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Biomedical subjects

Y B Yang

Publications and source records attributed to Y B Yang.

At least 19 recordsLinked to original sources
PubMed2002

Mathematical modelling of MSW incineration on a travelling bed.

The rising popularity of incineration of municipal solid waste (MSW) calls for detailed mathematical modelling and understanding of the incineration process. In this paper, governing equations for mass, momentum and heat transfer for both solid and gaseous phases in a moving bed in a solid-waste incineration furnace are described and relevant sub-models are presented. The burning rates of volatile hydrocarbons in the moving bed of solids are limited not only by the reaction kinetics but also the mixing of the volatile fuels with the under-fire air. The mixing rate is averaged across a computation cell and correlated to a number of parameters including local void fraction of the bed, gas velocity and a length scale comparable to the particle size in the bed. A correlation equation is also included to calculate the mixing in the freeboard area immediately next to the bed surface. A small-scale fixed bed waste incinerator was built and test runs were made in which total mass loss from the bed, temperature and gas composition at different locations along the bed height were measured. A 2-D bed-modelling program (FLIC) was developed which incorporates the various sub-process models and solves the governing equations for both gases and solids. Thermal and chemical processes are mainly confined within a layer about 5-9 times in thickness of the averaged particle size in the burning bed. For a large part of the burning process, the total mass loss rate was constant until the solid waste was totally dried out and a period of highly rising CO emission followed. The maximum bed temperature was around 1200 K. The whole burning process ended within 60 min. Big fluctuations in species concentration were observed due to channelling and subsequent 'catastrophic' changes in the local bed conditions. Reasonably good agreement between modelling and measurements has been achieved. Yet the modelling work is complicated by the channelling phenomenon in the bed. Numerical simulations without consideration of the channelling effect produced very good agreement with experiments concerning the total mass loss, but significant discrepancy exists for temperature and gas composition profiles. Transient phenomena such as the breaking of waste particles and the "catastrophic" creation of new burning channels occurring during waste incineration is a vital area requiring further investigation at the fundamental level. The underlying theory of bed behaviour must be extended to include these transient events.

Disaster Planning↗
PubMed2001

Functional signal peptides bind a soluble N-terminal fragment of SecA and inhibit its ATPase activity.

The selective recognition of pre-secretory proteins by SecA is essential to the process of protein export from Escherichia coli, yet very little is known about the requirements for recognition and the mode of binding of precursors to SecA. The major reason for this is the lack of a soluble system suitable for biophysical study of the SecA-precursor complex. Complicating the development of such a system is the likelihood that SecA interacts with the precursor in a high affinity, productive manner only when it is activated by binding to membrane and SecYEG. A critical aspect of the precursor/SecA interaction is that it is regulated by various SecA ligands (nucleotide, lipid, SecYEG) to facilitate the release of the precursor, most likely in a stepwise fashion, for translocation. Several recent reports show that functions of SecA can be studied using separated domains. Using this approach, we have isolated a proteolytically generated N-terminal fragment of SecA, which is stably folded, has high ATPase activity, and represents an activated version of SecA. We report here that this fragment, termed SecA64, binds signal peptides with significantly higher affinity than does SecA. Moreover, the ATPase activity of SecA64 is inhibited by signal peptides to an extent that correlates with the ability of these signal peptides to inhibit either SecA translocation ATPase or in vitro protein translocation, arguing that the interaction with SecA64 is functionally significant. Thus, SecA64 offers a soluble, well defined system to study the mode of recognition of signal peptides by SecA and the regulation of signal peptide release.

Adenosine Triphosphatases↗
PubMed2000

[Inhibition of transmembrane K(+) currents in mammalian ventricular myocytes by antiarrhythmic agent RP62719].

The effect of RP62719 on the inward rectifier K(+) current (I(K1)),transient outward K(+) current (I(to)) and delayed outward K(+) current (I(K)) in isolated cardiac myocytes was determined using the whole cell patch clamp technique in guinea pig and dog. RP62719 decreased I(K1) with an inhibitory concentration 50 (IC(50) ) of 5.0+/-1.0 micromol/L at -100 mV in guinea pig ventricular cells. In dog ventricular myocytes, RP62719 inhibited Ito by 84+/-4.4% with an IC(50) of 1.2+/-0.51 micromol/L at +40 mV. In guinea pig ventricular cells, RP62719 decreased I(K): I(Kstep) by 50.0+/-8.3%%and I(Ktail) by 56.0+/-4.9% at +40 mV, respectively. RP62719 inhibited I(Kstep) with an IC(50) of 4.2+/-0.8 micromol/L and I(Ktail) with an IC(50) of 3.3+/-0.75 micromol/L. Thus it is suggested that the ionic mechanism of antiarrhymic effect by RP62719 may be due to its inhibition of I(K1),I(to) and I(K).

Animals↗
PubMed1997

SecE-depleted membranes of Escherichia coli are active. SecE is not obligatorily required for the in vitro translocation of certain protein precursors.

Membrane vesicles were prepared from Escherichia coli cells in which SecE was depleted to 2% of wild-type membranes. SecE depletion had pleiotropic effects; SecD, SecF, SecG, and SecY were decreased 4-6-fold, whereas SecA was increased about 16-fold over that of wild-type membranes. These membranes were substantially active in the in vitro translocation of proOmpA, which was mediated by the SecA pathway since it was inhibited by azide. Similar substantial translocation activities were observed for proLamB and proLpp in the SecE-depleted membranes. However, the translocation of proPhoA was more severely impaired. These data indicate that SecE may enhance but is not obligatorily required for the translocation of at least certain precursors, and suggest that the effects of the SecE depletion on protein translocation may be precursor-dependent.

Adenosine Triphosphatases↗
PubMed1997

Differential translocation of protein precursors across SecY-deficient membranes of Escherichia coli: SecY is not obligatorily required for translocation of certain secretory proteins in vitro.

SecY, a component of the protein translocation system in Escherichia coli, was depleted at a nonpermissive temperature in a strain which had a temperature-sensitive polar effect on the expression of its secY. Membrane vesicles prepared from these cells, when grown at the nonpermissive temperature, contained about 5% SecY and similarly low levels of SecG. As expected, translocation of alkaline phosphatase precursors across these SecY-deficient membranes was severely impaired and appeared to be directly related to the decrease of SecY amounts. However, despite such a dramatic reduction in SecY and SecG levels, these membranes exhibited 50 to 70% of the wild-type translocation activity, including the processing of the signal peptide, of OmpA precursor (proOmpA). This translocation activity in SecY-deficient membranes was still SecA and ATP dependent and was not unique to proOmpA, as lipoprotein and lambda receptor protein precursors were also transported efficiently. Membranes that were reconstituted from these SecY-depleted membranes contained undetectable amounts of SecY yet were also shown to possess substantial translocation activity for proOmpA. These results indicate that the requirement of SecY for translocation is not obligatory for all secretory proteins and may depend on the nature of precursors. Consequently, it is unlikely that SecY is the essential core channel through which all precursors traverse across membranes; rather, SecY probably contributes to efficiency and specificity.

Alkaline Phosphatase↗
PubMed1996

Influence of column type and chromatographic conditions on the ion-exchange chromatography of immunoglobulins.

Immunoglobulins are often purified by affinity chromatography. However, this technique is costly, can result in poor resolution for subclasses (or is only group specific), and leads to possible leaching of contaminants into the purified products. Ion-exchange chromatography has shown great potential and has found an increased usage in the purification of immunoglobulins. The aim of this study is to further understand the separation mechanism with emphasis on the influence of column type and chromatographic conditions on the peak shape, selectivity and changes in the elution patterns. Included are strong cation-exchange, strong anion-exchange and weak anion-exchange columns. Five immunoglobulin G antibodies were used as test probes. Some sera and ascites were also used in the study. Among the chromatographic conditions examined were mobile phase pH, buffer type, buffer concentration, gradient rate, and column temperature. Significant differences in the chromatographic behavior (elution pattern, peak shape and selectivity) of the test samples are discussed in regard to the column type and the chromatographic conditions.

Animals↗
PubMed1994

Purification and characterization of L-aminoacylase from Alcaligenes denitrificans DA181.

The L-aminoacylase produced intracellularly by Alcaligenes denitrificans DA181 was purified to homogeneity. This enzyme had an apparent molecular weight of 80,000, and was composed of two subunits of identical molecular weight. Its isoelectric point was pH 5.1. The optimal reaction temperature and pH were 65 degrees C and 8.0, respectively. This enzyme showed specificity toward N-acetyl-derivative of hydrophobic L-amino acids with N-acetyl-L-valine as the favored substrate, followed by N-acetyl-L-alanine.

Alcaligenes↗
PubMed1993

Fimbriated stationary phases for proteins.

This paper describes synthetic procedures for preparing fimbriated stationary phases on poly(styrene-divinylbenzene) (PS-DVB) packing materials. The synthesis consists of a five-step procedure in which the order in which the steps are carried out may be varied. These steps are (i) polymerization of monomers to form an amphiphilic copolymer or oligomer, (ii) adsorption of either monomers, or polymer onto the PS-DVB surface, (iii) solvent induction of functional group orientation at the PS-DVB-polymer interface and polymer-water interface, (iv) a cross-linking reaction that forms a hydrophilic surface layer, and (v) derivatization of the surface layer with stationary phase.

Chromatography, Affinity↗
PubMed1993

Heat-shock inhibits protein synthesis and eIF-2 activity in cultured cortical neurons.

Stress, such as heat-shock, hypoxia and hypoglycemia, inhibits the initiation of protein synthesis. The effects of heat-shock on protein synthesis, eucaryotic initiation factor 2 (eIF-2) activity, protein kinase C (PKC), and casein kinase II (CKII) activities were studied in primary cortical neuronal cultures. In neurons exposed to heat-shock at 44 degrees C for 20 min, protein synthesis is inhibited by more than 80%, and is accompanied by a 60% decrease in eIF-2 activity. Steady state PKC and CK II activities were not affected by heat-shock. Vanadate (200 microM), a protein phosphotyrosine phosphatase inhibitor, partially prevented the depression of eIF-2 activity during heat-shock, and increased CKII activity by 90%. In contrast, staurosporine (62nM), a protein kinase C inhibitor, did not affect eIF-2 activity. We conclude that heat-shock causes a change in the phosphorylation/dephosphorylation of regulatory proteins leading to a depressed eIF-2 activity and protein synthesis in neurons.

Alkaloids↗
PubMed1993

Depression of neuronal protein synthesis initiation by protein tyrosine kinase inhibitors.

Growth factors stimulate cellular protein synthesis, but the intracellular signaling mechanisms that regulate initiation of mRNA translation in neurons have not been clarified. A rate-limiting step in the initiation of protein synthesis is the formation of the ternary complex among GTP, eukaryotic initiation factor 2 (eIF-2), and the initiator tRNA. Here we report that genistein, a specific tyrosine kinase inhibitor, decreases tyrosine kinase activity and the content of phosphotyrosine proteins in cultured primary cortical neurons. Genistein inhibits protein synthesis by > 80% in a dose-dependent manner (10-80 micrograms/ml) and concurrently decreases ternary complex formation by 60%. At the doses investigated, genistein depresses tyrosine kinase activity and concomitantly stimulates PKC activity. We propose that a protein tyrosine kinase participates in the initiation of protein synthesis in neurons, by affecting the activity of eIF-2 directly or through a protein kinase cascade.

Analysis of Variance↗
PubMed1993

Effects of acute hypoxia on intracranial dynamics in unanesthetized goats.

The effects of acute hypoxia on intracranial dynamics were studied in six unanesthetized goats that were subjected to hypobaric hypoxia at a simulated 4,000-m high altitude for 2 h. Another six nonhypoxic goats served as control group for cerebral water content (CWC). In this study, we measured cerebral blood flow (CBF) by electromagnetic flowmeter, intracranial pressure (ICP) through an intraventricular catheter, and CWC by a wet-to-dry weight technique for a given volume of brain tissue. Intracranial compliance was calculated from a pressure-volume index derived from the change in intraventricular pressure after a subarachnoid injection of saline. During acute hypoxia CBF increased from 146.0 +/- 10.60 to 185.3 +/- 7.83 ml x min-1.100 g-1, ICP increased from 15.4 +/- 1.82 to 27.4 +/- 3.84 cmH2O, and CWC increased from 78.7 +/- 0.30 to 79.5 +/- 0.16%, but intracranial compliance decreased from 0.65 +/- 0.28 to 0.36 +/- 0.19 ml/cmH2O. The data suggest that acute hypoxia results in a significant disturbance of intracranial dynamics, which may be one of the important pathophysiological mechanisms of high-altitude cerebral edema and acute mountain sickness.

Acute Disease↗
PubMed1992

Factors affecting the separation and loading capacity of proteins in preparative gradient elution high-performance liquid chromatography.

The optimum conditions for the purification of proteins by gradient elution in reversed-phase liquid chromatography were studied, with emphasis on the column length. Because of the strong dependence of the retention of proteins on the mobile phase composition, very short columns can be used successfully to perform analytical separations. A similar conclusion is extended to preparative separations. Columns with different lengths and diameters were used. The dependence of the loading capacity for touching band separation on the column length, diameter and volume was studied, in addition to the regeneration time between successive runs, the starting mobile phase composition and the necessary column efficiency.

Animals↗
PubMed1992

Characterization of D-aminoacylase from Alcaligenes denitrificans DA181.

The D-aminoacylase produced by Alcaligenes denitrificans DA181 was a new type of aminoacylase which had both high stereospecificity and specific activity. The molecular weight and isoelectric point of this enzyme were 58,000 and 4.4, respectively. The apparent Km and kcat values of this enzyme for N-acetyl-D-methionine were estimated to be 0.48 mM and 6.24 x 10(4) min-1, respectively. The optimum temperature was 45 degrees C. The enzyme was stable up to 55 degrees C for 1 hr in the presence of 0.2 mg/ml bovine serum albumin. The enzyme was stable in the pH range of 6.0 to 11.0 with an optimum pH of 7.5. This enzyme contained about 2.1 g atom of zinc per mole of enzyme. Enzyme activity was inhibited by incubation with EDTA. The inhibition by EDTA was fully reversed by Co2+ and partially by Zn2+.

Alcaligenes↗
PubMed1991

Coated hydrophilic polystyrene-based packing materials.

A very hydrophilic high-performance liquid chromatographic base support was created from microparticulate, macroporous poly(styrene-divinylbenzene) beads. An organic monomer containing cross-linking functionalities was coated on the poly(styrene-divinylbenzene), followed by a catalyzed cross-linking reaction. The coatings formed contain only stable chemical bonds (e.g., C-C, C-O-C), and easily-derivatized hydroxyl moieties. This coated base support was evaluated for hydrophilicity, chemical stability, solvent compatibility, rigidity, and irreversible adsorption. Derivatives of the coated base support were made and applied in various modes of chromatography.

Cations↗
PubMed1991

Purification and Characterization of d-Aminoacylase from Alcaligenes faecalis DA1.

A d-aminoacylase from Alcaligenes faecalis DA1 has been purified to homogeneity by a simple purification procedure with two columns, Fractogel DEAE-650 and HW-50. The specific activity of the purified enzyme was found to be 580 U/mg of protein with N-acetyl-dl-methionine as the reaction substrate. The apparent molecular weight and isoelectric point of this enzyme were determined to be 55,000 and 5.4, respectively.

Journal Article↗
PubMed1990

Flow-through particles for the high-performance liquid chromatographic separation of biomolecules: perfusion chromatography.

This paper reports a new technique for reducing resistance to stagnant mobile phase mass transfer without sacrificing high adsorbent capacity or necessitating extremely high pressure operation. The technique involves the flow of liquid through a porous chromatographic particle, and has thus been termed "perfusion chromatography". This is accomplished with 6000-8000 A pores which transect the particle. Data from electron microscopy, column efficiency, frontal analysis and theoretical modelling all suggest that mobile phase will flow through these large pores. In this manner, solutes enter the interior of the particles through a combination of convective and diffusional transport, with convection dominating for Peclet numbers greater than one. The implications of flow through particles on bandspreading, resolution and dynamic loading capacity are examined. It is shown that the rate of solute transport is strongly coupled to mobile phase velocity such that bandspreading, resolution of proteins and dynamic loading capacity are unaffected by increases in mobile phase velocity up to several thousand centimeters per hour. The surface area of this very large-pore diameter material is enhanced by using a network of smaller, 500-1500 A interconnecting pores between the throughpores. Scanning electron micrographs show that the pore network is continuous and that no point in the matrix is more than 5000-10,000 A from a through-pore. As a consequence, diffusional path lengths are minimized and the large porous particles take on the transport characteristics of much smaller particles but with a fraction of the pressure drop. Capacity and resolution studies show that these materials bind and separate an amount of protein equivalent to that of conventional high-performance liquid chromatography as well as low performance agarose-based media at greater than 10-100 times higher mobile phase velocity with no loss in resolution.

Chromatography, High Pressure Liquid↗
PubMed1990

[Detection of natural killer cells activity in patients with laryngeal carcinoma using the 51Cr release assay].

The NK activity has been examined in a 4-hour 51Cr release assay in 59 untreated patients with laryngeal carcinoma, 46 untreated patients with nasopharyngeal carcinoma and 18 untreated patients with other head and neck carcinomas. The kinetic changes of NK activity in 10 patients with laryngeal carcinoma before and after surgery was also observed. The results showed that the NK activity of patients was significantly lower than that of the normal control and decreased progressively in the advanced cases. A significant decrease in NK activity in laryngeal carcinoma patients was found 3-5 days after surgery, and an increase in activity was seen more than one week later. These results suggest that the test of NK activity in untreated laryngeal carcinoma patients might be a parameter to predict the course and prognosis of the disease.

Adult↗