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Biomedical subjects

H Tong

Publications and source records attributed to H Tong.

At least 37 records · Page 2Linked to original sources
PubMed1998

A low temperature flotation method to rapidly isolate lipoproteins from plasma.

To minimize oxidative modification, a low temperature, sequential flotation method was developed to isolate plasma lipoproteins in 18 h using a benchtop ultracentrifuge. The protein distributions were characterized using agarose and SDS-polyacrylamide gel electrophoresis, and an SDS-Lowry protein assay. The lipid distributions were assessed using a gas chromatography-mass spectrometric assay for cholesterol and an enzymatic assay for triglycerides. To validate the rapid flotation method, lipoproteins were also isolated from the same plasma samples using a modified Havel et al. flotation method (J. Clin. Invest. 34: 1345-1353, 1955). The same lipoproteins and apolipoproteins were present in fractions of comparable density, and the summed recoveries of protein, cholesterol, and triglyceride were also identical for the Havel et al. and rapid flotation procedures. Likewise, the amount of cholesterol and triglyceride in corresponding very low, intermediate, and low density lipoprotein (VLDL/IDL and LDL) fractions was the same for the two flotation procedures. The triglyceride and cholesterol levels in high density lipoprotein (HDL) isolated by rapid flotations, however, were 9-12% higher than in the HDL as isolated by Havel et al. Because a 9-12% increase in the HDL fraction reflects only 1-4% of the total triglyceride and cholesterol in plasma, we conclude that, while maintained at 4 degrees C, lipoproteins were quantitatively isolated from human plasma in 1 day.

Adult↗
PubMed1997

Crystal structure of murine/human Ubc9 provides insight into the variability of the ubiquitin-conjugating system.

Murine/human ubiquitin-conjugating enzyme Ubc9 is a functional homolog of Saccharomyces cerevisiae Ubc9 that is essential for the viability of yeast cells with a specific role in the G2-M transition of the cell cycle. The structure of recombinant mammalian Ubc9 has been determined from two crystal forms at 2.0 A resolution. Like Arabidopsis thaliana Ubc1 and S. cerevisiae Ubc4, murine/human Ubc9 was crystallized as a monomer, suggesting that previously reported hetero- and homo-interactions among Ubcs may be relatively weak or indirect. Compared with the known crystal structures of Ubc1 and Ubc4, which regulate different cellular processes, Ubc9 has a 5-residue insertion that forms a very exposed tight beta-hairpin and a 2-residue insertion that forms a bulge in a loop close to the active site. Mammalian Ubc9 also possesses a distinct electrostatic potential distribution that may provide possible clues to its remarkable ability to interact with other proteins. The 2-residue insertion and other sequence and structural heterogeneity observed at the catalytic site suggest that different Ubcs may utilize catalytic mechanisms of varying efficiency and substrate specificity.

Amino Acid Sequence↗
PubMed1997

Empirical parameterization of a model for predicting peptide helix/coil equilibrium populations.

A modification of the Lifson-Roig formulation of helix/coil transitions is presented; it (1) incorporates end-capping and coulombic (salt bridges, hydrogen bonding, and side-chain interactions with charged termini and the helix dipole) effects, (2) helix-stabilizing hydrophobic clustering, (3) allows for different inherent termination probabilities of individual residues, and (4) differentiates helix elongation in the first versus subsequent turns of a helix. Each residue is characterized by six parameters governing helix formation. The formulation of the conditional probability of helix initiation and termination that we developed is essentially the same as one presented previously (Shalongo W, Stellwagen, E. 1995. Protein Sci 4:1161-1166) and nearly the mathematical equivalent of the new capping formulation incorporated in the model presented by Rohl et al. (1996. Protein Sci 5:2623-2637). Side-chain/side-chain interactions are, in most cases, incorporated as context dependent modifications of propagation rather than nucleation parameters. An alternative procedure for converting [theta]221 values to experimental fractional helicities ( ) is presented. Tests of the program predictions suggest this method may have some advantages both for designed peptides and for the analysis of secondary structure preferences that could drive the formation of molten-globule intermediates on protein folding pathways. The model predicts the fractional helicity of 385 peptides with a root-mean-square deviation (RMSD) of 0.050 and locates (with precise definition of the termini in many cases) helices in proteins as well as competing methods. The propagation and nucleation parameters were derived from NMR data and from the CD data for a 79 peptide "learning set" for which an excellent fit resulted (RMSD = 0.0295). The current set of parameter corrections for capping boxes, helix dipole interactions, and side-chain/side-chain interactions (coulombic, hydrogen bonding and hydrophobic clustering), although still under development provide a significant improvement in both helix/coil equilibrium prediction for peptides and helix location in protein sequences. This is clearly evident in the rms deviations between CD measures and calculated values of fractional helicity for different classes of peptides before and after applying the corrections: for peptides lacking capping boxes and i/i + 3 and i/i + 4 side-chain/side-chain interactions RMSD = 0.044 (n = 164) versus RMSD = 0.054 (0.172 without the corrections, n = 221) for peptides that required context-dependent corrections of the parameters. If we restrict the analysis to N-acylated peptides with helix stabilizing side-chain/side-chain interactions (including N-capping boxes), the degree to which our corrections account for the stabilizing interaction can be judged from the change in helicity underestimation, ( calc- CD): -0.15 +/- 0.10, which is reduced to -0.018 +/- 0.048 (n = 191) upon applying the corrections.

Algorithms↗
PubMed1996

[Lectin-binding histochemical study on developing corneal epithelium and stroma of normal human being].

PURPOSE: To study glycoconjugates in developing cornea of normal human. METHODS: Lectin-binding histochemical method with five biotin-coupled lectins (conA, LCA, PNA, WGA, RCA). RESULTS: 1. WGA mainly stained the membrane of epithelial cells, especially in surface cells. It also stained anterior stroma of embryo. 2. All the layers of epithelium could be stained by conA, uneven stain was seen in stroma of embryo corneas. 3. RCA, LCA receptor distributed in base of cornea epithelium. 4. PNA can only stain cornea epithelium of 16 embryo. CONCLUSIONS: 1. Glycoconjugates of binding conA, WGA, PNA, LCA, RCA are in the developing corneas of human. 2. The distribution of these lectin receptors may change with corneal developing. It may be related to the maturity of corneas.

Corneal Stroma↗
PubMed1996

[The changes of striatal dopamine levels in the brain in rat models for Parkinson's disease after gene therapy].

OBJECTIVE: To examine the levels of striatal dopamine and metabolic products in rat model of Parkinson's disease (PD) by intracerebral grafting of the genetically modified muscle cells expressing tyrosine hydroxylase (TH). METHODS: The levels of striatal dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid in the striatum of rat models for PD were determined by using high performance liquid chromatography (HPLC). The tests were done in 2, 4, 12, 20 and 23 weeks after intracerebral transplantation of cultared muscle cells (myotubes) expressing TH gene. RESULTS: The levels of dopamine and its metabolites in the striatum increased significantly after transplantation of TH-expressed cultured muscle cells. For example, striatal dopamine level increased from 30.53 pg/mg to 876.25 pg/mg in brain tissue at 12 weeks after grafting. The striatal dopamine level was the highest at this time and then decreased gradually. In the control that received transplantation of muscle cells expressing LacZ gene, the striatal dopamine level showed no changes before and after grafting. CONCLUSIONS: The transplantation of TH gene-expressed cultured muscle cells is useful in the treatment of PD rat model, because it can increase striatal dopamine level.

Animals↗
PubMed1995

Overexpression, purification, and crystallization of the DNA binding and dimerization domains of the Epstein-Barr virus nuclear antigen 1.

The Epstein-Barr virus nuclear antigen (EBNA) 1 binds to and activates DNA replication from the latent origin of Epstein-Barr virus. Six different fragments of EBNA1 that retain DNA binding activity were expressed in bacteria, purified, and crystallized. Two fragments, EBNA470-619 and EBNA470-607, formed well ordered crystals that diffracted beyond 2.5-A resolution. Two different EBNA470-619 crystals were grown from sodium formate, pH 6-6.5. One crystal belonged to the trigonal space group P3 with unit cell dimensions a = b = 86.5 A and c = 31.8 A and with two molecules in the asymmetric unit. The other crystal, which appeared only twice and was likely related to the P3 crystal form, belonged to the trigonal space group P312 with cell dimensions a = b = 86.7 A and c = 31.8 A. Crystals of EBNA470-607 were grown by lowering the salt concentration to 0-100 mM NaCl at pH 6.0. These crystals belonged to the orthorhombic space group P2(1)2(1)2(1) and had cell dimensions a = 59 A, b = 66.9 A, and c = 69.8 A with two molecules in the asymmetric unit.

Antigens, Viral↗
PubMed1995

2-Amino-3-ketobutyrate-CoA ligase from beef liver mitochondria: an NMR spectroscopic study of low-barrier hydrogen bonds of a pyridoxal 5'-phosphate-dependent enzyme.

A study of protons associated with low-barrier hydrogen bonds in 2-amino-3-ketobutyrate-CoA ligase (AKB-ligase, EC 2.3.1.29) by NMR is reported. Three resonances are observed in the range of delta H = 15-20 ppm when the NMR spectrum of AKB-ligase is recorded at 600 MHz. These low-barrier hydrogen bonds are associated respectively with a side chain proton, the PLP pyridinium ring nitrogen proton, and the PLP Schiff base proton at the active site of the ligase. The pyridinium proton has been assigned a chemical shift of 19.10 ppm and the Schiff base proton 14.90 ppm. The third low-barrier hydrogen bond associated proton resonating at 16.20 ppm is assigned to a proton of a side chain group. All three resonances disappear when pyridoxal phosphate is removed from the ligase. Consistent with NOE coupling, the side chain group proton should be close to the proton of the Schiff base nitrogen of the pyridoxal 5'-phosphate. The effects of temperature, pH, substrate, and NOE on the three resonances are also studied, in order to assign the protons. The three low-barrier hydrogen bonds described in this report may serve to anchor the cofactor in the active site of 2-amino-3-ketobutyrate-CoA ligase.

Acetyltransferases↗
PubMed1995

Milk electrolyte content of Dahl hypertensive and normotensive rats.

Milk samples were collected from lightly anesthetized lactating female rats of the Dahl hypertension-sensitive (SS/Jr) and Dahl hypertension-resistent (SR/Jr) inbred strains on postnatal days 8, 14 and 18. These milk samples were stored at 4 degrees C until analyzed for content of calcium (Ca++), magnesium (Mg++), chloride (Cl-), potassium (K+), sodium (Na+), and total protein. Our findings revealed that milk samples from females of the two strains were remarkably similar in content of electrolytes and total protein at each of the three sampling times. The only significant difference from among a total of 18 strain comparisons across the three sample times was that milk levels of Ca++ were significantly higher in SS/Jrs compared to SR/Jrs on postnatal day 14. These findings clearly indicate that these milk constituents do not differ between mothers of the two Dahl strains. Thus, milk concentrations of electrolytes and protein do not appear to serve as an environmental stimulus during the preweanling period for the dramatic age-related increases in arterial pressure characteristic of the SS/Jr strain.

Animals↗
PubMed1995

Development of hypertension in spontaneously hypertensive rats: role of milk electrolytes.

1. Milk samples were collected from lactating spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) normotensive females at 8, 14 or 18 days postpartum. 2. Milk samples were later analysed for content of calcium (Ca++), sodium (Na+), potassium (K+), chloride (Cl-) and total protein. 3. Milk samples from SHR females had significantly higher concentrations of Na+ (at 8 and 14 days) and Cl- and significantly lower concentrations of Ca++, K+ (at 8 and 14 days) and total protein compared to milk samples from WKY females. 4. Preweanling dietary content of several electrolytes implicated in the development of hypertension differed dramatically between SHR and WKY strains. This altered diet early in life may serve as an environmental trigger for progressive age-related increases in arterial pressure in SHR.

Animals↗
PubMed1995

[Potassium metabolism in warm heart surgery].

The dynamic balance of potassium getting in and out the body during and after operation was surveyed in 26 adult patients who underwent cardiac surgery with normothermic cardiopulmonary bypass (CPB). We found that during CPB, because of the continous perfusion of warm oxygenated blood cardioplegia solution, a large volume of potassium entered the body; meanwhile, only a little potassium removed from the body in urine. At the moment of CPB finish, the volume of residual potassium in the body was as large as 74.5mmol (mean), that was 1.2mmol/kg body weight. The mean plasma potassium level at the points of aorta cross clamp release and CPB finish were 6.8mmol/L and 5.5mmol/L respectively, and all in normal range at several times measured from the end of operation (usually 12-2pm) to the second postoperative day 7am. The volume of potassium replenishment according to urine output in this group was much less than those in hypothermic CPB patients from the end of operation to the first postoperative day 7am. During CPB extracellular potassium shifted into intracellular fluid, but the shifting became opposite around the operative day 6pm, and the final equilibrium of potassium metabolism was achieved at the second postoperative day 7am. The remarkable hyperkalemia at the point of aorta cross clamp release may delay the heart rebeating ( > 5min, 35%), but the automatic rebeating rate was still very high (96%). With regard to the prevention of hyperkalemia during CPB, we also give some suggestions in this paper.

Adolescent↗
PubMed1994

2-Amino-3-ketobutyrate-CoA ligase from beef liver mitochondria. Purification and partial sequence.

2-Amino-3-ketobutyrate-CoA ligase (EC 2.3.1.29), or aminoacetone synthetase, has been purified by a nine-step procedure from 1.0 kg of beef liver to yield 8.8 mg of homogeneous enzyme. The homogeneous form of the enzyme, a monomer of M(r) = 44,000, shows unusually high absorption at 430 nm, with a ratio of absorbance at 280 and 430 nm of 2.6. On storage a species with an additional absorption peak at 332 is formed. Neither the 430-nm peak nor the 332-430 ratio is affected by pH or substrates. The peak at 430 nm and enzyme activity are both reduced by borohydride reduction and treatment with cysteine. The first 21 amino acids at the NH2-terminal of the ligase occur in the sequence Ser-Ala-Leu-Ala-Gln-Leu-Arg-Gly-Ile-Leu-Glu-Glu-Glu-Leu-Glu-Ser-Ile-Arg- Gly-Ala - Gly. No homology is detectable in the first 20 amino acids of the Escherichia coli and beef liver mitochondria enzymes. However, homology is found around the lysine residue to which the pyridoxal 5'-phosphate is attached in the two enzymes. A very hydrophobic peptide containing pyridoxal phosphate having the following sequence Leu-Leu-Gly-Val-Met-Asp-Gln-Val-Thr-Ile-Ile-Asn-Ser-Thr-Leu-Gly-Lys(P xy)-Ala- Leu-Gly-Gly-Ala-Ser-Gly-Gly-Tyr-Thr-Thr-Gly-Pro-Gly-Ala-Leu-Val has been isolated from the ligase. Fourteen residues around the lysine to which the pyridoxal 5'-phosphate is bound are completely identical with the pyridoxal 5'-phosphate containing peptide isolated from the E. coli 2-amino-3-ketobutyrate-CoA ligase.

Acetyltransferases↗
PubMed1993

Cofactor identification of threonine-serine dehydratase from sheep liver.

L-Threonine-serine dehydratase catalyzes the conversion of L-threonine and L-serine to alpha-ketobutyric acid and pyruvate, respectively. The enzyme has been purified to homogeneity from extracts of sheep liver. In the past, various cofactors have been suggested for threonine dehydratase from both prokaryotic and eukaryotic tissue. While some direct evidence for the presence of pyriodoxal 5'-phosphate in impure preparations is present in the literature no direct evidence for the cofactor in homogeneous dehydrogenase from mammalian tissue has been reported. The threonine dehydratase of sheep liver has been obtained in a homogeneous form and a spectral study provides clear evidence for the presence of pyridoxal 5'-phosphate. Both the physical properties of homogeneous threonine dehydratase and a study of spectral properties of its cofactor are reported in this communication.

Amino Acids↗
PubMed1993

Altered gustatory development in Na(+)-restricted rats is not explained by low Na+ levels in mothers' milk.

Placing pregnant rats on a sodium-deficient diet (0.03% NaCl) very early in gestation and then weaning the offspring (sodium-restricted rats) to the same diet precludes development of amiloride-sensitive sodium taste transduction pathways in the offspring. However, normal amiloride-sensitive sodium taste responses can be restored by permitting sodium ingestion by sodium-restricted rats. The present study tested the hypothesis that the concentration of sodium in sodium-restricted mothers' milk must be abnormally low in order to preserve altered gustatory function in the offspring. Other milk electrolyte and total protein concentrations were determined as well. Milk sodium was similar between sodium-restricted and control rat mothers at 10-13 and 16-20 days postpartum, as were levels of potassium and chloride. At 10-13 days postpartum, total protein was higher in milk from sodium-restricted mothers. Sodium-restricted mothers' milk calcium concentrations were higher versus controls at 16-20 days postpartum. These results indicate that the lack of gustatory amiloride sensitivity in sodium-restricted rats cannot be attributed to deficient dietary sodium levels during the suckling period.

Amiloride↗
PubMed1993

Endogenous adenosine, A1 adenosine receptor, and pertussis toxin sensitive guanine nucleotide binding protein mediate hypoxia induced AV nodal conduction block in guinea pig heart in vivo.

OBJECTIVE: The aim was to study the mechanism of global hypoxia induced atrioventricular (AV) nodal conduction block in vivo using a guinea pig model. METHODS: Animals subjected to 10 min periods of global hypoxia, induced by decremental changes in O2 content in the inhaled gas mixture, were randomly divided into three groups: group I = control; group II = animals treated with a new selective A1 adenosine receptor antagonist, N0861 (N6-endo-norbornan-2-yl-9-methyladenine); and group III-animals treated with pertussis toxin, an irreversible inhibitor of specific guanine nucleotide binding proteins (G protein). RESULTS: Progressive incremental hypoxia was associated with progressive AV nodal conduction delay culminating in a complete block. In addition, there was an inverse relationship between the severity of hypoxia and time to AV nodal block. Both N0861 and pertussis toxin treatment significantly reduced the degree of hypoxia induced AV nodal conduction block and delayed its appearance. CONCLUSIONS: (1) Endogenous adenosine released during acute global hypoxia causes AV nodal conduction block observed under these conditions in the guinea pig heart in vivo; and (2) this action of adenosine in the AV node is mediated by an A1 adenosine receptor and a pertussis toxin sensitive G protein.

Adenine↗
PubMed1993

Ethanol-induced aspartate and taurine release from primary astrocyte cultures.

Exposure of primary astrocyte cultures to isosmotic ethanol from 10-100 mM led to both swelling of the cells and release of [3H]taurine and D-[3H]aspartate. Exposure to hyperosmotic ethanol, in the same concentration range, caused neither swelling nor release. Release was inhibited by the anion transport blocker L-644,711, already shown to inhibit amino acid release evoked by hypoosmotic or high-potassium medium, conditions that also cause astrocytic swelling. Ethanol-induced release generally showed a decline in response to successive exposures to ethanol, and release was not dependent on extracellular calcium. Thus, the characteristics of swelling-induced release of amino acids by isosmotic ethanol seem to correspond to those of swelling-induced release from astrocytes due to exposure to hypotonic or high-K+ media. We discuss whether such effects may contribute to CNS damage after head injury and stroke.

Animals↗
PubMed1993

The study of relationships between the damage of visual function and hemorrheology, ocular rheography, as well as other related factors in patients with primary open angle glaucoma (POAG).

Researches of glaucoma visual function damage, hemorheology, ocular rheography and other related multiplex factors, with computed multifactorial stepwise regression analysis, indicate that the elevation of intraocular pressure (IOP) is not the only factor to induce visual impairment. POAG patients are shown to have markedly reduced diastolic perfusion pressure in ophthalmic artery, besides prolonged filling time of the retinal artery and vein, diminished erythrocyte deformability and increased platelet adhesion rate. The characteristic of ocular rheographic changes in POAG patients of controlled IOP suggest that the abnormal changes of hemorheology be responsible for the damage of visual function. The abnormal changes of plasma viscosity, blood apparent viscosity, blood viscoelasticity, hematocrit, mean red cell volume etc could be found in addition to the reduction of PO2, SO2% and trace elements (Cr, Zn, Mg, Fe) in patients with POAG. Computed multiply stepwise regression analysis and Bayes discrimination were made among the deteriorate rate of quantitative visual field and 28 factors from the combined investigation. Only the hemorrheological index could be introduced into the multiply regression equations. The more the damage of visual function in POAG patients, the more accurate the discriminatory effect. Based upon the results, and the reports of articles published as well as the search of 1982-1989 Compact Combridge MEDLINE Medical data base, the new clinical concept could be presented that POAG might belong to the scope of blood hyperviscosity syndromes. When IOP elevation and the decrease of ocular artery pressure existed, the damage of visual function in POAG patients should be deteriorated.

Adult↗
PubMed1992

Characterization of dibromopolychlorodibenzo-p-dioxins and dibromopolychlorodibenzofurans in municipal waste incinerator fly ash using gas chromatography/mass spectrometry.

Several fly ash samples from different countries were analyzed for dibromopolychlorodibenzo-p-dioxins and -dibenzofurans (DBPCDD/Fs) using capillary column gas chromatography/high-resolution mass spectrometry. Concentrations of these compounds ranged from low to high parts-per-trillion (pptr) depending on the location where the samples were collected. The mass spectrometer, operated at greater than 10,000 mass resolution in combination with the high resolving power of capillary column GC, permitted characterization of DBPCDD/Fs in municipal waste incinerator fly ash for the first time. The isomer patterns of tetra- through octa-congener groups of DBPCDD/Fs are remarkably similar regardless of where the fly ash originated, indicating common mechanisms of formation of these compounds in the incinerators.

Benzofurans↗