Use of Single-Dose Dexamethasone in Patients With Diabetes Undergoing Surgery: A Systematic Review and Meta-Analysis.

The purpose of this review was to examine the effect of single-dose dexamethasone on perioperative blood glucose in diabetic patients. We used PubMed, Cochrane Library, MEDLINE, CINAHL, Google Scholar, and grey literature for our search. Only randomized controlled trials were included. Risk ratio (RR) and mean difference (MD) were used to estimate outcomes with suitable effect models. Quality of evidence was assessed using the Risk of Bias and GRADE systems. We analyzed seven trials involving 1,321 patients. Diabetic patients treated with single-dose dexamethasone had statistically significant changes in blood glucose levels from baseline by 33.61 mg/dL (MD, 33.61; 95% CI, 17.59 to 49.63; P < .0001). Dexamethasone increased blood glucose levels 1-4 hours (MD, 29.02; 95% CI, 7.09 to 50.94; P = .010), 8-24 hours (MD, 30.81; 95% CI, 9.21 to 52.41; P = .005) after administration and increased risks of hyperglycemia. However, there was no difference in surgical site infection (SSI) (RR, 0.81; 95% CI, 0.59 to 1.11; P = .19). Effect size imprecision, substantial heterogeneity, and publication bias was the study's limitations. We found that single-dose dexamethasone increased glucose concentration 24 hours after surgery with little to no effect on SSI. Extrapolation of these findings to clinical settings must take into consideration the review's limitations.

Integrating single-molecule FRET and biomolecular simulations to study diverse interactions between nucleic acids and proteins.

The conformations of biological macromolecules are intimately related to their cellular functions. Conveniently, the well-characterized dipole-dipole distance-dependence of Förster resonance energy transfer (FRET) makes it possible to measure and monitor the nanoscale spatial dimensions of these conformations using fluorescence spectroscopy. For this reason, FRET is often used in conjunction with single-molecule detection to study a wide range of conformationally dynamic biochemical processes. Written for those not yet familiar with the subject, this review aims to introduce biochemists to the methodology associated with single-molecule FRET, with a particular emphasis on how it can be combined with biomolecular simulations to study diverse interactions between nucleic acids and proteins. In the first section, we highlight several conceptual and practical considerations related to this integrative approach. In the second section, we review a few recent research efforts wherein various combinations of single-molecule FRET and biomolecular simulations were used to study the structural and dynamic properties of biochemical systems involving different types of nucleic acids (e.g., DNA and RNA) and proteins (e.g., folded and disordered).

A microchip-based proteolytic digestion system driven by electroosmotic pumping.

Microchip-based proteomic analysis requires proteolytic digestion of proteins in microdevices. Enzyme reactors in microdevices, fabricated in glass, silicon, and PDMS substrates, have recently been demonstrated for model protein digestions. The common approach used for these enzyme reactors is employment of a syringe pump(s) to generate hydrodynamic flow, driving the proteins through the reactors. Here we present a novel approach, using electroosmotic flow (EOF) to electrokinetically pump proteins through a proteolytic system. The existence of EOF in the proteolytic system packed with immobilized trypsin gel beads was proven by imaging the movement of a neutral fluorescent marker. Digestions of proteins were subsequently carried out for 12 min, and the tryptic peptides were analyzed independently using capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MS). The results from CE analysis of the tryptic peptides from the EOF-driven proteolytic system and a conventional water bath digestion were comparable. MALDI-TOF MS was used to identify the parent protein and the tryptic peptides using MS-Fit database searching. The potential utility of the EOF-driven proteolytic system was demonstrated by direct electro-elution of proteins from an acrylamide gel into the proteolytic system, with elution and tryptic digestion achieved in a single step. The EOF-driven proteolytic system, thus, provides a simple way to integrate protein digestion into an electrophoretic micro total analysis system for protein analysis and characterization.

Laser-induced fluorescence detection on multichannel electrophoretic microchips using microprocessor-embedded acousto-optic laser beam scanning.

An improved method for fast scanning and fluorescence detection on multimicrochannel microchips is presented using acousto-optic-deflection-driven laser-beam scanning. A microprocessor embedded subsystem used in conjunction with LabView program as the human-machine interface for control of laser-beam scanning and data preprocessing allowed faster scanning and addressing speeds to be attained and improved attenuation calibration and the data sampling speed. This system allows for flexible, high-resolution fluorescence detection for multimicrochannel electrophoresis in a manner that can be applied to a number of high-throughput analysis applications. Incorporating an F-theta focusing lens into the optical set-up allowed for a laser spot as small as 10 microm to accurately be addressed to the center of microchannels. With this spot size, it will be possible to further increase the channel density in the scanning range without encountering crosstalk. Using a six-channel microchip (four separation channels, two alignment channels), the simultaneous separation and fluorescence detection of amino acids and DNA digest samples in four channels is illustrated. User-friendly interpretation of the separation data is facilitated not only by a peak alignment/normalization routine developed within the software, but also through improved signal-to-noise ratios obtained through exploitation of signal processing.

A simple PDMS-based electro-fluidic interface for microchip electrophoretic separations.

High voltage electrodes for electrophoresis have been integrated into a polymer layer that can be reversibly bound to glass microchips for electrophoretic separations. By using the liquid precursor to the polymer polydimethylsiloxane (PDMS), platinum electrodes and reservoirs can be positioned prior to solidification, providing a simple and flexible method for electrode interface construction. Field strengths up to 875 V cm(-1) over an 8 cm separation channel can be applied to the system without any loss in performance of the interface. The interface can function as an electro-fluidic interface between the high voltage power supply and the separation channel and, when reversibly sealed to an etched glass plate, functions as a cover plate establishing a hybrid PDMS-glass microchip in which the electrodes are directly integrated onto the device. The versatility of this approach is not only demonstrated by separating DNA fragments in a novel buffer sieving matrix, but also with the molecular diagnostic analysis of a variety of DNA samples for Duschenne Muscular Dystrophy and cytomegalovirus (CMV) infection, using both microchip interface configurations.

Hydroxypropyl cellulose as an adsorptive coating sieving matrix for DNA separations: artificial neural network optimization for microchip analysis.

Effective DNA separations in microelectrophoretic systems are complicated by the need to passivate the surface dynamically or covalently. We describe the optimization and utilization of a novel buffer system for fast DNA separations by capillary and microchip electrophoresis without the need for any surface modification or conditioning prior to separation. At concentrations as high as 5%, hydroxypropyl cellulose (HPC) has a relatively low viscosity, allowing for microchip channel filling to be performed with ease. A MES/TRIS buffer system at pH 6.1 eliminates the need for surface preconditioning procedures due to the promotion of hydrogen bonding of HPC with the wall. An additional benefit with this buffer system is the low current observed at high fields when compared to other common DNA separation buffers. An artificial neural network (ANN) was used to model the data and to predict the optimum conditions. Utility of the ANN-optimized system for molecular diagnostic testing was demonstrated by performing microchip separations on DNA samples from patients suspected of having genetic mutations associated with Duchenne muscular dystrophy (DMD). Microchip analysis easily allowed for the patient samples positive for DMD mutations to be distinguished from patient samples negative for the disease.