4-Dimethylaminoantipyrine as a Broad Electrochemical Indicator for Immunosensors Platform.

Here, we describe 4-dimethylaminoantipyrine (4-DMAA)-mediated interfacing as a broad biochemical indicator to stabilize and promote the higher response of electrodes for immunological detection. We hypothesized that the improved biological interactions of 4-DMAA with electrodes and biological samples may be due to the interaction properties of the benzene and pyrazole chemical groups with graphite and proteins, respectively. In order to demonstrate that 4-DMAA could be used as a general indicator in electrochemical immunoassays, we used peptides as probes for the diagnosis of four neglected tropical infectious diseases Tegumentary leishmaniasis, Visceral leishmaniasis, Strongyloidiasis, and Leprosy on commercial graphite screen-printed electrodes. 4-DMAA oxidation was used to indicate specific biological recognition between the epitope-based peptide and serum immunoglobulin G (IgG) from infected patients. We demonstrated that 4-DMAA should be incorporated into the electrodes prior to serum application, which avoids interference with its sensitivity and specificity. In addition, 4-DMAA oxidizes at a low anodic potential, and the oxidation peak is useful for detecting proteins in biological fluids. In summary, we have successfully demonstrated the broad application of 4-DMAA as a general indicator for the specific diagnosis of four infectious diseases in electrochemical immunosensors. Such a strategy is quite advantageous for indirect detection of proteins that lack electrochemical activities or are spatially inaccessible on the electrode surface. This new indicator opens a new avenue for monitoring biological recognition, especially for immunosensors.

Strongyloidiasis Serological Analysis with Three Different Biological Probes and Their Electrochemical Responses in a Screen-Printed Gold Electrode.

(1) Background: The validation of biological antigens is the study's utmost goal in biomedical applications. We evaluated three different probes with single and multiple epitopes through electrochemical detection of specific IgG in serum for human strongyloidiasis diagnosis. (2) Methods: Screen-printed gold electrodes were used and probes consisting of two single-epitope synthetic peptides (D3 and C10) with different sequences, and a multi-epitope antigen [detergent phase (DP)-hydrophobic membrane proteins]. Human serum samples from three populations were used: Strongyloides stercoralis positive, positive for other parasitic infections and negative controls. To test the immobilization of probes onto a screen-printed gold electrode and the serum IgG detection, electrochemical analyses were carried out through differential pulse voltammetry (DPV) and the electrode surface analyses were recorded using atomic force microscopy. (3) Results: The electrochemical response in screen-printed gold electrodes of peptides D3 and C10 when using positive serum was significantly higher than that when using the DP. Our sensor improved sensitivity to detect strongyloidiasis. (4) Conclusions: Probes' sequences are critical factors for differential electrochemical responses, and the D3 peptide presented the best electrochemical performance for strongyloidiasis detection, and may efficiently substitute whole antigen extracts from parasites for strongyloidiasis diagnosis in electrochemical immunosensors.

Application of ZnO Nanocrystals as a Surface-Enhancer FTIR for Glyphosate Detection.

Glyphosate detection and quantification is still a challenge. After an extensive review of the literature, we observed that Fourier transform infrared spectroscopy (FTIR) had practically not yet been used for detection or quantification. The interaction between zinc oxide (ZnO), silver oxide (Ag2O), and Ag-doped ZnO nanocrystals (NCs), as well as that between nanocomposite (Ag-doped ZnO/AgO) and glyphosate was analyzed with FTIR to determine whether nanomaterials could be used as signal enhancers for glyphosates. The results were further supported with the use of atomic force microscopy (AFM) imaging. The glyphosate commercial solutions were intensified 10,000 times when incorporated the ZnO NCs. However, strong chemical interactions between Ag and glyphosate may suppress signaling, making FTIR identification difficult. In short, we have shown for the first time that ZnO NCs are exciting tools with the potential to be used as signal amplifiers of glyphosate, the use of which may be explored in terms of the detection of other molecules based on nanocrystal affinity.

Bioelectrode applied to diagnosis of cardiac disease.

This paper describes the assembly of a bioelectrode based on poly(3-aminophenol) and anti-troponin T antibody for recognition of troponin T, which is a specific biomarker for diagnosis of acute myocardial infarction. This disease causes loss of cellular components, allowing the output of molecules such as troponin T. This proteic component acts as biomarker for diagnosis of acute myocardial infarction due to their high sensitivity and specificity. Poly(3-aminophenol) was electrodeposited onto fluorine doped tin oxide (FTO) coated glass and characterized by spectroscopic methods (UV-Visible, fluorescence, infrared), electrochemical methods (cyclic voltammetry and electrochemical impedance spectroscopy) and morphological methods (laser interferometry, field emission scanning electronic microscopy, and atomic force microscopy). UV/Vis analysis indicated that poly(3-aminophenol) presents extension of conjugation, in according with fluorescence studies. Electrochemical studies indicated that poly(3-aminophenol) electrodeposited in FTO is a material with passivating characteristics for anions and capacity of retaining cationic compounds. Laser interferometry showed that poly(3-aminophenol) covers the FTO surface with a thickness off 375 ± 75 nm. Surface images by FE-SEM and AFM have shown a full coverage on the FTO by the polymer film. The incorporation of anti-troponin T antibody on FTO electrode modified with poly(3-aminophenol) allowed effective and selective detection of cardiac biomarker troponin T, by electrochemical impedance spectroscopy (label free) and by photoluminescence, based on CdSe/ZnS quantum dots. This research shows the step by step assembly of the bioelectrode, used for detection of troponin T by impedimetric and fluorescence methods, opening the opportunity for its use in the diagnosis of others diseases.

Functional epitope core motif of the Anaplasma marginale major surface protein 1a and its incorporation onto bioelectrodes for antibody detection.

Anaplasmosis, a persistent intraerythrocytic infection of cattle by Anaplasma marginale, causes severe anemia and a higher rate of abortion, resulting in significant loss to both dairy and beef industries. Clinical diagnosis is based on symptoms and confirmatory laboratory tests are required. Currently, all the diagnostic assays have been developed with whole antigens with indirect ELISA based on multiple epitopes. In a pioneer investigation we demonstrated the use of critical motifs of an epitope as biomarkers for immunosensor applications. Mimotopes of the MSP1a protein functional epitope were obtained through Phage Display after three cycles of selection of a 12-mer random peptide library against the neutralizing monoclonal antibody 15D2. Thirty-nine clones were randomly selected, sequenced, translated and aligned with the native sequence. The consensus sequence SxSSQSEASTSSQLGA was obtained, which is located in C-terminal end of the 28-aa repetitive motif of the MSP1a protein, but the alignment and sequences' variation among mimotopes allowed us to map the critical motif STSSxL within the consensus sequence. Based on these results, two peptides were chemically synthesized: one based on the critical motif (STSSQL, Am1) and the other based on the consensus sequence aligned with the native epitope (SEASTSSQLGA, Am2). Sera from 24 infected and 52 healthy animals were tested by ELISA for reactivity against Am1 and Am2, which presented sensitivities of 96% and 100%, respectively. The Am1 peptide was incorporated onto a biolectrode (graphite modified with poly-3-hydroxyphenylacetic acid) and direct serum detection was demonstrated by impedance, differential pulse voltammetry, and atomic force microscopy. The electrochemical sensor system proved to be highly effective in discriminating sera from positive and negative animals. These immunosensors were highly sensitive and selective for positive IgG, contaminants did not affect measurements, and were based on a simple, fast and reproducible electrochemical system.

Translation, adaptation and inter-rater reliability of the administration manual for the Fugl-Meyer assessment.

BACKGROUND: Recently, the reliability of the Brazilian version of the Fugl-Meyer Assessment (FMA) was assessed through the scoring given according to observations made by a single evaluator who applied the test. When different raters apply the scale, the reliability may depend on the interpretation given to the assessment sheet. In such cases, a clear administration manual is essential for ensuring homogeneity of application. OBJECTIVES: To translate and adapt the French Canadian version of the FMA administration manual into Brazilian Portuguese and to evaluate the inter-rater reliability when different evaluators apply the FMA on the basis of the information contained in the manual. METHODS: Eighteen adults (59±10 years) with chronic hemiparesis (38±35 months after a stroke) took part in this study. Eight patients participated in the first part of the study and 10 in the second part. Based on analyzing the results from part 1, an adapted version was developed, in which information and photos were added to illustrate the positions of the patient and evaluator. The inter-rater reliability was assessed using the intraclass correlation coefficient (ICC). RESULTS: The reliability of the FMA based on the adapted version of the manual was excellent for the total motor scores for the upper limbs (ICC=0.98) and lower limbs (ICC=0.90), as well as for movement sense (ICC=0.98) and upper and lower-limb passive range of motion (ICC=0.84 and 0.90, respectively). The reliability was moderate for tactile sensitivity (0.75). The joint pain assessment presented low reliability. CONCLUSIONS: The results showed that, except for pain assessment, application of the FMA based on the adapted version of the application manual for Brazilian Portuguese presented adequate inter-rater reliability.

Tradução, adaptação e confiabilidade interexaminadores do manual de administração da escala de Fugl-Meyer / Translation, adaptation and inter-rater reliability of the administration manual for the Fugl-Meyer assessment

CONTEXTUALIZAÇÃO: Recentemente a confiabilidade da versão brasileira da Escala de Fugl-Meyer (EFM) foi avaliada pela pontuação dada pela observação de um único examinador que aplicou a escala. Quando diferentes examinadores aplicam a escala, a confiabilidade pode depender da interpretação dada à ficha de avaliação. Nesse caso, um manual de administração claro é fundamental para garantir homogeneidade na aplicação. OBJETIVOS: Traduzir e adaptar para o português-Brasil a versão do Manual de Administração em francês-canadense da EFM e avaliar a confiabilidade interexaminadores quando diferentes examinadores aplicam a EFM com base nas informações contidas no manual. MÉTODOS: Participaram do estudo 18 adultos (59±10 anos) com hemiparesia crônica (38±35 meses pós-Acidente Vascular Encefálico). Oito sujeitos participaram da primeira parte do estudo e dez, da segunda parte. Baseada na análise dos resultados da parte 1, desenvolveu-se uma versão adaptada à qual foram adicionadas informações e fotos para ilustrar a posição do paciente e do examinador. A confiabilidade interexaminadores foi avaliada com o Coeficiente de Correlação Intraclasse (CCI). RESULTADOS: A confiabilidade da EFM baseada na versão adaptada do manual foi excelente para o escore motor total do membro superior (MS, CCI=0,98) e membro inferior (MI, CCI=0,90), sentido de movimento (CCI=0,98), amplitude de movimento (ADM) passiva do MS (CCI=0,84) e do MI (CCI=0,90) e moderada para a sensibilidade tátil (0,75). A avaliação da dor articular apresentou baixa confiabilidade. CONCLUSÃO: Os resultados mostram que, com exceção da avaliação da dor, a aplicação da EFM com base na versão adaptada do manual de aplicação em português-Brasil apresenta adequada confiabilidade interexaminadores.

BACKGROUND: Recently, the reliability of the Brazilian version of the Fugl-Meyer Assessment (FMA) was assessed through the scoring given according to observations made by a single evaluator who applied the test. When different raters apply the scale, the reliability may depend on the interpretation given to the assessment sheet. In such cases, a clear administration manual is essential for ensuring homogeneity of application. OBJECTIVES: To translate and adapt the French Canadian version of the FMA administration manual into Brazilian Portuguese and to evaluate the inter-rater reliability when different evaluators apply the FMA on the basis of the information contained in the manual. METHODS: Eighteen adults (59±10 years) with chronic hemiparesis (38±35 months after a stroke) took part in this study. Eight patients participated in the first part of the study and 10 in the second part. Based on analyzing the results from part 1, an adapted version was developed, in which information and photos were added to illustrate the positions of the patient and evaluator. The inter-rater reliability was assessed using the intraclass correlation coefficient (ICC). RESULTS: The reliability of the FMA based on the adapted version of the manual was excellent for the total motor scores for the upper limbs (ICC=0.98) and lower limbs (ICC=0.90), as well as for movement sense (ICC=0.98) and upper and lower-limb passive range of motion (ICC=0.84 and 0.90, respectively). The reliability was moderate for tactile sensitivity (0.75). The joint pain assessment presented low reliability. CONCLUSIONS: The results showed that, except for pain assessment, application of the FMA based on the adapted version of the application manual for Brazilian Portuguese presented adequate inter-rater reliability.