RESUMO
Ready-to-use therapeutic food (RUTF) is a shelf-stable, low moisture, energy dense medicinal food composed of peanut butter, vegetable oils, milk powder, a multiple micronutrient premix and sugar. RUTF is used by millions of children annually to treat malnutrition. After mixing, RUTF is a semisolid covered with oil. To produce a homogenous RUTF, hydrogenated vegetable oils are incorporated in small quantities. This study utilized a benchtop methodology to test the effect of RUTF ingredients on oil separation. An acceptable oil separation was <4%. This method compared 15 different vegetable oil stabilizers with respect to oil separation. The dynamic progression of oil separation followed a Michaelis-Menten pattern, reaching a maximum after 60 days when stored at 30 °C. Hydrogenated vegetable oils with triglyceride or 50% monoglycerides reduced the oil separation to acceptable levels. The additive showing the largest reduction in oil separation was used in an industrial trial, where it also performed acceptably. In conclusion, fully hydrogenated soybean and rapeseed oil added as 1.5% controlled oil separation in RUTF.
RESUMO
High Campylobacter prevalence during early childhood has been associated with stunting and environmental enteric dysfunction (EED), especially in low resource settings. This study assessed the prevalence, diversity, abundance, and co-occurrence of Campylobacter spp. in stools from children in a rural area of eastern Ethiopia and their association with microbiome, diarrhea, and EED in children. Stool samples (n = 100) were collected from randomly selected children (age range: 360-498 days) in five kebeles in Haramaya District, Ethiopia. Diarrhea, compromised gut permeability, and gut inflammation were observed in 48, 45, and 57% of children, respectively. Campylobacter prevalence and species diversity were assessed using PCR and meta-total RNA sequencing (MeTRS). The prevalence of Campylobacter spp. in the children's stools was 50% (41-60%) by PCR and 88% (80-93.6%) by MeTRS (P < 0.01). Further, seven Campylobacter species (Campylobacter jejuni, Campylobacter upsaliensis, Campylobacter hyointestinalis, Campylobacter coli, Campylobacter sp. RM6137, uncultured Campylobacter sp., and Campylobacter sp. RM12175) were detected by MeTRS in at least 40% of children stools in high abundance (>1.76-log read per million per positive stool sample). Four clusters of Campylobacter species (5-12 species per cluster) co-occurred in the stool samples, suggesting that Campylobacter colonization of children may have occurred through multiple reservoirs or from a reservoir in which several Campylobacter species may co-inhabit. No associations between Campylobacter spp., EED, and diarrhea were detected in this cross-sectional study; however, characteristic microbiome profiles were identified based on the prevalence of Campylobacter spp., EED severity, and diarrhea. Forty-seven bacterial species were correlated with Campylobacter, and 13 of them also correlated with gut permeability, gut inflammation and/or EED severity. Forty-nine species not correlated with Campylobacter were correlated with gut permeability, gut inflammation, EED severity and/or diarrhea. This study demonstrated that (1) in addition to C. jejuni and C. coli, multiple non-thermophilic Campylobacter spp. (i.e., Campylobacter hyointestinalis, Campylobacter fetus, and Campylobacter concisus) were frequently detected in the children's stools and (2) the Campylobacter, gut permeability, gut inflammation, EED severity, and diarrhea were associated with characteristic microbiome composition. Additional spatial and longitudinal studies are needed to identify environmental reservoirs and sources of infection of children with disparate Campylobacter species and to better define their associations with EED in low-income countries.
Assuntos
Infecções por Campylobacter , Campylobacter , Microbiota , Infecções por Campylobacter/epidemiologia , Criança , Pré-Escolar , Estudos Transversais , Diarreia/epidemiologia , Etiópia/epidemiologia , HumanosRESUMO
OBJECTIVES: Environmental enteric dysfunction (EED), a clinically asymptomatic condition characterized by inflammation of the small bowel mucosa, villous atrophy, and increased gut permeability, is common among children in developing countries. Because of abnormal gut mucosa and altered gut microbiome, EED could potentially affect the metabolism and enterohepatic circulation of bile acids. METHODS: In 313 children, aged 12 to 59 months, EED was assessed by the dual sugar absorption test. Serum bile acids were measured using stable-isotope liquid chromatography-tandem mass spectrometry. RESULTS: In the overall study population, serum cholic acid and chenodeoxycholic acid were lower, whereas glycocholic acid, taurodeoxycholic acid, glycodeoxycholic acid, glycolithocholic acid, and glycoursodeoxycholic acid were significantly higher at older ages. Independent of age, serum taurochenodeoxycholic acid, tauromuricholic acid, and glycoursodeoxycholic acid were significantly different between 244 children with EED and 69 children without EED. Total serum bile acids (median, interquartile range) were 4.51 (2.45, 7.51) and 5.10 (3.32, 9.01) µmol/L in children with and without EED, respectively (age-adjusted, Pâ=â0.0009). The proportion of bile acids conjugated with taurine instead of glycine was higher in children with EED (Pâ<â0.0001). CONCLUSIONS: EED is associated with altered bile acid metabolism in young children in rural Malawi. Further work is needed to determine the generalizability of these findings in other study populations.
Assuntos
Ácidos e Sais Biliares/sangue , Síndromes de Malabsorção/diagnóstico , Biomarcadores/sangue , Estudos de Casos e Controles , Pré-Escolar , Cromatografia Líquida , Estudos Transversais , Países em Desenvolvimento , Feminino , Humanos , Lactente , Intestino Delgado/metabolismo , Síndromes de Malabsorção/sangue , Malaui , Masculino , Saúde da População Rural , Espectrometria de Massas em TandemRESUMO
OBJECTIVE: Environmental enteric dysfunction (EED) can be assessed by the lactulose:mannitol (L:M) test. Our objective was to determine if selected host fecal transcripts were correlated with EED, and whether transcripts and clinical characteristics could be used to predict EED in rural African children. METHODS: Demographic and sanitation characteristics, along with L:M testing and host fecal transcript analyses from 798 asymptomatic Malawian children aged 12 to 61 months were compared with linear growth over the subsequent 3 months. Fecal host mRNA analysis included quantification of expression of 18 transcripts associated with L:M. Permeability was categorized as normal (L:M ≤ 0.15), moderate (0.15Assuntos
Enteropatias/diagnóstico
, Mucosa Intestinal/metabolismo
, Lactulose/metabolismo
, Manitol/metabolismo
, RNA Mensageiro/metabolismo
, África
, Desenvolvimento Infantil
, Pré-Escolar
, Meio Ambiente
, Fezes/química
, Feminino
, Humanos
, Lactente
, Enteropatias/complicações
, Masculino
, População Rural
RESUMO
RF2a is a bZIP transcription factor that regulates expression of the promoter of rice tungro bacilliform badnavirus. RF2a is predicted to include three domains that contribute to its function. The results of transient assays with mutants of RF2a from which one or more domains were removed demonstrated that the acidic domain was essential for the activation of gene expression, although the proline-rich and glutamine-rich domains each played a role in this function. Studies using fusion proteins of different functional domains of RF2a with the 2C7 synthetic zinc finger DNA-binding domain showed that the acidic region is a relatively strong activation domain, the function of which is dependent on the context in which the domain is placed. Data from transgenic plants further supported the conclusion that the acidic domain was important for maintaining the biological function of RF2a. RF2a and TBP (TATA-binding protein) synergistically activate transcription in vitro (Zhu, Q., Ordiz, M. I., Dabi, T., Beachy, R. N., and Lamb, C. (2002) Plant Cell 14, 795-803). In vitro and in vivo assays showed that RF2a interacts with TBP through the glutamine-rich domain but not the acidic domain. Functional analysis of such interactions indicates that the acidic domain activates transcription through mechanisms other than via the direct recruitment of TBP.
Assuntos
Oryza/metabolismo , Proteínas de Plantas , Transativadores/química , Transativadores/genética , Fatores de Transcrição de Zíper de Leucina Básica , Eletroforese em Gel de Poliacrilamida , Regulação da Expressão Gênica , Genes Reporter , Glutamina/química , Immunoblotting , Modelos Genéticos , Mutação , Fenótipo , Plantas Geneticamente Modificadas , Plasmídeos/metabolismo , Prolina/química , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/metabolismo , Rhizobium/metabolismo , Nicotiana/genética , Nicotiana/microbiologia , Fatores de Transcrição/química , Fatores de Transcrição/genética , Transcrição Gênica , Dedos de ZincoRESUMO
TATA binding protein (TBP) and transcription factor IIB (TFIIB) are key factors for the assembly of eukaryotic transcription initiation complexes. We used a rice whole-cell extract in vitro transcription system to characterize the functional interactions of recombinant plant TBP and TFIIB. Bacterially expressed rice TBP (OsTBP2) bound to the TATA box of the rice pal gene encoding phenylalanine ammonia-lyase, caused DNA bending, and enhanced basal transcription from the pal promoter in a TATA box-dependent manner. Recombinant rice TFIIB (OsTFIIB) stimulated the DNA binding and bending activities of OsTBP2 and synergistically enhanced OsTBP2-mediated transcription from the pal promoter and the promoter of Rice tungro bacilliform virus but not from the barley pr1 promoter. We also demonstrate a physical interaction between OsTBP2 and RF2a, a rice bZIP transcription factor that bound to the box II cis element of the promoter of Rice tungro bacilliform virus, resulting in enhanced transcription from the viral promoter. Enhancement of rice whole-cell extracts with recombinant transcription factors thus provides a powerful tool for the in vitro determination of plant gene regulation mechanisms. We conclude that OsTBP2 undergoes promoter-specific functional interactions with both the basal transcription factor OsTFIIB and the accessory transcription factor RF2a.