Droplet digital PCR quantifies host inflammatory transcripts in feces reliably and reproducibly

AbstractThe gut is the most extensive, interactive, and complex interface between the human host and the environment and therefore a critical site of immunological activity. Non-invasive methods to assess the host response in this organ are currently lacking. Feces are the available analyte which have been in proximity to the gut tissue.We applied a method of concentrating host transcripts from fecal specimens using a existing bead-based affinity separation method for nucleic acids and quantified transcripts using droplet digital PCR (ddPCR) to determine the copy numbers of a variety of key transcripts in the gut immune system. ddPCR compartmentalizes the reaction in a small aqueous droplet suspended in oil, and counts droplets as either fluorescent or non-fluorescent. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used to normalize transcript concentration.This method was applied to 799 fecal samples from rural Malawian children, and over 20,000 transcript concentrations were quantified. Host mRNA was detected in >99% samples, a threshold for target detection was established at an average expression of 0.02 copies target/GAPDH, above which correlation coefficient between duplicate measurements is >0.95. Quantities of transcript detected using ddPCR were greater than standard qPCR. Fecal sample preservation at the time of collection did not require immediate freezing or the addition of buffers or enzymes. Measurements of transcripts encoding immunoactive proteins correlated with a measure of gut inflammation in the study children, thereby substantiating their relevance. This method allows investigators to interrogate gene expression in the gut

Supplementary feeding in the care of the wasted HIV infected patient

Wasting and food insecurity are commonly seen in patients receiving antiretroviral treatment (ART) programs in sub-Saharan Africa and south Asia, and supplementary feeding is often offered in conjunction with ART. Evidence for the effectiveness of such supplementary feeding is scant. A randomised, investigatorblinded, controlled clinical trial of two types of supplementary food, corn/ soy blended flour and a ready-to-use peanut butterbased lipid paste, in wasted adults in Blantyre, Malawi is describedand the results summarised. A historical control group who did not receive supplementary food is described as well. Provision of about half of the daily energy requirement as a supplementary food for 14 weeks resulted in more rapid restoration of a normal BMI; and the energy-dense, ready-to-use paste was associated with more rapid weight gain than the blended flour. Survival was similar among the 3 groups. The strong association between lower BMI and survival indirectly suggests that there may well beclinical benefit from supplementary feeding in this population. No differences were seen in ART adherence or quality of life with more rapid restoration of BMI. Further research is urgently needed concerning the widespread practice of supplementary feeding in HIV/ AIDS care to most effectively utilize this intervention

Child Survival in sub Sahara Africa: the role of CAPGAN and regional child health practitioners & scientists

Close to a decade ago the Millennium Development Goals (MDG) were developed and unanimously accepted by the General Assembly of the United Nations. The think tank behind the MDGs, the Earth Institute at ColumbiaUniversity in New York led by the economist Jeffry Sachs , developed 8 overarching goals, which -if achieved- would liberate the world from povert

Effect of cowpea flour processing on the chemical properties and acceptability of a novel cowpea blended maize porridge

Childhood growth stunting is a pervasive problem in Malawi and is in large part due to low quality complementary foods and chronic gut inflammation. Introducing legumes such as cowpea (Vigna unguiculata) into the complementary diet has the potential to improve childhood growth by improving diet quality through improvements in macro- and micronutrients and also by reducing gut inflammation. However, cowpea is relatively underutilized in complementary feeding in Malawi due to its strong taste, long processing time, and high energy requirements for processing. Effective utilization of cowpea in complementary feeding requires processing which may affect chemical composition as well as sensory quality. The present study evaluated the effect of processing on the retention of zinc, crude fibre, and flavonoid in roasted, boiled, and dehulled cowpea flours, and assessed the acceptability of maize porridge (70%) enriched with one of the three cowpea flours (30%). Roasting, dehulling, and boiling did not have any effect on zinc content. Crude fibre content increased after processing by all methods. Processing had no effect on measurable flavonoids. Roasted, boiled, and dehulled cowpea blended maize porridges were acceptable to children with mean quantities of leftover food of less than 3g from the given 100g. Caregivers also rated the blended flours to be highly acceptable to them as well, with maize porridge blended with dehulled cowpea flour the most acceptable to both children and caregivers. These results demonstrate that cowpea flour, processed by any of these three different methods, could serve as a useful addition to maize porridge for complementary feeding of children in sub-Saharan Africa

Consumption of animal-source protein is associated with improved height-for-age z scores in rural Malawian children aged 12–36 months

Linear growth faltering, caused by insufficient diet, recurrent infections and environmental enteric dysfunction (EED), continues to plague young children in low- and middle-income countries (LMICs). Diets in LMICs are primarily plant based, and thus have poor-quality protein and low levels of essential micronutrients. The aim of this study was to assess the association of the type and protein quality of food consumed with stunting, EED and acute malnutrition in children aged 6⁻36 months in Limera and Masenjere, two rural Southern Malawian communities. This is a secondary analysis of two randomized controlled trials that tested the effects of common bean and cowpea flour on stunting in children aged 6⁻36 months. We used data from two interactive 24-h dietary recalls conducted 12 weeks after enrolment into each trial. Food intakes were compared between the regions using Chi-square and Student’s t-test. There were 355 children that participated in the dietary recalls. The diets of children were of poor quality, but the children from Limera consumed more fish (54% vs. 35%, p = 0.009) and more bioavailable protein (26.0 ± 10.3 g/day vs. 23.1 ± 8.1 g/day, p = 0.018, respectively) than children in Masenjere. Food type and protein quality were not associated with any of the outcomes except an association between animal protein consumption and improvement in height-for-age z scores in children aged 12⁻36 months (p = 0.047). These findings support the notion that animal-source food (ASF) consumption in this vulnerable population promotes linear growth

Environmental enteric dysfunction and the fecal microbiota in malawian children

Environmental enteric dysfunction (EED) is often measured with a dual sugar absorption test and implicated as a causative factor in childhood stunting. Disturbances in the gut microbiota are hypothesized to be a mechanism by which EED is exacerbated, although this supposition lacks support. We performed 16S ribosomal RNA gene sequencing of fecal samples from 81 rural Malawian children with varying degrees of EED to determine which bacterial taxa were associated with EED. At the phyla level, Proteobacteria abundance is reduced with severe EED. Among bacterial genera, Megasphaera, Mitsuokella, and Sutterella were higher in EED and Succinivibrio, Klebsiella, and Clostridium_XI were lower in EED. Bacterial diversity did not vary with the extent of EED. Though EED is a condition that is typically believed to affect the proximal small bowel, and our focus was on stool, our data do suggest that there are intraluminal microbial differences that reflect, or plausibly lead to, EED

Factors affecting tether use and misuse

This project investigated factors relating to tether use and misuse. Volunteer testing was performed with 37 subjects on 16 different vehicles using 2 forward-facing child restraints (Britax Marathon 70 or the Evenflo Triumph), with each subject performing 8 child restraint installations on a set of four vehicles. Vehicles were selected to provide a variety of general tether locations (filler panel, upper seatback, lower seatback, floor, or roof), as well as a variety of recommended tether routings with respect to the head restraint: under, over, around. Simple instruction regarding the LATCH system was provided after the fourth trial. Subjects used the tether in 89% of the 294 forward-facing trials and attached the tether correctly in 57% of installations. Subjects were more likely to use the tether if they were less than age 40, had previous tether experience, if the tether was located on a filler panel, and if the vehicle did not have any potentially confusing hardware. In addition, tether use was 83% in the first four trials and increased to 95% in the last four trials after instruction was provided. Subjects had the greatest difficulty in the pickup truck, which use loops of webbing as a router for the installed position and the tether anchor for the adjacent position; the tether was attached correctly in only 11 percent of installations. Tethers were more likely to be used when the tether anchor was located on the filler panel of sedans, which had a use rate of 95 percent, compared to when the anchor was located on the floor, roof, or seatback, which had use rates ranging from 79 to 89 percent. Tethers were less likely to be attached correctly when there was potentially confusing hardware present, 47 percent, compared to 70 percent. In addition, tether anchors located on the filler panel or mid seatback had higher rates of correct attachment, 60 and 69 percent , respectively, than those on the floor, roof, or lower seatback, which all had correct attachment rates lower than 50 percent . No vehicle tether hardware characteristics or vehicle manual directions were associated specifically with correct tether routing and head restraint position. Installations involving the single tether strap were 10 times as likely to have the tether attached correctly and 1.7 times as likely to be routed correctly and have the head restraint positioned correctly, compared with installations with the v-shaped tether. Lack of instruction in most vehicle owner’s manuals regarding the routing of a V-style tether more challenging to use. With the single strap-style tether, it was more straightforward to have the tether strap flat and pull it tight, as well as to route it as directed. With the V-style tether, the adjustment hardware was often located underneath or close to the head restraint when installed in the vehicle, making it difficult to tighten. Recommendations to reduce tether misuse include labeling tether anchors, eliminating confusing hardware, allowing any head restraint position (including removal), providing instruction for routing V-style tethers, allowing options in tether routing, and redesigning tether anchors/routers found in pickup trucks.Insurance Institute for Highway Safetyhttp://deepblue.lib.umich.edu/bitstream/2027.42/106031/1/102990.pd

Antibiotics as part of the management of severe acute malnutrition

BACKGROUND: Severe acute malnutrition contributes to 1 million deaths among children annually. Adding routine antibiotic agents to nutritional therapy may increase recovery rates and decrease mortality among children with severe acute malnutrition treated in the community. METHODS: In this randomized, double-blind, placebo-controlled trial, we randomly assigned Malawian children, 6 to 59 months of age, with severe acute malnutrition to receive amoxicillin, cefdinir, or placebo for 7 days in addition to ready-to-use therapeutic food for the outpatient treatment of uncomplicated severe acute malnutrition. The primary outcomes were the rate of nutritional recovery and the mortality rate. RESULTS: A total of 2767 children with severe acute malnutrition were enrolled. In the amoxicillin, cefdinir, and placebo groups, 88.7%, 90.9%, and 85.1% of the children recovered, respectively (relative risk of treatment failure with placebo vs. amoxicillin, 1.32; 95% confidence interval [CI], 1.04 to 1.68; relative risk with placebo vs. cefdinir, 1.64; 95% CI, 1.27 to 2.11). The mortality rates for the three groups were 4.8%, 4.1%, and 7.4%, respectively (relative risk of death with placebo vs. amoxicillin, 1.55; 95% CI, 1.07 to 2.24; relative risk with placebo vs. cefdinir, 1.80; 95% CI, 1.22 to 2.64). Among children who recovered, the rate of weight gain was increased among those who received antibiotics. No interaction between type of severe acute malnutrition and intervention group was observed for either the rate of nutritional recovery or the mortality rate. CONCLUSIONS: The addition of antibiotics to therapeutic regimens for uncomplicated severe acute malnutrition was associated with a significant improvement in recovery and mortality rates. (Funded by the Hickey Family Foundation and others; ClinicalTrials.gov number, NCT01000298.