Socio-economic inequalities in the effect of public policies and the COVID-19 pandemic on exclusive breastfeeding in Chile.

OBJECTIVE: To assess the impact of the addition of 12 maternity leave (ML) weeks (2011), a pay for performance (P4P) exclusive breastfeeding (EBF) promotion strategy (2015), and the COVID-19 pandemic in EBF inequalities in Chile. STUDY DESIGN: Interrupted time-series analyses (ITSAs). METHODS: Aggregated national EBF data by municipality and month were collected from 2009 to 2020. We assess the impact of the three events in EBF inequalities using two procedures: 1. ITSA stratified by municipal SES quintiles (Q1-Q5); 2. Calculating the EBF slope index of inequality (SII). RESULTS: The EBF prevalence was higher in lower SES municipalities before and after the three time-events. No impact in EBF inequalities was observed after the extended ML. The P4P strategy increased EBF at six months in all SES quintiles (effect size between 4% and 5%), but in a higher level in poorer municipalities (SII: -0.36% and -1.05%). During COVID-19, wealthier municipalities showed a slightly higher EBF at six months prevalence (SII: 1.44%). CONCLUSION: The null impact of the extended ML in EBF inequalities could be explained by a low access to ML among affiliated to the public health system (20%). The P4P strategy includes multiple interventions that seemed effective in increasing EBF across all SES quintiles, but further in lower quintiles. The restrictions in healthcare access in poorer municipalities could explain EBF inequalities during COVID-19.

Glass-coated ferromagnetic microwire-induced magnetic hyperthermia for in vitro cancer cell treatment.

Limitations in effectiveness and the invasive nature of current cancer treatment options emphasize the need for further clinical advancements. Among other approaches, targeted hyperthermia is as a new strategy aimed at targeting cancerous cells to improve the efficacy of radiotherapy or cytotoxic drugs. However, the testing of magnetic vehicles has mainly focused on the use of nanoparticles. In this work, Fe77B10Si10C3 glass-coated amorphous magnetic microwires were assessed for the first time as magnetic vehicles with high potential for the localized heating of osteosarcoma cells by means of an AC magnetic field. The results from the in vitro assays performed inside a microfluidic device demonstrated the ability of these magnetic microwires to induce malignant cell death. Exposing the system to different magnetic fields for less than 1 h provoked a reduction up to 89% of the osteosarcoma cell population, whereas healthy myoblastoma cells remained nearly unaffected. The proposed technology demonstrates in vitro the effectiveness of these microwires as vehicles for targeted magnetic hyperthermia.

Antibody biosensors for spoilage yeast detection based on impedance spectroscopy.

Brettanomyces is a yeast species responsible for wine and cider spoilage, producing volatile phenols that result in off-odors and loss of fruity sensorial qualities. Current commercial detection methods for these spoilage species are liable to frequent false positives, long culture times and fungal contamination. In this work, an interdigitated (IDE) biosensor was created to detect Brettanomyces using immunological reactions and impedance spectroscopy analysis. To promote efficient antibody immobilization on the electrodes' surface and to decrease non-specific adsorption, a Self-Assembled Monolayer (SAM) was developed. An impedance spectroscopy analysis, over four yeast strains, confirmed our device's increased efficacy. Compared to label-free sensors, antibody biosensors showed a higher relative impedance. The results also suggested that these biosensors could be a promising method to monitor some spoilage yeasts, offering an efficient alternative to the laborious and expensive traditional methods.

Effectiveness of nanoencapsulated methotrexate against osteosarcoma cells: in vitro cytotoxicity under dynamic conditions.

Cancer is a leading cause of mortality in the world, with osteosarcoma being one of the most common types among children between 1 and 14 years old. Current treatments including preoperative chemotherapy, surgery and postoperative chemotherapy produce several side effects with limited effectiveness. The use of lipid nanoparticles as biodegradable shells for controlled drug delivery shows promise as a more effective and targeted tumor treatment. However, in vitro validation of these vehicles is limited due to fluid stagnation in current techniques, in which nanoparticles sediment onto the bottom of the wells killing the cells by asphyxiation. In the current series of experiments, results obtained with methotrexate-lipid nanoparticles under dynamic assay conditions are presented as a promising alternative to current free drug based therapies. Effects on the viability of the U-2 OS osteosarcoma cell line of recirculation of cell media, free methotrexate and blank and methotrexate containing lipid nanoparticles in a 11 µM concentration were successfully assessed. In addition, several designs for the microfluidic platform used were simulated using COMSOL-Multiphysics, optimized devices were fabricated using soft-lithography and simulated parameters were experimentally validated. Nanoparticles did not sediment to the bottom of the platform, demonstrating the effectiveness of the proposed system. Moreover, encapsulated methotrexate was the most effective treatment, as after 72 h the cell population was reduced nearly 40% while under free methotrexate circulation the cell population doubled. Overall, these results indicate that methotrexate-lipid nanoparticles are a promising targeted therapy for osteosarcoma treatment.

Development of a biological protocol for endotoxin detection using quartz crystal microbalance (QCM).

In this paper, a biological protocol for endotoxin detection has been developed and optimized by quartz crystal microbalance (QCM). The parameters involved in the formation of the self-assembled monolayer (SAM) have been analyzed, and a study of the pH of the ligand buffer has been performed in order to find the best condition for the ligand immobilization and, in consequence, for the endotoxin detection. The detection limit obtained with the characterized biological protocol corresponds to 1.90 µg/ml. The effectiveness of the optimized biological protocol has been analyzed by cyclic voltammetry analysis.

Screening and selection of synthetic peptides for a novel and optimized endotoxin detection method.

The current validated endotoxin detection methods, in spite of being highly sensitive, present several drawbacks in terms of reproducibility, handling and cost. Therefore novel approaches are being carried out in the scientific community to overcome these difficulties. Remarkable efforts are focused on the development of endotoxin-specific biosensors. The key feature of these solutions relies on the proper definition of the capture protocol, especially of the bio-receptor or ligand. The aim of the presented work is the screening and selection of a synthetic peptide specifically designed for LPS detection, as well as the optimization of a procedure for its immobilization onto gold substrates for further application to biosensors.

Single-cell trapping and selective treatment via co-flow within a microfluidic platform.

Lab on a chip (LOC) systems provide interesting and low-cost solutions for key studies and applications in the biomedical field. Along with microfluidics, these microdevices make single-cell manipulation possible with high spatial and temporal resolution. In this work we have designed, fabricated and characterized a versatile and inexpensive microfluidic platform for on-chip selective single-cell trapping and treatment using laminar co-flow. The combination of co-existing laminar flow manipulation and hydrodynamic single-cell trapping for selective treatment offers a cost-effective solution for studying the effect of novel drugs on single-cells. The operation of the whole system is experimentally simple, highly adaptable and requires no specific equipment. As a proof of concept, a cytotoxicity study of ethanol in isolated hepatocytes is presented. The developed microfluidic platform controlled by means of co-flow is an attractive and multipurpose solution for the study of new substances of high interest in cell biology research. In addition, this platform will pave the way for the study of cell behavior under dynamic and controllable fluidic conditions providing information at the individual cell level. Thus, this analysis device could also hold a great potential to easily use the trapped cells as sensing elements expanding its functionalities as a cell-based biosensor with single-cell resolution.

Smart central venous port for early detection of bacterial biofilm related infections.

Central venous catheters (CVC) are commonly used in clinical practice to improve a patient's quality of life. Unfortunately, there is an intrinsic risk of acquiring an infection related to microbial biofilm formation inside the catheter lumen. It has been estimated that 80 % of all human bacterial infections are biofilm-associated. Additionally, 50 % of all nosocomial infections are associated with indwelling devices. Bloodstream infections account for 30-40 % of all cases of severe sepsis and septic shock, and are major causes of morbidity and mortality. Diagnosis of bloodstream infections must be performed promptly so that adequate antimicrobial therapy can be started and patient outcome improved. An ideal diagnostic technology would identify the infecting organism(s) in a timely manner, so that appropriate pathogen-driven therapy could begin promptly. Unfortunately, despite the essential information it provides, blood culture, the gold standard, largely fails in this purpose because time is lost waiting for bacterial or fungal growth. This work presents a new design of a venous access port that allows the monitoring of the inner reservoir surface by means of an impedimetric biosensor. An ad-hoc electronic system was designed to manage the sensor and to allow communication with the external receiver. Historic data recorded and stored in the device was used as the reference value for the detection of bacterial biofilm. The RF communication system sends an alarm signal to the external receiver when a microbial colonization of the port occurs. The successful in vitro analysis of the biosensor, the electronics and the antenna of the new indwelling device prototype are shown. The experimental conditions were selected in each case as the closest to the clinical working conditions for the smart central venous catheter (SCVC) testing. The results of this work allow a new generation of this kind of device that could potentially provide more efficient treatments for catheter-related infections.

Immunohistochemical evidence for myofibroblast-like cells associated with liver injury induced by aflatoxin B1 in rainbow trout (Oncorhynchus mykiss).

In mammalian species, profibrogenic cells are activated to become myofibroblasts in response to liver damage. Few studies have examined hepatic myofibroblasts and their role in liver damage in teleosts. The aim of the present study was to investigate the involvement of myofibroblast-like cells in rainbow trout (Oncorhynchus mykiss) with hepatic damage induced by aflatoxin B1 (AFB1). Histopathological and immunohistochemical analyses characterized alterations in the liver stroma during the carcinogenic process. Anti-human α-smooth muscle actin (SMA) and anti-human desmin primary antibodies were used in immunohistochemistry. Only the anti-SMA reagent labelled cells in trout liver. In the livers of control fish, only smooth muscle in blood vessels and around bile ducts was labelled. In the livers from AFB1-treated fish, SMA-positive cells were present in the stroma surrounding neoplastic lesions and in areas of desmoplastic reaction. These observations indicate that in teleosts, as in mammals, the myofibroblast-like cell is involved in fibrosis associated with liver injury. Chronic liver injury induced in trout by aflatoxin may provide a useful model system for study of the evolution of such mechanisms.

Trends in overweight among women differ by occupational class: results from 33 low- and middle-income countries in the period 1992-2009.

OBJECTIVE: There has been an increase in overweight among women in low- and middle-income countries but whether these trends differ for women in different occupations is unknown. We examined trends by occupational class among women from 33 low- and middle-income countries in four regions. DESIGN: Cross-national study with repeated cross-sectional demographic health surveys. SUBJECTS: Height and weight were assessed at least twice between 1992 and 2009 in 248,925 women aged 25-49 years. Interviews were conducted to assess occupational class, age, place of residence, educational level, household wealth index, parity, age at first birth and breastfeeding. We used logistic and linear regression analyses to assess the annual percent change in overweight (body mass index >25 kg m(-2)) by occupational class. RESULTS: The prevalence of overweight ranged from 2.2% in Nepal in 1992-1997 to 75% in Egypt in 2004-2009. In all the four regions, women working in agriculture had consistently lower prevalence of overweight, while women from professional, technical, managerial as well as clerical occupational classes had higher prevalence. Although the prevalence of overweight increased in all the occupational classes in most regions, women working in agriculture and production experienced the largest increase in overweight over the study period, while women in higher occupational classes experienced smaller increases. To illustrate, overweight increased annually by 0.5% in Latin America and the Caribbean and by 0.7% in Sub-Saharan Africa among women from professional, technical and managerial classes, as compared with 2.8% and 3.7%, respectively, among women in agriculture. CONCLUSION: The prevalence of overweight has increased in most low- and middle-income countries, but women working in agriculture and production have experienced larger increases than women in higher occupational classes.

Trends in overweight by educational level in 33 low- and middle-income countries: the role of parity, age at first birth and breastfeeding.

This study examined trends in overweight among women of reproductive age by educational level in 33 low- and middle-income countries, and estimated the contribution of parity, age at first birth and breastfeeding to these trends. We used repeated cross-sectional Demographic Health Surveys of 255,828 women aged 25-49 years interviewed between 1992 and 2009. We applied logistic regression to model overweight (>25 kg m(-2) ) as a function of education, reproductive variables and time period by country and region. The prevalence of overweight ranged from 3.4% in South and Southeast Asia to 73.7% in North Africa West/Central Asia during the study period. The association between education and overweight differed across regions. In North Africa West/Central Asia and Latin American, lower education was associated with higher overweight prevalence, while the inverse was true in South/Southeast Asia and Sub-Saharan Africa. In all regions, there was a consistent pattern of increasing overweight trends across all educational groups. Older age at first birth, longer breastfeeding and lower parity were associated with less overweight, for differences by educational level in overweight prevalence and trends.

Real time monitoring of the impedance characteristics of Staphylococcal bacterial biofilm cultures with a modified CDC reactor system.

Detection of device-associated infectious processes is still an important clinical challenge. Bacteria grow adhered to the device surfaces creating biofilms that are resistant to antimicrobial agents, increasing mortality and morbidity. Thus there is need of a surgical procedure to remove the indwelling infected device. The elevated cost of these procedures, besides patients discomfort and increased risks, highlights the need to develop more efficient, accurate and rapid detection methods. Biosensors integrated with implantable devices will provide an effective diagnostic tool. In vivo, rapid and sensitive detection of bacteria attached to the device surfaces will allow efficient treatments. Impedance spectroscopy technique would be an adequate tool to detect the adherence and the growth of the microorganism by monitoring the impedance characteristics. In this work a label-free interdigitated microelectrode (IDAM) biosensor has been developed to be integrated with implantable devices. Impedance characterization of Staphylococcus epidermidis biofilms has been performed achieving electrical monitoring of the bacterial growths in a few hours from the onset of the infection. This pathogen represents the most common microorganism related to intravascular catheters associated infections. The experimental setup presented in this work, a modified CDC biofilm reactor, simulates the natural environment conditions for bacterial biofilm development. The results prove that the low range of frequency is the most suitable setting for monitoring biofilm development. Our findings prove the effectiveness of this technique which shows variations of 59% in the equivalent serial capacitance component of the impedance.

Phylogenetic and host-parasite relationship analysis of Henneguya multiplasmodialis n. sp. infecting Pseudoplatystoma spp. in Brazilian Pantanal wetland.

A new species of the genus Henneguya (Henneguya multiplasmodialis n. sp.) was found infecting the gills of three of 89 specimens (3.3%) of Pseudoplatystoma corruscans and two of 79 specimens (2.6%) of Pseudoplatystoma reticulatum from rivers in the Pantanal wetland, Brazil. Partial sequencing of the 18S rDNA gene of the spores obtained from one plasmodium from the gills of P. corruscans and other one from the gills of P. reticulatum, respectively, resulted in a total of 1560 and 1147 base pairs. As the spores of H. multiplasmodialis n. sp. resemble those of Henneguya corruscans, which is also a parasite of P. corruscans, sequencing of the 18S rDNA gene of the spores of H. corruscans found on P. corruscans caught in the Brazilian Pantanal wetland was also provided to avoid any taxonomic pendency between these two species, resulting in 1913 base pairs. The sequences of H. multiplasmodialis n. sp. parasite of P. corruscans and P. reticulatum and H. corruscans did not match any of the Myxozoa available in the GenBank. The similarity of H. multiplasmodialis n. sp. obtained from P. corruscans to that from P. reticulatum was of 99.7%. Phylogeny revealed a strong tendency among Henneguya species to form clades based on the order and/or family of the host fish. H. multiplasmodialis n. sp. clustered in a clade with Henneguya eirasi and H. corruscans, which are also parasites of siluriforms of the family Pimelodidae and, together with the clade composed of Henneguya spp. parasites of siluriforms of the family Ictaluridae, formed a monophyletic clade of parasites of siluriform hosts. The histological study revealed that the wall of the plasmodia of H. multiplasmodialis n. sp. were covered with a stratified epithelium rich in club cells and supported by a layer of connective tissue. The interior of the plasmodia had a network of septa that divided the plasmodia into numerous compartments. The septa were composed of connective tissue also covered on both sides with a stratified epithelium rich in club cells. Inflammatory infiltrate was found in the tissue surrounding the plasmodia as well as in the septa.

GMR sensors: magnetoresistive behaviour optimization for biological detection by means of superparamagnetic nanoparticles.

An immunomagnetic method for the selective and quantitative detection of biological species by means of a magnetoresistive biosensor and superparamagnetic particles has been optimized. In order to achieve this, a giant magnetoresistive [Co (5.10nm)/Cu (2.47 nm)](20) multilayer structure has been chosen as the sensitive material, showing a magnetoresistance of 3.60% at 215 Oe and a sensitivity up to 0.19 Ω/Oe between 145 Oe and 350 Oe. The outward gold surface of the sensor is biofunctionalized with a Self-Assembled Monolayer (SAM). In addition, three different types of magnetic labels have been tested. 2 µm diameter magnetic carriers (7.68 pg ferrite/particle) have shown the best response and they have induced a shift in the magnetoresistive hysteresis loops up to 9% at 175 Oe.

Host-parasite-environment relationship, morphology and molecular analyses of Henneguya eirasi n. sp. parasite of two wild Pseudoplatystoma spp. in Pantanal Wetland, Brazil.

A new myxosporean species, Henneguya eirasi n. sp., is described parasitizing the gill filaments of Pseudoplatystoma corruscans and Pseudoplatystoma fasciatum (Siluriformes: Pimelodidae) caught in the Patanal Wetland of the state of Mato Grosso, Brazil. The parasite formed white, elongated plasmodia measuring up to 3mm. Mature spores were ellipsoidal in the frontal view, measuring 37.1 ± 1.8 µm in total length, 12.9 ± 0.8 µm in body length, 3.4 ± 0.3 µm in width, 3.1 ± 0.1 µm in thickness and 24.6 ± 2.2 µm in the caudal process. Polar capsules were elongated and equal in size, measuring 5.4 ± 0.5 µm in length and 0.7 ± 0.1 µm in width. Polar filaments had 12-13 coils. Histopathological analysis revealed that the parasite developed in the sub-epithelial connective tissue of the gill filaments and the plasmodia were surrounded by a capsule of host connective tissue. The plasmodia caused slight compression of the adjacent tissues, but no inflammatory response was observed in the infection site. Ultrastructure analysis revealed a single plasmodial wall connected to the ectoplasmic zone through numerous pinocytotic canals. The plasmodial wall exhibited numerous projections and slightly electron-dense material was found in the ectoplasm next to the plasmodial wall, forming a line just below the wall. Partial sequencing of the 18S rDNA gene of H. eirasi n. sp. obtained from P. fasciatum resulted in a total of 1066 bp and this sequence did not match any of the Myxozoa available in the GenBank. Phylogenetic analysis revealed the Henneguya species clustering into clades following the order and family of the host fishes. H. eirasi n. sp. clustered alone in one clade, which was the basal unit for the clade composed of Henneguya species parasites of siluriform ictalurids. The prevalence of the parasite was 17.1% in both fish species examined. Parasite prevalence was not influenced by season, host sex or host size.

Henneguya pseudoplatystoma n. sp. causing reduction in epithelial area of gills in the farmed pintado, a South American catfish: histopathology and ultrastructure.

The present study is part of an ongoing investigation into the characteristics of Myxozoan parasites of Brazilian freshwater fish and was carried out using morphology, histopathology and electron microscopy analysis. A new Myxosporea species (Henneguya pseudoplatystoma) is described causing an important reduction in gill function in the farmed pintado (a hybrid fish from a cross between Pseudoplatystoma corruscans and Pseudoplatystoma fasciatum), which is a commercially important South American catfish. From a total of 98 pintado juveniles from fish farms in the states of São Paulo and Mato Grosso do Sul (Brazil), 36 samples (36.7%) exhibited infection of the gill filaments. Infection was intense, with several plasmodia occurring on a same gill filament. The plasmodia were white and measured up to 0.5mm in length; mature spores were ellipsoidal in the frontal view, measuring 33.2+/-1.9 microm in total length, 10.4+/-0.6 microm in body length, 3.4+/-0.4 microm in width and 22.7+/-1.7 microm in the caudal process. The polar capsules were elongated, measuring 3.3+/-0.4 microm in length and 1.0+/-0.1 microm in width and the polar filaments had six to seven turns. Histopathological analysis revealed the parasite in the connective tissue of the gill filaments and lamella. No inflammatory process was observed, but the development of the plasmodia reduced the area of functional epithelium. Ultrastructural analyses revealed a single plasmodial wall, which was in direct contact with the host cells and had numerous projections in direction of the host cells as well as extensive pinocytotic canals. A thick layer (2-6 microm) of fibrous material and numerous mitochondria were found in the ectoplasm. Generative cells and the earliest stage of sporogenesis were seen more internally. Advanced spore developmental stages and mature spores were found in the central portion of the plasmodia.

Light, electron microscopy and histopathology of Myxobolus salminus n. sp., a parasite of Salminus brasiliensis from the Brazilian Pantanal.

In this report, we describe the morphology and histopathology of Myxobolus salminus n. sp., a parasite of the gill filaments of wild Salminus brasiliensis (dourado) from the Brazilian Pantanal. The small polysporic plasmodia were approximately 100 microm in diameter and the development was asynchronous. The mature spores were oval to pear shaped and had a smooth wall. The spore measurements were (mean+/-S.D., with range in parentheses): length 10.1+/-0.4 microm (9.6-10.5), width 6.1+/-0.4 microm (5.8-6.6) and thickness 5.0+/-0.6 microm (4.7-5.3). The polar capsules were elongated and of equal size: length 4.6+/-0.2 microm (4.3-4.8) and width 1.7+/-0.1 microm (1.5-1.9). The histological analysis revealed numerous plasmodia in the blood vessels of the gill filaments. The site of parasite development was the wall of the large-caliber blood vessel of the gill filament, with progressive growth towards the lumen, resulting in the obstruction of blood flow, congestion and perivascular edema. The ultrastructural study revealed that the plasmodial wall was composed of two membranes, had numerous pinocytic canals and was in direct contact with the basement membrane of the vessel. The development of the parasite was asynchronous, with mature spores, immature spores and young developmental stages randomly distributed throughout the plasmodium. The prevalence of the parasite was 4.4%, with male and female fish being infected.

Myxobolus cordeiroi n. sp., a parasite of Zungaro jahu (Siluriformes: Pimelodiade) from Brazilian Pantanal: morphology, phylogeny and histopathology.

This work is part of an ongoing investigation into the characteristics of Myxozoan parasites of freshwater fish in Brazil and was carried out using morphology, histopathology and molecular analysis. A new Myxosporea species (Myxobolus cordeiroi) is described infecting the jaú catfish (Zungaro jahu). Fifty jaú specimens were examined and 78% exhibited plasmodia of the parasite. The plasmodia were white and round, measuring 0.3-2.0mm in diameter and the development occurred in the gill arch, skin, serosa of the body cavity, urinary bladder and eye. The spores had an oval body and the spore wall was smooth. Partial sequencing of the 18S rDNA gene resulted in a total of 505bp and the alignment of the sequences obtained from samples in different organs revealed 100% identity. In the phylogenetic analysis, the Myxobolus species clustered into two clades-one primarily parasites of freshwater fish and the other primarily parasites of marine fish. M. cordeiroi n. sp. was clustered in a basal position in the freshwater fish species clade. The histological analysis revealed the parasite in the connective tissue of the different infected sites, thereby exhibiting affinity to this tissue. The plasmodium was surrounded by an outer collagen capsule of fibers with distinct orientation from the adjacent connective tissue and an inner layer composed of delicate collagen fibrils-more precisely reticular fibers. The development of the parasite in the cornea and urinary bladder caused considerable stretching of the epithelium.

Hemoglobin concentration is inversely associated with erythrocyte folate concentrations in Colombian school-age children, especially among children with low vitamin B12 status.

BACKGROUND: While the majority of cases of nutritional anemia in developing countries are caused by iron deficiency, other micronutrient deficiencies may also be involved. In Colombia, it was recently reported that 38% of school children were anemic; yet, the rate of iron deficiency was only 3.6%. OBJECTIVE: To determine if micronutrients other than iron were responsible for low hemoglobin concentrations in Colombian school children. METHODS: We examined hemoglobin concentrations in relation to plasma ferritin, vitamin A, vitamin B12, and erythrocyte folate levels in a representative sample of 2812 low- and middle-income children (5-12 years) from Bogotá, Colombia. RESULTS: In multivariate analysis, hemoglobin concentration was positively associated with child's age, mother's age, household's socioeconomic stratum, and family income. Low ferritin was related to 3.6 g/l lower hemoglobin concentration (95% confidence interval=-6.0, -1.3). Unexpectedly, we found an inverse trend in hemoglobin concentration by quartiles of erythrocyte folate; the adjusted hemoglobin concentration difference between the highest and lowest folate quartiles was -6.0 g/l (95% confidence interval=-7.2, -4.9; P for trend <0.0001). This difference was greatest among children with vitamin B12 concentration <148 pmol/l (-11.5 g/l), followed by children with vitamin B12 concentration 148-221 pmol/l (-7.7 g/l), and smallest in children with vitamin B12 concentration >221 pmol/l (-5.7 g/l); P for interaction=0.04. CONCLUSIONS: Hemoglobin concentration is inversely related to erythrocyte folate concentrations in a setting where folate fortification was adopted more than a decade ago. The impact of improving vitamin B12 status on this inverse relationship should be examined.

Magnetoresistive immunosensor for the detection of Escherichia coli O157:H7 including a microfluidic network.

A hand held device has been designed for the immunomagnetic detection and quantification of the pathogen Escherichia coli O157:H7 in food and clinical samples. In this work, a technology to manufacture a Lab on a Chip that integrates a 3D microfluidic network with a microfabricated biosensor has been developed. With this aim, the sensing film optimization, the design of the microfluidic circuitry, the development of the biological protocols involved in the measurements and, finally, the packaging needed to carry out the assays in a safe and straightforward way have been completed. The biosensor is designed to be capable to detect and quantify small magnetic field variations caused by the presence of superparamagnetic beads bound to the antigens previously immobilized on the sensor surface via an antibody-antigen reaction. The giant magnetoresistive multilayer structure implemented as sensing film consists of 20[Cu(5.10nm)/Co(2.47 nm)] with a magnetoresistance of 3.20% at 235Oe and a sensitivity up to 0.06 Omega/Oe between 150Oe and 230Oe. Silicon nitride has been selected as optimum sensor surface coating due to its suitability for antibody immobilization. In order to guide the biological samples towards the sensing area, a microfluidic network made of SU-8 photoresist has been included. Finally, a novel packaging design has been fabricated employing 3D stereolithographic techniques. The microchannels are connected to the outside using standard tubing. Hence, this packaging allows an easy replacement of the used devices.