Early Life Child Micronutrient Status, Maternal Reasoning, and a Nurturing Household Environment have Persistent Influences on Child Cognitive Development at Age 5 years : Results from MAL-ED

Funding Information: The Etiology, Risk Factors and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development Project (MAL-ED) is carried out as a collaborative project supported by the Bill & Melinda Gates Foundation, the Foundation for the NIH, and the National Institutes of Health/Fogarty International Center. This work was also supported by the Fogarty International Center, National Institutes of Health (D43-TW009359 to ETR). Author disclosures: BJJM, SAR, LEC, LLP, JCS, BK, RR, RS, ES, LB, ZR, AM, RS, BN, SH, MR, RO, ETR, and LEM-K, no conflicts of interest. Supplemental Tables 1–5 and Supplemental Figures 1–3 are available from the “Supplementary data” link in the online posting of the article and from the same link in the online table of contents at https://academic.oup.com/jn/. Address correspondence to LEM-K (e-mail: [email protected]). Abbreviations used: HOME, Home Observation for Measurement of the Environment inventory; MAL-ED, The Etiology, Risk Factors, and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development Project; TfR, transferrin receptor; WPPSI, Wechsler Preschool Primary Scales of Intelligence.Peer reviewe

Early life child micronutrient status, maternal reasoning, and a nurturing household environment have persistent influences on child cognitive development at age 5 years: Results from MAL-ED

Background: Child cognitive development is influenced by early-life insults and protective factors. To what extent these factors have a long-term legacy on child development and hence fulfillment of cognitive potential is unknown. Objective: The aim of this study was to examine the relation between early-life factors (birth to 2 y) and cognitive development at 5 y. Methods: Observational follow-up visits were made of children at 5 y, previously enrolled in the community-based MAL-ED longitudinal cohort. The burden of enteropathogens, prevalence of illness, complementary diet intake, micronutrient status, and household and maternal factors from birth to 2 y were extensively measured and their relation with the Wechsler Preschool Primary Scales of Intelligence at 5 y was examined through use of linear regression. Results: Cognitive T-scores from 813 of 1198 (68%) children were examined and 5 variables had significant associations in multivariable models: mean child plasma transferrin receptor concentration (β: −1.81, 95% CI: −2.75, −0.86), number of years of maternal education (β: 0.27, 95% CI: 0.08, 0.45), maternal cognitive reasoning score (β: 0.09, 95% CI: 0.03, 0.15), household assets score (β: 0.64, 95% CI: 0.24, 1.04), and HOME child cleanliness factor (β: 0.60, 95% CI: 0.05, 1.15). In multivariable models, the mean rate of enteropathogen detections, burden of illness, and complementary food intakes between birth and 2 y were not significantly related to 5-y cognition. Conclusions: A nurturing home context in terms of a healthy/clean environment and household wealth, provision of adequate micronutrients, maternal education, and cognitive reasoning have a strong and persistent influence on child cognitive development. Efforts addressing aspects of poverty around micronutrient status, nurturing caregiving, and enabling home environments are likely to have lasting positive impacts on child cognitive development.publishedVersio

DiagnÃstico microbiolÃgico, imunoenzimÃtico e molecular e perfil de genes associados à virulÃncia de Campylobacter

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgicoCampylobacter sp. à uma importante causa de enterite de origem alimentar com alta incidÃncia na populaÃÃo infantil de paÃses em desenvolvimento. No entanto, o diagnÃstico especÃfico de sua etiologia segue como um desafio, visto que mÃtodos moleculares e imunoenzimÃticos tÃm se mostrado mais sensÃveis. Postulamos que o conhecimento de sua virulÃncia e o diagnÃstico especÃfico possam ajudar na identificaÃÃo e potencial controle da campilobacteriose na infÃncia. Foi determinada a etiologia de diarreia por Campylobacter sp., em um estudo transversal sobre diarreia em crianÃas de 0-36 meses residentes da Ãrea urbana de Fortaleza, CearÃ, Brasil, que necessitaram de atendimento mÃdico de urgÃncia por causa de doenÃa diarreica. ApÃs a aprovaÃÃo Ãtica do estudo, um questionÃrio foi aplicado para qualificar as condiÃÃes clÃnicas apresentadas por cada crianÃa no momento da admissÃo. O DNA foi extraÃdo diretamente de amostras fecais coletadas de 226 crianÃas. Para a detecÃÃo do agente etiolÃgico, utilizamos diagnÃstico molecular (PCR e RT-PCR) e diagnÃstico imunoenzimÃtico (ELISA), alÃm da detecÃÃo de genes associados à virulÃncia de C. jejuni (PCR). Campylobacter sp. foi encontrado em 8,9% (20/225) das amostras, por diagnÃstico microbiolÃgico convencional. C. jejuni e C. coli foram detectados em 19,5% (44/226) e 1,3% (3/226) das amostras diarreicas, respectivamente, por PCR. Os diagnÃsticos por RT-PCR e ELISA alcanÃaram 26,7% (60/225) e 37,9% (58/153), respectivamente. Quando considerada a combinaÃÃo de diagnÃsticos (positividade no diagnÃstico microbiolÃgico ou no imunoenzimÃtico e ao menos em um dos testes moleculares) a prevalÃncia encontrada foi de 16,4% (37/226). A concordÃncia entre os testes para diagnÃstico utilizados foi de moderada a regular, de acordo com o Ãndice de Kappa. Genes associados à virulÃncia foram detectados nas seguintes proporÃÃes de amostras positivas para C. jejuni: flaA, 79,5% (35/44); racR, 97,7% (43/44) e dnaJ, 88,6% (39/44) â relacionados à adesÃo bacteriana e colonizaÃÃo; ciaB, 97,7% (43/44); pldA, 45,4% (20/44) e pVir 0% (0/44) â relacionados à invasÃo; e cdtABC em 95,4% (42/44) das amostras, operon relacionado à produÃÃo da toxina citoletal distensora (CDT). Sinais e sintomas especÃficos, tais como sangue nas fezes, vÃmito, febre e/ou dor abdominal, apesar de bastante frequentes, nÃo foram associados com a detecÃÃo de C. jejuni. O perfil de distribuiÃÃo dos genes de virulÃncia de C. jejuni nÃo apresentou correlaÃÃo com a apresentaÃÃo clÃnica da doenÃa, mesmo quando tal perfil foi categorizado de acordo com a funÃÃo das proteÃnas codificadas pelos genes, o que nos leva a crer que outros fatores, talvez relacionados à susceptibilidade do hospedeiro, possam ser mais importantes do que a variabilidade genÃtica do micro-organismo. ConcluÃmos que Campylobacter sp. foi detectado em percentual relevante da populaÃÃo estudada, principalmente quando os mÃtodos diagnÃsticos foram utilizados de forma combinada. Em geral, os genes de virulÃncia foram detectados em uma alta proporÃÃo das amostras positivas para C. jejuni, embora os genes relacionados à invasÃo tenham sido menos frequentemente encontrados. Corroboramos dados de outros grupos sobre a necessidade de revisÃo do diagnÃstico para Campylobacter sp. em prol da inclusÃo de metodologias mais sensÃveis e espÃcie-especÃficas, alÃm da busca por marcadores para inflamaÃÃo intestinal e fatores preditivos de cultura negativa.Campylobacter sp. is an important cause of food-borne gastroenteritis with high incidence in children living in developing countries. However, the specific diagnosis of its etiology remains as a challenge, since conventional diagnosis by culture is now challenged by molecular and immunoenzymatic methods, which have greater sensitivity. We postulate that the knowledge of its virulence and specific diagnosis may assist in identifying and potentially controling campylobacteriosis in childhood. We determined the etiology of Campylobacter sp. associated diarrhea, in a cross-sectional study of diarrhea in children aged 0-36 months from the urban area of Fortaleza, CearÃ, Brazil, who required emergency medical care because of diarrheal disease. After ethical approval of the study, a questionnaire was applied to describe the clinical conditions presented by each child at the time of admission. DNA was extracted directly from fecal samples collected from 226 children. For the determination of the etiologic agent we used molecular diagnostics (PCR and RT-PCR) and diagnostic immunoassay (ELISA), besides the detection of virulence associated genes of C. jejuni (PCR). Campylobacter sp. was found in 8.9% (20/225) of the samples by conventional microbiological diagnosis. C. jejuni and C. coli were detected in 19.5% (44/226) and 1.3% (3/226) of the diarrheic samples, respectively. The diagnostic RT-PCR and ELISA reached 26.7% (60/225) and 37.9% (58/153) of positivity, respectively. When considering the combination of diagnostic (positive in microbiological diagnosis or immunoassay and at least one of the molecular tests) the prevalence was 16.4% (37/226). The agreement between the tests used for diagnosis was moderate to regular, according to Kappa index. The presence of C. jejuniÂs virulence-associated genes that encode proteins related to the pathogenesis of micro-organism were detected in the following proportions of C. jejuni-positive DNA samples: flaA, 79.5% (35/44); racR, 97.7% (43/44) and dnaJ, 88.6% (39/44) â related to bacterial adhesion and colonization; ciaB, 97.7 % (43/44); pldA, 45.4% (20/44) and pVir 0% (0/44) â related to invasion, and cdtABC in 95.4% (42/44) of samples related to citoletal distending toxin (CDT). Specific signs and symptoms such as blood in the stool, vomiting, fever and/or abdominal pain, although quite frequent, were not associated with the detection of C. jejuni. The distribution profile of C. jejuniÂs virulence genes was not correlated with the clinical presentation of the disease, even when this profile was categorized according to the function of the proteins encoded by the genes, which leads us to believe that other factors, perhaps related to host susceptibility, may be more important than genetic variability of the microorganism. We conclude that Campylobacter sp. was detected in a significant percentage of the children 0-36 months with diarrhea, especially when the diagnostic methods were used in combination. In general, the virulence genes were detected in a high proportion of C. jejuni-positive samples, although the invasion-related genes have been found less frequently. Our data corroborates findings from other groups on the need to revise the diagnostic for Campylobacter sp. towards the inclusion of more sensitive and species-specific methods, as well as search for extra markers for intestinal inflammation and predictors of negative culture

Preliminary results of Lean method implementation in a pathology lab from Northeastern Brazil

Oncologic care shows a growing and unmet demand, and requires the search for alternatives that allow the efficient use of limited resources, the building of autonomy, and the endeavour for continuous improvement of processes. In the present work, we present the implementation of Lean philosophy at a pathology laboratory of an oncology hospital. Among the preliminary results, we highlight the redefinition of the dynamics of the staff, and the physical reorganization of the area. Such important changes culminated in an expressive reduction of lead time, even with a significant increase in the monthly load of exams

MEDI3902 Correlates of Protection against Severe Pseudomonas aeruginosa Pneumonia in a Rabbit Acute Pneumonia Model.

Pseudomonas aeruginosa is among the most formidable antibiotic-resistant pathogens and is a leading cause of hospital-associated infections. With dwindling options for antibiotic-resistant infections, a new paradigm for treatment and disease resolution is required. MEDI3902, a bispecific antibody targeting the P. aeruginosa type III secretion (T3S) protein PcrV and Psl exopolysaccharide, was previously shown to mediate potent protective activity in murine infection models. With the current challenges associated with the clinical development of narrow-spectrum agents, robust preclinical efficacy data in multiple animal species are desirable. Here, we sought to develop a rabbit P. aeruginosa acute pneumonia model to further evaluate the activity of MEDI3902 intervention. In the rabbit model of acute pneumonia, prophylaxis with MEDI3902 exhibited potent dose-dependent protection, whereas those receiving control IgG developed fatal hemorrhagic necrotizing pneumonia between 12 and 54 h after infection. Blood biomarkers (e.g., partial pressure of oxygen [pO2], partial pressure of carbon dioxide [pCO2], base excess, lactate, and creatinine) were grossly deranged for the vast majority of control IgG-treated animals but remained within normal limits for MEDI3902-treated animals. In addition, MEDI3902-treated animals exhibited a profound reduction in P. aeruginosa organ burden and a marked reduction in the expression of proinflammatory mediators from lung tissue, which correlated with reduced lung histopathology. These results confirm that targeting PcrV and Psl via MEDI3902 is a promising candidate for immunotherapy against P. aeruginosa pneumonia

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative <it>Escherichia coli</it>

Abstract Background Enteroaggregative Escherichia coli (EAEC) causes diarrhea, malnutrition and poor growth in children. Human breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. Goat milk with and without human lysozyme (HLZ) may improve the repair of intestinal barrier function damage induced by EAEC. This work investigates the effect of the milks on intestinal barrier function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC. Methods Rat intestinal epithelial cells (IEC-6, ATCC, Rockville, MD) were used for proliferation, migration and viability assays and human colon adenocarcinoma (Caco-2, ATCC, Rockville, MD) and human larynx carcinoma (HEp-2, ATCC, Rockville, MD) cells were used for bacterial adhesion assays. Goats expressing HLZ in their milk were generated and express HLZ in milk at concentration of 270 μg/ml . Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the presence and absence of EAEC strain 042 (O44:H18). Results Cellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat milk compared to cells with no milk. Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the presence of milk. Milk from HLZ transgenic goats had significantly more migration compared to control milk. Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in less colonization than control milk using a HEp-2 assay. Both milks had significantly increased cellular viability as well as less apoptosis in both the absence and presence of EAEC. Conclusions These data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC and that goat milk with a higher concentration of lysozyme offers additional protection.</p

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

AbstractBackgroundEnteroaggregative Escherichia coli (EAEC) causes diarrhea, malnutrition and poor growth in children. Human breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. Goat milk with and without human lysozyme (HLZ) may improve the repair of intestinal barrier function damage induced by EAEC. This work investigates the effect of the milks on intestinal barrier function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC.MethodsRat intestinal epithelial cells (IEC-6, ATCC, Rockville, MD) were used for proliferation, migration and viability assays and human colon adenocarcinoma (Caco-2, ATCC, Rockville, MD) and human larynx carcinoma (HEp-2, ATCC, Rockville, MD) cells were used for bacterial adhesion assays. Goats expressing HLZ in their milk were generated and express HLZ in milk at concentration of 270 μg/ml . Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the presence and absence of EAEC strain 042 (O44:H18).ResultsCellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat milk compared to cells with no milk. Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the presence of milk. Milk from HLZ transgenic goats had significantly more migration compared to control milk. Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in less colonization than control milk using a HEp-2 assay. Both milks had significantly increased cellular viability as well as less apoptosis in both the absence and presence of EAEC.ConclusionsThese data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC and that goat milk with a higher concentration of lysozyme offers additional protection

Antigenicity of a whole parasite vaccine as promising candidate against canine leishmaniasis.

Human visceral leishmaniasis, one of the most important zoonoses, is caused by the protozoaLeishmania chagasi(syn. L. infantum ) and is present as a fatal disease common in South America and Europe where dogs and wild canids are the main reservoirs. A vaccine against visceral leishmaniasis would be an important tool in the control of this disease in dogs. Although the current strategies for vac-cination against leishmaniasis are based on the use of recombinant antigens, killed vaccines are still attractive in terms of stability of their biochemical composition and antigenicity, cost, and safety. Here we evaluate the immunogenicity of a whole parasite vaccine as a prom-ising candidate against canine leishmaniasis, demonstrated by cellular reactivity, changes in the cellular profile of the peripheral blood and by the differential production of immunoglobulins. Our results showed that immunization elicited mainly a strong cellular reactivity and increase in T-lymphocytes, particularly the subpopulation CD8 + that would be related to the control of tissue parasitism. In addi-tion, a higher production of anti- Leishmania total IgG, characterized by mixed isotypes profile (IgG1 and IgG2), was demonstrated

Antigenicity of a whole parasite vaccine as promising candidate against canine leishmaniasis.

Human visceral leishmaniasis, one of the most important zoonoses, is caused by the protozoaLeishmania chagasi(syn. L. infantum ) and is present as a fatal disease common in South America and Europe where dogs and wild canids are the main reservoirs. A vaccine against visceral leishmaniasis would be an important tool in the control of this disease in dogs. Although the current strategies for vac-cination against leishmaniasis are based on the use of recombinant antigens, killed vaccines are still attractive in terms of stability of their biochemical composition and antigenicity, cost, and safety. Here we evaluate the immunogenicity of a whole parasite vaccine as a prom-ising candidate against canine leishmaniasis, demonstrated by cellular reactivity, changes in the cellular profile of the peripheral blood and by the differential production of immunoglobulins. Our results showed that immunization elicited mainly a strong cellular reactivity and increase in T-lymphocytes, particularly the subpopulation CD8 + that would be related to the control of tissue parasitism. In addi-tion, a higher production of anti- Leishmania total IgG, characterized by mixed isotypes profile (IgG1 and IgG2), was demonstrated

Evaluation of 16S rRNA qPCR for detection of <i>Mycobacterium leprae</i> DNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases

<p><i>Mycobacterium leprae</i> bacilli are mainly transmitted by the dissemination of nasal aerosols from multibacillary (MB) patients to susceptible individuals through inhalation. The upper respiratory tract represents the main entry and exit routes of <i>M. leprae</i>. Therefore, this study aimed to evaluate the sensitivity and specificity of real-time quantitative polymerase chain reaction (qPCR) in detecting <i>M. leprae</i> in nasal secretion (NS) and skin biopsy (SB) samples from MB and paucibacillary (PB) cases. Fifty-four NS samples were obtained from leprosy patients at the Dona Libânia National Reference Centre for Sanitary Dermatology in Ceará, Brazil. Among them, 19 MB cases provided both NS and SB samples. Bacilloscopy index assays were conducted and qPCR amplification was performed using specific primers for <i>M. leprae</i> 16S rRNA gene, generating a 124-bp fragment. Primer specificity was verified by determining the amplicon melting temperature (<i>T</i>_<i>m</i> = 79.5 °C) and detection limit of qPCR was 20 fg of <i>M. leprae</i> DNA. Results were positive for 89.7 and 73.3% of NS samples from MB and PB cases, respectively. SB samples from MB patients were 100% positive. The number of bacilli detected in NS samples were 1.39 × 10³–8.02 × 10⁵, and in SB samples from MB patients were 1.87 × 10³–1.50 × 10⁶. Therefore, qPCR assays using SYBR Green targeting <i>M. leprae</i> 16S rRNA region can be employed in detecting <i>M. leprae</i> in nasal swabs from leprosy patients, validating this method for epidemiological studies aiming to identify healthy carriers among household contacts or within populations of an endemic area.</p