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51.
John P. Hilton ThaiHuu Nguyen Mihaela Barbu Renjun Pei Milan Stojanovic Qiao Lin 《Microfluidics and nanofluidics》2012,13(5):749-760
We present a bead-based approach to microfluidic polymerase chain reaction (PCR), enabling fluorescent detection and sample conditioning in a single microchamber. Bead-based PCR, while not extensively investigated in microchip format, has been used in a variety of bioanalytical applications in recent years. We leverage the ability of bead-based PCR to accumulate fluorescent labels following DNA amplification to explore a novel DNA detection scheme on a microchip. The microchip uses an integrated microheater and temperature sensor for rapid control of thermal cycling temperatures, while the sample is held in a microchamber fabricated from (poly)dimethylsiloxane and coated with Parylene. The effects of key bead-based PCR parameters, including annealing temperature and concentration of microbeads in the reaction mixture, are studied to achieve optimized device sensitivity and detection time. The device is capable of detecting a synthetically prepared section of the Bordetella pertussis genome in as few as 10 temperature cycles with times as short as 15?min. We then demonstrate the use of the procedure in an integrated device; capturing, amplifying, detecting, and purifying template DNA in a single microfluidic chamber. These results show that this method is an effective method of DNA detection which is easily integrated in a microfluidic device to perform additional steps such as sample pre-conditioning. 相似文献
52.
It has been found that using a segmented grinding wheel with a fluid chamber can significantly minimise the quantity of coolant while improving the ground surface integrity. The present investigation aims to explore the fluid flow mechanism in such a wheel system. To this end, the Weber theory for Newtonian jet instability was applied to quantitatively determine the contribution of coolant flow rate to mist and ligament modes. A semi-analytical model was then developed to predict the mist flow rate by taking into account both the grinding parameters and fluid properties. It was shown that the model prediction was in good agreement with experimental measurements. Because of the comprehensive integration of variables in the formulation, the model provides a good fundamental understanding of the mist formation and offers a practical guideline for the selection and use coolant in minimising the mist flow rate. 相似文献
53.
Cassidy E. Wideman James Nguyen Sean D. Jeffries Boyer D. Winters 《International journal of molecular sciences》2021,22(1)
Reminder cues can destabilize consolidated memories, rendering them modifiable before they return to a stable state through the process of reconsolidation. Older and stronger memories resist this process and require the presentation of reminders along with salient novel information in order to destabilize. Previously, we demonstrated in rats that novelty-induced object memory destabilization requires acetylcholine (ACh) activity at M1 muscarinic receptors. Other research predominantly has focused on glutamate, which modulates fear memory destabilization and reconsolidation through GluN2B- and GluN2A-containing NMDARs, respectively. In the current study, we demonstrate the same dissociable roles of GluN2B- and N2A-containing NMDARs in perirhinal cortex (PRh) for object memory destabilization and reconsolidation when boundary conditions are absent. However, neither GluN2 receptor subtype was required for novelty-induced destabilization of remote, resistant memories. Furthermore, GluN2B and GluN2A subunit proteins were upregulated selectively in PRh 24 h after learning, but returned to baseline by 48 h, suggesting that NMDARs, unlike muscarinic receptors, have only a temporary role in object memory destabilization. Indeed, activation of M1 receptors in PRh at the time of reactivation effectively destabilized remote memories despite inhibition of GluN2B-containing NMDARs. These findings suggest that cholinergic activity at M1 receptors overrides boundary conditions to destabilize resistant memories when other established mechanisms are insufficient. 相似文献
54.
Nguyen Khanh Toan Nguyen Chi Tai Soo-A Kim Sang-Gun Ahn 《International journal of molecular sciences》2021,22(1)
Salivary gland dysfunction induces salivary flow reduction and a dry mouth, and commonly involves oral dysfunction, tooth structure deterioration, and infection through reduced salivation. This study aimed to investigate the impact of aging on the salivary gland by a metabolomics approach in an extensive aging mouse model, SAMP1/Klotho -/- mice. We found that the salivary secretion of SAMP1/Klotho -/- mice was dramatically decreased compared with that of SAMP1/Klotho WT (+/+) mice. Metabolomics profiling analysis showed that the level of acetylcholine was significantly decreased in SAMP1/Klotho -/- mice, although the corresponding levels of acetylcholine precursors, acetyl-CoA and choline, increased. Interestingly, the mRNA and protein expression of choline acetyltransferase (ChAT), which is responsible for catalyzing acetylcholine synthesis, was significantly decreased in SAMP1/Klotho -/- mice. The overexpression of ChAT induced the expression of salivary gland functional markers (α–amylase, ZO-1, and Aqua5) in primary cultured salivary gland cells from SAMP1/Klotho +/+ and -/- mice. In an in vivo study, adeno-associated virus (AAV)-ChAT transduction significantly increased saliva secretion compared with the control in SAMP1/Klotho -/- mice. These results suggest that the dysfunction in acetylcholine biosynthesis induced by ChAT reduction may cause impaired salivary gland function 相似文献
55.
Thi Kieu Oanh Nguyen Thi Luong Vu Minh Quan Nguyen Huynh Kim Khanh Ta Kyoung Sun Park Soo Hyeon Kim Chong Jai Kim Yeon Jin Jang Han Choe 《International journal of molecular sciences》2021,22(10)
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a member of the colony-stimulating factor (CSF) family, which functions to enhance the proliferation and differentiation of hematopoietic stem cells and other hematopoietic lineages such as neutrophils, dendritic cells, or macrophages. These proteins have thus generated considerable interest in clinical therapy research. A current obstacle to the prokaryotic production of human GM-CSF (hGM-CSF) is its low solubility when overexpressed and subsequent complex refolding processes. In our present study, the solubility of hGM-CSF was examined when combined with three N-terminal fusion tags in five E. coli strains at three different expression temperatures. In the five E. coli strains BL21 (DE3), ClearColi BL21 (DE3), LOBSTR, SHuffle T7 and Origami2 (DE3), the hexahistidine-tagged hGM-CSF showed the best expression but was insoluble in all cases at each examined temperature. Tagging with the maltose-binding protein (MBP) and the b′a′ domain of protein disulfide isomerase (PDIb′a′) greatly improved the soluble overexpression of hGM-CSF at 30 °C and 18 °C. The solubility was not improved using the Origami2 (DE3) and SHuffle T7 strains that have been engineered for disulfide bond formation. Two conventional chromatographic steps were used to purify hGM-CSF from the overexpressed PDIb′a′-hGM-CSF produced in ClearColi BL21 (DE3). In the experiment, 0.65 mg of hGM-CSF was isolated from a 0.5 L flask culture of these E. coli and showed a 98% purity by SDS-PAGE analysis and silver staining. The bioactivity of this purified hGM-CSF was measured at an EC50 of 16.4 ± 2 pM by a CCK8 assay in TF-1 human erythroleukemia cells. 相似文献
56.
Chi Nguyen Quynh Ho Minh Thi Tran Chung Chinh Doan Son Nghia Hoang Diem Hong Tran Long Thanh Le 《International journal of molecular sciences》2021,22(9)
Simulated microgravity (SMG) induced the changes in cell proliferation and cytoskeleton organization, which plays an important factor in various cellular processes. The inhibition in cell cycle progression has been considered to be one of the main causes of proliferation inhibition in cells under SMG, but their mechanisms are still not fully understood. This study aimed to evaluate the effects of SMG on the proliferative ability and cytoskeleton changes of Chang Liver Cells (CCL-13). CCL-13 cells were induced SMG by 3D clinostat for 72 h, while the control group were treated in normal gravity at the same time. The results showed that SMG reduced CCL-13 cell proliferation by an increase in the number of CCL-13 cells in G0/G1 phase. This cell cycle phase arrest of CCL-13 cells was due to a downregulation of cell cycle-related proteins, such as cyclin A1 and A2, cyclin D1, and cyclin-dependent kinase 6 (Cdk6). SMG-exposed CCL-13 cells also exhibited a downregulation of α-tubulin 3 and β-actin which induced the cytoskeleton reorganization. These results suggested that the inhibited proliferation of SMG-exposed CCL-13 cells could be associate with the attenuation of major cell cycle regulators and main cytoskeletal proteins. 相似文献
57.
T. Nguyen I. Zarudi L.C. Zhang 《International Journal of Machine Tools and Manufacture》2007,47(1):97-106
This paper studies an innovative development of a steel grinding–hardening technology using an inert cryogen—liquid nitrogen. It was found that phase transformations took place during grinding with the application of liquid nitrogen and resulted in hardened surface layer in a ground component. The layer had a fine laths martensite structure which gave rise to a remarkably high hardness. It was also shown that the treatment can produce superior surface integrity, with compressive surface residual stresses and without surface oxidation. Due to the inert nature of the liquid nitrogen, the grinding process becomes environmentally conscious. 相似文献
58.
The molecular orientation at the outer surface of injection–stretch–blow‐molded bottles made from poly(ethylene terephthalate) was characterized and quantified by means of front‐surface reflection infrared spectroscopy based on a method developed previously. Results were obtained for two different bottle shapes (cylindrical and rectangular) molded at different injection mold temperatures (16, 38, and 60°C). For the cylindrical bottles, the preferred molecular chain orientation was found to be in the axial direction, with the Hermans orientation function near 0.3 for all three mold temperatures. For the less symmetrical rectangular bottles, a significant difference was observed between the large and small faces. For the large face, the orientation was mainly in the hoop direction; the Hermans orientation function was in the range of 0.3–0.5 and was essentially the same at all mold temperatures and positions along the bottle height. For the small face, on the other hand, the preferred orientation changed from the hoop direction near the bottom to the axial direction near the top, and the variation was more pronounced at lower mold temperatures. The utility of the front‐surface reflection technique was clearly demonstrated. It was also applied, with the use of an infrared microscope, to examine the orientation gradient across the wall thickness. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 1319–1327, 2007 相似文献
59.
Xiaohong Gu Tinh Nguyen Mounira Oudina David Martin Bouchra Kidah Joan Jasmin Aziz Rezig Lipiin Sung Eric Byrd Jonathan W. Martin Derek L. Ho Y. C. Jean 《Journal of Coatings Technology and Research》2005,2(7):547-556
Atomic force microscopy (AFM) has been used to study the morphology and microstructure of an amine-cured epoxy before and
after outdoor exposure. Measurements were made from samples prepared in an essentially CO2-free, H2O-free glove box and from samples prepared in ambient conditions. For those prepared in a CO2-free glove box, AFM imaging was conducted on (1) an unexposed air/coating surface, (2) an unexposed coating bulk, (3) an
unexposed coating/substrate interface, and (4) a field exposed air/coating surface. For samples prepared in ambient conditions,
only the unexposed air/coating surface was investigated. The same regions of the exposed samples were scanned periodically
by the AFM to monitor changes in the surface morphology of the coating as UV exposure progressed. Small angle neutron scattering
and Fourier transform infrared spectroscopy (FTIR) studies were performed to verify the microstructure and to follow chemical
changes during outdoor exposure, respectively. The results have shown that amine blushing, which occurs only under ambient
conditions, had a significant effect on the surface morphology and microstructure of the epoxy. The surface morphology of
the samples prepared under CO2-free, dry conditions was generally smooth and homogeneous. However, the interface and the bulk samples clearly revealed a
two-phase structure consisting of bright nodular domains and dark interstitial regions, indicating an inhomogeneous microstructure.
Such heterogeneous structure of the bulk was in good agreement with results obtained by small angle neutron scattering of
unexposed samples and by AFM phase imaging of the degraded sample surface. The relationship between submicrometer physical
changes and molecular chemical degradation is discussed.
Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, October 27–29, 2004, in Chicago,
IL. 相似文献
60.
We consider the classic problem of pole placement by state feedback. We offer an eigenstructure assignment algorithm to obtain a novel parametric form for the pole-placing feedback matrix that can deliver any set of desired closed-loop eigenvalues, with any desired multiplicities. This parametric formula is then exploited to introduce an unconstrained nonlinear optimisation algorithm to obtain a feedback matrix that delivers the desired pole placement with optimal robustness and minimum gain. Lastly we compare the performance of our method against several others from the recent literature. 相似文献