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This study aimed to evaluate the HerbValo method in comparison to conventional field measurement techniques as a tool for estimating the herbage utilization and dry matter (DM) intake by grazing dairy cows under subtropical conditions. It was carried out during 18 months in a commercial farm of Southern Brazil. The herbage utilization was estimated monthly in two to four randomly selected paddocks (total n = 40) using the rising-plate meter or the double-sampling technique. In parallel, the herbage utilization was estimated by the HerbValo method, which is based on simple descriptions of the herd, supplements, pasture and grazing management. Values of herbage utilization estimated by field techniques were linearly (P < 0.01) related to those estimated by HerbValo, with no effect of pasture type (tropical vs. temperate) on the origin or on the slope of the regression (slope = 0.97; origin = -0.1; R 2 = 0.81; rsd = 0.17 t DM/ha). At cow × day level, values of herbage intake estimated by field techniques were also linearly related to those estimated by HerbValo (P < 0.01; R 2 = 0.82; rsd = 1.30 kg DM/cow/day). A negative linear relationship (P < 0.01) between herbage and supplement intake was obtained for both field (slope = -1.06; R 2 = 0.72; rsd = 1.64) and HerbValo (slope = -0.92; R 2 = 0.82; rsd = 1.08) approaches. Herbage utilization and intake by a dairy herd in a subtropical grazing-based system can be reliably estimated by the HerbValo method with the advantage of not requiring direct field pasture measurements., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Introduction: Burnout syndrome results from a response to chronic work stress and is responsible for causing symptoms related to three dimensions: emotional exhaustion, reflecting work overload; depersonalization, characterized by professional detachment and cynicism; and reduced professional accomplishment, related to low productivity at work. Burnout is often associated with jobs that require professionals to have direct contact with users, such as health professionals. Primary Health Care is the assistance level that greatest contact with the community and requires teamwork, thus exposing workers to potential psychosocial stressors., Objectives: To identify the prevalence of symptoms of burnout syndrome among Primary Health Care professionals in Toledo, state of Paraná, Brazil., Methods: This was a descriptive, quantitative, cross-sectional study. A sociodemographic questionnaire and the Maslach Burnout Inventory, Human Services Survey, were used to assess the outcomes., Results: The prevalence of high risk for the development of burnout syndrome was 10.6%, and, when dimensions were evaluated separately, it was found that 29.8, 52.1 and 22.3% of participants showed a high level of symptoms in the dimensions emotional exhaustion, reduced professional accomplishment, and depersonalization, respectively. Previous use of psychiatric medication due to another condition had a significant correlation with high risk for burnout., Conclusions: The results of this research corroborated other similar studies, contributing to knowledge on the subject in a region of the state of Paraná where there was still no research on the syndrome., Competing Interests: Conflicts of interest: None

The membrane of methicillin-resistant Staphylococcus aureus (MRSA) contains penicillin-binding proteins (PBPs) in the phospholipidic bilayer, with the protein PBP2a being linked with the resistance mechanism. In this work we confirm the role of PBP2a with molecular-level information obtained with Langmuir monolayers as cell membrane models. The MRSA cell membrane was mimicked with a mixed monolayer of dipalmitoyl phosphatidyl glycerol (DPPG) and cardiolipin (CL), also incorporating PBP2a. The surface pressure-area isotherms and the Brewster angle microscopy (BAM) images for these mixed monolayers were significantly affected by the antibiotic meropenem, which is PBP2a inhibitor. The meropenem effects were associated with the presence of PBP2a, as they were absent in the Langmuir monolayers without PBP2a. The relevance of PBP2a was confirmed with results where the antibiotic methicillin, known to be unsuitable to kill MRSA, had the same effects on mixed DPPG/CL and DPPG/CL-PBP2a monolayers since it prevented PBP2a from incorporating in the monolayer. The biological implication of the findings presented here is that a successful antibiotic against MRSA should be able to interact with PBP2a, but in the membrane., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Alzheimer's disease (AD) is the most common form of dementia and affect more than 50 million people worldwide. Thus, there is a high demand by non-invasive methods for an early diagnosis. This work explores the AD diagnostic using the amyloid beta 1-40 (Aβ40) peptide encapsulated into dipalmitoyl phosphatidyl glycerol (DPPG) liposomes and immobilized on polyethylene imine previously deposited on screen-printed carbon electrodes to detect autoantibodies against Aβ40, a potential biomarker found in plasma samples., Methods: The immunosensor assembly was accompanied by atomic force microscopy (AFM) images that showed globular aggregates from 20 to 200 nm corresponding liposomes and by cyclic voltammetry (CV) through increase of the voltammogram area each material deposited. After building the immunosensor, when it was exposed to antibody anti-Aβ40, there was an increase in film roughness of approximately 9 nm, indicating the formation of the immunocomplex., Results: In the detection by CV, the presence of specific antibody, in the range of 0.1 to 10 μg/ml, resulted in an increase in the voltammograms area and current in 0.45 V reaching 3.2 µA.V and 5.7 μA, respectively, in comparison with the control system, which remained almost unchanged from 0.1 μg/ml. In patient samples, both cerebrospinal fluid (CSF) and plasma, was possible separated among positive and negative samples for AD using CV profile and area, with a difference of 0.1 μA.V from the upper error bar of healthy samples for CSF sample and 0.6 μA.V for plasma sample., Conclusions: These results showed the feasibility of the method employed for the non-invasive diagnostic of Alzheimer's disease detecting natural autoantibodies that circulate in plasma through a simple and easy-to-interpret method., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Coronavirus disease 2019 (COVID-19) is a viral respiratory disease that spreads rapidly, reaching pandemic status, causing the collapse of numerous health systems, and a strong economic and social impact. The treatment so far has not been well established and there are several clinical trials testing known drugs that have antiviral activity, due to the urgency that the global situation imposes. Drugs with specific mechanisms of action can take years to be discovered, while vaccines may also take a long time to be widely distributed while new virus variants emerge. Thus, drug repositioning has been shown to be a good strategy for defining new therapeutic approaches. Studies of the effect of enriched heparin in the replication of severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) in vitro assays justify the advance for clinical tests., Methods and Analysis: A phase I/II triple-blind parallel clinical trial will be conducted. Fifty participants with radiological diagnosis of grade IIA pneumonia will be selected, which will be allocated in 2 arms. Participants allocated in Group 1 (placebo) will receive nebulized 0.9% saline. Participants allocated in Group 2 (intervention) will receive nebulized enriched heparin (2.5 mg/mL 0.9% saline). Both groups will receive the respective solutions on a 4/4 hour basis, for 7 days. The main outcomes of interest will be safety (absence of serious adverse events) and efficacy (measured by the viral load).Protocols will be filled on a daily basis, ranging from day 0 (diagnosis) until day 8., Competing Interests: The authors declare that there is no conflict of interest., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)

Spatial location is a factor that may facilitate the consumption of processed foods due to the ease of access, especially in highly urbanized, high density populations. This study presents the differences in food consumption in populations with different densities and urban and rural characteristics, and income above and below ten dollars a day. A sample of 2,130 subjects was used from 9 populations that included the Capital of Colombia, outlying medium-sized municipalities, metropolitan-area municipalities and small villages. The results confirm that processed and ultra-processed foods are consumed more in cities and urban areas than in smaller and rural populations, and that there are differences in consumption generated by income.

Venous thromboembolism (VTE) is a serious clinical condition which early and accurate diagnosis may contribute to the reduction of associated morbidity and mortality. VTE occurs when a blood clot (thrombus) blocks the vein blood flow causing deep vein thrombosis (DVT) and, when it migrates to the lungs, it may clog the pulmonary arteries characterizing pulmonary embolism (PE). Analysis using fibrin degradation products or D-dimer and coagulation factor VIII may assist early diagnosis of VTE. Thus, two immunosensors were built using layer-by-layer (LbL) films technique, one containing the anti-D-dimer immobilized on polyethylene imine (PEI) and another the anti-FVIII on silk fibroin (SF). Immunosensor response, the antigen-antibody specific interaction, was investigated using cyclic voltammetry. When immunosensors, PEI/anti-D-dimer and SF/anti-FVIII, were exposed to antigens, D-dimer and Factor VIII, the voltammograms area and current were significantly increased with increasing specific antigen concentration. The specific interaction was confirmed with control experiments, electrodes containing only PEI or SF, that no significant changes in the voltammogram responses were observed and principal component analysis confirmed these results. The films formation and response were verified using scanning electronic microscopy (SEM). The developed immunosensor seems to be a promising and effective early complementary exam to assist in the VTE diagnosis, through the combined response of two biomarkers very sensible., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The growing restriction of antibiotic growth promoters (AGP) use in farming animals has raised a concern regarding the viability of the animal production system. In this new context, feed additives with proven positive impact on intestinal health may be used as strategy to avoid losses on performance. The aim of this study was to evaluate the effects of a protected blend of organic acids and essential oils [P(OA+EO)] on growth performance, nutrient digestibility, and intestinal health of broiler chickens. A total of 1,080 Cobb × Cobb 500 male broilers were randomly distributed in four treatments with 10 replicates (27 birds/each). Treatments were as follow: non-challenged control; challenged control; AGP (enramycin at 10 g/t); and P(OA+EO) at 300 g/t. All birds on challenged groups were challenged with Eimeria spp. at 1 day and with Clostridium perfringens at 11, 12, and 13 days. Body weight gain (BWG), feed intake and feed conversion ratio (FCR) were evaluated until 42 days. At 17 days, one bird per pen was orally gavaged with fluorescein isothiocyanate-dextran (FITC-d) and blood samples were collected for FITC-d detection to assess intestinal permeability. At 21 days, apparent ileal nutrient and energy (IDE) digestibility, intestinal macroscopic and histologic alterations (ISI) and, expression of mucin2 (MUC2), claudin1 (CLDN1), and occludin (OCLN) genes in the jejunum were evaluated. From 1 to 42 days, birds from the non-challenged and P(OA+EO) groups had greater ( P < 0.001) BWG compared to challenged control and AGP groups. The challenged control group presented the worst FCR ( P < 0.001). IDE was 106 kcal/kg greater when broilers were fed P(OA+EO) compared to the challenged control group. Broilers supplemented with P(OA+EO) had improved intestinal integrity with lower blood FITC-d concentration and ISI scores, and greater expression of MUC2, CLDN1, and OCLN genes compared to the challenged control group ( P < 0.05). In conclusion, the P(OA+EO) and the AGP led to increased growth performance, nutrient digestibility and intestinal health of challenged broilers. A marked difference occurred in favor of the P(OA+EO), suggesting that this blend may be used to improve intestinal health and broiler growth performance in AGP free programs., (Copyright © 2020 Stefanello, Rosa, Dalmoro, Segatto, Vieira, Moraes and Santin.)

The diagnostics of the autoimmune hemolytic anemia (AIHA), a rare disease caused by autoantibody-induced hemolysis, is still prone to false positives for it is based on visual observation in the so-called Direct Coombs test. In this study, we developed a specific IgG hemolysis immunosensor produced with layer-by-layer (LbL) films containing a monoclonal antibody against human immunoglobulin (mAbIMUG) deposited along with a layer of silk fibroin (SF) derived from Bombyx mori cocoons. Adsorption of mAbIMUG on a SF layer was confirmed by the fluorescence emission band at 326 nm. Immunosensors were prepared with LbL films deposited on interdigitated gold electrodes for impedance spectroscopy and on screen printed carbon electrodes for electrochemical measurements. When the SF/mAbIMUGLbL film was exposed to healthy red blood cells (RBCs), no cell binding was observed by the optical microscopy images. In addition, no major changes were observed in the signals of the square wave voltammogram and in the impedance spectra. In contrast, the electrochemical signal was significantly increased and the dielectric loss curve shifted for the sensing units containing RBCs with the antibody attached on the surface (" sick cells "). Furthermore, cell attachment was so strong that optical images still showed covered electrodes even after washing in PBS buffer. The detection with two distinct methods seems promising for an effective diagnosis of AIHA.

Two experiments were conducted to compare the immunocompetence of Cobb high performance and rustic Label Rouge broilers and the influence of reduced growth rates subsequent to feed restriction on the IgY anti-bovine serum albumin (BSA) response. In the first experiment (EXP), 360 broilers were assigned to 36 cages from 1 to 42 days of age. A completely randomized design was applied in a 3 × 2 factorial arrangement, with 3 groups (Label Rouge, Cobb ad libitum, and Cobb Restricted Intake), and 2 levels of energy (3,100 and 2,800 kcal/kg); there were 6 replicates per treatment. In the second EXP, 384 Cobb 500 male broilers were randomly assigned to the following feed restriction programs from day 8 to 16: Control, fed ad libitum; Quantitative (80% of the control amount); By Time (fed for 8 h/d), and Qualitative (80% limiting nutrients) restriction. Blood samples were collected on days 35 and 42 (EXP 1) and weekly from day 7 to 42 (EXP 2) for IgY anti-BSA quantification. In EXP 1, the production of IgY anti-BSA was lower in the Cobb groups (P < 0.0001) than in the Label Rouge group, and higher in the Cobb Restricted Intake group (P < 0.0001) compared with the same genetic strain fed ad libitum. Birds fed the low energy diet presented lower (P ≤ 0.06) IgY anti-BSA, independent of genetics. In EXP 2, no difference (P > 0.05) was observed 1 wk after the first BSA inoculation. However, at day 28, birds in all feed restriction programs had higher (P < 0.05) IgY anti-BSA than the Control group fed ad libitum. At day 35, the greatest residual effect of IgY anti-BSA was observed in the Quantitative restriction group. No differences (P > 0.05) were observed between groups after 42 d. The 3 early feed restriction programs had beneficial effects on the humoral immune response. Overall, Quantitative restriction promoted a longer lasting IgY anti-BSA response. Lower growth rate, due to feed restriction or genetic potential, improves humoral immunity in broiler chickens.

In this paper, we show that chitosan may induce conformation changes in silk fibroin (SF) in layer-by-layer (LbL) films, which were used as matrix for immobilization of the enzyme phytase to detect phytic acid. Three chitosan (CH) samples possessing distinct molecular weights were used to build CH/SF LbL films, and a larger change in conformation from random coils to β-sheets for SF was observed for high molecular weight chitosan (CHH). The CHH/SF LbL films deposited onto interdigitated gold electrodes were coated with a layer of phytase, with which phytic acid could be detected down to 10 -9 M using impedance spectroscopy as the principle of detection and treating the data with a multidimensional projection technique. This high sensitivity may be ascribed to the suitability of the CHH/SF matrix, thus indicating that the molecular-level interactions between chitosan and SF may be exploited in other biosensors and biodevices., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We report an electrochemical glucose biosensor made with layer-by-layer (LbL) films of functionalized reduced graphene oxide (rGO) and glucose oxidase (GOx). The LbL assembly using positively and negatively charged rGO multilayers represents a simple approach to develop enzymatic biosensors. The electron transport properties of graphene were combined with the specificity provided by the enzyme. rGO was obtained and functionalized using chemical methods, being positively charged with poly(diallyldimethylammonium chloride) to form GPDDA, and negatively charged with poly(styrene sulfonate) to form GPSS. Stable aqueous dispersions of GPDDA and GPSS are easily obtained, enabling the growth of LbL films on various solid supports. The use of graphene in the immobilization of GOx promoted Direct Electron Transfer, which was evaluated by Cyclic Voltammetry. Amperometric measurements indicated a detection limit of 13.4μmol·L(-1) and sensitivity of 2.47μA·cm(-2)·mmol(-1)·L for glucose with the (GPDDA/GPSS)1/(GPDDA/GOx)2 architecture, whose thickness was 19.80±0.28nm, as determined by Surface Plasmon Resonance (SPR). The sensor may be useful for clinical analysis since glucose could be detected even in the presence of typical interfering agents and in real samples of a lactose-free milk and an electrolyte solution to prevent dehydration., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Layer-by-Layer (LbL) films containing silk fibroin (SF) and the 40 aminoacid-long amyloid-β peptide (Aβ1-40) were prepared with the purpose of developing a new prototype of an electrochemical immunosensor. The film showed a satisfactory growth in quartz substrate and screen-printed carbon electrodes, as observed by UV-vis spectroscopy and cyclic voltammetric, respectively. The peptide immobilized in LbL films in junction with SF shows secondary structure induced, as shown by circular dichroism measurements, favoring the interaction SF/peptide LbL film with the specific antibody. Immunosensor showed a linear response in the presence of the antibody with concentrations from 0 to 10ngmL(-1) both analyzed by current changes in 0.3V and voltammogram area. This system can be applied as a new prototype for preliminary diagnosis of Alzheimer's disease., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Hymenaea stigonocarpa is a deciduous and monoecious Neotropical tree species pollinated by bats. Due to overexploitation and habitat destruction, the population size has drastically diminished in nature. No previous study has investigated Mendelian inheritance, linkage, and genotypic disequilibrium in the available microsatellite markers in this species. So, our aim was to estimate these parameters using six microsatellite loci in a sample of 470 adults and 219 juveniles from two populations of H. stigonocarpa. In addition, 30 seeds per tree from 35 seed-trees were collected. Each seed was kept record of the seed-trees and fruit origin. Based on the six microsatellite loci, we found that only 10.6% of the cases showed significant deviations from Mendelian segregation and 15.3% showed linkage. We detected no evidence of genotypic disequilibrium between the loci in the adult trees or juveniles. Thus, our results suggest that these loci can be used with great accuracy in future genetic analyses of H. stigonocarpa populations.

The application of Design of Experiments (DoE) to the determination of optimum conditions for an extraction process relies on the correct selection of mathematical models. The linear model is the one typically used; however, in some cases it does not always have superior performance, ignoring the real nature of the data and its appropriate descriptive model. In order to evaluate the extraction efficiency of isorhamnetin-3-O-rutinoside from flowers of Calendula officinalis L. a multivariate factorial analysis was used. Simulations were conducted using linear, quadratic, full cubic and special cubic models. A Simplex-Centroid design was chosen as it delivered greater precision with only minor errors versus other models tested. Analyses were performed by capillary zone electrophoresis using sodium tetraborate buffer (40mmolL(-1), pH 9.4) containing 10% methanol. The detection was linear over a range of 8.0-50.0mgL(-1) (r(2)=0.996), and the limits of detection (LOD) and quantification (LOQ) for isorhamnetin-3-O-rutinoside were 3.44mgL(-1) and 11.47mgL(-1), respectively. The full cubic model showed the best extraction results, with an error of 3.40% compared to analysis of variance, and a determination coefficient of 0.974. The difference between the responses at the reference point, calculated by the model, and the experimental response, varies around 2.72% for full cubic model. Comparison of the four models showed the full cubic model was the most appropriate one, allowing greater efficiency in the extraction of isorhamnetin-3-O-rutinoside. Selection of the model made it possible to obtain a 60% increase in sensitivity compared to the linear model., (Copyright © 2016. Published by Elsevier B.V.)

Objective: To assess the relationship between periodontal treatment needs by elderly Brazilians and contextual as well as individual variables., Methods: A cross-sectional study was carried out to assess the need for clinical periodontal treatment, based on National Oral Health Survey (SB Brasil 2010) data on the presence of dental calculus, shallow (3-5 mm) and deep (≥ 6 mm) periodontal pockets, and gingival bleeding in elderly people (n = 7,619). The contextual variables included the Municipal Human Development Index (MHDI), income inequality (Gini Index) and coverage of the municipal population by the Family Health Strategy (FHS) program oral health teams.<0} The individual variables were sex, income, education level and self-reported skin color. Multilevel logistic regression models were used to calculate the odds ratio (OR) and 95% confidence intervals (CI95%) between periodontal treatment needs and the contextual as well as individual variables., Results: Gingival bleeding was found in 20.7% of the elderly analyzed (n = 1,577), dental calculus in 34% (n = 2,590), shallow periodontal pockets in 15.6% (n = 1,189), and deep periodontal pockets in 4.2% (n = 320). Individual factors were correlated with all the outcomes assessed. Sex was a protective factor in regard to gingival bleeding (OR = 0.87; CI95% 0.76-1.00), dental calculus (OR = 0.86; CI95% 0.75-0.99), shallow periodontal pockets (OR = 0.69; CI95% 0.60-0.80) and deep periodontal pockets (OR = 0.58; CI95% 0.45-0.74). It was found that fewer women needed treatment. Elderly people who self-reported having nonwhite skin had higher chances of needing periodontal treatment. Skin color was a risk factor for gingival bleeding (OR = 1.32; CI95% 1.14-1.53), dental calculus (OR = 1.32; CI95%1.14-1.54) and shallow periodontal pockets (OR = 1.27; CI95% 1.09-1.49). Education level was associated with the presence of dental calculus (OR = 0.77; CI95% 0.66-0.89), shallow periodontal pockets (OR = 0.86; CI95% 0.73-1.00) and deep periodontal pockets (OR = 0.74; CI95% 0.57-0.97), thus acting as a risk factor for undereducated elderly people. There was a correlation between population coverage by the Family Health Strategy (FHS) program oral health teams and the presence of gingival bleeding (OR = 0.67; CI95% 0.52-0.88), shallow periodontal pockets (OR = 0.76; CI95% 0.58-0.98) and deep periodontal pockets (OR = 0.62; CI95% 0.44-0.89), making these teams act as a protective factor., Conclusions: This study showed evidence of the sociocontextual as well as individual sociodemographic characteristics influencing periodontal treatment needed by elderly Brazilians, based on the clinical features of periodontal disease. The results suggest the existence of inequality related to periodontal treatment needs among elderly Brazilians, especially in regard to sex and ethnicity, in addition to a potentially positive impact from the expansion of the Family Health Strategy (FHS) program oral health teams.

The effects of lutein and conjugated linoleic acid (CLA) on growth performance and immune response of broiler chickens were evaluated in the presence and absence of Salmonella lipopolysaccharide (LPS) immune challenge. Cobb chicks (360; 1 to 22 d of age) were used in a 3 × 2 factorial arrangement of CLA (0, 1, and 2%) and lutein (0 and 50 mg/kg) dietary levels. At d 8 and 15, birds were injected with BSA to assess IgY production. At d 20, birds were injected with LPS. Samples of liver, spleen, and duodenum were collected at 3 and 16 h post-LPS challenge for RT-qPCR analysis of RXRα, RXRγ, PPARα, PPARγ, TLR-4, IL-1β, IL-2, IL-10, and IL-12 gene expression. CLA decreased BW, BW gain (BWG), and G:F from d 1 to 20, but these effects were reversed when lutein was included in the 1% CLA diet (P < 0.001). The production of IgY anti-BSA increased following a 2% CLA supplementation (P < 0.01). LPS increased the liver:BW ratio at 3 h post-injection (P < 0.001) and decreased BWG at 3, 16, and 40 h (P < 0.001). Lutein decreased plasmatic nitric oxide levels (P < 0.01). LPS downregulated PPARα mRNA in the duodenum (P = 0.02) and liver (P = 0.04), and PPARγ (P = 0.01) and RXRα (P = 0.08) in the spleen; these effects were not reversed by CLA or lutein as initially hypothesized. Although LPS upregulated IL-1β (P = 0.02) and IL-12 (P = 0.07) expression, lutein downregulated these pro-inflammatory cytokines in the liver (P = 0.03 and P = 0.07, respectively). Lutein decreased splenic (P = 0.09) but increased hepatic (P = 0.06) TLR-4 mRNA. A dietary CLA supplementation of 2% increased hepatic RXRα (P = 0.10). In conclusion, CLA decreased broiler chicken growth performance, but lutein could prevent this negative effect (depending on CLA dose). Lutein had an anti-inflammatory effect, and a 2% CLA supplementation improved the humoral immune response., (© 2015 Poultry Science Association Inc.)

In this work nanostructured film composites of the monoamine oxidase B (MAO-B) enzyme, free or encapsulated in liposomes, were fabricated by the layer-by-layer (LbL) self-assembly technique, employing polyethylene imine (PEI) as polycation. Initially, the MAO-B enzyme was incorporated into liposomes in order to preserve its enzymatic structure ensuring their activity and catalytic stability. The LbL film growth was monitored by surface plasmon resonance (SPR) by gold resonance angle shift analysis after each bilayer deposition. Subsequently, the films were applied as amperometric biosensors for dopamine detection using Prussian Blue (PB) as the electron mediator. The biosensor fabricated by MAO-B incorporated into liposomes composed of DPPG:POPG in the ratio (1:4) (w/w) showed the best performance with a sensitivity of 0.86 (μA cm(-2))/(mmol L(-1)) and a detection limit of 0.33 mmol L(-1).

This work reports the production of an exo-polygalacturonase (exo-PG) by Rhizomucor pusillus A13.36 in submerged cultivation (SmC) in a shaker at 45°C for 96 h. A single pectinase was found and purified in order to analyze its thermal stability, by salt precipitation and hydrophobic interaction chromatography. The pectinase has an estimated Mw of approximately 43.5-47 kDa and optimum pH of 4.0 but is stable in pH ranging from 3.5 to 9.5 and has an optimum temperature of 61°C. It presents thermal stability between 30 and 60°C, has 70% activation in the presence of Ca 2+ , and was tested using citrus pectin with a degree of methyl esterification (DE) of 26%. E a ( d ) for irreversible denaturation was 125.5 kJ/mol with positive variations of entropy and enthalpy for that and Δ G ( d ) values were around 50 kJ/mol. The hydrolysis of polygalacturonate was analyzed by capillary electrophoresis which displayed a pattern of sequential hydrolysis (exo). The partial identification of the primary sequence was done by MS MALDI-TOF and a comparison with data banks showed the highest identity of the sequenced fragments of exo-PG from R. pusillus with an exo-pectinase from Aspergillus fumigatus . Pectin hydrolysis showed a sigmoidal curve for the Michaelis-Menten plot., Competing Interests: The authors declare that there are no competing interests regarding the publication of this paper.

Immunosensors based on electrical impedance spectroscopy (EIS) are increasingly being used as a fast and potentially low cost method for clinical diagnostics. In this work we fabricated immunosensors by depositing layer-by-layer (LbL) films made with an antigenic peptide (p17-1) sequence (H2N-LSGGELDRWEKIRLRPGG-OH) and lignin on interdigitated gold electrodes, which could detect anti-p17 (HIV, human immune deficiency virus) antibodies (Ab) in phosphate buffered solutions (PBS). The molecular recognition interaction between the peptide (p17-1) and the specific Ab (anti-p17) yielded substantial changes in morphology of the with LbL films, with increased roughness according to atomic force microscopy data. This interaction is behind the high sensitivity of the immunosensor. Indeed, from the EIS results, we noted that the capacitance increased significantly with the specific Ab concentration, before getting close to saturation of available peptide sites at high concentrations. Concentrations of specific antibodies as low as 0.1 ng/mL could be detected and the immunosensors had their activity preserved for two months at least. The selectivity of the immunosensor was confirmed with two types of control experiments. First, no changes in impedance were observed when the lignin/peptide LbL immunosensor was immersed into a PBS solution containing the non-specific Ab (anti-HCV for Hepatitis C) antibodies. Furthermore, for sensing units made LbL films of lignin only, the electrical response was not affected by adding specific antibodies into the PBS buffer. The successful immunosensing for HIV with antigenic peptides in a lignin matrix is also relevant for valorization of lignin, which is an important biomass component in the sugar and ethanol industry, and brings the prospect for all-organic, biocompatible sensors if implantation is ever required., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The rubber tree [Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell. Arg.] is the only plant species worldwide that is cultivated for the commercial production of natural rubber. This study describes the genetic diversity of the Hevea spp. complex that is available in the main ex situ collections of South America, including Amazonian populations that have never been previously described. Genetic data were analyzed to determine the genetic structure of the wild populations, quantify the allelic diversity and suggest the composition of a core collection to capture the maximum genetic diversity within a minimal sample size. A total of 1,117 accessions were genotyped with 13 microsatellite markers. We identified a total of 408 alleles, 319 of which were shared between groups and 89 that were private in different groups of accessions. In a population structure and principal component analysis, the level of clustering reflected a primary division into the following two subgroups: cluster 1, which consisted of varieties from the advanced breeding germplasm that originated from the Wickham and Mato Grosso accessions; and cluster 2, which consisted of the wild germplasm from the Acre, Amazonas, Pará and Rondônia populations and Hevea spp. The analyses revealed a high frequency of gene flow between the groups, with the genetic differentiation coefficient (GST) estimated to be 0.018. Additionally, no distinct separation among the H. brasiliensis accessions and the other species from Amazonas was observed. A core collection of 99 accessions was identified that captured the maximum genetic diversity. Rubber tree breeders can effectively utilize this core collection for cultivar improvement. Furthermore, such a core collection could provide resources for forming an association panel to evaluate traits with agronomic and commercial importance. Our study generated a molecular database that should facilitate the management of the Hevea germplasm and its use for subsequent genetic and genomic breeding.

Genipa americana is a tropical tree species that is widely distributed in the humid tropical and subtropical regions of Central and South America. This study investigated Mendelian inheritance, genetic linkage, and genotypic disequilibrium at six microsatellite loci developed for G. americana. Adult trees (188) and regenerants (163) were sampled and genotyped in a fragmented population of the species. We also genotyped open-pollinated seeds from 12 seed-trees during reproductive events in 2010 and 2011. Significant deviations from the expected 1:1 Mendelian segregation were detected in 29.5% of the tests. Significant genetic linkage between pairwise loci was detected in 54.4% of the tests, but no genotypic disequilibrium was detected between pairwise loci for adult trees and regenerants. Overall, the results indicate that the six loci analyzed may be used in studies of G. americana's genetic diversity and structure, its mating system, and in parentage analyses.

We report the infant feeding experiences in the first month of life for 2,053 infants participating in "Malnutrition and Enteric Infections: Consequences for Child Health and Development" (MAL-ED). Eight sites (in Bangladesh, India, Nepal, Pakistan, Brazil, Peru, South Africa, Tanzania), each followed a cohort of children from birth (by day 17), collecting detailed information on infant feeding practices, diet and illness episodes. Mothers were queried twice weekly regarding health status, breastfeeding and the introduction (or no) of non-breast milk liquids and foods. Here, our goal is to describe the early infant feeding practices in the cohort and evaluate factors associated with termination of exclusive breastfeeding in the first month of life. With data from enrollment to a visit at 28-33 days of life, we characterized exclusive, predominant or partial breastfeeding (using a median of 6-9 visits per child across the sites). Only 6 of 2,053 infants were never breastfed. By one month, the prevalences of exclusive breastfeeding were < 60% in 6 of 8 sites, and of partial breastfeeding (or no) were > 20% in 6 of 8 sites. Logistic regression revealed that prelacteal feeding (given to 4-63% of infants) increased the likelihood of partial breastfeeding (Odds Ratio (OR): 1.48 (95% confidence interval (CI): 1.04, 2.10), as did the withholding of colostrum (2-16% of infants) (OR: 1.63:1.01, 2.62), and being a first-time mother (OR: 1.38:1.10, 1.75). Our results reveal diversity across these sites, but an overall trend of early transition away from exclusive breastfeeding in the first month of life. Interventions which introduce or reinforce the WHO/UNICEF Ten Steps for Successful Breastfeeding are needed in these sites to improve breastfeeding initiation, to reinforce exclusive breastfeeding and delay introduction of non-breast milk foods and/or liquids.

The objective was to identify behaviors and sexual practices of men who have sexual relations with other men in the context of vulnerability to HIV/AIDS. This was a cross-sectional, exploratory and descriptive study. It was carried out in a gay sociability place in Fortaleza, Ceará, Brazil, between November 2010 and March 2011, through interviews with 189 men who have sex with men. The ethical aspects were respected. We found a sample consisting mostly by young, single, and highly educated men. The sexual history demonstrated the early onset of sexual activity, with a high prevalence of sexual intercourse with a partner of the opposite sex. There was also a high prevalence of HIV testing. Sexual practices revealed high prevalence of performing oral and anal sex, as well as high levels of protection in anal sex, despite the low protection in oral sex. A greater incorporation of prevention practices was found compared to the national scene in the beginning of the disease outbreak.

The overall goal of The Etiology, Risk Factors and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development (MAL-ED) cohort study is to evaluate the roles of repeated enteric infection and poor dietary intakes on the development of malnutrition, poor cognitive development, and diminished immune response. The use of 8 distinct sites for data collection from Latin America, sub-Saharan Africa, and South Asia allow for an examination of these relationships across different environmental contexts. Key to testing study hypotheses is the collection of appropriate data to characterize the dietary intakes and nutritional status of study children from birth through 24 months of age. The focus of the current article is on the collection of data to describe the nature and adequacy of infant feeding, energy and nutrient intakes, and the chosen indicators to capture micronutrient status in children over time., (© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

In this work we developed an immunosensor for HIV-1 diagnostics that exploits the biorecognition between the antibody anti-p24 and the antigenic peptide p24-3 (AMATLRAEQASQEVKNWMTETL- LVQNA) derived from the HIV-1 p24 protein. p24-3 was encapsulated in phospholipid liposomes and immobilized in layer-by-layer (LbL) films produced with polyethyleneimine (PEI). The incorporation of p24-3 into liposomes was investigated using circular dichroism (CD) spectroscopy, from which an increase in the alpha helix conformation could be noted. The maximum fluorescence emission for p24-3 occurred at 340 nm in solution, compatible with the tryptophan residue being exposed to the solvent, and at 335 and 322 nm when in liposomes and PEI/p24-3-liposome LbL films, respectively. This blue shift is consistent with the tryptophan being in a partially buried environment. With the preserved structure in the LbL films, p24-3 could recognize the anti-p24 antibody in impedance spectroscopy measurements. Therefore, LbL films containing p24-3 may be suitable for detecting HIV-1 in a low-cost, easy-to-use immunosensing assay.

Iron, copper, and zinc are key micronutrients that play an important role in the immune response to Mycobacterium tuberculosis. The present study aimed to evaluate the association between serum levels of those micronutrients, inflammatory markers, and the smear and culture conversion of M. tuberculosis during 60 days of tuberculosis treatment. Seventy-five male patients with pulmonary tuberculosis (mean age, 40.0 ± 10.7 years) were evaluated at baseline and again at 30 and 60 days of tuberculosis treatment. Serum levels of iron, copper, zinc, albumin, globulin, C-reactive protein, and hemoglobin, and smear and cultures for M. tuberculosis in sputum samples were analyzed. Compared to healthy subjects, at baseline, patients with PTB had lower serum iron levels, higher copper levels and copper/zinc ratio, and similar zinc levels. During the tuberculosis treatment, no significant changes in the serum levels of iron, zinc, and copper/zinc were observed. Lower serum copper levels were associated with bacteriological conversion in tuberculosis treatment (tuberculosis-negative) at 30 days but not at 60 days (tuberculosis-positive). C-reactive protein levels and the C-reactive protein/albumin ratio were lower in tuberculosis-negative patients than in tuberculosis-positive patients at 30 and 60 days after treatment. Albumin and hemoglobin levels and the albumin/globulin ratio in patients with pulmonary tuberculosis increased during the study period, regardless of the bacteriological results. High serum globulin levels did not change among pulmonary tuberculosis patients during the study. Serum copper levels and the C-reactive protein/albumin ratio may be important parameters to evaluate the persistence of non-conversion after 60 days of tuberculosis treatment, and they may serve as predictors for relapse after successful treatment.

A direct, low-cost method to determine the concentration of lactose is an important goal with possible impact in various types of industry. In this study, a biosensor is reported that exploits the specific interaction between lactose and the enzyme β-galactosidase (β-Gal) normally employed to process lactose into glucose and galactose for lactose-intolerant people. The biosensor was made with β-Gal immobilized in layer-by-layer (LbL) films with the polyelectrolyte poly(ethylene imine) (PEI) and poly(vinyl sufonate) (PVS) on an indium tin oxide (ITO) electrode modified with a layer of Prussian Blue (PB). With an ITO/PB/(PEI/PVS)1(PEI/β-Gal)30 architecture, lactose could be determined with an amperometric method with sensitivity of 0.31 μA mmol(-1) cm(-2) and detection limit of 1.13 mmol L(-1), which is sufficient for detecting lactose in milk and for clinical exams. Detection occurred via a cascade reaction involving glucose oxidase titrated as electrolytic solution in the electrochemical cell, while PB allowed for operation at 0.0 V versus saturated calomel electrode, thus avoiding effects from interfering species. Sum-frequency generation spectroscopy data for the interface between the LbL film and a buffer containing lactose indicated that β-Gal lost order, which is the first demonstration of structural effects induced by the molecular recognition interaction with lactose.

Objective: To determine whether serum selenium levels are associated with the conversion of bacteriological tests in patients diagnosed with active pulmonary tuberculosis after eight weeks of standard treatment., Methods: We evaluated 35 healthy male controls and 35 male patients with pulmonary tuberculosis, the latter being evaluated at baseline, as well as at 30 and 60 days of antituberculosis treatment. For all participants, we measured anthropometric indices, as well as determining serum levels of albumin, C-reactive protein (CRP) and selenium. Because there are no reference values for the Brazilian population, we used the median of the serum selenium level of the controls as the cut-off point. At 30 and 60 days of antituberculosis treatment, we repeated the biochemical tests, as well as collecting sputum for smear microscopy and culture from the patients., Results: The mean age of the patients was 38.4 ± 11.4 years. Of the 35 patients, 25 (71%) described themselves as alcoholic; 20 (57.0%) were smokers; and 21 (60.0%) and 32 (91.4%) presented with muscle mass depletion as determined by measuring the triceps skinfold thickness and arm muscle area, respectively. Of 24 patients, 12 (39.2%) were classified as moderately or severely emaciated, and 15 (62.5%) had lost > 10% of their body weight by six months before diagnosis. At baseline, the tuberculosis group had lower serum selenium levels than did the control group. The conversion of bacteriological tests was associated with the CRP/albumin ratio and serum selenium levels 60 days after treatment initiation., Conclusions: Higher serum selenium levels after 60 days of treatment were associated with the conversion of bacteriological tests in pulmonary tuberculosis patients.

An important step in several bioanalytical applications is the immobilization of biomolecules. Accordingly, this procedure must be carefully chosen to preserve their biological structure and fully explore their properties. For this purpose, we combined the versatility of the layer-by-layer (LbL) method for the immobilization of biomolecules with the protective behavior of liposome-encapsulated systems to fabricate a novel amperometric glucose biosensor. To obtain the biosensing unit, an LbL film of the H2O2 catalyst polypeptide microperoxidase-11 (MP-11) was assembled onto an indium-tin oxide (ITO) electrode followed by the deposition of a liposome-encapsulated glucose oxidase (GOx) layer. The biosensor response toward glucose detection showed a sensitivity of 0.91±0.09 (μA/cm2)/mM and a limit of detection (LOD) of 8.6±1.1 μM, demonstrating an improved performance compared to similar biosensors with a single phospholipid-liposome or even containing a non-encapsulated GOx layer. Finally, glucose detection was also performed in a zero-lactose milk sample to demonstrate the potential of the biosensor for food analysis., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Premise of the Study: Microsatellite primers were developed for the tree species Genipa americana (Rubiaceae) for further population genetic studies. •, Methods and Results: We identified 144 clones containing 65 repeat motifs from a genomic library enriched for (CT)8 and (GT)8 motifs. Primer pairs were developed for 32 microsatellite loci and validated in 40 individuals of two natural G. americana populations. Seventeen loci were polymorphic, revealing from three to seven alleles per locus. The observed and expected heterozygosities ranged from 0.24 to 1.00 and from 0.22 to 0.78, respectively. •, Conclusions: The 17 primers identified as polymorphic loci are suitable to study the genetic diversity and structure, mating system, and gene flow in G. americana.

Layer-by-layer (LbL) films have been exploited in drug delivery systems that may be used in the form of patches, but the encapsulation of poor water soluble drugs and their release with a controlled rate are still major challenges to be faced. In this paper, we demonstrate the controlled release of aloin (barbaloin), an important component of the widely used Aloe vera, encapsulated into liposomes and immobilized in LbL films with a polyelectrolyte. With a systematic study using fluorescence spectroscopy of aloin release from solutions and from LbL films with different phospholipid liposomes, we inferred that optimized release was achieved with aloin incorporated into palmitoyl oleyl phosphatidyl glycerol (POPG) or dipalmitoyl phosphatidyl glycerol (DPPG) liposomes immobilized in LbL films. Significantly, with this optimized system aloin was almost completely released within 30 h, with a small release rate at the end, which followed a sharp release in the first 5h. Upon comparing the rates of the distinct systems, we conclude that the main factors controlling the release are the electrostatic interactions involving the negatively charged phospholipids. Because these interactions can be tuned in LbL films, the approach used here opens the way for new drug delivery systems to be developed with fine control of the drug release., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The peptide NS5A-1 (PPLLESWKDPDYVPPWHG), derived from hepatitis C virus (HCV) NS5A protein, was immobilized into layer-by-layer (LbL) silk fibroin (SF) films. Deposition was monitored by UV-vis absorption measurements at each bilayer deposited. The interaction SF/peptide film induced secondary structure in NS5A-1 as indicated by fluorescence and circular dichroism (CD) measurements. Voltammetric sensor (SF/NS5A-1) properties were observed when the composite film was tested in the presence of anti-HCV. The peptide-silk fibroin interaction studied here showed new architectures for immunosensors based on antigenic peptides and SF as a suitable immobilization matrix.

Unlabelled: Temporomandibular disorders (TMD) are characterized by the presence of temporomandibular joint (TMJ) and/or masticatory muscle pain and dysfunction. Low-level laser is presented as an adjuvant therapeutic modality for the treatment of TMD, especially when the presence of inflammatory pain is suspected., Objective: To systematically review studies that investigated the effect of low level laser therapy (LLLT) on the pain levels in individuals with TMD., Material and Methods: The databases Scopus, embase, ebsco and PubMed were reviewed from January/2003 to October/2010 with the following keywords: laser therapy, low-level laser therapy, temporomandibular joint disorders, temporomandibular joint dysfunction syndrome, temporomandibular joint, temporomandibular, facial pain and arthralgia, with the inclusion criteria for intervention studies in humans. exclusion criteria adopted were intervention studies in animals, studies that were not written in english, Spanish or Portuguese, theses, monographs, and abstracts presented in scientific events., Results: After a careful review, 14 studies fit the criteria for inclusion, of which, 12 used a placebo group. As for the protocol for laser application, the energy density used ranged from 0.9 to 105 J/cm², while the power density ranged from 9.8 to 500 mW. The number of sessions varied from 1 to 20 and the frequency of applications ranged from daily for 10 days to 1 time per week for 4 weeks. A reduction in pain levels was reported in 13 studies, with 9 of these occurring only in the experimental group, and 4 studies reporting pain relief for both the experimental group and for the placebo., Conclusion: Most papers showed that LLLT seemed to be effective in reducing pain from TMD. However, the heterogeneity of the standardization regarding the parameters of laser calls for caution in interpretation of these results. Thus, it is necessary to conduct further research in order to obtain a consensus regarding the best application protocol for pain relief in patients with TMD.

Objective: To identify the frequency of signs and symptoms of temporomandibular disorder (TMD) and its severity in individuals with headache., Study Design: 60 adults divided into three groups of 20 individuals: chronic daily headache (CDH), episodic headache (EH) and a control group without headache (WH). Headache diagnosis was performed according to the criteria of International Headache Society and the signs and symptoms of TMD were achieved by using a clinical exam and an anamnestic questionnaire. The severity of TMD was defined by the temporomandibular index (TMI)., Results: The TMD signs and symptoms were always more frequent in individuals with headache, especially report of pain in TMJ area (CDH, n=16; EH, n=12; WH, n=6), pain to palpation on masseter (CDH, n=19; EH, n=16; WH, n=11) which are significantly more frequent in episodic and chronic daily headache. The mean values of temporomandibular and articular index (CDH patients) and muscular index (CDH and EH patients) were statistically higher than in patients of the control group, notably the articular (CDH=0.38; EH=0.25;WH=0.19) and muscular (CDH=0.46; EH=0.51; WH=0.26) indices., Conclusions: These findings allow us to speculate that masticatory and TMJ pain are more common in headache subjects. Besides, it seems that the TMD is more severe in headache patients.

An experiment was conducted to evaluate the effect of CLA on the immune response and performance of piglets when subjected to an immune challenge. A total of 32 weanling pigs (17 to 23 d of age) with an initial BW of 8.9 kg were allotted to a 3 × 2 factorial arrangement of treatments. There were 3 levels of dietary CLA (0%, 1%, and 2%) and 2 levels of lipopolysaccharide (LPS) challenge (unchallenged and challenged). Challenged pigs were challenged on d 7 and 21. On d 4 and 18, all pigs were inoculated with BSA for assessment of IgG production. There was no difference in growth performance among piglets receiving different CLA supplementation levels. However, LPS-challenged piglets had poorer BW (P < 0.05), ADFI (P < 0.01), and ADG (P < 0.001) compared with the control group at d 35 postweaning. Lipopolysaccharide-challenged piglets also had increased respiratory rate (P < 0.001) and rectal temperature (P < 0.001), and decreased plasma proteins, hematocrit, and white blood cell counts (P < 0.05). Production of IgG against BSA was increased in the 1% CLA supplementation group (P < 0.001), indicating that CLA has an immunomodulatory effect. Supplementation with CLA did not affect lymphocyte proliferation, percentage of CD4(+) and CD8(+) cells, plasma proteins, red and white blood cell count, respiratory rate, or rectal temperature after LPS challenge. Although CLA supplementation did not influence growth performance or certain immune system measurements, the increased IgG titers with 1% CLA dietary supplementation indicate that it has a beneficial effect on the humoral immune system of weaned piglets.

Campylobacter is an important cause of foodborne gastroenteritis. We determined the occurrence of Campylobacter jejuni and Campylobacter coli, using culture-based methods and PCRs targeting virulence-associated genes (VAGs) among children aged ≤14 years who were treated for diarrhoea at emergency rooms in northeastern Brazil. Genomic DNA was extracted directly from stool samples collected from 366 children. A questionnaire was also applied to qualify the clinical conditions presented by each child at the time of admission. C. jejuni and C. coli were detected in 16.4 % (60/366) and 1.4 % (5/366) of the diarrhoeal samples, respectively, by PCR, a much higher proportion than that detected by conventional methods. C. jejuni VAGs were detected in the following proportions of hipO-positive samples: ciaB, 95 % (57/60); dnaJ, 86.7 % (52/60); racR, 98.3 % (59/60); flaA, 80 % (48/60); pldA, 45 % (27/60); cdtABC, 95 % (57/60); and pVir 0 % (0/60). Particular symptoms, such as blood in faeces, vomiting, fever, and/or abdominal pain, were not associated with detection of C. jejuni nor were they associated with any particular VAG or combination of VAGs (P>0.05). C. jejuni and its VAGs were detected in a substantial proportion of the children admitted. Further efforts shall be directed towards elucidating whether these genetic factors or their expressed proteins play a role in Campylobacter pathogenesis.

The placenta is fundamental for fetal development. The aim of this study was to determine Ca, Fe, Cu, and Zn content in the fetal and maternal portions of the placentas of teenage and adult women. Measurement of the minerals was conducted using Synchrotron radiation total reflection X-ray fluorescence. Forty samples from the fetal portion of teenagers and adults and 40 samples from the maternal portion of teenagers and adults were analyzed. There were significant differences in the Ca and Cu concentrations of the placenta's maternal portion when compared to the fetal portion, for both teenagers and adults. There were differences in Fe and Zn concentrations only when comparing the maternal portion of placenta with the fetal portion of the adults. These results suggest important differences in mineral content based on the placental portion. No significant difference was observed between the minerals studied of the maternal portion of teenagers and adults; however, in the fetal portion, mineral concentrations were greater in adults than in teenagers. Therefore, the mineral concentration of the fetal portion of the placenta is influenced by the mother's age. If there is mineral's competition between the mother and fetus during pregnancy in adolescence due to the importance of these minerals in growth and development, then the mechanism and reason for it should be elucidated in future research. In addition, we believe that further research should be carried out on transporters of these minerals in the same portions of the placentas analyzed by our group, in teenagers and adults.

Antigenic peptides may be immobilized in nanostructured films in order to build highly specific immunosensors and other devices that require molecular recognition, with no need to use complex molecules. A major challenge for such endeavors, however, is to preserve the secondary structure of the peptides after immobilization. In this study, we show that the peptide p17-1 (LSGGELDRWEKIRLRPGG), derived from the HIV-1 p17 protein, may be immobilized in Layer-by-Layer (LbL) films made with polyelectrolytes. Its structure was preserved only if incorporated into phospholipid liposomes, according to fluorescence and circular dichroism (CD) spectroscopy. The lack of secondary structure for the peptide in the LbL film may be associated with the film-forming procedure in which p17-1 was adsorbed from an aqueous solution, where it does not form alpha helices. The importance of structure preservation was clear in the attempts to produce electrochemical immunosensors with the p17-1 peptide without being protected in liposomes in an LbL film. There was no detectable influence of the presence of anti-p17 antibodies, though some molecular interaction could be inferred from the voltammograms. In contrast, for p17-1 incorporated in liposomes electrochemical immunosensors could be obtained with the voltamogramms showing strong molecular recognition with the antibodies. These results indicated that phospholipids serve as a suitable matrix for immobilization of peptides, and confirmed the importance of structure preservation in electrochemical immunosensors.

The development of new methods and concepts to visualize massive amounts of data holds the promise to revolutionize the way scientific results are analyzed, especially when tasks such as classification and clustering are involved, as in the case of sensing and biosensing. In this paper we employ a suite of software tools, referred to as PEx-Sensors, through which projection techniques are used to analyze electrical impedance spectroscopy data in electronic tongues and related sensors. The possibility of treating high dimension datasets with PEx-Sensors is advantageous because the whole impedance vs. frequency curves obtained with various sensing units and for a variety of samples can be analyzed at once. It will be shown that non-linear projection techniques such as Sammon's Mapping or IDMAP provide higher distinction ability than linear methods for sensor arrays containing units capable of molecular recognition, apparently because these techniques are able to capture the cooperative response owing to specific interactions between the sensing unit material and the analyte. In addition to allowing for a higher sensitivity and selectivity, the use of PEx-Sensors permits the identification of the major contributors for the distinguishing ability of sensing units and of the optimized frequency range. The latter will be illustrated with sensing units made with layer-by-layer (LbL) films to detect phytic acid, whose capacitance data were visualized with Parallel Coordinates. Significantly, the implementation of PEx-Sensors was conceived so as to handle any type of sensor based on any type of principle of detection, representing therefore a generic platform for treating large amounts of data for sensors and biosensors., (© The Royal Society of Chemistry 2011)

The purpose of this study was to trace the contraceptive history of sterilized women and identify the associations between educational, sexual and obstetric variables and the women's age when they underwent the procedure for tubal ligation (TL). This is a retrospective documentary study performed at the Lígia Barros Costa Natural Birthing Center in Fortaleza, Ceará, with 1423 records, dating from 2005 to 2008, 277 of which referred to sterilized women. Data analysis involved applying the calculation of frequencies, Pearson's chi-square test and correlation of Pearson/Spearman. Sterilized women represented a population with low education, marital union, and a history of infrequent use of other contraceptive methods other than condoms and the pill. Numbers of pregnancies and abortions/miscarriages were related with the age of TL, unlike the variables of education and the age of the first sexual intercourse. With this knowledge at hand, nurses can improve their look towards women looking forward to TL, and thus strengthen education strategies and promote greater diversity in the alternatives for contraception.

Liposomes have been applied to many fields as nanocarriers, especially in drug delivery as active molecules may be entrapped either in their aqueous interior or onto the hydrophobic surface. In this paper we describe the fabrication of layer-by-layer (LbL) films made with liposomes incorporating the anti-inflammatory ibuprofen. The liposomes were made with dipalmitoyl phosphatidyl choline (DPPC), dipalmitoyl phosphatidyl glycerol (DPPG) and palmitoyl oleoyl phosphatidyl glycerol (POPG). LbL films were assembled via alternate adsorption of the polyamidoamine dendrimer (PAMAM), generation 4, and liposomes containing ibuprofen. According to dynamic light scattering measurements, the incorporation of ibuprofen caused DPPC and DPPG liposomes to become more stable, with a decrease in diameter from 140 to 74 nm and 132 to 63 nm, respectively. In contrast, liposomes from POPG became less stable, with an increase in size from 110 to 160 nm after ibuprofen incorporation. These results were confirmed by atomic force microscopy images of LbL films, which showed a large tendency to rupture for POPG liposomes. Film growth was monitored using nanogravimetry and UV-Vis spectroscopy, indicating that growth stops after 10 bilayers. The release of ibuprofen obtained with fluorescence measurements was slower for the liposomes, with decay times of 9.2 and 8.5 h for DPPG and POPG liposomes, respectively, than for the free drug with a decay time of 5.2 h. Ibuprofen could also be released from the LbL films made with DPPG and POPG liposomes, which is promising for further uses in patches.

Adolescence is marked by intensive growth and development. When pregnancy occurs during this period of the mother's growth, there is an increase in her nutritional needs. The aim of this study was to determine the levels of calcium, iron, copper, and zinc in maternal plasma, the placenta, and in the cord plasma of pregnant teenagers and adults. A total of 80 sets of maternal plasma, placentas, and cord plasma (40 from teenagers and 40 from adults) were analyzed using synchrotron radiation total reflection X-ray fluorescence. The levels of calcium, copper, and zinc in the maternal and cord plasma from teenagers were not significantly different than those from adults. Iron levels in the teenagers maternal and cord plasma were higher than in the adults. All of the minerals analyzed were present at higher levels in the placentas from adults than those from teenagers. However, the low quantities of placental calcium, iron, copper, and zinc in the teenagers do not compromise the levels of these minerals in the cord plasma. Future research regarding the placental transport of these minerals is recommended to investigate the efficiency of mechanisms of transfer of these minerals in pregnant teenagers.

Over the past century, the Brazilian Atlantic forest has been reduced to small, isolated fragments of forest. Reproductive isolation theories predict a loss of genetic diversity and increases in inbreeding and spatial genetic structure (SGS) in such populations. We analysed eight microsatellite loci to investigate the pollen and seed dispersal patterns, genetic diversity, inbreeding and SGS of the tropical tree Copaifera langsdorffii in a small (4.8 ha), isolated population. All 112 adult trees and 128 seedlings found in the stand were sampled, mapped and genotyped. Seedlings had significantly lower levels of genetic diversity (A=16.5±0.45, mean±95% s.e.; H(e)=0.838±0.006) than did adult trees (A=23.2±0.81; H(e)=0.893±0.030). Parentage analysis did not indicate any seed immigration (m(seeds)=0) and the pollen immigration rate was very low (m(pollen)=0.047). The average distance of realized pollen dispersal within the stand was 94 m, with 81% of the pollen travelling <150 m. A significant negative correlation was found between the frequency and distance of pollen dispersal (r=-0.79, P<0.01), indicating that short-distance pollinations were more frequent. A significant SGS for both adults (∼50 m) and seedlings (∼20 m) was also found, indicating that most of the seeds were dispersed over short distances. The results suggested that the spatial isolation of populations by habitat fragmentation can restrict seed and pollen gene flow, increase SGS and affect the genetic diversity of future generations.

The use of melanin in bioinspired applications is mostly limited by its poor stability in solid films. This problem has been addressed here by incorporating melanin into dipalmitoyl phosphatidyl glycerol (DPPG) liposomes, which were then immobilized onto a solid substrate as an LbL film. Results from steady-state and time-resolved fluorescence indicated an increased stability for melanin incorporated into DPPG liposomes. If not protected by liposomes, melanin looses completely its fluorescence properties in LbL films. The thickness of the liposome-melanin layer obtained from neutron reflectivity data was 4.1+/-0.2 nm, consistent with the value estimated for the phospholipid bilayer of the liposomes, an evidence of the collapse of most liposomes. On the other hand, the final roughness indicated that some of the liposomes had their structure preserved. In summary, liposomes were proven excellent for encapsulation, thus providing a suitable environment, closer to the physiological conditions without using organic solvents or high pHs., (Copyright 2010 Elsevier Inc. All rights reserved.)