High prevalence of selected viruses and parasites and their predictors in Malawian children.

Enteric pathogens have been related to child undernutrition. Whereas there are lots of data on enteric bacterial microbiota and infections, much less is known about the incidence of prevalence of intestinal colonisation with viruses or important parasitic species. This study assessed the presence of selected viruses and parasites in stools of 469, 354, 468 Malawian children at 6, 12 and 18 months. We also assessed environmental predictors of the presence of viruses and parasites among 6-month infants. Microbial presence was documented using real-time polymerase chain reaction (PCR). Enteroviruses were identified in 68%, 80% and 81% of the stool samples at 6, 12 and 18 months children, rhinovirus in 28%, 18% and 31%, norovirus in 24%, 22% and 16%, parechovirus in 23%, 17% and 17%, rotavirus in 3%, 1% and 0.6%, Giardia lamblia in 9.6%, 23.5% and 26%, and Cryptosporidium (spp.) in 6%, 8% and 2% of the 6, 12 and 18 months stool samples. Dry season (May-October) was associated with a low infection rate of enterovirus, norovirus and Cryptosporidium (spp.). Higher father's education level, less number of person in the household and higher sanitation were associated with a low infection rate of enterovirus, norovirus and rotavirus, respectively. The results suggest that the prevalence of asymptomatic viral and parasitic infections is high among Malawian children and that the family's living conditions and seasonality influence the rate of infections.

Periapical infection may affect birth outcomes via systemic inflammation.

OBJECTIVES: Maternal dental periapical infections are associated with preterm birth and intrauterine growth restriction. This study investigates whether the association is mediated through bacterial spread from periapical lesions to placenta (direct pathway) or systemic inflammatory reaction (indirect pathway). MATERIALS AND METHODS: We compared birth outcomes in Malawian mothers with and without periapical infection. As markers of a direct pathway, we identified placental bacteria using a 16S rDNA approach and assessed histological evidence of inflammation in the placenta and amniotic membranes. We measured C-reactive protein, alpha-1-acid glycoprotein, and salivary cortisol as markers of an indirect pathway. We used regression models to associate the predictor variables with duration of pregnancy and newborn size. RESULTS: Of 1,024 women, 23.5% had periapical infection. There was no association of periapical infection with either bacterial DNA or histological inflammation in placenta or membranes. Periapical infection was associated with C-reactive protein, alpha-1-acid glycoprotein, and cortisol concentrations in a dose-dependent manner at 36 weeks. Addition of alpha-1-acid glycoprotein or cortisol concentration into regression models attenuated the association between periapical infection and pregnancy outcomes. CONCLUSION: There was no evidence of direct spread of periapical bacteria to the placenta. Periapical infections and adverse pregnancy outcomes are in part mediated through systemic inflammation.

Effect of 12-month intervention with lipid-based nutrient supplement on the physical activity of Malawian toddlers: a randomised, controlled trial.

Physical activity is beneficial for children's well-being. The effect of dietary supplementation on children's physical activity in food-insecure areas remains little studied. We examined the effects of a lipid-based nutrient supplement (LNS) on children's objectively measured physical activity in a randomised, controlled, outcome-assessor-blinded trial. Mothers of the children received one capsule daily of Fe-folic acid (IFA), one capsule containing eighteen micronutrients (MMN) or one 20 g sachet of LNS (containing twenty-two MMN, protein, carbohydrates, essential fatty acids and 494 kJ (118 kcal)) during pregnancy and for 6 months thereafter. Children in the IFA and MMN groups received no supplementation, and these groups were collapsed into a single control group; children in the LNS group received 20 g LNS from 6 to 18 months. We measured physical activity with accelerometers over 1 week at 18 months. The main outcome was mean vector magnitude counts/15 s. Of the 728 children at the beginning of child intervention at 6 months, 570 (78 %) provided sufficient data for analysis. The mean accelerometer counts for the 190 children in the LNS group and for the 380 children in the control group were 303 (sd 59) and 301 (sd 56), respectively (P for difference=0·65). LNS, given to mothers during pregnancy and 6 months postpartum and to their infants from 6 to 18 months of age, did not increase physical activity among 18-month-old children.

Malawian parents' perceptions of physical activity and child development: a qualitative study.

BACKGROUND: In scientific studies, physical activity is measured by the amount of bodily movement, but lay perceptions of physical activity might be different. Parental influence is important for the development of children's physical activity behaviour, and parental perceptions of facilitators of physical activity are context specific. We aimed to investigate how parents of young Malawian children conceptualize physical activity in childhood, situate it in child development and understand its facilitators. METHODS: We used convenience sampling to identify parents of young children from different socio-economic backgrounds and age groups in semi-rural area of Malawi. We conducted in-depth interviews with 16 parents, a focus group discussion with six parents and key informant interviews with two nurses in Malawi. Six of the participants were fathers. We analysed the data with conventional qualitative content analysis by inductive approach. RESULTS: The parents emphasized practical skills, education and proper behaviour as goals for their children. They viewed activity as encompassing both mental and physical qualities and they perceived it as a positive attribute of children. The parents discussed skills acquisition, social competence, health and bodily movement as signs for being active. As facilitators of physical activity the parents mentioned balanced diet, good health and stimulation. The main concerns of the parents in regard to facilitators of physical activity and good child development were the availability of food and the child being healthy. CONCLUSIONS: Malawian parents' concept of children's physical activity is more comprehensive than scientific definition and includes aspects of both physical and mental activity.

Postnatal growth velocity and overweight in early adolescents: a comparison of rural and urban African boys and girls.

OBJECTIVES: To compare growth velocity of two African child cohorts and examine the relationship between postnatal growth velocity in infancy/early childhood and the risk of overweight/stunting in early adolescence. METHODS: The study used data from two child cohorts from urban (Birth to Twenty Cohort, South Africa) and rural (Lungwena Child Survival Study, Malawi) African settings. Mixed effect modelling was used to derive growth and peak growth velocities. T-tests were used to compare growth parameters and velocities between the two cohorts. Linear and logistic regression models were used to determine the relationship between growth velocity and early adolescent (ages 9-11 years) body mass index and odds of being overweight. RESULTS: Children in the BH cohort were significantly taller and heavier than those in the Lungwena cohort, and exhibited faster weight and height growth velocity especially in the first year of life (P < 0.05). No significant association was shown between baseline weight (αw ) and overweight in early adolescence (OR = 1.25, CI = 0.67, 2.34). The weight growth velocity parameter ßw was highly associated with odds of being overweight. Association between overweight in adolescence and weight velocity was stronger in infancy than in early childhood (OR at 3 months = 4.80, CI = 2.49, 9.26; OR at 5 years = 2.39, CI = 1.65, 3.47). CONCLUSION: High weight and height growth velocity in infancy, independent of size at birth, is highly associated with overweight in early adolescence. However, the long term effects of rapid growth in infancy may be dependent on a particular population's socio-economic status and level of urbanization.

Feasibility and validity of the ActiGraph GT3X accelerometer in measuring physical activity of Malawian toddlers.

AIM: To test the feasibility and validity of the ActiGraph GT3X accelerometer in measuring physical activity of rural Malawian toddlers. METHODS: Fifty-six children aged 16.0-18.5 months wore the accelerometer on their right hip for 7 days. We analysed days with a minimum of 600 min of wear time, excluding night time and periods when the unit registered zero for 20 consecutive minutes. The first and last days were excluded as they were incomplete. Accelerometer counts were compared with coded free play video recordings to define median accelerometer counts for sedentary, light, moderate and vigorous activity. Count cut points were defined for moderate to vigorous physical activity, with predictive validity assessed using a second set of video recordings. RESULTS: Median wear time was 797 min/day, with 79% of participants completing at least four eligible days. Accelerometer counts were significantly higher for observed moderate to vigorous physical activity, than lighter activity, with cut points of 208 counts/15 sec for vector magnitude and 35 counts/15 sec for vertical axis, showing sensitivity of 94.2% and 84.1% and specificity of 90.9% and 84.6%, respectively. CONCLUSION: The accelerometer proved a feasible and valid method of assessing physical activity among Malawian toddlers.

Continuation of linear growth failure and its association with cognitive ability are not dependent on initial length-for-age: a longitudinal study from 6 months to 11 years of age.

AIM: To examine linear growth and its association with cognitive ability at age 11 years among full-term singletons with varying degree of stunting or without stunting at age 6 months. METHODS: A total of 1516 Filipino term-born singletons were followed bimonthly from age 6 to 24 months and took cognitive and academic achievement tests at 11 years. The tests were factor-analysed to give a summary cognitive ability score. Linear regression was used to examine associations and adjust for covariates. RESULTS: Mean height-for-age Z-score (HAZ) was -1.0 at age 6 months and -2.4 at 24 months. HAZ at 6 months, change in HAZ from 6 to 24 months and change in HAZ from 24 months to 11 years were positively associated with cognitive ability at 11 years (each p < 0.001). The association was seen in all categories of HAZ at 6 months. CONCLUSION: In this setting where linear growth retardation was common, association between linear growth after age 6 months and cognitive ability in adolescence was not dependent on initial HAZ. Prevention of growth stunting may benefit all children regardless of their initial HAZ.

Seasonality of gestational weight gain and foetal growth in rural Malawi.

OBJECTIVE: To study seasonal variation of maternal anthropometry and newborn weights. DESIGN: A retrospective descriptive study from 1995 to 1998. SETTING: Lungwena, a rural health centre in Malawi, southeast Africa. SUBJECTS: One thousand and thirty two women with singleton pregnancy, a minimum of eight weeks antenatal follow-up and a baby who was weighed in the first month of life. Maternal data were sought from a health centre antenatal register and linked with infant information collected from a newborn clinic. RESULTS: The mean (SD) gestational weight and fundal height gains among the pregnant women were 240 (200) grams and 0.9 (0.2) cm/week and the mean (SD) newborn weight 3360 (530) grams. Gestational weight gains followed a seasonal pattern and they were typically highest among women delivering in the third quarter of the year (mean gain 250-300 g/week) and lowest among those delivering in January-May (mean gain 100-200 g/week) (p < 0.001 for seasonality, ANOVA). For maternal fundal height gains and newborn weights, seasonality was less obvious (but statistically significant, p < 0.05, ANOVA) and its pattern was influenced by year of study. On average, newborn weights peaked in the last quarter (mean 3350-3400 grams) and nadired during the second quarter (mean 3200-3300 grams). There was only a modest correlation between maternal weight gain in pregnancy and the weight of her newborn (Pearson's correlation coefficient 0.13). CONCLUSION: In rural Malawi, maternal weight gains during pregnancy are more strongly associated with season than fundal height gains or newborn weights. In adverse environmental conditions, foetal growth ismaintained at least partially at the expense of mother's nutritional status.

Lectin binding affinities of human milk fat globule (HMFG) membrane antigens.

Six biotinylated lectins with differing specificities and two monoclonal antibodies (III D 5 and III H 2) were used to characterize the sugar-residues in human milk fat globule (HMFG) membrane antigens. Immunoblotting analysis revealed that most of the antigens contain several sugars. However, the molecules exclusively reacting with anti-HMFG III D 5, a monoclonal antibody previously shown to detect antigen(s) positively correlating with the expression of estrogen receptors in mammary and gynaecological carcinomas, could only be stained with peanut agglutinin and Ricinus communis-lectins. One of these antigens, a 42-57 kDa molecule, was shown to have a complexed quaternary structure with galactose determining the antigenic specificity. It is suggested that the production of this glycoprotein in estrogen sensitive tissues results from activation of galactosyl-transferase-enzyme at the same time as the expression of estrogen receptors.

Breast milk from women living near Lake Malawi is high in docosahexaenoic acid and arachidonic acid.

Adequate long-chain polyunsaturated fatty acid (LCPUFA) intake is critical during the fetal and infant periods. We quantified fatty acid content of breast milk (n=718) and plasma from six month old infants (n=412) in southern Malawi, and in usipa (n=3), a small dried fish from Lake Malawi. Compared to global norms, Malawian breast milk fatty acid content (% of total fatty acids) was well above average levels of arachidonic acid [ARA] (0.69% vs. 0.47%) and docosahexaenoic acid [DHA] (0.73% vs. 0.32%). Average Malawian infant plasma ARA (7.5%) and DHA (3.8%) levels were comparable to those reported in infants consuming breast milk with similar fatty acid content. The amounts (mg) of DHA, EPA and ARA provided by a 3 oz (85 g) portion of dried usipa (1439, 659 and 360, respectively) are considerably higher than those for dried salmon. Usipa may be an important source of LCPUFA for populations in this region.

Effect of 12-month intervention with lipid-based nutrient supplements on physical activity of 18-month-old Malawian children: a randomised, controlled trial.

BACKGROUND/OBJECTIVES: This study measured the effects of dietary supplementation with lipid-based nutrient supplements (LNSs) on 18-month-old children's physical activity. SUBJECTS/METHODS: In a randomised, controlled, outcome-assessor blinded trial 1932 six-month-old children from Malawi received one of five interventions daily from 6-18 months of age: 10-g milk-LNS, 20-g milk-LNS, 20-g non-milk-LNS, 40-g milk-LNS or 40-g non-milk-LNS, or received no intervention in the same period (control). The control group received delayed intervention with corn-soy blend from 18-30 months. Physical activity was measured over 1 week by ActiGraph GT3X+ accelerometer at 18 months. Main outcome was mean vector magnitude accelerometer counts/15 s. Analyses were restricted to children with valid accelerometer data on at least 4 days with minimum 6 h of wearing time per day. RESULTS: Of the 1435 children recruited to this substudy, 1053 provided sufficient data for analysis. The mean (s.d.) vector magnitude accelerometer counts in the total sample were 307 (64). The difference (95% CI) in mean accelerometer counts, compared with the control group, was 8 (-6 to 21, P=0.258) in 10-g milk-LNS, 3 (-11 to 17, P=0.715) in 20-g milk-LNS, 5 (-8 to 19, P=0.445) in 20-g non-milk-LNS, 10 (-3 to 23, P=0.148) in 40-g milk-LNS and 2 (-12 to 16, P=0.760) in 40-g non-milk-LNS groups. CONCLUSIONS: Provision of 10-40 g doses of LNS daily for 12 months did not increase physical activity of Malawian toddlers.

Activity of CD4-Pseudomonas exotoxin against cells expressing diverse forms of the HIV and SIV envelope glycoproteins.

CD4(178)-PE40 is a genetically engineered hybrid toxin containing a portion of human CD4 linked to the translocation and ADP-ribosylation domains of Pseudomonas exotoxin A. In vitro, the molecule has been shown to selectively kill cells expressing the envelope glycoproteins of human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV), and to inhibit HIV spread. In this report we examine the activity of the hybrid toxin against cells expressing diverse forms of the HIV and SIV envelope glycoproteins, encoded by recombinant vaccinia virus vectors. The activity of CD4(178)-PE40 was found to be unaffected by mutations in the HIV-1 or HIV-2 envelope glycoprotein genes, which prevent normal proteolytic processing of the corresponding gp160 precursor molecules. Cells expressing a mutant HIV-1 envelope glycoprotein lacking most of the cytoplasmic tail of the gp41 transmembrane subunit were also sensitive to the hybrid toxin. Most interestingly, HIV-1, HIV-2, and SIVmac envelope glycoprotein molecules known to have widely differing affinities for CD4 were found to be comparably effective at mediating sensitivity to CD4(178)-PE40. By virtue of its ability to kill infected cells, the hybrid toxin inhibited the spread of SIVmac in vitro. These results indicate that CD4(178)-PE40 is active against cells expressing HIV and SIV envelope glycoproteins with a diverse array of structural differences.

No evidence for true HTLV-I or HIV-1 antibodies in Finnish Lapps.

Previous seroepidemiologic studies have suggested that in addition to certain subtropical and tropical parts of the world, human T cell leukemia virus type I (HTLV-I) may be endemic in the arctic regions, too. We studied 111 sera collected from original inhabitants of Finnish Lapland with ELISA and Western blot analysis for antibodies to HTLV-I and human immunodeficiency virus type 1 (HIV-1). No true positive sera for either virus were found in the confirmatory Western blot assays, albeit 6 and 2%, respectively, were positive in the screening ELISA assays. Despite the small sample size this survey does not support the hypothesis that HTLV-I would be endemic in the Arctic.

Predictive factors associated with significant urinary tract abnormalities in infants with pyelonephritis.

BACKGROUND: Major urinary tract abnormalities are detected in 20 to 40% of infants with acute pyelonephritis (APN). Early detection of structural defects is essential for protecting the kidneys from reinfection and subsequent scarring. The purpose of this study was to investigate whether any factors present during the acute phase of infection could predict the presence of existing significant urinary tract abnormalities in infants. METHODS: A prospective study of 180 infants, aged 1 to 24 months, with APN was conducted. Blood and urine samples were collected. Renal ultrasound (US) was performed within 0 to 6 days from admission. Final diagnosis of the urinary tract anatomy was elucidated using the results of two or more radiologic imaging studies. RESULTS: Risk factors for the presence of significant urinary tract abnormalities in infants were pathogens other than Escherichia coli in urine [relative risk (RR) 3.4, 95% confidence interval (CI) 2.2 to 5.3; P = 0.001], positive blood culture (RR 2.3, 95% CI 1.3 to 4.0; P = 0.039), young age (1 to 6 months) (RR 2.2, 95% CI 1.3 to 3.9; P = 0.004), lack of papG adhesin genes of E. coli in urine (RR 2.1, 95% CI 1.2 to 3.9; P = 0.016) and abnormal renal US (RR 2.0, 95% CI 1.2 to 3.4; P = 0.008). CONCLUSIONS: Infants 1 to 6 months of age with APN caused by bacteria other than E. coli or by papG-negative E. coli strain, positive blood culture and abnormal renal US carry an increased risk for significant urinary tract abnormalities and need enforced follow-up.

Use of immunohistochemical staining panel for characterisation of ovarian neoplasms.

Eighty five ovarian epithelial and non-epithelial tumours were studied by peroxidase histochemical staining for their reactivity with six monoclonal human milk fat globule (HMFG) antibodies, peanut agglutinin (PNA) lectin, and a monoclonal cytokeratin antibody. HMFG IIIC12 and cytokeratin antibodies distinguished epithelial from non-epithelial tumours. The staining patterns of mucinous and serous tumours were essentially different from each other; poorly differentiated anaplastic carcinomas showed similar antigenic content to that of the serous cystadenocarcinomas. Furthermore, staining with PNA lectin and HMFG antibodies was useful in distinguishing clear cell carcinomas from other malignant epithelial tumours of the ovary.

Anti-HIV effects of CD4-Pseudomonas exotoxin on human lymphocyte and monocyte/macrophage cell lines.

CD4(178)-PE40 is a recombinant protein consisting of the HIV envelope glycoprotein-binding region of human CD4 linked to active domains of Pseudomonas aeruginosa exotoxin A. The hybrid toxin selectively kills HIV-infected human T-cell lines and protects against HIV spread in mixtures of uninfected and infected cells. We now report that CD4(178)-PE40 also selectively kills chronically HIV-1-infected cells of monocyte/macrophage lineage. The results provide further support for therapeutic use of this hybrid toxin in the treatment of HIV-infected individuals.

Antenatal and perinatal predictors of infant mortality in rural Malawi.

BACKGROUND: The slow pace in the reduction of infant mortality in sub-Saharan Africa has partially been attributed to the epidemic of human immunodeficiency virus (HIV) infection. To facilitate early interventions, antenatal and perinatal predictors of 1st year mortality were identified in a rural community in southern Malawi. METHODS: A cohort of 733 live born infants was studied prospectively from approximately 24 gestation weeks onwards. Univariate analysis was used to determine relative risks for infant mortality after selected antenatal and perinatal exposures. Multivariate modelling was used to control for potential confounders. FINDINGS: The infant mortality rate was 136 deaths/1000 live births. Among singleton newborns, the strongest antenatal and perinatal predictors of mortality were birth between May and July, maternal primiparity, birth before 38th gestation week, and maternal HIV infection. Theoretically, exposure to these variables accounted for 22%, 22%, 17%, and 15% of the population attributable risk for infant mortality, respectively. INTERPRETATION: The HIV epidemic was an important but not the main determinant of infant mortality. Interventions targetting the offspring of primiparous women or infants born between May and July or prevention of prematurity would all have considerable impact on infant survival.

The expression of CEA, CA 19-9 and HMFG antigens in ovarian clear-cell and endometrioid carcinomas.

The tumour antigen expression of ovarian and endometrial endometrioid carcinomas, ovarian clear-cell carcinomas as well as endometrial and cervical clear-cell carcinomas were immunohistochemically compared. Of special interest were potential differences between the endometrioid and clear-cell carcinomas of the ovary. The expression of CEA and CA 19-9 tumour antigens in all these tumour types was heterogeneous, with 10-20% of the cases being positive for CEA and 40-75% being positive for CA 19-9. In contrast, HMFG IIIC 12, a monoclonal antibody originally directed against human milk fat globule (HMFG) membrane antigens, invariably detected a corresponding antigen on every case of these tumour types. Another HMFG antibody, SM IF 3, on the other hand, detected antigenic material on all clear-cell tumour types, but only rarely on endometrioid tumours of the ovary or endometrium. While HMFG IIIC 12 detects an antigen present on all ovarian, endometrial and mammary carcinomas, antibody SM IF 3 thus appears to be more restricted in its staining patterns. Our results with both of these antibodies indicate that ovarian clear-cell carcinomas and ovarian endometrioid carcinomas have antigenic differences, which provides further evidence that they belong to different tumour entities.

Verbal autopsy as a technique to establish causes of infant and child mortality.

In order to develop a method for retrospectively establishing causes of paediatric deaths in Mangochi District, Malawi, we translated a verbal autopsy questionnaire into Chichewa and Chiyao. This form was used to interview guardians of 82 children who died in hospital between June 1993 and May 1994. Based on the guardians' answers, we were able to determine the causes of death for these children. According to our experience, verbal autopsy appeared simple to administer and relatively easy to interpret. To test the validity of this method in southern Malawi, we compared its results to hospital diagnosis, using a sample of 36 children who had died in hospital and whose records could be located. In this preliminary validation, we found a high sensitivity and specificity for the most common childhood illnesses in the area. Thus, verbal autopsy appeared a promising method of collecting population-based information about paediatric mortality, the full applicability of which should be tested in larger studies in Malawi.