A quantitative PCR-electrochemical genosensor test for the screening of biotech crops

The design of screening methods for the detection of genetically modified organisms (GMOs) in food would improve the efficiency in their control. We report here a PCR amplification method combined with a sequence-specific electrochemical genosensor for the quantification of a DNA sequence characteristic of the 35S promoter derived from the cauliflower mosaic virus (CaMV). Specifically, we employ a genosensor constructed by chemisorption of a thiolated capture probe and p-aminothiophenol gold surfaces to entrap on the sensing layer the unpurified PCR amplicons, together with a signaling probe labeled with fluorescein. The proposed test allows for the determination of a transgene copy number in both hemizygous (maize MON810 trait) and homozygous (soybean GTS40-3-2) transformed plants, and exhibits a limit of quantification of at least 0.25% for both kinds of GMO lines

Nrf2-interacting nutrients and COVID-19 : time for research to develop adaptation strategies

There are large between- and within-country variations in COVID-19 death rates. Some very low death rate settings such as Eastern Asia, Central Europe, the Balkans and Africa have a common feature of eating large quantities of fermented foods whose intake is associated with the activation of the Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) anti-oxidant transcription factor. There are many Nrf2-interacting nutrients (berberine, curcumin, epigallocatechin gallate, genistein, quercetin, resveratrol, sulforaphane) that all act similarly to reduce insulin resistance, endothelial damage, lung injury and cytokine storm. They also act on the same mechanisms (mTOR: Mammalian target of rapamycin, PPAR gamma:Peroxisome proliferator-activated receptor, NF kappa B: Nuclear factor kappa B, ERK: Extracellular signal-regulated kinases and eIF2 alpha:Elongation initiation factor 2 alpha). They may as a result be important in mitigating the severity of COVID-19, acting through the endoplasmic reticulum stress or ACE-Angiotensin-II-AT(1)R axis (AT(1)R) pathway. Many Nrf2-interacting nutrients are also interacting with TRPA1 and/or TRPV1. Interestingly, geographical areas with very low COVID-19 mortality are those with the lowest prevalence of obesity (Sub-Saharan Africa and Asia). It is tempting to propose that Nrf2-interacting foods and nutrients can re-balance insulin resistance and have a significant effect on COVID-19 severity. It is therefore possible that the intake of these foods may restore an optimal natural balance for the Nrf2 pathway and may be of interest in the mitigation of COVID-19 severity

Cabbage and fermented vegetables : From death rate heterogeneity in countries to candidates for mitigation strategies of severe COVID-19

Large differences in COVID-19 death rates exist between countries and between regions of the same country. Some very low death rate countries such as Eastern Asia, Central Europe, or the Balkans have a common feature of eating large quantities of fermented foods. Although biases exist when examining ecological studies, fermented vegetables or cabbage have been associated with low death rates in European countries. SARS-CoV-2 binds to its receptor, the angiotensin-converting enzyme 2 (ACE2). As a result of SARS-CoV-2 binding, ACE2 downregulation enhances the angiotensin II receptor type 1 (AT(1)R) axis associated with oxidative stress. This leads to insulin resistance as well as lung and endothelial damage, two severe outcomes of COVID-19. The nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is the most potent antioxidant in humans and can block in particular the AT(1)R axis. Cabbage contains precursors of sulforaphane, the most active natural activator of Nrf2. Fermented vegetables contain many lactobacilli, which are also potent Nrf2 activators. Three examples are: kimchi in Korea, westernized foods, and the slum paradox. It is proposed that fermented cabbage is a proof-of-concept of dietary manipulations that may enhance Nrf2-associated antioxidant effects, helpful in mitigating COVID-19 severity.Peer reviewe

Targeting Helicase-Dependent Amplification Products with an Electrochemical Genosensor for Reliable and Sensitive Screening of Genetically Modified Organisms

Cultivation of genetically modified organisms (GMOs) and their use in food and feed is constantly expanding; thus, the question of informing consumers about their presence in food has proven of significant interest. The development of sensitive, rapid, robust, and reliable methods for the detection of GMOs is crucial for proper food labeling. In response, we have experimentally characterized the helicase-dependent isothermal amplification (HDA) and sequence-specific detection of a transgene from the Cauliflower Mosaic Virus 35S Promoter (CaMV35S), inserted into most transgenic plants. HDA is one of the simplest approaches for DNA amplification, emulating the bacterial replication machinery, and resembling PCR but under isothermal conditions. However, it usually suffers from a lack of selectivity, which is due to the accumulation of spurious amplification products. To improve the selectivity of HDA, which makes the detection of amplification products more reliable, we have developed an electrochemical platform targeting the central sequence of HDA copies of the transgene. A binary monolayer architecture is built onto a thin gold film where, upon the formation of perfect nucleic acid duplexes with the amplification products, these are enzyme-labeled and electrochemically transduced. The resulting combined system increases genosensor detectability up to 10⁶-fold, allowing Yes/No detection of GMOs with a limit of detection of ∼30 copies of the CaMV35S genomic DNA. A set of general utility rules in the design of genosensors for detection of HDA amplicons, which may assist in the development of point-of-care tests, is also included. The method provides a versatile tool for detecting nucleic acids with extremely low abundance not only for food safety control but also in the diagnostics and environmental control areas

Acessibilidade geográfica à atenção primária à saúde em distrito sanitário do município de Salvador, Bahia The geographical accessibility of primary health care in a sanitary district of the municipality Salvador, in the Brazilian State of Bahia

OBJETIVOS: avaliar a acessibilidade geográfica à atenção primária à saúde da população coberta pelos Programa de Agentes Comunitários de Saúde (PACS) e Programa de Saúde da Família (PSF) de um distrito sanitário de Salvador. MÉTODOS: estudo avaliativo transversal. Os dados foram coletados a partir de entrevistas realizadas junto aos profissionais das onze equipes de PACS e PSF. A análise dos dados contemplou duas dimensões: acesso da população e lógica de territorialização. RESULTADOS: o distrito apresentava relevo bastante acidentado e todas as equipes informaram a existência de barreiras geográficas na área, em especial escadarias, morros e ladeiras. Não foi observada diferença significativa na acessibilidade às equipes de PACS e PSF. O processo de territorialização esteve dissociado da concepção de território-processo. Algumas áreas de abrangência das equipes situavam-se em locais afastados das UBS, constituindo-se em importante barreira ao acesso da população. CONCLUSÕES: o processo de implantação da estratégia PACS/PSF no distrito e o modelo de territorialização adotado não atenderam aos princípios que devem nortear a organização da atenção primária à saúde, resultando em restrições na acessibilidade geográfica da população coberta. Recomenda-se que seja realizada uma revisão do desenho atual da territorialização, visando uma melhor adequação às diretrizes do PSF e à promoção de uma maior acessibilidade à atenção primária.<br>OBJECTIVES: to assess the geographical accessibility of primary health care for the population served by the Community Health Agents Program (PACS) and the Family Health Program (PSF) in one sanitary district in the city of Salvador, Brazil. METHODS: a evaluative cross-sectional study was carried out. The data were collected using interviews with health workers from the eleven PACS and PSF teams. Analysis of the data took two factors into account: access of the population and the logic of territorialization. RESULTS: the district has a relatively rugged landscape and all the teams reported the existence of geographical barriers in the area, in particular steps and steep hills. There was no difference in accessibility between the PACS and the PSF. The process of territorialization was unrelated to the concept of territory as a process. Some areas covered by the teams were located some distance from the UBS and this constituted a significant obstacle to access. CONCLUSIONS: the implantation of the PACS/PSF strategy in the district and the model of territorialization were not in line with the principles that should guide primary health care, resulting in geographical restrictions on access for the population served. It is recommended that the current territorial design be reviewed to ensure that it is more line with PSF guidelines and to enable better access to primary care

Atenção primária à saúde: estrutura das unidades como componente da atenção à saúde Primary health care: the structure of units as a component of health care

OBJETIVOS: avaliar a estrutura das unidades de saúde da atenção primária de dois municípios, comparando as unidades de saúde da família (USF) e as unidades de saúde convencionais (USC). MÉTODOS: estudo transversal, quantitativo, utilizou entrevistas estruturadas e observação da rotina de trabalho das equipes como técnica de coleta de dados. Foram avaliados três componentes: ambiente físico, recurso material e pessoal. RESULTADOS: os achados evidenciaram deficiências estruturais nos três componentes, mais, onde os itens avaliados foram observados em menos de 70,0% das USF. Nos municípios, menos de 50% das unidades possuíam equipamentos básicos. No município A, todas ESF contavam com médicos e enfermeiros e, no município B, 27,3% das ESF estavam sem médico. Ressaltam-se as deficiências que comprometem a biossegurança dos procedimentos e a ausência de estruturas que favorecem o desenvolvimento de intervenções coletivas, necessárias à mudança do modelo assistencial na perspectiva da vigilância da saúde. CONCLUSÕES: embora as redes causais que ligam a estrutura dos serviços de saúde aos seus efeitos sejam complexas e envolvam diversos fatores, os achados do estudo reiteram a necessidade de valorização dos componentes estruturais na avaliação e gestão de saúde.<br>OBJECTIVES: to assess the structure of the primary health care units in two municipalities, comparing the Family Health Units(FHU) and the conventional units. METHODS: a quantitative cross-sectional study was carried out using structured interviews and observation of the work routine of the teams as data collection techniques. Three components were evaluated: the physical environment, material resources, and staff. RESULTS: the findings provided evidence of structural shortcomings in all three components, these being more serious in the Family Health Units. The shortcomings in terms of physical structure were more serious in municipality B, where the items under evaluation were fund in less than 70% of FHUs. In both municipalities, less than 50% of units had basic equipment. In municipality A, all the conventional units had doctors and nurses, while in municipality B, 27.3% of FHUs had no doctor. Shortcomings were revealed that jeopardize biosafety and there was an absence of structures that favor the development of the kind of collective intervention that is necessary for changing the model of care by placing more emphasis on health surveillance. CONCLUSIONS: although the chains of causation that link the structure of health services to their effects are complex and involve various factors, the study findings reiterate the need for more attention to be paid to structural components when evaluating health management

Avaliação de iniciativas e programas intersetoriais em saúde: desafios e aprendizados Evaluation of initiatives and intersectorial programs in health: challenges and learning

O objetivo deste artigo é contribuir para a avaliação das iniciativas voltadas para a construção de arranjos intersetoriais no âmbito das políticas públicas de saúde e de proteção social. São tomados como focos analíticos a iniciativa de Desenvolvimento Local Integrado e Sustentável de Manguinhos (DLIS-Manguinhos) - uma proposta de base territorial e comunitária associada à estratégias de colaboração e negociação bottom up- e a experiência de implementação do programa federal de transferência condicionada de renda Bolsa Família (PBF). Nesta perspectiva, aspectos ligados às redes de mobilização social, perfil dos atores envolvidos, tipos de incentivos e níveis de integração institucional são tratados como elementos cruciais para a análise do alcance de programas e iniciativas que articulam propostas intersetoriais. Conclui-se que a interface e o diálogo entre pesquisa, avaliação e acompanhamento de processos decisórios constituem eixos centrais para o maior aprendizado social e institucional na área.<br>The objective of his article is to contribute to the evaluation of the initiatives aimed at the creation of intersectorial arrangements in the scope of public health policies and social protection. The focus is on the Integrated and Sustainable Local Development of Manguinhos (DLIS-Manguinhos) - a territory and community based proposal associated to bottom up cooperation and negotiation strategies - and the experience of establishing the conditioned cash transference federal program called Bolsa Família (Family Grant Program - PBF). In this perspective, the aspects related to social mobilization networks, profile of the players involved, types of incentive and levels of institutional integration are treated as crucial elements in the analysis of the programs and initiatives that articulate intersectorial proposals. It is concluded that the interface and dialog among research, evaluation and follow up of decision processes constitute the central axes for better social and institutional learning in the area