Biomedical subjects

Liang Guo

Publications and source records attributed to Liang Guo.

24 records · Page 2Linked to original sources

PubMed2004

Rapid isolation of high-quality total RNA from taxus and ginkgo.

An easy and efficient protocol was developed for isolating good-quality total RNA from various tissues including fruits, leaves, stems, and roots of ancient gymnosperm species, taxus and ginkgo. The protocol was developed based on the CTAB method with modifications, including higher-strength CTAB to help the lysis of plant cells, more PVP, and beta-mercaptoethanol to prevent oxidation of phenolic complexes, and higher-centrifugation force to get rid of most cell debris and to ensure RNA quality. In RNA isolation, chloroform/isoamyl alcohol was used to remove proteins, genomic DNA, and secondary metabolites and lithium chloride was subsequently adopted to concentrate total RNA away from most of the cytoplasmic components. Good-quality total RNA from various tissues of native taxus and ginkgo could be easily isolated within 24 hr by this protocol which avoided the limitation of plant materials and the usage of dangerous chemicals, such as phenol, and could provide total RNA for all kinds of further molecular studies.

Ginkgo biloba↗

PubMed2003

Effects of cytochrome c on the mitochondrial apoptosis-induced channel MAC.

Recent studies indicate that cytochrome c is released early in apoptosis without loss of integrity of the mitochondrial outer membrane in some cell types. The high-conductance mitochondrial apoptosis-induced channel (MAC) forms in the outer membrane early in apoptosis of FL5.12 cells. Physiological (micromolar) levels of cytochrome c alter MAC activity, and these effects are referred to as types 1 and 2. Type 1 effects are consistent with a partitioning of cytochrome c into the pore of MAC and include a modest decrease in conductance that is dose and voltage dependent, reversible, and has an increase in noise. Type 2 effects may correspond to "plugging" of the pore or destabilization of the open state. Type 2 effects are a dose-dependent, voltage-independent, and irreversible decrease in conductance. MAC is a heterogeneous channel with variable conductance. Cytochrome c affects MAC in a pore size-dependent manner, with maximal effects of cytochrome c on MAC with conductance of 1.9-5.4 nS. The effects of cytochrome c, RNase A, and high salt on MAC indicate that size, rather than charge, is crucial. The effects of dextran molecules of various sizes indicate that the pore diameter of MAC is slightly larger than that of 17-kDa dextran, which should be sufficient to allow the passage of 12-kDa cytochrome c. These findings are consistent with the notion that MAC is the pore through which cytochrome c is released from mitochondria during apoptosis.

Animals↗

PubMed2002

Dynamic coating ion-exchange chromatography of cations on an octadecyl-bonded silica stationary phase.

It was found that common cations (Na+, NH4+, K+, Mg2+ and Ca2+) could be strongly retained on an ODS stationary phase when aqueous solutions of carboxylic acids were used as eluents. The chromatographic conditions used in this work were the same as in common cation-exchange chromatography on a cation-exchange resin and the retention behavior of the above-mentioned cations on the ODS column was quite similar to that on a cation-exchange column. The retention behavior and mechanism have been investigated using a number of carboxylic acids as eluents. The retention mechanism of the cations in these experiments was considered to be a dynamic coating ion-exchange mechanism. The carboxylic acids in the mobile phase were coated onto the surface of the ODS stationary phase and formed a dynamic carboxylic acid functional layer which could act like the functional group layer of a carboxylic group cation exchanger.

Cation Exchange Resins↗

PubMed2002

Effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets.

Two experiments were conducted to investigate the effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets. In Experiment I 72 crossbred piglets were allotted to three treatments, i.e. 1) control (basal) diet, 2) alpha-Gal1 and 3) alpha-Gal5, the basal diet supplemented with 0.01% and 0.05% alpha-galactosidase preparation (alpha-Gal), respectively. Average daily gain, average daily feed intake and feed conversion ratio were not influenced by alpha-Gal supplementation. In Experiment II 72 crossbred weanling piglets were randomly assigned to four treatments with a complete factorial design, i.e. 1) basal diet, 2) basal diet with 0.01% alpha-Gal supplementation, 3) basal diet with 1% stachyose supplementation and 4) basal diet with 1% stachyose and 0.01% alpha-Gal supplementation. Average daily gain and feed conversion ratio were influenced by alpha-Gal and stachyose supplementation. The ileal digestibility of stachyose, raffinose, gross energy and crude protein was improved significantly by alpha-Gal supplementation. The microbial populations in the intestine were modified by both alpha-Gal and stachyose supplementation. The data suggested that stachyose supplementation had an adverse effect on the growth performance of piglets and alpha-Gal supplementation could ameliorate it, especially in younger pigs.

Age Factors↗

PubMedDate not supplied

Automated electrophysiology in the preclinical evaluation of drugs for potential QT prolongation.

INTRODUCTION: The human ether-a-go-go-related gene (hERG) potassium channel plays a major role in the electrical conductances involved in human heart repolarization. Drugs that decrease hERG K(+) currents are at risk to produce a prolongation of the cardiac action potential, resulting in an increase of the QT interval. Drug-induced QT prolongation or acquired long QT (aLQT) can lead to a fatal arrhythmia known as Torsade de Pointes (TdP). Electrophysiological methods are the best approach to evaluate potential drug candidates for hERG current inhibition. Here we identify limitations with the PatchXpress 7000A automated electrophysiology instrument and describe hERG protocol optimizations necessary for reliable preclinical assessment. METHODS: The PatchXpress 7000A automated electrophysiology system was used to evaluate a group of drugs with known hERG activity under voltage clamp conditions. We used a recombinant cell line expressing hERG, and assessed the inhibition of hERG K(+) currents at different drug concentrations. These data were used to determine hERG IC(50) values and compare assay parameters under different recording conditions. RESULTS: We found that due to limitations with the PatchXpress 7000A instrument, repeated compound additions were critical for achieving steady state drug concentrations that generated data comparable to standard patch clamp methods, particularly when similar voltage pulse protocols were implemented. Some discrepancies were observed between the PatchXpress 7000A and standard patch clamp techniques including shifts in IC(50) values for very hydrophobic compounds. Most hERG IC(50) values were within 3-fold of standard patch clamp IC(50) values. DISCUSSION: Automation of electrophysiology technologies has greatly improved the throughput of assessing lead drug candidates for hERG liability. To maintain hERG data quality comparable to standard patch clamp techniques, the PatchXpress 7000A instrument limitations should be recognized and protocols optimized accordingly to ensure accuracy.

Animals↗

PubMedDate not supplied

[An intron-free methyl jasmonate inducible geranylgeranyl diphosphate synthase gene from Taxus media and its functional identification in yeast].

Geranylgeranyl diphosphate synthase (GGPPS, EC: 2.5.1.29) catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for diterpenes including Taxol, one of the most potent antitumor drugs. In order to investigate the role of GGPP synthase in taxol biosynthesis, we cloned, characterized and functionally expressed the GGPP synthase gene from Taxus media. A 3743-bp genomic sequence of T. media was isolated by genome walking strategy which contained an 1182-bp open reading frame (ORF) encoding a 393-amino acid polypeptide that showed high similarity to other plant GGPPSs. Subsequently the full-length cDNA of the GGPPS gene of T. media (designated TmGGPPS) was amplified by RACE. Bioinformatic analysis showed that TmGGPPS was an intron-free gene and its deduced polypeptide contained all the five conserved domains and functional aspartate-rich motifs of the prenyltransferases. By constructing the phylogenetic tree of plant GGPPSs, it was found that plant-derived GGPPSs could be divided into two classes, angiosperm and gymnosperm classes, which might have evolved in parallel from the same ancestor. To our knowledge this was the first report that the geranylgeranyl diphosphate synthase genes were free of intron and evolved in parallel between angiosperms and gymnosperms. The coding sequence of TmGGPPS was expressed in yeast mutant (SFNY368) lacking of GGPP synthase activity through functional complementation, and the transgenic yeast showed to have activity of GGPP synthase. This was also the first time to use SFNY368 to identify the function of plant-derived GGPPSs. Furthermore, investigation of the impact of methyl jasmonate (MeJA) on the expression of TmGGPPS revealed that MeJA-treated T. media cultured cells had much higher expression of TmGGPPS than untreated cells.

Acetates↗