Multifunctional Biosensor Based on Localized Surface Plasmon Resonance for Monitoring Small Molecule–Protein Interaction

We report an optical sensor based on localized surface plasmon resonance (LSPR) to study small-molecule protein interaction combining high sensitivity refractive index sensing for quantitative binding information and subsequent conformation-sensitive plasmon-activated circular dichroism spectroscopy. The interaction of α-amylase and a small-size molecule (PGG, pentagalloyl glucose) was log concentration-dependent from 0.5 to 154 μM. In situ tests were additionally successfully applied to the analysis of real wine samples. These studies demonstrate that LSPR sensors to monitor small molecule–protein interactions in real time and in situ, which is a great advance within technological platforms for drug discovery.

Protein–polyphenol interaction on silica beads for astringency tests based on eye, photography or reflectance detection modes

Astringency is an organoleptic property of beverages and food products resulting mainly from the interaction of salivary proteins with dietary polyphenols. It is of great importance to consumers, but the only effective way of measuring it involves trained sensorial panellists, providing subjective and expensive responses. Concurrent chemical evaluations try to screen food astringency, by means of polyphenol and protein precipitation procedures, but these are far from the real human astringency sensation where not all polyphenol–protein interactions lead to the occurrence of precipitate. Here, a novel chemical approach that tries to mimic protein–polyphenol interactions in the mouth is presented to evaluate astringency. A protein, acting as a salivary protein, is attached to a solid support to which the polyphenol binds (just as happens when drinking wine), with subsequent colour alteration that is fully independent from the occurrence of precipitate. Employing this simple concept, Bovine Serum Albumin (BSA) was selected as the model salivary protein and used to cover the surface of silica beads. Tannic Acid (TA), employed as the model polyphenol, was allowed to interact with the BSA on the silica support and its adsorption to the protein was detected by reaction with Fe(III) and subsequent colour development. Quantitative data of TA in the samples were extracted by colorimetric or reflectance studies over the solid materials. The analysis was done by taking a regular picture with a digital camera, opening the image file in common software and extracting the colour coordinates from HSL (Hue, Saturation, Lightness) and RGB (Red, Green, Blue) colour model systems; linear ranges were observed from 10.6 to 106.0 μmol L−1. The latter was based on the Kubelka–Munk response, showing a linear gain with concentrations from 0.3 to 10.5 μmol L−1. In either of these two approaches, semi-quantitative estimation of TA was enabled by direct eye comparison. The correlation between the levels of adsorbed TA and the astringency of beverages was tested by using the assay to check the astringency of wines and comparing these to the response of sensorial panellists. Results of the two methods correlated well. The proposed sensor has significant potential as a robust tool for the quantitative/semi-quantitative evaluation of astringency in wine.

A expansão sistematizada de oportunidades de escolha como estratégia de suporte à participação

Baseado nos pressupostos subjacentes às práticas pedagógicas centradas na criança, este estudo tem por objetivo analisar a perspetiva dos educadores de infância sobre a promoção de oportunidades de escolha em contextos de ensino pré-escolar, tendo por referência o processo de inclusão de crianças com necessidades adicionais de suporte, bem como examinar os efeitos de um programa de intervenção, centrado na expansão de oportunidades de escolha, na participação de uma criança em situação de incapacidade. A fim de responder ao primeiro objetivo foi auscultada a opinião de 71 educadores de infância, através de uma pesquisa por inquérito. Já para o escrutínio dos efeitos do programa de intervenção, foi desenvolvido um estudo de caso único, tipo AB, avaliando o seu impacte na participação de uma criança de 4 anos com Paralisia Cerebral, através do uso de grelhas de observação e de uma entrevista semiestruturada à educadora. Os resultados sugerem que existe uma diferença significativa na facilidade de implementação de oportunidades de escolha, quando consideradas crianças com desenvolvimento típico e com necessidades adicionais de suporte, que parece também variar em função das diferentes rotinas do contexto pré-escolar e correlacionar-se com o nível de severidade da incapacidade. As características das crianças e as exigências curriculares são entendidas como as principais barreiras à implementação destas práticas. Os resultados sugerem, ainda, que o programa de intervenção – baseado no planeamento de oportunidades de escolha embebidas nos diferentes momentos da rotina do contexto pré-escolar - teve impacte positivo na participação da criança, com expansão dos indicadores de envolvimento, interação/comunicação e satisfação

Aumentar as oportunidades de comunicação nos momentos de brincadeira - descrição de um programa de intervenção

Objetivo: Partindo da análise da literatura realizada, em que é salientada a ideia de que o desenvolvimento e aprendizagem ocorrem a partir das experiências e influência dos contextos em que as crianças estão incluídas e a partir das interações que estabelecem nesses mesmos contextos, pretende-se compreender, neste estudo, se partindo do contexto natural, com diferentes materiais e atores, é possível aumentar as oportunidades de comunicação e participação de uma criança com restrições ao nível da linguagem, nos momentos de jogo. Método: Foi delineado e implementado um programa de intervenção, num estabelecimento de ensino, na valência de pré-escolar, onde estiveram envolvidas uma criança de 4 anos com restrições ao nível da linguagem oral, o grupo de crianças em que estava incluída e a educadora do grupo. Para monitorização do programa e avaliação da sua eficácia realizou-se um pré e pós teste, onde foi definida a linha de base da criança. Ao longo do programa foram registados, a partir de uma grelha de observação da frequência dos atos comunicativos da criança, nas situações de jogo planificadas. Os comportamentos e modelagem realizada por parte educador, foram igualmente registados e analisados. Recorreu-se a realização de entrevistas semiestruturadas para a recolha de dados relativos à criança e perceção do educador. Resultados/Discussão: Os dados recolhidos demonstram um aumento significativo das tomadas de iniciativa verbal e alternância entre turnos, ao longo da intervenção por parte da criança alvo. A presença do adulto e dos pares parecem revelar-se como facilitador dos atos comunicativos da criança com restrições ao nível da linguagem

Fatal disseminated paracoccidioidomycosis in a two-year-old child

A two year-old female child was admitted at the Pediatric Intensive Care Unit in a septic shock associated with a lymphoproliferative syndrome, with history of fever, adynamia and weight loss during the last two months. On admission, the main clinical and laboratory manifestations were: pallor, jaundice, disseminated enlarged lymph nodes, hepatosplenomegaly, crusted warts on face, anemia, eosinophilia, thrombocytopenia, increased direct and indirect bilirubin, alkaline phosphatase, and gammaglutamyl transpeptidase. A parenteral administration of fluids, dobutamine and mechanical ventilation was started, without improvement of the clinical conditions. A direct examination of exsudate collected from cervical lymph node revealed numerous oval-to-around cells with multiple budding, like a "pilot wheel" cell, suggesting Paracoccidioides brasiliensis. Even though treatment with intravenous sulfamethoxazole-trimethoprine was soon started, the child died 36 hours after hospital admission. Disseminated paracoccidioidomycosis was confirmed in the autopsy. This is the youngest case of paracoccidioidomycosis in children reported in the literature.

Lupus Myelopathy in a Child

A 5-year-old female developed, after a 7-month period of fever, anorexia, weight loss, and a transitory cutaneous erythematous eruption, a severe acute transverse myelopathy, with a partial recovery of motor and sensory function. She had positive antinuclear and antidouble-stranded DNA antibodies but no antiphospholipid antibodies. Six months later she had massive proteinuria and restarted treatment with steroids and cyclophosphamide. Our patient is one of the youngest reported with lupus myelopathy. We discuss the clinical presentation, the magnetic resonance imaging findings, and other relevant laboratory studies of this rare but serious complication of systemic lupus erythematosus.

Cardiac Hydatid Cyst in a Child

Cardiac hydatid cyst is a rare disease, especially in children. An 11-year-old boy with a previous anaphylactic reaction and episodes of abdominal pain was admitted for workup of an acquired long systolic murmur. Echocardiographic investigation disclosed a tumor of the right ventricular anterior wall, with multiple loculations. Magnetic resonance imaging characterized it as a multilobular tumor with cyst formation and disclosed another cyst in the right pulmonary artery. With a positive ELISA reaction the child was admitted for surgery with the diagnosis of cardiac and pulmonary hydatid cysts. Cardiac surgery was performed with good results, followed by medical treatment with albendazole.

Detection of EBV-DNA in serum samples of an immunosuppressed child during a three years follow-up: association of clinical and PCR data with active infection

Twenty-four whole blood and serum samples were drawn from an eight year-old heart transplant child during a 36 months follow-up. EBV serology was positive for VCA-IgM and IgG, and negative for EBNA-IgG at the age of five years old when the child presented with signs and symptoms suggestive of acute infectious mononucleosis. After 14 months, serological parameters were: positive VCA-IgG, EBNA-IgG and negative VCA-IgM. This serological pattern has been maintained since then even during episodes suggestive of EBV reactivation. PCR amplified a specific DNA fragment from the EBV gp220 (detection limit of 100 viral copies). All twenty-four whole blood samples yielded positive results by PCR, while 12 out of 24 serum samples were positive. We aimed at analyzing whether detection of EBV-DNA in serum samples by PCR was associated with overt disease as stated by the need of antiviral treatment and hospitalization. Statistical analysis showed agreement between the two parameters evidenced by the Kappa test (value 0.750; p < 0.001). We concluded that detection of EBV-DNA in serum samples of immunosuppressed patients might be used as a laboratory marker of active EBV disease when a Real-Time PCR or another quantitative method is not available.

Mother-to-child transmission of different HIV-1 subtypes among ARV Naïve infected pregnant women in Nigeria

The rate of mother-to-child transmission (MTCT) of HIV as well as the implications of the circulating multiple subtypes to MTCT in Nigeria are not known. This study was therefore undertaken to determine the differential rates of MTCT of HIV-1 subtypes detected among infected pregnant women before ARV intervention therapy became available in Nigeria. Twenty of the HIV-positive women who signed the informed consent form during pregnancy brought their babies for follow-up testing at age 18-24 months. Plasma samples from both mother and baby were tested for HIV antibody at the Department of Virology, UCH, Ibadan, Nigeria. All positive samples (plasma and peripheral blood mononuclear cells - PBMCs) were shipped to the Institute of Tropical Medicine, Antwerp, Belgium, where the subtype of the infecting virus was determined using the HMA technique. Overall, a mother-to-child HIV transmission rate of 45% was found in this cohort. Specifically, 36.4%, 66.7% and 100% of the women infected with HIV-1 CRF02 (IbNg), G and B, respectively, transmitted the virus to their babies. As far as it can be ascertained, this is the first report on the rate of MTCT of HIV in Nigeria. The findings reported in this paper will form a useful reference for assessment of currently available therapeutic intervention of MTCT in the country.

Toxoplasma-IgM and IgG-avidity in single samples from areas with a high infection rate can determine the risk of mother-to-child transmission

Anti-Toxoplasma IgG-avidity was determined in 168 serum samples from IgG- and IgM-positive pregnant women at various times during pregnancy, in order to evaluate the predictive value for risk of mother-to-child transmission in a single sample, taking the limitations of conventional serology into account. The neonatal IgM was considered the serologic marker of transmission. Fluorometric tests for IgG, IgM (immunocapture) and IgG-avidity were performed. Fifty-one of the 128 pregnant women tested gave birth in the hospital and neonatal IgM was obtained. The results showed 32 (62.75%) pregnant women having high avidity, IgM indexes between 0.6 and 2.4, and no infected newborn. Nineteen (37.25%) had low or inconclusive avidity, IgM indexes between 0.6 and 11.9, and five infected newborns and one stillbirth. In two infected newborns and the stillbirth maternal IgM indexes were low and in one infected newborn the only maternal parameter that suggested fetal risk was IgG-avidity. In the present study, IgG-avidity performed in single samples from positive IgM pregnant women helped to determine the risk of transmission at any time during pregnancy, especially when the indexes of the two tests were analysed with respect to gestational age. This model may be less expensive in developing countries where there is a high prevalence of infection than the follow-up of susceptible mothers until childbirth with monthly serology, and it creates a new perspective for the diagnosis of congenital toxoplasmosis.

Fatal septicemia caused by Chromobacterium violaceum in a child from Colombia

A 4-year old child living in Colombia presented with a history of fever and severe abdominal pain for four days. The patient developed pneumonia, septic shock, multiple organ failure and died on the fifth day of hospitalization. Chromobacterium violaceum was isolated from admission blood cultures and was resistant to ampicillin, cephalosporins, carbapenems and aminoglycosides.

(Un)suitability of the use of pH buffers in biological, biochemical and environmental studies and their interaction with metal ions – a review

The use of buffers to maintain the pH within a desired range is a very common practice in chemical, biochemical and biological studies. Among them, zwitterionic N-substituted aminosulfonic acids, usually known as Good’s buffers, although widely used, can complex metals and interact with biological systems. The present work reviews, discusses and updates the metal complexation characteristics of thirty one commercially available buffers. In addition, their impact on biological systems is also presented. The influences of these buffers on the results obtained in biological, biochemical and environmental studies, with special focus on their interaction with metal ions, are highlighted and critically reviewed. Using chemical speciation simulations, based on the current knowledge of the metal–buffer stability constants, a proposal of the most adequate buffer to employ for a given metal ion is presented.

Effects of Chlorophenoxy Herbicides and Their Main Transformation Products on DNA Damage and Acetylcholinesterase Activity

Persistent pesticide transformation products (TPs) are increasingly being detected among different environmental compartments, including groundwater and surface water. However, there is no sufficient experimental data on their toxicological potential to assess the risk associated with TPs, even if their occurrence is known. In this study, the interaction of chlorophenoxy herbicides (MCPA, mecoprop, 2,4-D and dichlorprop) and their main transformation products with calf thymus DNA by UV-visible absorption spectroscopy has been assessed. Additionally, the toxicity of the chlorophenoxy herbicides and TPs was also assessed evaluating the inhibition of acetylcholinesterase activity. On the basis of the results found, it seems that AChE is not the main target of chlorophenoxy herbicides and their TPs. However, the results found showed that the transformation products displayed a higher inhibitory activity when compared with the parent herbicides. The results obtained in the DNA interaction studies showed, in general, a slight effect on the stability of the double helix. However, the data found for 4-chloro-2-methyl-6-nitrophenol suggest that this transformation product can interact with DNA through a noncovalent mode.

The relationship of cardiorespiratory fitness, birth weight and parental BMI on adolescents’ obesity status

The aims of this study were as follows: (1) to analyze differences in cardiorespiratory fitness (CRF), parents' body mass index (BMI) and birth weight (BW) between non-overweight (NOW) and overweight/obese (OV/OB) adolescents, and (2) to investigate the association of those variables with the risk of their biological offspring being OV/OB. This study comprised 788 adolescents (477 girls and 311 boys), aged between 12 and 18 years. CRF was predicted by maximal multistage 20-m shuttle-run test according to the procedures described in FITNESSGRAM. Children's BMI was classified according to the International Obesity Task Force. Adolescents' BW was assessed from each child's pediatric record at birth. Parents' OV/OB status was defined and classified according to the World Health Organization. Socioeconomic status was defined by parental education. The prevalence OV/OB was 21.4 and 5.3%, respectively, and there were no gender differences. The OV/OB adolescents (girls and boys) had significantly (P

Aumento da Linguagem: Um estudo de caso

Objetivo: O autismo é uma perturbação global do desenvolvimento infantil que se prolonga por toda a vida e evolui com a idade. Carateriza-se pela presença de um desenvolvimento acentuadamente atípico na interação social e na comunicação, por um repertório de atividades e interesses marcadamente restritos, por dificuldades de flexibilidade de pensamento e de comportamento, que se exibem em comportamentos estereotipados e rígidos, associados a dificuldades na aceitação de alterações de rotinas. Dentro da grande variação possível na severidade do autismo, encontram-se crianças sem linguagem verbal e com dificuldades na comunicação por qualquer outra via, assim como se encontram crianças que apresentam linguagem verbal com um vocabulário restrito, repetitivo e não comunicativo. O objetivo principal deste trabalho é analisar e potenciar o desenvolvimento da linguagem, a aquisição de palavras e frases na criança autista, tendo em vista a generalização dos resultados. Para isso, foi realizado um estudo de caso de uma criança de 4 anos de idade com comportamentos que sugerem uma Perturbação Pervasiva do Desenvolvimento. Utilizou-se o recurso aos instrumentos de observação e entrevistas, procedendo-se à sua análise e ao desenho da intervenção a realizar. Foi implementada a intervenção “Ensinar crianças com autismo a fazer perguntas sobre objetos escondidos” na qual são utilizados brinquedos de interesse para a criança, motivacionais para a concretização do objetivo do estudo.