Biomedical subjects

H Liu

Publications and source records attributed to H Liu.

At least 991 records · Page 55Linked to original sources

PubMed1995

Rapid endocytosis of a G protein-coupled receptor: substance P evoked internalization of its receptor in the rat striatum in vivo.

Studies on cultured cells have shown that agonists induce several types of G protein-coupled receptors to undergo internalization. We have investigated this phenomenon in rat striatum, using substance P (SP)-induced internalization of the SP receptor (SPR) as our model system. Within 1 min of a unilateral striatal injection of SP in the anesthetized rat, nearly 60% of the SPR-immunoreactive neurons within the injection zone display massive internalization of the SPR--i.e., 20-200 SPR+ endosomes per cell body. Within the dendrites the SPR undergoes a striking translocation from the plasma membrane to endosomes, and these dendrites also undergo a morphological reorganization, changing from a structure of rather uniform diameter to one characterized by large, swollen varicosities connected by thin fibers. In both cell bodies and dendrites the number of SPR+ endosomes returns to baseline within 60 min of SP injection. The number of neurons displaying substantial endosomal SPR internalization is dependent on the concentration of injected SP, and the SP-induced SPR internalization is inhibited by the nonpeptide neurokinin 1 receptor antagonist RP-67,580. These data demonstrate that in the central nervous system in vivo, SP induces a rapid and widespread SPR internalization in the cell bodies and dendrites and a structural reorganization of the dendrites. These results suggest that many of the observations that have been made on the internalization and recycling of G protein-coupled receptors in in vitro transfected cell systems are applicable to similar events that occur in the mammalian central nervous system in vivo.

Analysis of Variance↗

PubMed1995

Increased density of cortical apolipoprotein E immunoreactive neurons in rabbit brain after dietary administration of cholesterol.

Rabbits were fed 2% cholesterol diet for 4, 6 and 8 weeks or control diet for 6 weeks. Frontal cortex and hippocampal formation were stained with apolipoprotein E (Apo-E) antibody using standard immunocytochemical methods. The number of neurons expressing the Apo-E epitope and the intensity of Apo-E immunoreactivity increased with increasing time on the cholesterol diet. Because Apo-E chaperones cholesterol in the brain, the data may suggest that elevated circulating levels of cholesterol eventually cause increased cerebral levels.

Animals↗

PubMed1995

Okadaic acid stimulates glucose transport in rat adipocytes by increasing the externalization rate constant of GLUT4 recycling.

GLUT4, the major insulin-responsive glucose transporter isoform in rat adipocytes, rapidly recycles between the cell surface and an intracellular pool with two first order rate constants, one for internalization (kin) and the other for externalization (kex). Insulin decreases kin by 2.8-fold and increases kex by 3.3-fold, thus increasing the steady-state cell surface GLUT4 level by approximately 8-fold (Jhun, B. H., Rampal, A. L., Liu, H., Lachaal, M., and Jung, C. (1992) J. Biol. Chem. 267, 17710-17715). To gain an insight into the biochemical mechanisms that modulate these rate constants, we studied the effects upon them of okadaic acid (OKA), a phosphatase inhibitor that exerts a insulin-like effect on glucose transport in adipocytes. OKA stimulated 3-O-methylglucose transport maximally 3.1-fold and increased the cell surface GLUT4 level 3.4-fold. When adipocytes were pulse-labeled with an impermeant, covalently reactive glucose analog, [3H]1,3-bis-(3-deoxy-D-glucopyranose-3-yloxy)-2-propyl 4-benzoylbenzoate, and the time course of labeled GLUT4 recycling was followed, the kex was found to increase 2.8-fold upon maximal stimulation by OKA, whereas the kin remained unchanged within experimental error. These findings demonstrate that OKA mimics the insulin effect on only GLUT4 externalization and suggest that insulin stimulates GLUT4 externalization by increasing the phosphorylation state of a serine/threonine phosphoprotein, probably by inhibiting protein phosphatase 1 or 2A.

Adipocytes↗

PubMed1995

IAM chromatography: an in vitro screen for predicting drug membrane permeability.

Fluid cell membranes are the main barrier to drug absorption when diffusion limits uptake. Immobilized artificial membranes (IAMs) are solid phase models of fluid membranes that predicted oral drug absorption in mice for a homologous set of cephalosporins. IAMs also predicted drug permeability through Caco-2 cells. Since drug permeability in Caco-2 cells is known to correlate with the oral absorption of drugs in humans, IAMs may also model drug absorption in humans. IAM analysis is experimentally simple, and large-volume screening of experimental compounds for drug absorption is possible.

Animals↗

PubMed1995

Membrane partition coefficients chromatographically measured using immobilized artificial membrane surfaces.

Immobilized artificial membranes (IAMs) are chromatographic surfaces prepared by covalently immobilizing cell membrane phospholipids. IAM surfaces mimic fluid cell membranes. Solute capacity factors (k'IAM) measured on IAM columns correlate very well with solute equilibrium partition coefficients (Km') measured in fluid liposome systems. For 23 structurally unrelated compounds, log-(k'IAM) correlates with log(Km') with a linear correlation coefficient r = 0.907. This indicates that solute partitioning between the IAM bonded phase and the aqueous mobile phase is similar to the solute partitioning between liposomes and the aqueous phase. Although both IAM chromatography and liposome partitioning can be used as in vitro methods to predict solute partitioning into cell membranes, IAM chromatography is experimentally convenient compared to liposome systems. To study the effect of lipid structure on drug binding to IAMs, IAMs were prepared from three different phosphatidylcholine ligands: (i) a diacylated phosphatidylcholine ligand, (ii) a single chain ether phosphatidylcholine ligand, and (iii) a single chain phosphatidylcholine ligand that lacks a glycerol backbone. Solute retention data were identical for all of these IAMs, and consequently, predictions of solute binding to fluid membranes were also identical. This indicates that the structure of the phosphatidylcholine ligand that is immobilized is not critical for the binding of solutes. Since the structure is not important, the binding of solutes to membranes is a bulk phase property, i.e., it is the interface created by the ligands that determines the solute binding properties, not the ligands themselves. Solute partitioning using octanol/water systems does not correlate with k'IAM unless a homologous series of hydrophobic solutes is being evaluated.

Binding Sites↗

PubMed1995

Molecular recognition of tRNA(Pro) by Escherichia coli proline tRNA synthetase in vitro.

In this study, we identify a subset of nucleotides that specify aminoacylation of tRNA(Pro) by Escherichia coli proline tRNA synthetase in vitro. Twenty-two tRNA(Pro) variants were prepared by in vitro transcription and their efficiency of aminoacylation with proline (kcat/KM) was measured. From this analysis, we conclude that recognition elements for tRNA(Pro) aminoacylation by ProRS are located in at least three domains of the tRNA molecule. The largest decreases in the kinetic parameters for aminoacylation resulted from single substitutions at position G72 of the acceptor stem and position G36 of the anticodon. Anticodon nucleotide G35 and position A73 in the acceptor stem were also identified as major recognition elements. Moreover, bases that are believed to be important for maintaining the tertiary structure of the tRNA (G15 and C48) appear to be important for efficient recognition of tRNA(Pro) by ProRS in vitro.

Amino Acyl-tRNA Synthetases↗

PubMed1995

Allotransplantation of whole spleen in patients with hepatic malignant tumors or hemophilia A. Operative technique and preliminary results.

BACKGROUND: It has been reported that the spleen performs a vital function in the fight against malignant tumors. The spleen is the primary producer of tuftsin, which can directly or indirectly kill tumor cells or inhibit their growth. The spleen is also believed to produce coagulating factor VIII. Therefore, allotransplantation of the spleen can be used in the treatment of patients with malignant tumors and hemophilia A. DESIGN: Heterotopic allotransplantation of whole spleen was performed in six patients with advanced hepatocellular carcinoma and hemophilia A. An adjuvant immunotherapy with interferon alfa was simultaneously administered in the patients with liver cancer. SETTING: Tongji Hospital, Tongji Medical University, Wuhan, China. RESULTS: Among six cases of allografting of whole spleen, five grafts were successful; one failed because of torsion of the splenic hilum. Three patients with hepatocellular carcinoma survived 9, 11, and 5 months after transplantation. Marked shrinkage of hepatic tumors and reduced serum alpha-fetoprotein levels were observed in these patients. On 5-year follow-up, three patients with hemophilia who had undergone splenic allografts were alive, and two had experienced substantial clinical improvement. In these two patients, when the grafts were functioning well and the recipients were free of acute rejection or graft-vs-host reaction, the mean plasma factor VIII activity remained between 30% and 36%, with peak factor VIII activities of 53.7% and 66.6%. We also evaluate operative technique and posttransplantation complications. CONCLUSION: Our results strongly imply that the spleen is one of the primary sites of synthesis of factor VIII and that the spleen has an inherent ability to fight malignant diseases. Allografting of whole spleen may be a promising technique for the treatment of patients with unresectable hepatocellular carcinoma and severe hemophilia A.

Adult↗

PubMed1995

Guinea pig and human cytomegaloviruses do not share cross-reactive neutralizing epitopes.

Human (HCMV) and guinea pig (gpCMV) cytomegaloviruses share biological similarities and DNA sequence homologies. Therefore, human and guinea pig sera were tested for cross-reactive antibodies. In eight human sere (four HCMV-seropositive and four HCMV-seronegative), low levels of neutralizing activity against gpCMV were not associated with antibodies to HCMV. The convalescent sera of one of three humans infected with either HCMV Towne vaccine or wild-type HCMV developed a fourfold increase in neutralizing titers to gpCMV after infection, but none of seven guinea pigs immunized with either HCMV Towne or purified HCMV gB developed neutralizing activity against gpCMV. Guinea pigs immunized with gpCMV did not develop antibodies to HCMV or human gB. Neither gpCMV, HCMV Towne stain or purified HCMV gB induced cross-reactive antibodies against the heterologous virus as detected by enzyme immunoassay. Our results indicate that gpCMV and HCMV share a very limited number, if any, of cross-reactive neutralizing epitopes.

Adult↗

PubMed1995

An in vivo 31P magnetic resonance spectroscopy study of uridine excess in rats fed orotic acid.

Spatially localized 31P NMR spectroscopy was used to assay in vivo the liver of intact rats fed orotic acid (OA) in a diet which produces hepatic steatosis. Twenty-three sets of multiple volume spectra were obtained from twenty-one 265- to 315-g female rats after 0-9 days of feeding either a 1% OA/64% sucrose diet (12 rats) or a 65% sucrose control diet (9 rats). The intensity of the in vivo diphosphodiester resonance ascribed to UDP-hexos(amin)es increased and the phosphomonoester resonance decreased in intensity prior to fatty infiltration. High resolution NMR spectroscopy of extracts of these livers indicated that the UDP-hexos(amin)e peak included four different UDP-sugars including UDP-N-acetylglucosamine (UDP-glcNAc), and that lower phosphocholine (P-Cho) accounted for the lower phosphomonoester resonance in vivo. Increased UDP-glcNAc is thought to reflect impaired lipoprotein glycosylation as a mechanism for hepatic steatosis in orotic acid feeding. P-Cho deficiency has been shown to be due to an increased rate of phosphatidylcholine synthesis. Low P-Cho concentration has been shown to be associated with lipid accumulation in a choline-deficient diet, but was not previously associated with hepatic steatosis in OA feeding. Changes in phosphorus metabolites were observed 2 days prior to development of fatty liver. HPLC assay of uridine nucleotides showed a good correlation between magnetic resonance spectroscopy and HPLC quantitation. In this study there were two biochemical correlates of impaired hepatic lipid secretion detectable by in vivo assay with 31P NMR spectroscopy. This method has application for noninvasive assays in ornithine transcarbamylase-deficient patients.

Animals↗

PubMed1995

Deriving accurate interproton distances from ROESY spectra with limited knowledge of scalar coupling constants via the CARNIVAL algorithm. An iterative complete-relaxation-matrix approach.

A method (termed CARNIVAL) for accurately determining distances from proton homonuclear rotating-frame Overhauser effect spectroscopy (ROESY) is described. The method entails an iterative calculation of the relaxation matrix using methodology introduced with the MARDIGRAS algorithm for analysis of two-dimensional nuclear Overhauser effect spectra (B. A. Borgias and T. L. James, J. Magn. Reson. 87, 475, 1990). The situation is complicated in the case of ROESY as spectral peak intensities are influenced by resonance offset and contributions from homonuclear Hartmann-Hahn (HOHAHA) transfer if the nuclear spins are related by scalar coupling. The effects of spin-locking field strength on distance determinations and the ensuing distance errors incurred when HOHAHA corrections are made with limited knowledge of scalar (J) coupling information have been evaluated using simulated ROESY intensities with a model peptide structure. It has been demonstrated that accurate distances can be obtained with little or no explicit knowledge of the homonuclear coupling constants over a moderate range of spin-locking field strengths. The CARNIVAL algorithm has been utilized to determine distances in a decapeptide using experimental ROESY data without measured coupling constants.

Algorithms↗

PubMed1995

Interproton distance bounds from 2D NOE intensities: effect of experimental noise and peak integration errors.

The effect of experimental and integration errors on the calculation of interproton distances from NOE intensities is examined. It is shown that NOE intensity errors can have a large impact on the distances determined. When multiple spin ('spin diffusion') effects are significant, the calculated distances are often underestimated, even when using a complete relaxation matrix analysis. In this case, the bias of distances to smaller values is due to the random errors in the NOE intensities. We show here that accurate upper and lower bounds of the distances can be obtained if the intensity errors are properly accounted for in the complete relaxation matrix calculations, specifically the MARDIGRAS algorithm. The basic MARDIGRAS algorithm has been previously described [Borgias, B.A. and James, T.L. (1990) J. Magn. Reson., 87, 475-487]. It has been shown to provide reasonably good interproton distance bounds, but experimental errors can compromise the quality of the resulting restraints, especially for weak cross peaks. In a new approach introduced here, termed RANDMARDI (random error MARDIGRAS), errors due to random noise and integration errors are mimicked by the addition of random numbers from within a specified range to each input intensity. Interproton distances are then calculated for the modified intensity set using MARDIGRAS. The distribution of distances that define the upper and lower distance bounds is obtained by using N randomly modified intensity sets. RANDMARDI has been used in the solution structure determination of the interstrand cross-link (XL) formed between 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) and the DNA oligomer d(5'-GCGTACGC-3')2 [Spielmann, H.P. et al. (1995) Biochemistry, 34, 12937-12953]. RANDMARDI generates accurate distances bounds from the experimental NOESY cross-peak intensities for the fixed (known) interproton distances in XL. This provides an independent internal check for the ability of RANDMARDI to accurately fit the experimental data. The XL structure determined using RANDMARDI-generated restraints is in good agreement with other biophysical data that indicate that there is no bend introduced into the DNA by the cross-link. In contrast, isolated spin-pair approximation calculations give distance restraints that, when applied in a restrained molecular dynamics protocol, produce a bent structure.

Algorithms↗

PubMed1995

Increased incidence of neurofibrillary tangles (NFT) in non-demented individuals with hypertension.

The incidence rates and numerical densities of argryophilic neurofibrillary tangles (NFT) and senile plaques (SP) were determined in non-demented individuals and subjects with Alzheimer's disease (AD). The non-AD subjects were grouped according to cardiac status; those individuals with critical coronary artery disease (cCAD), those hypertensive individuals without cCAD (HyperT), and those without heart disease (non-HD). The incidence and densities of SP and NFT were significantly greater in AD than any of the non-demented groups. The prevalence of SP was increased in both HyperT and cCAD compared to non-HD controls, while NFT occurrence was accentuated in non-demented HyperT subjects only. The densities of SP and NFT in HyperT were elevated compared to cCAD or both cCAD and non-HD controls; NFT densities were similar in cCAD and non-HD. NFT density increased with increasing age in only the non-HD and cCAD groups, suggesting a possible relationship between disease process and NFT formation in the AD and HyperT populations.

Age Factors↗

PubMed1995

Determination of amphotericin B in cerebrospinal fluid by solid-phase extraction and liquid chromatography.

A highly sensitive and reproducible liquid chromatography (LC) method for the determination of Amphotericin B in cerebrospinal fluid has been developed and validated. This LC-based method involves using nystatin as an internal standard and solid-phase extraction for sample preparation, followed by reversed-phase separation monitored by absorbance at 410 nm. The method has a limit of quantification of less than 1 ng ml-1 and excellent precision and accuracy, with both percentage relative standard deviation and percentage relative error less than 10%. The established linearity range was 1-10 ng ml-1 (r2 > 0.99). The extraction recovery of Amphotericin B from the cerebrospinal fluid is higher than 90% over the entire linear range. The method has been successfully employed for studying the penetration of Amphotericin B into the central nervous system in dogs and human.

Amphotericin B↗

PubMed1995

Cell layer-specific expression of cyclic nucleotide phosphodiesterases in rat arteries: molecular cloning using isolated cell layers from paraformaldehyde-fixed tissues.

We describe a method for the isolation of small quantities of large poly (A)+ mRNA from blood vessels of the rat as well as from distinct cell layers of the rat aorta. The poly (A)+ mRNA isolated by this method is suitable for use in reverse transcription polymerase chain reaction (RT-PCR) amplification of low abundance messages. In this method, anesthetized rats are perfused with ice-cold phosphate-buffered paraformaldehyde to allow for the in situ fixation of many of the main arteries of the rat. Following the in situ fixation of the rat vasculature, selected blood vessels can be removed, cleaned, and poly (A)+ mRNA purified. In addition, the distinct cell layers of the paraformaldehyde-fixed aorta can be mechanically separated and poly (A)+ mRNA purified selectively from each. The application of this method to the study of enzymes involved in cyclic nucleotide-mediated cell-signaling is illustrated by the cloning of two cyclic AMP phosphodiesterases from rat arteries, and from the selective amplification of message for these enzymes from different cell layers isolated from the rat aorta. This method should be applicable to determine if selected mRNAs are present in selected blood vessels of the rat, or within distinct cell layers of particular large blood vessels.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1995

Determination of optical properties and blood oxygenation in tissue using continuous NIR light.

In this article, we introduce a simple method to characterize optical properties and blood oxygenation in tissue using spatially resolved, steady-state reflectance. The method considers multiple source-detector separations larger than 2 cm, i.e. 20 times the optical mean free path in tissue, and makes an approximation to linearize the relationship between the separation and reflectance. Simulation results show that errors of the algorithm due to the approximation are less than 10%. Using a calibration sample, we calculate from the slope and intercept the absorption and reduced scattering coefficients, mu a and mu's, of a tissue-like solution, and experimental results confirm the usefulness of the method for quantitation of haemoglobin saturation in tissue.

Algorithms↗

PubMed1995

A dynamic phantom brain model for near-infrared spectroscopy.

This report describes the construction, fluid dynamics and optical properties of an in vitro model of the neonatal brain for testing near-infrared spectroscopy (NIRS) instruments. The brain model is a solid plastic structure containing a vascular network perfused with blood equilibrated with O2, N2 and CO2 in a closed circuit. The oxygenation state and haemoglobin concentration of the perfusate can be regulated and measured with a co-oximeter, providing a means to compare NIRS measurements of oxy-, deoxy- and total haemoglobin concentrations and haemoglobin O2 directly with a validated standard method. Fluid dynamic experiments revealed that the model's vasculature remains stable over time with minimal haemolysis. The model's optical properties were characterized by time-resolved and continuous wave NIRS between 670 and 850 nm as perfusate saturation was varied in the range 0-100%. Optical properties of the neonatal piglet brain were also determined by similar methods. No significant differences were found between the model and piglet brain in absorption coefficients, reduced scattering coefficients and optical pathlengths, indicating that the model optically simulates the piglet brain over a wide range of oxygenation states. These results demonstrate the potential utility of this dynamic phantom brain for testing NIRS instruments for accuracy and reliability.

Animals↗

PubMed1995

Storage temperature effect on the stability of morphine and codeine in urine.

Urine samples collected from one laboratory volunteer and five alleged heroin addicts are prepared (without preservatives) in 5-mL aliquots in glass culture tubes and stored at room, refrigerator, and freezer temperatures. Total morphine, total codeine, free morphine, and free codeine in these samples are analyzed at 30-day intervals for an 11-month period. Total morphine and total codeine concentration decreases are observed for all specimens in all storage conditions. For samples stored in the refrigerator and freezer, similar concentration decrease patterns are observed for total morphine and total codeine, and the decreases range from approximately 10 to 40%. The concentrations of free morphine and free codeine show slight but steady increases. For samples stored at room temperature, large decreases of total morphine are observed for three out of 10 specimens, and total codeine and total morphine concentrations (in seven other specimens) show a decrease pattern similar to that observed for the freezer and refrigerator storage conditions. Three concentration change patterns are observed for free morphine: The type I pattern follows the same decrease pattern observed for freezer and refrigerator storage conditions; the type II pattern shows free morphine increases (after 30-90 days of storage) that remain relatively high for the entire 11-month period; and the type III pattern shows initial increases, followed by gradual decreases to levels that are comparable with the specimens' respective initial concentrations. Free codeine concentrations show slight and steady increases for the entire 11-month period in all specimens.

Codeine↗

PubMed1995

Interactions of valproic acid with carbamazepine and its metabolites' concentrations, concentrations ratios, and level/dose ratios in epileptic children.

In two groups of epileptic children receiving carbamazepine (CBZ) therapy with or without valproic acid (VPA) comedication, we investigate the drug interactions of VPA on serum CBZ and its metabolites' concentrations, concentration ratios, and level/dose ratios. Serum total and free CBZ-10, 11-epoxide (CBZ-E) concentrations are significantly increased in patients taking CBZ plus VPA, together with higher CBZ-E/CBZ concentration ratios and CBZ-E level/dose ratios. These results reflect the accumulation of CBZ-E. The decreased concentration ratios of trans-10, 11-dihydroxy-10, 11-dihydro-CBZ (CBZ-H)/CBZ-E observed in patients taking CBZ plus VPA suggest an inhibition in the biotransformation from CBZ-E to CBZ-H. Significant negative correlations are found between serum VPA level and CBZ-H/CBZ-E concentration ratios, indicating that the inhibition of CBZ-E hydrolysis by VPA may depend on the concentration of VPA (total or free CBZ-H/CBZ-E concentration ratio = [formula: see text], respectively). VPA concentration also shows significant positive correlations with CBZ-E and CBZ level/dose ratios. Patients taking CBZ plus VPA have significant higher free fractions of CBZ and CBZ-E than do patients on CBZ alone, suggesting a protein-binding displacement by VPA.

Carbamazepine↗