Applications of peptide and protein-based materials in bionanotechnology

In this critical review we highlight recent advances in the use of peptide- and protein-related materials as smart building blocks in nanotechnology. Peptides and proteins can be very practical for new material synthesis and device fabrications. For example, peptides and proteins have superior specificity for target binding as seen in the antibody recognition and this biological recognition function can be used to assemble them into specific structures and shapes in large scale, as observed in the S-layer protein assembly. Collagens are assembled from triple helix peptides in micron-size with precise recognition between peptides and these biological assemblies can undergo smart structural change with pH, ionic strength, temperature, electric/magnetic fields. In addition, assemblies of peptides can template complex 3D crystallization processes with catalytic function, thus enabling to grow various materials in physiological conditions at low temperature in aqueous solution. The biomimetic growth of nanomaterials in aqueous solution is extremely useful when they are applied to therapeutics and medical imaging in vivo since these nanomaterials will be well dispersed in bodies. Peptides also play significant roles in signal transduction pathways in cells. For example, neuropeptides are used as neurotransmitters between synapses and these peptides bind receptors on the surface of cells to cascade the signal transduction. These versatile functions of peptides are extremely practical and here we discuss them with examples of relevant applications such as nanoreactors, sensors, electronics, and stimulus-responsive materials. It should be noted that peptide/protein assemblies can be applied to build up micron-scale materials that still feature excellent nano-scale ensembles, which essentially bridges the nano-world and the micro-world (86 references)

Avaliação anatômica das vértebras cervicais em ratos Wistar por meio de radiografias digitais e sua correlação com os estágios da maturação das vértebras cervicais em humanos

O presente trabalho visa identificar e evidenciar a curva de crescimento em animais utilizados em pesquisa laboratorial. Em muitos casos a idade biológica se torna fundamental para resultados precisos e confiáveis, em especial na ortodontia, pois a maturação óssea é avaliada por meio de radiografias, e as mudanças observadas indicam em qual estágio se encontra o indivíduo. A idade cronológica sem dúvida é a mais utilizada, sendo observada mesmo antes do nascimento, durante a gestação e até os últimos dias de vida deste indivíduo. Contudo a idade cronológica em alguns casos não evidencia a exatidão da fase de maturação; um dos métodos utilizados é a avaliação da maturação das vértebras cervicais. A método que foi utilizada na avaliação das alterações das vértebras em animais foi a mesma que os autores utilizam em humanos. Sendo assim, esse trabalho tem como objetivo principal a identificação da idade óssea em animais para limitar a distribuição dos mesmos, contribuindo com a eficiência nos experimentos quando a idade e fase do desenvolvimento pode influenciar nos resultados das pesquisas. Os 35 ratos Wistar foram observados por um período de 160 dias, iniciando pelo 22º dia (desmame), com cortes transversais periódicos para pesagem, medição do comprimento e radiografias digitais obtidas com os animais imobilizados por anestésicos. As imagens radiográficas das vértebras cervicais (C2 e C3) foram sumetidas a densitometria ótica e mensuradas por meio de um programa de computador. As variáveis foram submetidas à análise estatística individual e relacionadas entre si obtendo resultados significantes para o crescimento (p<0,001). O grupo masculino apresenta médias significantes maiores que as do grupo feminino em todas as avaliações realizadas (p<0,001). Para a densidade houve aumento conforme...The aim of this study was to identify and evidence the growth curve of animals used for laboratorial research purposes. The biological age is important for precise and reliable results, especially in orthodontics, since radiographs are used to evaluate the bone maturation and the stage in which is the patient according to the changes that are observed. The chronological age is certainly the most utilized, been observed even before birth, during pregnancy until the person's last days of life. However, the chronological age, in some cases, does not evidence exactly the maturation phase. Thus, the evaluation of bone age or maturation of cervical vertebras is a commonly used method.The methodology used for the evaluation of the vertebral alterations in animals was the same that used in humans. Therefore, the main objective of this study was the identification of the bone age in animals to limit indiscriminate use, contributing with the experiences efficiency when the age and development phase can influence the research results. The 35 Wistar rats were observed for a period of 260 days, beginning in the 22nd day (weaning). Periodical transversal cuts for weight analysis, length measurement and digital radiographs were taken with the animals immobilized by anesthetics. The radiographic images of cervical vertebras (C2 and C3) were submitted to optical densitometry and measured by computer software. The variables were submitted to individual and each other related statistical analysis. The results were significant for growth (p< 0.001). The male group showed significantly higher mediums, than that showed by the female group in all evaluations. Conclusion: There exist a growth outbreak and the growth curves of weight and length, and the alterations... (Complete abstract click electronic access below)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Biomimetic conformation-specific assembly of proteins at artificial binding sites nanopatterned on silicon

Biomolecules such as enzymes and antibodies possess binding sites where the molecular architecture and the physicochemical properties are optimum for their interaction with a particular target, in some cases even differentiating between stereoisomers. Here, we mimic this exquisite specificity via the creation of a suitable chemical environment by fabricating artificial binding sites for the protein calmodulin (CaM). By downscaling well-known surface chemical modification methodologies to the nanometer scale via silicon nanopatterning, the Ca(2+)-CaM conformer was found to selectively bind the biomimetic binding sites. The methodology could be adapted to mimic other protein-receptor interactions for sensing and catalysis

Self-assembled peptide nanotubes for electronics and sensor devices

Non-lithographic fabrications of devices such as electronics and sensor have been studied extensively by assembling nanometer-sized building blocks into the device configurations. While various nanowires and nanoparticles with superior physical properties have been synthesized as the building blocks, more reproducible methods to assemble them onto precise positions are desirable to construct nanodevices. We developed peptide nanotubes as multifunctional smart building blocks. We designed these nanotubes to incorporate biomolecular recognition components (antibody), and our strategy is to use those functionalized peptide nanotubes, which can recognize and selectively bind a well-defined region on antigen-patterned substrates, as building blocks to assemble nanoscale architectures at uniquely defined positions. In order for the application in electric device fabrications, after configuring device geometries with these nanotubes by the biomolecular recognition, we turned on the biomineralization function of peptides on the nanotube sidewall to develop various material coatings such as metals and semiconductors for electronics and sensor applications. It should be noted that the coating morphology such as particle-domain size and inter-particle distance on the nanotubes could be tuned by peptide sequences and conformations. Due to these peptides' catalytic function, some semiconductor coatings could be developed at room temperature on the nanotube

Assemblies of functional peptides and their applications in building blocks for biosensors

We highlight our recent applications of functional peptide nanotubes, self-assembled from short peptides with recognition elements, as building blocks to develop sensors. Peptide nanotubes with high aspect ratios are excellent building blocks for directed assembly into device configurations, and their combining structures with the nanometric diameters and the micrometric lengths enables to bridge the nano-world and the micro-worl

Single-cell pathogen detection with a reverse-phase immunoassay on impedimetric transducers

The risk of infectious diseases has compelled some industries to establish a zero-tolerance standard for the presence of microorganisms in a given sample. Here, we address this issue with a novel reverse-phase immunoassay on impedimetric transducers for the specific detection of extremely low numbers of pathogens (less than 10 cells). After simply spotting the sample onto the electrodes, physisorbed analytes were targeted with urease-labeled antibodies, and the urease on the pathogens hydrolyzed urea to ionic species with a concomitant decrease of the resistivity of the solution. By this methodology, the limit of detection (LOD) based on the 3 sigma criterion was 1 Escherichia coli cell with an assay time under 1 h. However, the precise number of cells present in highly diluted samples is uncertain, making it difficult to assess the final LOD of the sensor. We overcome this problem by using an atomic force microscope to deposit and image in situ the exact number cells on the transducer. After performing the immunoassay, a single E. coli cell was successfully detected without ambiguity in the number of cells even in the presence of a 10(4) excess of a competing microorganism, thus demonstrating the outstanding LOD and selectivity of the proposed reverse-phase immunoassay

Label-free cancer cell detection with impedimetric transducers

While cancer is still an implacable disease, many cancers can be cured if they are diagnosed in an early stage. Recently, it was reported that the transformation from normal cells to cancer cells can change their mechano-elastic properties to become softer and more deformable. If some cancer cells are more deformable, then a progressive increase of the volume of softer cancer cells should be induced as an abrupt change in osmolarity is applied. On the basis of this hypothesis, we developed a sensor that can electronically monitor the volume increase of cancer cells under hyposmotic pressure. By this methodology, K:Molv NIH 3T3 cells, 786-O human kidney carcinoma cells, and MPSC-1 ovarian cancer cells were successfully detected within 30 min using on the order of 10 cells. These cancer cells could be detected with the same sensitivity even in the presence of a vast excess of the respective noncancerous cells [NIH 3T3 cells, human embryonic kidney (HEK) 293 cells, ovarian surface epithelial (OSE) cells]. Since the proposed impedimetric sensor could be useful for detecting cancer cells fast and reliably, it could be further implemented in the screening of large populations of tissue samples and the detection of circulating tumor cells for point-of-care applications

Anatomical evaluation of the cervical vertebrae of Wistar rats by means of digital radiographs and its correlation with the maturation stages of human cervical vertebrae

INTRODUCTION: Biological age is an important parameter for growth and development assessment. It can be evaluated through the observation of radiographic changes in skeletal maturation of cervical vertebrae. OBJECTIVE: This study aims to: a) verify if there is correlation between growth curve and the stages of bone age of animals used in laboratories, by evaluating radiographs of the cervical vertebrae; b) correlate these stages with their correspondents in humans. METHODS: 35 Wistar rats were evaluated for a period of 160 days, starting at day 22nd (weaning), with cross sections for periodic weighing, length measurement and digital radiography. Radiographs of the cervical vertebrae (C2 and C3) were measured by means of a computer program (Radio IMP). Data were submitted to statistical analysis (ANOVA) and Pearson correlation. RESULTS: Growth spurt was characterized by fast increasing in weight and length. Through ANOVA, differences were observed in the cervical measurements between days 22, 97, 127, 157, 187 and 217 (p <0.001). A high correlation was found between increasing in body length and weight, as well as in cervical vertebrae height (r = 0.86). Increments in concavities of vertebrae were also observed, similar to humans. CONCLUSIONS: There is correlation between body growth and maturation of cervical vertebrae in rats. Despite the continuous development of concavities, it was not possible to clearly identify the 5/6 stages as in studies of cervical vertebrae maturation in humans.INTRODUÇÃO: a idade biológica é um parâmetro importante na avaliação do crescimento e desenvolvimento, podendo ser avaliada por meio da observação de alterações na maturação óssea das vértebras. OBJETIVO: o presente estudo visa descrever e relacionar a curva de crescimento de ratos utilizados em pesquisas laboratoriais com os estágios de idade óssea, avaliados por radiografias de vértebras cervicais, e correlacionar esses estágios com estudos correspondentes em humanos. MÉTODOS: foram avaliados 35 ratos Wistar em um período de 160 dias, iniciando no 22º dia de vida (desmame), com cortes transversais periódicos para pesagem, medição do comprimento e radiografias digitais. As radiografias das vértebras cervicais (C2 e C3) foram mensuradas por meio de um programa de computador (Radio IMP). Os dados foram submetidos à análise estatística de variância (ANOVA). RESULTADOS: o surto de crescimento caracterizou-se por aumento rápido de peso e comprimento, seguido por um período de crescimento lento e de estabilidade. Uma alta correlação (r = 0,86) foi verificada entre o aumento de peso e o comprimento do corpo, bem como o comprimento das vértebras cervicais. Incrementos nas concavidades das vértebras dos ratos foram observados, semelhantemente aos resultados obtidos em estudos em humanos. CONCLUSÕES: existe correlação entre o crescimento corporal de ratos e a maturação das vértebras cervicais. Apesar da detecção de desenvolvimento contínuo de concavidades das vértebras, não foi possível identificar claramente os 5 ou 6 estágios de maturação óssea descritos em seres humanos