration but are rich in iron absorption inhibitors, phytic acid (PA), and polyphenols (PP). To evaluate the potential of beans to combat iron deficiency, three iron absorption studies were carried out in 61 Rwandese women with low iron status. Studies 1 and 2 compared iron absorption from high and low PP beans, similar in PA and iron, fed as bean puree in a double meal design or with rice and potatoes as multiple meals. Study 3 compared iron absorption from high and normal iron beans with similar PP levels and a PA:iron molar ratio, fed with potatoes or rice in multiple meals. Iron absorption was measured as erythrocyte incorporation of stable iron isotopes. In study 1, iron absorption from the high PP bean (3.4%) was 27% lower (P < 0.01) than from low PP bean (4.7%), but when fed in multiple meals (study 2), there was no difference (7 and 7.4%, respectively; P > 0.05). In study 3, iron absorption from the high iron bean (3.8%) was 40% lower (P < 0.001) than from the normal iron bean (6.3%), resulting in equal amounts of iron absorbed. When beans were combined with other meal components in multiple meals, high PP concentration had no negative impact on iron absorption. However, the quantity of iron absorbed from composite meals with high iron beans was no higher than with normal iron beans, indicating that efficacious iron biofortification may be difficult to achieve in beans rich in PA and PP.

Distribution and Determinants of Serum Zinc, Copper, and Selenium Levels among Children under Five Years from Popokabaka, Democratic Republic of Congo: A Cross-Sectional Study.

Information about essential trace elements among children in many African countries, including the Democratic Republic of Congo (DRC), is limited. We aimed to measure the distribution and determinants of serum zinc (Zn), copper (Cu), and selenium (Se) concentrations in a representative sample of children under five years old. We conducted a community-based cross-sectional study in Popokabaka, DRC. Blood samples were drawn from 412 children. The serum concentrations of minerals were measured using inductively coupled plasma−mass spectrometry. The median concentrations (P25−P75) of Zn, Cu, and Se were 61.9 µg/dL (52.8−70.2), 145.5 (120.0−167.0) µg/dL and 5.3 (4.3−6.3) µg/dL. The CRP-adjusted prevalence of serum Se deficiency was 84.1% (95% confidence interval [CI] 81.4−87.0) and of Zn deficiency was 64.6% (95% CI 59.8−69.1%). Only a few children were Cu deficient [1.5% (0.6−3.2)]. Evidence of inflammation (C-reactive protein, >5 mg/L) was associated with a lower Se concentration and higher Cu concentration. Furthermore, serum Se concentration was positively associated with linear growth. The average Cu/Zn molar ratio (2:1) was twice that recommended. Children in western Popokabaka had higher Zn and Se levels than their eastern neighbors. Zinc and selenium deficiencies are common among children in Popokabaka and require attention and prioritization.

Micronutrient intake and prevalence of micronutrient inadequacy among women (15-49 y) and children (6-59 mo) in South Kivu and Kongo Central, Democratic Republic of the Congo (DRC).

Iron biofortified beans and carotenoid enriched cassava are proposed as a solution to combat iron and vitamin A deficiencies, respectively, in the Democratic Republic of Congo (DRC). To inform the need for biofortified foods, we conducted a survey in 2014 in two provinces of the DRC, South Kivu and Kongo Central. Unexpectedly, women of reproductive age (WRA; 15-49 y) and their children (6-59 m) had a low prevalence of biochemical iron and vitamin A deficiency, based on ferritin and retinol binding protein, respectively. To better understand the lack of biochemical deficiency of these nutrients, we examined the prevalence of inadequate intake for these and other select nutrients. Dietary intake was assessed using 24-hour recalls among 744 mother-child dyads. Repeat recalls on a non-consecutive day were conducted with a subsample of the study population to account for intra-individual variation and estimate usual intake. In WRA, the prevalence of inadequate iron intakes were 33% and 29% in South Kivu and Kongo Central, respecitvely. The prevalence of inadequate vitamin A intakes among WRA was low in South Kivu (18%) and negligible in Kongo Central (1%). Iron inadequacy was highest in infants (6-11 m) at 82% and 64% in South Kivu and Kongo Central, respectively. Among older children (12-59 m) in both provinces, the prevalence of iron inadequacy was similar at ~20%. There was a high prevalence of inadequate zinc intake in women and children (i.e. 79-86% among WRA and 56-91% among children 6-59 m) consistent with our findings of a high prevalence of low serum zinc in the same sample. Dietary data here corroborate the low prevalence of biochemical vitamin A deficiency but not iron. However, any change to the supply of red palm oil (primary source of vitamin A) would dramatically reduce population vitamin A intakes, thus a carotenoid enriched cassava program may be beneficial as a safety net measure. Crops biofortified with zinc also appear warranted. We caution that our findings cannot be extrapolated to the entire Congo where diverse agro-ecological landscape exist or when political and environmental shocks occur which challenge food production.

A new turbidimetric method for assaying serum C-reactive protein based on phosphocholine interaction.

BACKGROUND: C-reactive protein (CRP) is able to bind phospholipids (mainly phosphocholine) in the presence of calcium ions. We investigated the use of this property for developing an affordable turbidimetrical CRP assay based on diluted soy oil. METHODS: Serum (or heparinized plasma) was mixed with Intralipid 20% in Tris-calcium buffer (pH 7.5). After 30 min of incubation at 37 degrees C, the CRP-phospholipids complexes were measured by turbidimetry (660 nm/700 nm) with a Cobas 6000 analyzer (Roche). Results were compared with those obtained using a typical immunoturbidimetric method (Roche). RESULTS: Good correlation (r(2)=0.931) was obtained between the functional and the immunoturbidimetric CRP assay. Within-run and between-run %CV values for the functional assay were 2.4% (100 mg/L); 6.0% (50 mg/L); 10% (20 mg/L), and 3.6% (100 mg/L); 8.0% (50 mg/L); 11% (20 mg/L), respectively. The limit of detection was 7 mg/L. Results were not affected by serum calcium, triglyceride, or phospholipid concentrations. CONCLUSIONS: The functional CRP assay allowed measurement of CRP in serum and plasma in the range of 7 mg/L-400 mg/L. The assay is particularly suited in conditions where resources are restricted. Since the assay is species independent, the described functional CRP assay could be used for veterinary purposes as well.

Anemia and Micronutrient Status of Women of Childbearing Age and Children 6-59 Months in the Democratic Republic of the Congo.

Little is known about the micronutrient status of women and children in the Democratic Republic of the Congo, which is critical for the design of effective nutrition interventions. We recruited 744 mother-child pairs from South Kivu (SK) and Kongo Central (KC). We determined hemoglobin (Hb), serum zinc, vitamin B12, folate, ferritin, soluble transferrin receptor (sTfR), retinol binding protein (RBP), C-reactive protein, and α-1 acid glycoprotein concentrations. Anemia prevalence was determined using Hb adjusted for altitude alone and Hb adjusted for both altitude and ethnicity. Anemia prevalence was lower after Hb adjustment for altitude and ethnicity, compared to only altitude, among women (6% vs. 17% in SK; 10% vs. 32% in KC), children 6-23 months (26% vs. 59% in SK; 25% vs. 42% in KC), and children 24-59 months (14% vs. 35% in SK; 23% vs. 44% in KC), respectively. Iron deficiency was seemingly higher with sTfR as compared to inflammation-adjusted ferritin among women (18% vs. 4% in SK; 21% vs. 5% in KC), children 6-23 months (51% vs. 14% in SK; 74% vs. 10% in KC), and children 24-59 months (23% vs. 4% in SK; 58% vs. 1% in KC). Regardless of indicator, iron deficiency anemia (IDA) never exceeded 3% in women. In children, IDA reached almost 20% when sTfR was used but was only 10% with ferritin. Folate, B12, and vitamin A (RBP) deficiencies were all very low (<5%); RBP was 10% in children. The prevalence of anemia was unexpectedly low. Inflammation-adjusted zinc deficiency was high among women (52% in SK; 58% in KC), children 6-23 months (23% in SK; 20% in KC), and children 24-59 months (25% in SK; 27% in KC). The rate of biochemical zinc deficiency among Congolese women and children requires attention.

A functional turbidimetric method to determine C-reactive protein in horses.

A turbidimetric method to determine serum C-reactive protein (CRP) concentration, based on soybean oil-phosphocholine interaction, was performed on horse serum samples to evaluate its potential diagnostic value in veterinary medicine. Intralipid 20% in 0.1 M Tris-calcium buffer (pH 7.5) was added to horse serum. After 30 min of incubation at 37 °C, the CRP-phosphocholine complexes were turbidimetrically, bichromatically (660 nm/700 nm) quantified on a commercial analyzer. Furthermore, comparison between CRP and other inflammatory markers, including white blood cell and neutrophil counts, was performed to evaluate the diagnostic value of both tests. Standardization of the assay was done using a commercial human CRP calibrator. The CRP measurements were performed on serum samples (296 patients and 34 controls). Reference values were found to be lower than 10 mg/l. The method was found to be linear between 1 and 400 mg/l. A moderate correlation was observed between CRP values and the relative neutrophil counts. Receiver-operating characteristics analysis demonstrated the area under the curve for CRP was 0.928, which was superior (P < 0.001) to the neutrophil count (0.804) and the leukocyte count (0.664) in detecting the presence of inflammation. This CRP assay showed reliable results as an acute phase test in horses, confirming its species-independent capability to detect CRP in various mammals, including horses.