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61.
Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array 总被引:1,自引:0,他引:1
Malhotra R Patel V Chikkaveeraiah BV Munge BS Cheong SC Zain RB Abraham MT Dey DK Gutkind JS Rusling JF 《Analytical chemistry》2012,84(14):6249-6255
Multiplexed biomarker protein detection holds unrealized promise for clinical cancer diagnostics due to lack of suitable measurement devices and lack of rigorously validated protein panels. Here we report an ultrasensitive electrochemical microfluidic array optimized to measure a four-protein panel of biomarker proteins, and we validate the protein panel for accurate oral cancer diagnostics. Unprecedented ultralow detection into the 5-50 fg·mL(-1) range was achieved for simultaneous measurement of proteins interleukin 6 (IL-6), IL-8, vascular endothelial growth factor (VEGF), and VEGF-C in diluted serum. The immunoarray achieves high sensitivity in 50 min assays by using off-line protein capture by magnetic beads carrying 400,000 enzyme labels and ~100,000 antibodies. After capture of the proteins and washing to inhibit nonspecific binding, the beads are magnetically separated and injected into the array for selective capture by antibodies on eight nanostructured sensors. Good correlations with enzyme-linked immunosorbent assays (ELISA) for protein determinations in conditioned cancer cell media confirmed the accuracy of this approach. Normalized means of the four protein levels in 78 oral cancer patient serum samples and 49 controls gave clinical sensitivity of 89% and specificity of 98% for oral cancer detection, demonstrating high diagnostic utility. The low-cost, easily fabricated immunoarray provides a rapid serum test for diagnosis and personalized therapy of oral cancer. The device is readily adaptable to clinical diagnostics of other cancers. 相似文献
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Cell transplantation into the seminiferous tubules is a useful technique for the study of physiological and pathological conditions affecting the testis. However, the precise three-dimensional organization and, particularly, the complex connectivity of the seminiferous network have not yet been thoroughly characterized. To date, the technical approaches to address these issues have included manual dissection under the stereomicroscope, reconstruction of histological serial sections, and injection of contrast dyes, but all of them have yielded only partial information. Here, using an approach based on the microinjection of a self-polymerizing resin followed by chemical digestion of the surrounding soft tissues, we reveal fine details of the seminiferous tubule scaffold and its connections. These replicas of the testis seminiferous network were studied by scanning electron microscopy. The present results not only establish a morphological basis for more precise microinjection into the mouse seminiferous tubules but also enable a more profound investigation of physiological and embryological features of the testis. 相似文献
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G. Munir G. Koller L. Di Silvio M. J. Edirisinghe W. Bonfield J. Huang 《Journal of the Royal Society Interface》2011,8(58):678-688
Bioactive hydroxyapatite (HA) with addition of silicon (Si) in the crystal structure (silicon-doped hydroxyapatite (SiHA)) has become a highly attractive alternative to conventional HA in bone replacement owing to the significant improvement in the in vivo bioactivity and osteoconductivity. Nanometre-scaled SiHA (nanoSiHA), which closely resembles the size of bone mineral, has been synthesized in this study. Thus, the silicon addition provides an extra chemical cue to stimulate and enhance bone formation for new generation coatings, and the next stage in metallic implantation design is to further improve cellular adhesion and proliferation by control of cell alignment. Topography has been found to provide a powerful set of signals for cells and form contact guidance. Using the recently developed novel technique of template-assisted electrohydrodynamic atomization (TAEA), patterns of pillars and tracks of various dimensions of nanoSiHA were achieved. Modifying the parameters of TAEA, the resolution of pattern structures was controlled, enabling the topography of a substrate to be modified accordingly. Spray time, flow rate and distance between the needle and substrate were varied to improve the pattern formation of pillars and tracks. The 15 min deposition time provided the most consistent patterned topography with a distance of 50 mm and flow rate of 4 µl min−1. A titanium substrate was patterned with pillars and tracks of varying widths, line lengths and distances under the optimized TAEA processing condition. A fast bone-like apatite formation rate was found on nanoSiHA after immersion in simulated body fluid, thus demonstrating its high in vitro bioactivity. Primary human osteoblast (HOB) cells responded to SiHA patterns by stretching of the filopodia between track and pillar, attaching to the apex of the pillar pattern and stretching between two. HOB cells responded to the track pattern by elongating along and between the track, and the length of HOB cells was proportional to the gaps between track patterns, but this relationship was not observed on the pillar patterns. The study has therefore provided an insight for future design of next generation implant surfaces to control and guide cellular responses, while TAEA patterning provides a controllable technique to provide topography to medical implants. 相似文献
68.
Susane Oshiro Marta Ramalho Paloma Cristina Durães Raissa Andrade Karolina Silva Flaviano Silvério 《Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment》2020,37(9):1510-1519
ABSTRACT
p-Cresol has been identified as a flavouring compound in cheeses; however, scientific studies have already identified p-cresol as a potential chemical contaminant in environmental matrices. Thus, the objective of this study was to evaluate four traditional methods for extracting p-cresol from cheese samples in order to validate the best method, and finally to apply it to five cheese samples with different origins, processing and ripeness times. The analyses were performed by gas chromatography-mass spectrometry after derivatisation of p-cresol with anhydride acetic and pyridine. Better results were achieved by the QuEChERS method, which showed recovery higher than 80%, relative standard deviation lower than 16%, limit of quantification of 5 μg kg?1 and linearity between 5 and 400 μg kg?1 with R2 0.99. p-Cresol was quantified in almost all of the samples analysed at different concentration levels, which were in an increasing order at μg kg?1: Cheddar (< LOQ), Parmesan (8 ± 0.7), Gorgonzola (103 ± 14), smoked Provolone (365 ± 28) and barbecue cheese (1001 ± 187). Although no maximum residue limit has been established for p-cresol in food, the results suggest that cheeses exposed to charcoal combustion notably increase the p-cresol levels and may represent a hazard to human health, especially in risk groups such as patients with chronic kidney disease who have serious problems with p-cresol. 相似文献
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The ability of foodborne pathogens to survive in certain foods mainly depends on stress response mechanisms. Insight into molecular properties enabling pathogenic bacteria to survive in food is valuable for improvement of the control of pathogens during food processing. Raw milk cheeses are a potential source for human infections with Shiga toxin-producing Escherichia coli (STEC). In this review, we focused on the stress response mechanisms important for allowing STEC to survive raw milk cheese production processes. The major components and regulation pathways for general, acid, osmotic, and heat shock stress responses in E. coli and the implications of these responses for the survival of STEC in raw milk cheeses are discussed. 相似文献