Development of ELISA Using Recombinant Proteins for the Diagnosis of Mycoplasma hyopneumoniae Infection.

In order to develop a more sensitive and reliable method for detection of serum antibodies against Mycoplasma hyopneumoniae infection in pigs, six recombinant proteins of M. hyopneumoniae (P102, P95, P46, P97 like, Lppt, and hypothetical P987) were used for the standardization of an indirect enzyme-linked immunosorbent assay (ELISA). The proteins were evaluated against 50 sera of the specific pathogen-free and 50 sera of pigs with lesions suggestive of infection. The sensitivity was 88%, 86%, 78%, 74%, 66%, and 60% for the proteins P102, P95, P46, P97 like, Lppt, and hypothetical protein P987, respectively. Moreover, the proteins were used to establish the seroprevalence in two different commercial herds (254 sera pigs from farm considered free of M. hyopneumoniae and 246 from farm with clinical signs of enzootic pneumonia and positive serology for M. hyopneumoniae) and the positive rate was 65.2% for P95, 54.6% for P102, 40.2% for P46, 37.2% for P97 like, 17.4% for the hypothetical P987, and 14% for Lppt protein. In addition, the ELISA with six recombinant proteins was compared to commercial HerdCheck kit using 118 random pig sera samples and the results showed that ELISA with recombinant proteins were more sensitive than the commercial test. These data show that the recombinant proteins P95 and P102 are potential targets to be used in diagnostic tests to detect antibodies against M. hyopneumoniae. Although more studies are necessary, this study provides insights that these recombinant proteins can be useful in epidemiological investigations and as potential biomarkers in differentiating infected animals from those vaccinated.

Uric acid is independent and inversely associated to glomerular filtration rate in young adult Brazilian individuals.

BACKGROUND AND AIMS: Uric acid, the end-product of human purine metabolism, is associated with hypertension, diabetes and obesity. It has also been independently associated with the onset of chronic kidney disease in several populations. In this study, the association between serum uric acid (SUA) level and estimated glomerular filtration rate (eGFR) was investigated in healthy individuals belonging to two Brazilian birth cohorts. METHODS AND RESULTS: Data from 3541 to 3482 individuals, aged 30 and 22-years old, respectively, was included. eGFR was calculated using Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation based on creatinine measurement. Regression analyses were sex-stratified due to interaction between SUA and sex (p < 0.001) and adjusted for perinatal, cardiometabolic and behavioral variables. We observed an inverse association between eGFR and SUA even after adjustment. In the highest tertile (3rd) of SUA, the eGFR coefficients at 30-years were-0.21 (95%CI -0.24;-0.18) for men and -0.20 (95%CI -0.23; -0.17) for women; at 22-years, were -0.09 (95%CI -0.12;-0.05) for men and -0.13 (95%CI -0.15; -0.10) for women. Higher differences among exponential means (95% CI) of eGFR between the 1st and the 3rd tertile of SUA were seen in older participants, being more pronounced in men. At 22-years, the highest difference was found in women. CONCLUSIONS: In young healthy individuals from a low-middle income country, SUA level was inversely associated with eGFR. Gender-related differences in eGFR according tertiles of SUA were higher in men at 30-years and in women at 22-years.

DNA microarray for forensic intelligence purposes: High-density SNP profiles obtained directly from casework-like samples with and without a DNA purification step.

SNP analyses from a forensic intelligence perspective have proven to be an important tool to restrict the number of suspected offenders and find missing persons. DNA microarray assays have been demonstrated as a potential feature in forensic analysis, like such as forensic genetic genealogy. The objective of this study was to describe the results from DNA microarray assay from saliva samples deposited on a glass surface collected from by a double swab technique, commonly applied in crime scenes. Eighteen samples from the same person were subjected to Infinium® Global Screening Array-24 v1.0 (~642.824 SNP markers) in two different protocols - with or without the DNA purification procedure. The measured genotype was compared with a Consensus Genotype, obtained from standard control samples, and the parameters such as Call Rate and GenCall Scores were evaluated. Results showed that the Call Rate parameter is enough to estimate the probability of obtaining a correct genotype in the SNP assay. Reliable genotypes with a confidence level of more than 90% (at least 90.15%) were observed in Call Rates above 69.41%, regardless of the experimental condition. Our data demonstrate that DNA Microarray from samples collected under conditions such as those found at crime scenes can generate high-density SNP genetic profiles with a confidence level greater than 90%. Enzymatic adjustments and protocol changes may enable DNA microarray assays for crime analysis and investigation purposes eliminating the purification step in the future. Our data suggest that DNA microarray can support criminal investigation teams from a forensic intelligence perspective.

Genetic variants in miRNAs differentially expressed during brain development and their relevance to psychiatric disorders susceptibility.

OBJECTIVES: MicroRNAs (miRNAs) play an important regulatory role in the expression of genes involved in brain functions during development. Genetic variants in miRNA genes may impact their regulatory function and lead to psychiatric disorders. To evaluate the role of genetic variants in genes of miRNAs differentially expressed during neurodevelopment on autism spectrum disorder (ASD), attention deficit/hyperactivity disorder (ADHD), schizophrenia (SCZ), and major depressive disorder (MDD). METHODS: The miRNAs were identified in the literature. Summary statistics from the most recent genome-wide association studies to date were used to evaluate the association between the selected polymorphisms and each disorder in a look-up approach. In a global analysis, we compared the standardised risk effect of variants in neurodevelopment-related miRNAs with those in the remaining miRNAs from miRBase. RESULTS: The global analysis showed that variants in neurodevelopment-related miRNAs had higher risk effects compared to the other miRNAs for SCZ (p = 0.010) and ADHD (p = 0.001). MIR33B, MIR29B2, MIR29C, MIR137, and MIR135A1 were significantly associated with SCZ, while 55.9% of the miRNAs were at least nominally associated with one or more psychiatric disorders (p < 0.05). CONCLUSIONS: Genetic variants in neurodevelopment-related miRNAs play an important role in the genetic susceptibility of psychiatric disorders, mainly SCZ and ADHD.

The Role of MIR9-2 in Shared Susceptibility of Psychiatric Disorders during Childhood: A Population-Based Birth Cohort Study.

BACKGROUND: It has been suggested that microRNAs (miRNAs; short non-protein-coding RNA molecules that mediate post-transcriptional regulation), including mir-9 and mir-34 families, are important for brain development. Current data suggest that mir-9 and mir-34 may have shared effects across psychiatric disorders. This study aims to explore the role of genetic polymorphisms in the MIR9-2 (rs4916723) and MIR34B/C (rs4938723) genes on the susceptibility of psychiatric disorders in children from the 2004 Pelotas Birth Cohort. METHODS: Psychiatric disorders were assessed in 3585 individuals using Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV), criteria through the application of standard semi-structured interviews (using the Development and Well-Being Assessment, DAWBA) at the six-years-of-age follow-up. The outcome was defined as the presence of any mental disorder. We also considered two broad groups of internalizing and externalizing disorders to further investigate the role of these variants in mental health. RESULTS: We observed an association between rs4916723 (MIR9-2) and the presence of any psychiatric disorder (odds ratios (OR) = 0.820; 95% CI = 0.7130-0.944; p = 0.006) and a suggestive effect on internalizing disorders (OR = 0.830; 95% CI = 0.698-0.987; p = 0.035). rs4938723 (MIR34B/C) was not associated with any evaluated outcome. CONCLUSION: The study suggests that MIR9-2 may have an important role on a broad susceptibility for psychiatric disorders and may be important mainly for internalization problems.

Clonal expansion across the seas as seen through CPLP-TB database: A joint effort in cataloguing Mycobacterium tuberculosis genetic diversity in Portuguese-speaking countries.

Tuberculosis (TB) remains a major health problem within the Community of Portuguese Language Speaking Countries (CPLP). Despite the marked variation in TB incidence across its member-states and continued human migratory flux between countries, a considerable gap in the knowledge on the Mycobacterium tuberculosis population structure and strain circulation between the countries still exists. To address this, we have assembled and analysed the largest CPLP M. tuberculosis molecular and drug susceptibility dataset, comprised by a total of 1447 clinical isolates, including 423 multidrug-resistant isolates, from five CPLP countries. The data herein presented reinforces Latin American and Mediterranean (LAM) strains as the hallmark of M. tuberculosis populational structure in the CPLP coupled with country-specific differential prevalence of minor clades. Moreover, using high-resolution typing by 24-loci MIRU-VNTR, six cross-border genetic clusters were detected, thus supporting recent clonal expansion across the Lusophone space. To make this data available to the scientific community and public health authorities we developed CPLP-TB (available at http://cplp-tb.ff.ulisboa.pt), an online database coupled with web-based tools for exploratory data analysis. As a public health tool, it is expected to contribute to improved knowledge on the M. tuberculosis population structure and strain circulation within the CPLP, thus supporting the risk assessment of strain-specific trends.

[Prevalence of antidepressant use and associated factors among adults in Pelotas, Rio Grande do Sul State, Brazil, 2006]. / Prevalência e fatores associados ao uso de antidepressivos em adultos de área urbana de Pelotas, Rio Grande do Sul, Brasil, em 2006.

The objective of this study was to estimate the prevalence and determinants of antidepressant use among adults in a city in southern Brazil. The cross-sectional study was based on interviews with adults 40 years or older living in the urban area of Pelotas, Rio Grande do Sul State, Brazil, 2006. The chi-squared test was used for comparisons between proportions. Non-conditional logistic regression was used for multivariate analysis, with robust adjustment for variance. A total of 1,327 adults were interviewed, of whom 9.3% had used antidepressants. Use of antidepressants was significantly associated with female gender (OR: 2.45; 95%CI: 1.50-4.02), higher socioeconomic status (OR: 2.07; 95%CI: 1.28-3.34), and unemployment (OR: 1.65; 95%CI: 1.06-2.55). The prevalence of antidepressant use was higher than in other studies. Identification of individual determinants of use can serve as the basis for strategies to decrease the consumption of antidepressants in these groups.

[Prevention of neural tube defects: prevalence of folic acid supplementation during pregnancy and associated factors in Pelotas, Rio Grande do Sul State, Brazil]. / Prevenção de defeitos do tubo neural: prevalência do uso da suplementação de ácido fólico e fatores associados em gestantes na cidade de Pelotas, Rio Grande do Sul, Brasil.

To determine folic acid use and associated factors, a cross-sectional population-based study was conducted in all five maternity hospitals in Pelotas, Rio Grande do Sul State, Brazil. Data were collected from April 1 to August 15, 2006 (n = 1,450 women). A standard questionnaire was applied in the hospitals. Statistical analysis used Poisson regression. Prevalence of folic acid consumption during pregnancy was 31.8%, and periconceptional use was 4.3%. The following were associated with folic acid use: white skin color, schooling > 9 years, family income > R$600, age > 30 years, planned pregnancy, > or =7 prenatal visits, knowledge on folic acid, and prenatal care in the private health system. In order to prevent neural tube defects, it is important to promote folic acid use among childbearing-age women and to supply folic acid to poor women.

[The process of detection and treatment of cases of tuberculosis in a prison]. / O processo de detecção e tratamento de casos de tuberculose em um presídio.

This study seeks to analyze the process of detection and treatment of cases of tuberculosis (TB) in a prison in the south of Brazil. An active and passive search for TB was conducted to estimate the scale of TB in a prison with 764 inmates. In conjunction with the detection strategies and clinical follow-up of the 41 TB cases, participant observation and records in field diaries were performed, making it possible to analyze the scope and limitations of detection and treatment of cases of TB in prison. The development of search strategies is discussed along with the use of questionnaires to detect symptomatic cases, as well as the inadequacy of the clinical follow-up of TB cases, involvement of different workers and coordination between prison and health services. There is clear potential for the control of TB using an active search to induce the passive detection and screening for symptoms that - even skewed by the perceptions of inmates regarding symptoms of TB - enabled an increase in detection. The functional dynamics of prison life hamper the inclusion of health routines and can restrict actions to control TB and other diseases. In the process of control of TB in prisons, the feasibility of effective detection methods is as important as planning based on disease conditions, network services and workers involved.

The BACTEC MGIT(tm) 320 system as a laboratory tool to diagnose tuberculosis in a Brazilian hospital with a high prevalence of HIV infection.

INTRODUCTION: The World Health Organization endorses the BACTEC Mycobacterial Growth Indicator Tube (MGIT)(tm) system as a rapid, sensitive, and specific method to diagnostic of tuberculosis. Here, we compared the performance of this system against Ogawa-Kudoh cultures and microscopy. METHODS: A total of 927 samples were obtained between December 2011 and December 2013 from 652 cases of suspected tuberculosis at the School Hospital of the Federal University of Rio Grande in Brazil. RESULTS: The MGIT system confirmed tuberculosis in more cases in less time. CONCLUSIONS: The MGIT system is an effective tool for early diagnosis of tuberculosis, especially in patients with HIV/AIDS.

Evaluation of direct microplate nitrate reductase assay as a rapid method for the detection of multiple and extensively tuberculosis drug resistance.

INTRODUCTION: Reports of Mycobacterium tuberculosis resistant to multiple drugs are increasing globally and laboratories are becoming increasingly aware of the need for drug susceptibility testing. In recent years, due to the long time required by conventional drug susceptibility testing, new approaches have been proposed for faster detection of drug resistance, such as the nitrate reductase assay, considered fast and inexpensive, making it a good diagnostic tool for low resource countries. OBJECTIVE: The present study proposed a fast direct colorimetric drug susceptibility testing method in a microplate format using solid medium. MATERIALS AND METHODS: The diagnostic accuracy was evaluated by comparing the proportion method with the direct nitrate reductase assay in plates. Frozen sputum samples, known to be positive, were decontaminated and processed by Petroff method. The decontaminated suspension was used to perform direct nitrate reductase assay in 7H11 medium using 1 µ g/ml rifampicin (RIF), 0.2 µg/ml isoniazid (INH), 2 µg/ml ofloxacin (OFX), 6 µg/ml kanamycin (KAN), 2 µg/ml amikacin (AMK) and 10 µg/ml capreomycin (CAP). Eighty-four samples were tested and the results for 69% of them were available within 21 days. RESULTS: The sensitivity and specificity compared to the proportion method, was 98.5% and 100% for INH, 98.3% and 96.2% for RIF, 91.7% and 100% for KAN, 78.8% and 97.3% for OFX, 100% and 100% for AMK and CAP, respectively. CONCLUSION: The results lead to the conclusion that direct nitrate reductase assay, in this new format, is an accurate, quick and inexpensive method to determine the susceptibility profile of M. tuberculosis and may become an alternative for countries with limited resources.

Clonal expansion across the seas as seen through CPLP-TB database: A joint effort in cataloguing Mycobacterium tuberculosis genetic diversity in Portuguese-speaking countries

Tuberculosis (TB) remains a major health problem within the Community of Portuguese Language Speaking Countries (CPLP). Despite the marked variation in TB incidence across its member-states and continued human migratory flux between countries, a considerable gap in the knowledge on the Mycobacterium tuberculosis population structure and strain circulation between the countries still exists. To address this, we have assembled and analysed the largest CPLP M. tuberculosis molecular and drug susceptibility dataset, comprised by a total of 1447 clinical isolates, including 423 multidrug-resistant isolates, from five CPLP countries. The data herein presented reinforces Latin American and Mediterranean (LAM) strains as the hallmark of M. tuberculosis populational structure in the CPLP coupled with country-specific differential prevalence of minor clades. Moreover, using high-resolution typing by 24-loci MIRU-VNTR, six cross-border genetic clusters were detected, thus supporting recent clonal expansion across the Lusophone space.

Immunological characterization of Mycoplasma hyopneumoniae recombinant proteins.

Mycoplasma hyopneumoniae, the primary pathogen of enzootic pneumonia, is highly prevalent worldwide and causes major economic losses to the pig industry. Commercial vaccines are widely used in the control of this disease, however, they provide only partial protection. The aim of this study was to evaluate 34 recombinant proteins of M. hyopneumoniae expressed in Escherichia coli. Antigenic and immunogenic properties of these proteins were analyzed. For this, the proteins were tested against hyperimmune and convalescent pig sera through ELISA and Western blot. Immunogenicity of the recombinant proteins was evaluated in BALB/c mice following intramuscular inoculation. Most antigens were able to induce a strong immune response and sera from inoculated mice were able to recognize native proteins by cell ELISA and Western blot. Several recombinant proteins were specifically recognized by convalescent pig sera, indicating they are expressed during infection. These data may help to develop more efficacious vaccines against M. hyopneumoniae.

O processo de detecção e tratamento de casos de tuberculose em um presídio / The process of detection and treatment of cases of tuberculosis in a prison

Resumo O estudo objetiva analisar o processo de detecção e tratamento de casos de tuberculose (TB) em um presídio do sul do Brasil. Realizou-se busca ativa e passiva para estimar a magnitude da TB entre uma população de 764 detentos. Simultaneamente às estratégias de detecção e ao seguimento clínico dos 41 casos confirmados, realizou-se observação participante e registros em diário de campo, o que possibilitou analisar potenciais e limitações das ações de detecção e tratamento da TB prisional. Discute-se o desenvolvimento das estratégias de busca, o uso de questionários para detecção de sintomáticos, a fragilidade do seguimento clínico dos casos de TB, o envolvimento de diferentes trabalhadores e a articulação entre presídio e serviços de saúde. São potencialidades para o controle da TB utilizar a busca ativa como indutora da detecção passiva e a triagem por sintomas que, mesmo interferida pelas percepções dos detentos sobre os sintomas de TB, permitiu aumentar a detecção. A dinâmica funcional do presídio dificulta a inserção de rotinas de saúde, podendo limitar ações de controle da TB e outras enfermidades. No processo de controle da TB em presídios, a exequibilidade de métodos de detecção eficazes é tão importante quanto planejá-lo a partir das condições de adoecimento, rede de serviços e trabalhadores envolvidos.

Abstract This study seeks to analyze the process of detection and treatment of cases of tuberculosis (TB) in a prison in the south of Brazil. An active and passive search for TB was conducted to estimate the scale of TB in a prison with 764 inmates. In conjunction with the detection strategies and clinical follow-up of the 41 TB cases, participant observation and records in field diaries were performed, making it possible to analyze the scope and limitations of detection and treatment of cases of TB in prison. The development of search strategies is discussed along with the use of questionnaires to detect symptomatic cases, as well as the inadequacy of the clinical follow-up of TB cases, involvement of different workers and coordination between prison and health services. There is clear potential for the control of TB using an active search to induce the passive detection and screening for symptoms that – even skewed by the perceptions of inmates regarding symptoms of TB – enabled an increase in detection. The functional dynamics of prison life hamper the inclusion of health routines and can restrict actions to control TB and other diseases. In the process of control of TB in prisons, the feasibility of effective detection methods is as important as planning based on disease conditions, network services and workers involved.

Locomotor and peripheral effects of sibutramine modulated by 5-HT2 receptors.

Sibutramine has been described as an anti-obesity drug with the ability to inhibit serotonin (5-HT), noradrenaline, and dopamine re-uptake, but without affinity to histamine and muscarinic receptors. On the other hand, cyproheptadine antagonizes serotonin 5-HT(2A), 5-HT(2B), and 5-HT(2C), histamine H1, and muscarinic (M) receptors. There are many reports concerning the influence of sibutramine on central serotoninergic pathways. In this study, we suggest that peripheral pathways may also be involved in the serotoninergic effects of sibutramine. In vivo experiments were undertaken to investigate the serotoninergic effects of sibutramine on body mass, the glycogen concentration in the diaphragm of rats, and locomotor behaviour. Rats were submitted to oral treatment with sibutramine, cyproheptadine, or sibutramine applied in combination with cyproheptadine, for a period of 2 months to investigate the 5-HT2 effects of sibutramine on these parameters. As the results demonstrated, the lower increase in body mass and the increased glycogen levels in the diaphragm muscle of rats treated with sibutramine seem to be modulated by 5-HT2 receptors, since these effects were completely antagonized by cyproheptadine in the group treated with the 2 drugs co-applied. Furthermore, the behavioural results also suggest that mechanisms modulated by 5-HT2 receptors are involved in the increase of locomotion in the rats treated with sibutramine, since the effect did not occur in the rats treated with sibutramine co-applied with the 5-HT2 receptor antagonist, cyproheptadine. The results suggest that sibutramine modifies energy-related parameters such as body mass, diaphragm glycogen, and locomotor behaviour in rats via 5-HT2 serotoninergic pathways.

Prevalência e fatores associados ao uso de antidepressivos em adultos de área urbana de Pelotas, Rio Grande do Sul, Brasil, em 2006 / Prevalence of antidepressant use and associated factors among adults in Pelotas, Rio Grande do Sul State, Brazil, 2006

Este trabalho determina a prevalência e fatores associados ao uso de antidepressivos em adultos residentes na área urbana da cidade de Pelotas, Rio Grande do Sul, Brasil. Foi realizado um estudo transversal em uma amostra representativa de adultos com idade igual ou superior a 40 anos, moradores na zona urbana de Pelotas, em 2006. Para as comparações entre proporções, utilizou-se o teste de qui-quadrado e, para a análise multivariada, a regressão logística não condicional. Foram entrevistados 1.327 adultos, dos quais 9,3 por cento tinham usado antidepressivos. O consumo desses medicamentos associou-se significativamente ao sexo feminino (OR: 2,45; IC95 por cento: 1,50-4,02); ao nível sócio-econômico mais elevado, classes A e B (OR: 2,07; IC95 por cento: 1,28-3,34); e a não estar exercendo uma atividade de trabalho (OR: 1,65; IC95 por cento: 1,06-2,55). A prevalência de uso de antidepressivos foi superior às encontradas em outros estudos. Os determinantes individuais de utilização poderão servir de embasamento às estratégias para diminuir o consumo de antidepressivos nesses grupos.

The objective of this study was to estimate the prevalence and determinants of antidepressant use among adults in a city in southern Brazil. The cross-sectional study was based on interviews with adults 40 years or older living in the urban area of Pelotas, Rio Grande do Sul State, Brazil, 2006. The chi-squared test was used for comparisons between proportions. Non-conditional logistic regression was used for multivariate analysis, with robust adjustment for variance. A total of 1,327 adults were interviewed, of whom 9.3 percent had used antidepressants. Use of antidepressants was significantly associated with female gender (OR: 2.45; 95 percentCI: 1.50-4.02), higher socioeconomic status (OR: 2.07; 95 percentCI: 1.28-3.34), and unemployment (OR: 1.65; 95 percentCI: 1.06-2.55). The prevalence of antidepressant use was higher than in other studies. Identification of individual determinants of use can serve as the basis for strategies to decrease the consumption of antidepressants in these groups.

Prevenção de defeitos do tubo neural: prevalência do uso da suplementação de ácido fólico e fatores associados em gestantes na cidade de Pelotas, Rio Grande do Sul, Brasil / Prevention of neural tube defects: prevalence of folic acid supplementation during pregnancy and associated factors in Pelotas, Rio Grande do Sul State, Brazil

Com o objetivo de determinar a prevalência do uso do ácido fólico e fatores associados na gestação e no período periconcepcional, realizou-se um estudo transversal de base populacional nas cinco maternidades da cidade de Pelotas, Rio Grande do Sul, Brasil. A coleta de dados ocorreu no período de 1° de abril a 15 de agosto de 2006, com 1.450 mulheres. As entrevistas foram realizadas em nível hospitalar por questionário padronizado. A análise estatística se realizou por regressão de Poisson. A prevalência do uso de ácido fólico na gestação foi de 31,8 por cento, e no período periconcepcional, foi de 4,3 por cento. Os fatores associados ao uso de ácido fólico foram: cor branca, escolaridade acima de nove anos, renda acima de 600 Reais, idade acima de trinta anos, gestação planejada, sete ou mais consultas de pré-natal, consultas na rede privada de saúde e conhecimento sobre o ácido fólico. Para diminuir a prevalência de defeitos do tubo neural, é importante promover-se o uso do ácido fólico nas mulheres em idade fértil, nas mulheres sócio-economicamente menos favorecidas e torná-lo disponível na rede pública de saúde.

To determine folic acid use and associated factors, a cross-sectional population-based study was conducted in all five maternity hospitals in Pelotas, Rio Grande do Sul State, Brazil. Data were collected from April 1 to August 15, 2006 (n = 1,450 women). A standard questionnaire was applied in the hospitals. Statistical analysis used Poisson regression. Prevalence of folic acid consumption during pregnancy was 31.8 percent, and periconceptional use was 4.3 percent. The following were associated with folic acid use: white skin color, schooling > 9 years, family income > R$600, age > 30 years, planned pregnancy, > 7 prenatal visits, knowledge on folic acid, and prenatal care in the private health system. In order to prevent neural tube defects, it is important to promote folic acid use among childbearing-age women and to supply folic acid to poor women.