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Biomedical subjects

S J Yao

Publications and source records attributed to S J Yao.

At least 19 recordsLinked to original sources

An improved procedure for production of human epidermal growth factor from recombinant E. coli.

An improved procedure for the fermentation and purification of human epidermal growth factor (hEGF) was developed. Recombinant Escherichia coli HB-101 [lacUV5omp08hEGF] harboring plasmid lacUV5omp08hEGF encoding hEGF was used in fermentation to increase levels of hEGF. Medium composition, and the levels of inoculum, inducer (isopropyl-beta-D-thiogalactoside) and ampicillin were optimized with respect to volumetric fermentation of hEGF. As a result, the hEGF concentration reached a high value of 242 mg l(-1) and the amount of heterogeneous protein decreased by 62% compared with that before optimization in batch fermentation. High-quality hEGF was purified from the fermentation culture by centrifugation, salting-out, resuspension, recentrifugation and finally gel chromatography on a Grad-iFrac System using Sephadex G-50 superfine. The purity of hEGF and the total yield were more than 94% and higher than 36%, respectively, and SDS-PAGE of the purified hEGF demonstrated a single band corresponding to an hEGF standard. In particular, a very important phenomenon was found, i.e. that the amount of heterogenous protein in fermentation broths cultured in media with high concentrations of lactose is far less than that cultured in media with high concentrations of glucose.

Biotechnology↗

[Genetical effect of different rye chromosomes on the acid phosphatase (Acph) secretion of common wheat roots under phosphorus starvation conditions].

The effects of different rye chromosomes on Acph secretion of common wheat roots under P starvation conditions were studied by using a set of Chinese Spring-Imperial (CS-IMP) alien addition lines as materials. The Acph activity measurement results showed that P starvation is an induction factor for Acph gene expression; Different chromosomes of Imperial rye in Chinese Spring background has different effects on the secretion of Acph by corresponding addition line roots. Among them, chromosome 1R had the strongest promoting effect; The IEF diagram of Acph isozymes clearly demonstrated that chromosome 1R in rye genome carries P starvation inducible Acph gene(s).

Acid Phosphatase↗

Voltage polarity relay--optimal control of electrochemical urea oxidation.

Voltage polarity relay (VPR) is shown to optimize the urea oxidation rate and urea current utilization under constant current conditions in direct electrochemical urea oxidation. Direct electrochemical urea oxidation is characterized by reversible deactivation of the working electrode due to oxidation products remaining on the surface and the requirement that the working electrode potential remain below about 1.1 V relative to Ag/AgCl in order to prevent undesirable secondary electrochemical oxidations. The VPR method monitors the potential of the working electrode relative to a suitable reference and changes system polarity when the upper potential set limit is reached. Thus, what was the working electrode becomes the counter electrode and vice versa. Since urea oxidation products are desorbed from the counter electrode when its potential drops below about -0.6 V relative to Ag/AgCl, alternating electrode functions between working and counter provides cyclic electrode regeneration and continuous urea oxidation. VPR is believed to optimize constant current control for any electrochemical system that exhibits behavior similar to direct electrochemical urea oxidation.

Electrochemistry↗

Electrochemical glucose sensing at low potentials.

An electrochemical sensing method was studied, employing cyclic voltammetry at smooth Pt-electrodes in the low potential region. It has certain advantages over methods that employ higher potentials or constant voltage amperometry. There are also advantages over methods employing active enzymes or other unstable elements. In addition to a specific scanning potential we have studied voltage-delay-pulsing techniques for electrode rejuvenation and innovative methods of creating analyte-selective covering membrane. At potentials below -0.6 V vs Ag/AgCl, peaks of adsorbed glucose species have been observed in cyclic voltammograms. These peaks at about -0.7 V (oxidation peak) and -0.8 V (reduction peak) have yielded reproducible current-glucose concentration relationships, linear through the region of clinically important glucose concentrations (50-300 mg/dl) and stable over time. Since these potentials are not in the redox range of many potentially interfering substances, selectivity is enhanced.

Biosensing Techniques↗

Orbital reconstruction with proplast.

Proplast was implanted in 15 cases of orbital reconstruction. Seven cases were orbital floor fractures, four cases were phthisical enophthalmos, and four cases were anophthalmic enophthalmos. In orbital floor fracture cases, thin (2-mm) Proplast was placed onto the fractured orbital floor after the incarcerated orbital tissue was released. To correct the enophthalmic socket, Proplast was shaped into small pieces (10 x 5 x 2 mm), then inserted into the subperiosteal space from all directions to fill up the orbital cavity. The results of these 15 cases were satisfactory. The average follow-up period is 13 months. The increased orbital volume seems less than the volume of the implanted Proplast; therefore, we suggest an 80-90% overcorrection rate in accordance with the computer measurements. In sum, Proplast serves as a very good material for orbital floor reconstruction and volume replacement.

Adolescent↗

Immobilized glucose oxidase in the potentiometric detection of glucose.

Previous work has shown that glucose oxidase can be immobilized on platinum to give an electrode that responds potentiometrically to glucose over the clinically useful range of about 10-250 mg glucose/100 mL. The present studies were carried out with electrochemically pretreated platinum and with gold or porous graphite substituted for the platinum support. The presence of the enzyme gave a significantly enhanced potentiometric response over that obtained with the bare support for both the pretreated platinum and the porous graphite, but not with gold. However, with platinum the potentiometric response became more negative with increasing glucose concentration. With porous graphite, the potential changed in the positive direction as the glucose concentration was increased. Hysteresis was demonstrated for the platinum-enzyme electrode. Mass transfer measurements with a rotating ring-disc electrode (RRDE) showed measurable diffusional resistances to the transport of a model electroactive compound (potassium ferrocyanide) through a matrix of immobilized enzyme attached to the disc of the RRDE. These results are part of a larger study to define the source of the potentiometric response by examining the roles of the support and the mass transfer resistances through the immobilized enzyme matrix.

Aspergillus niger↗

Immobilized enzyme electrodes for the potentiometric measurement of glucose concentration: immobilization techniques and materials.

Glucose oxidase, catalase, and bovine serum albumin were co-immobilized with glutaraldehyde around a platinum screen or around a single platinum-iridium wire. The potential difference between this dual enzyme electrode and a Ag/AgCL reference electrode was proportional to the logarithm of the glucose concentration over the range from 10 to about 150 mg glucose per 100 ml in buffered solution at pH 7.4 and 37 degrees C. The enzyme electrode responded in serum only if coated with a semipermeable film, such as cellulose acetate, to exclude serum macromolecules. The potentiometric results were similar to those obtained with the two enzymes co-immobilized in polyacrylamide gel around a platinum screen or with only one of the enzymes, glucose oxidase, covalently coupled to a platinum screen. The results so far suggest that glucose for development of a continuous in vivo glucose sensor.

Catalase↗

Hyaluronidase-bound membrane as a biomaterial for implantable fuel cells.

A new biomaterial containing covalently bound hyaluronidase was prepared. An application of this enzyme membrane is to improve the performance of an implantable fuel cell. Hyaluronic acid is a contributor to the viscosity of tissue fluids but can be a potential fuel source because of its sugar content. The incorporation of immobilized hyaluronidase would not only contribute to a more available fuel supply by splitting hyaluronic acid but, perhaps more importantly, enhance the rate of mass transport of fuel, O2, and reaction products by reducing the viscosity near the electrode membranes. Hyaluronidase was bound to Sepharose gel and its thermoplastic membrane after activation by cyanogen bromide. Fourteen and 22% of the activities were recovered from the gel and membrane, respectively. The activity of the bound enzyme was stable for six months at 0 degrees C. The addition of hyaluronic acid, 1 mg/ml, to a typical implantable type bioautofuel cell in vitro increased external solution viscosity from 1.1 to 2.5-2.8 cP and reduced voltage output under 10 komega by 60% in 3 hr. When the hyaluronidase bound membrane was placed at the anode, viscosity of the glucose-hyaluronic acid solution was lowered to 1.8 cP and the cell output increased to the original level of a glucose-fueled cell in 3 hr. Glucosamine-equivalent released from hyaluronic acid at the electrode was 3.1 mg after 22.5 hr. This represents 90% of the theoretical consumption. Restoration of the cell output was probably a combination of the enhanced transport of fuel, O2 and products, and/or appearance of a new fuel, glucosamine-equivalent.

Biocompatible Materials↗

A thin-film glucose electrode system with compensation for drift.

Thin-film platinum electrodes have been prepared for use in a system for measurement of glucose. Cyclic voltammetry was used to test the electrodes in phosphate and bicarbonate solutions, and in the dialysate of human serum. The peak current output for glucose concentrations between 50-300 mg/dl in a phosphate buffer, and 100-250 mg/dl in the dialysate of human serum, were linear. Two of the three inflection points occurring in the voltammograms are invariant with changes in scan rate, pH, CO2, O2, and glucose. A method for using these points to compensate for voltage drift and current shift is discussed.

Blood Glucose↗

A micro carbon electrode for nitric oxide monitoring.

A nitric oxide (NO) probe, consisting of a micro carbon fiber working electrode, 10 microns diameter, a platinum counter electrode, and a silver/silver chloride (Ag/AgCl) reference electrode, has been developed. The carbon fiber working electrode is covered with a Nafion cation exchange membrane. Using differential pulse voltammetry (DPV), we found the NO to N2O reduction current peak at approximately -1.35 V versus Ag/AgCl. This has been reported by others. The DPV current outputs are linearly related to dissolved NO concentrations [NO] in the 2-10 microM range. Catecholamines were found not to interfere with the reduction signal. The Nafion membrane also prevents interference by NO2-, NO3-, and amino acids at normal physiologic pH (pH 7.4). The effects of O2 are accounted for through sampling and subtracting background currents from the peak current. To increase sensitivity and shorten response time, a method of integrated pulse amperometry (IPA) was used for the study. The IPA charge outputs (delta C) are linear to the dissolved [NO] in the 50-350 nM range. The carbon fiber electrode has the potential of being miniaturized to a smaller electrode, allowing detection of NO released from the subendothelial space.

Carbon↗