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Z W Yu

Publications and source records attributed to Z W Yu.

At least 19 recordsLinked to original sources

On the normalization method in two-dimensional correlation spectra when concentration is used as a perturbation parameter.

Data pretreatment is of importance in two-dimensional (2D) correlation analysis when composition is used as a perturbation parameter. For composition-oriented studies, different normalization methods based on both external parameters (i.e., concentration) and internal parameters (i.e., absorbance from individual components) have been compared. It was found that when there is no overlapping between absorption bands of interest, no normalization is needed for data pretreatment. When overlapped bands must be used for 2D correlation analysis, the mean-centered normalization method could be used to obtain correct signs in synchronous spectra for a transformation process in the specific form of A-->kC. The intensity of the 2D spectrum, however, may not accurately reflect quantitative information of the overall extent of spectral intensity variation observed during experiments.

Algorithms↗

Kinetic phase behavior of distearoylphosphatidylethanolamine dispersed in glycerol.

Phase behavior of distearoylphosphatidylethanolamine dispersed in excess glycerol has been examined by differential scanning calorimetry. Transformation from lamellar-gel to lamellar crystalline phase was found to take place at temperatures near 74.9 degrees C upon cooling and near 76.3 degrees C during heating scans. The transition can also be observed under isothermal conditions at temperature in this range. The kinetics of the transformation from lamellar-gel to lamellar-crystal phase was analyzed by the well-known Avrami equation. The apparent Avrami exponents were found to be approximately 1.6. The effective dimensionality of the growth pattern can then be set as 1, after taking into account the contribution of nucleation at the examination temperatures. The activation energy of the phase transition was estimated as approximately 255 kJ mol(-1). The data are discussed in terms of development of successful cryoprotective strategies using glycerol.

Calorimetry, Differential Scanning↗

Insulin can enhance GLUT4 gene expression in 3T3-F442A cells and this effect is mimicked by vanadate but counteracted by cAMP and high glucose--potential implications for insulin resistance.

UNLABELLED: It is well-established that high levels of cAMP or glucose can produce insulin resistance. The aim of this study was to characterize the interaction between these agents and insulin with respect to adipose tissue/muscle glucose transporter isoform (glucose transporter 4, GLUT4) gene regulation in cultured 3T3-F442A adipocytes and to further elucidate the GLUT4-related mechanisms in insulin resistance. Insulin (10(4) microU/ml) treatment for 16 h clearly increased GLUT4 mRNA level in cells cultured in medium containing 5.6 mM glucose but not in cells cultured in medium with high glucose (25 mM). 8-Bromo-cAMP (1 or 4 mM) or N(6)-monobutyryl cAMP, a hydrolyzable and a non-hydrolyzable cAMP analog, respectively, markedly decreased the GLUT4 mRNA level irrespective of glucose concentrations. In addition, these cAMP analogs also inhibited the upregulating effect of insulin on GLUT4 mRNA level. Interestingly, the tyrosine phosphatase inhibitor vanadate (1-50 microM) clearly increased GLUT4 mRNA level in a time- and concentration-dependent manner. Furthermore, cAMP-induced inhibition of the insulin effect was also prevented by vanadate. In parallel to the effects on GLUT4 gene expression, both insulin, vanadate and cAMP produced similar changes in cellular GLUT4 protein content and cAMP impaired the effect of insulin to stimulate (14)C-deoxyglucose uptake. In contrast, insulin, vanadate or cAMP did not alter insulin receptor (IR) mRNA or the cellular content of IR protein. IN CONCLUSION: (1) Both insulin and vanadate elicit a stimulating effect on GLUT4 gene expression in 3T3-F442A cells, but a prerequisite is that the surrounding glucose concentration is low. (2) Cyclic AMP impairs the insulin effect on GLUT4 gene expression, but this is prevented by vanadate, probably by enhancing the tyrosine phosphorylation of signalling peptides and/or transcription factors. (3) IR gene and protein expression is not altered by insulin, vanadate or cAMP in this cell type. (4) The changes in GLUT4 gene expression produced by cAMP or vanadate are accompanied by similar alterations in GLUT4 protein expression and glucose uptake, suggesting a role of GLUT4 gene expression for the long-term regulation of cellular insulin action on glucose transport.

3T3 Cells↗

The effect of dimethyl sulphoxide on the structure and phase behaviour of palmitoleoylphosphatidylethanolamine.

The thermotropic phase behaviour and structure of a nonbilayer-forming lipid, 1-palmitoyl-2-oleoyl-phosphatidylethanolamine, dispersed in water and in aqueous solutions of up to 50 wt% dimethyl sulphoxide (DMSO) have been characterised using synchrotron X-ray diffraction methods. It was found that the presence of DMSO in the solvent induced an increase in the temperature of lamellar-gel to lamellar-liquid-crystal phase transition and a decrease in the temperature of the lamellar-liquid-crystal to inverted-hexagonal phase transition of the phospholipid. The presence of DMSO also caused a decrease in the X-ray repeat spacings of all the phases studied. Electron density profiles of the phospholipid dispersed in water and 50 wt% DMSO in the bilayer gel state were calculated. The presence of 50 wt% DMSO caused the apparent disappearance of the solvent layer separating phospholipid bilayers in the gel state. The results suggest that DMSO contributes to the bilayer electron density profile and that the amphiphilic solvent molecules partition into the interfacial region.

Dimethyl Sulfoxide↗

The upregulating effect of insulin and vanadate on cell surface insulin receptors in rat adipocytes is modulated by glucose and energy availability.

The aim of this study was to further characterize the rapid effects of insulin and the tyrosine phosphatase inhibitor vanadate to amplify cell surface insulin binding capacity in isolated rat adipocytes. The effect of 20 min insulin treatment (1000 microU/ml) was 2- to 3-fold (p < 0.01) when cells were treated in medium containing 5.6 mM D-glucose, but it was totally absent in glucose-free medium. Other carbon energy sources, such as fructose and pyruvate, could only partly substitute for D-glucose, with an approximately 1.5-fold insulin effect. Moreover, inhibiting transmembrane glucose transport with cytochalasin B completely blocked the effect of insulin to enhance cell surface binding. The effect of vanadate was only partly glucose-dependent, since a submaximal effect (1.5- to 2-fold, p<0.05) was seen also in the absence of glucose. The tyrosine kinase inhibitor genistein markedly blunted the effect of vanadate (from 3- to 4-fold to approximately 2-fold, p < 0.05) also indicating the importance of tyrosine phosphorylation-related mechanisms in the upregulation of cell surface insulin binding. Glycosylation of insulin receptors as a mechanism for this effect appears unlikely since neither the effect of insulin nor that of vanadate was altered by the glycosylation inhibitor tunicamycin. The time course for the insulin effect displayed a long duration (at least 6 h), suggesting a maintenance role of insulin keeping its receptors accessible for ligand binding at the cell surface. In conclusion, the effect of insulin and vanadate to upregulate cell-surface insulin receptors is energy-dependent and to some extent specifically glucose-dependent.

Adipocytes↗

[Chiral separation of glycerin monosulfides and bisulfides with a stationary phase of amylose-tris(3,5-dimethylphenylcarbamate)].

Amylose-tris(3,5-dimethylphenylcarbamate) (ADMPC) and aminopropylated silica gel(APS) were prepared after the reported methods. ADMPC was immobilized on APS from tetrahydrofuran solution with a coating amount of 15%. The chiral stationary phase was packed in a stainless-steel column(150 mm x 4.6 mm i.d.) by slurry method. The column was used for the enantioseparation of 6 glycerin monosulfides and 4 glycerin bisulfides. Mixtures of hexane and 2-propanol were used as mobile phases. The enantiomers of the monosulfides could be separated quite well, while those of the bisulfides could not at all. This phenomenon shows that the ether oxygen atom of the monosulfides plays a key role in the chiral discrimination process. The retention time of the solutes increases significantly as the amount of 2-propanol decreases. This shows the main interaction between the solutes and the chiral stationary phase is hydrogen bonding. A dynamic model is presented to account for the chiral discrimination mechanism.

Amylose↗

[Direct optical resolution of acidic biphenyl drugs by high performance liquid chromatography on tris(3,5-dimethylphenylcarbamate) of cellulose].

A chiral stationary phase was prepared by coating cellulose-tris(3, 5-dimethylphenylcarbamate) onto aminopropylated silica gel. A series of enantiomeric acidic biphenyl drugs were directly resolved on the chiral stationary phase (CSP) by normal-phase high performance liquid chromatography (HPLC). A hexane-2-propanol eluting system containing 1% of trifluoroacetic acid was used as mobile phase. Efficient optical resolution of the acidic biphenyl drugs has been attained. The factors that influence chiral discrimination such as structural characeristic of the samples and mobile phase were investigated. An interaction model between the stationary phase and the samples was discussed. The results showed that efficient optical resolution of racemic carboxylic acids could be attained by normal-phase HPLC on CSP using a hexane-2-propanol eluting system containing 1% of trifluoroacetic acid.

Carbamates↗

Molecular forces between membranes displaying neutral glycosphingolipids: evidence for carbohydrate attraction.

The surface force apparatus was used to determine the fundamental forces governing the adhesion between mixed bilayer membranes comprising lactosyl ceramide (LacCer) and di-tridecanoyl-phosphatidyl choline. Forces between membranes were quantified as a function of the glycolipid surface densities, which ranged from 0 to 30 mol %. Control measurements of the forces between pure phosphatidylcholine membranes and mixed bilayers of lactosyl ceramide with phosphocholine showed that the steric thickness of the carbohydrate headgroups increased from 19 to 25 A when the glycolipid density increased from 10 to 20 mol %. The layer compressibility also decreased with increasing carbohydrate coverage, but the corresponding adhesion between lactosyl ceramide-containing membranes increased with increasing amounts of glycosphingolipid in them. The nonspecific van der Waals forces accounted for the attraction measured in the control experiments and that between identical 10 mol % LacCer bilayers. However, the increase in the adhesion with increasing glycolipid density was 2-4 times greater than predicted by Lifschitz theory. Additionally, the forces measured during separation of membranes containing 20 and 30 mol % glycosphingolipid indicated that the headgroups bind and rearrange during bilayer detachment. The interactions between the carbohydrates are weak and apparently dynamic, and they generate an additional density-dependent intermembrane attraction that is on the order of the van der Waals force.

Adsorption↗

Solvation effects of dimethyl sulphoxide on the structure of phospholipid bilayers.

The effect of dimethylsulphoxide (DMSO), a widely used solvent in life sciences to stabilise biological membrane structures at low temperatures, on the structure of model membranes has been examined using X-ray diffraction methods. It was found that increasing concentrations of DMSO in water caused a progressive decrease in lamellar repeat spacings of multilamellar dispersions of both saturated and mono-unsaturated phosphatidylcholines. The lamellar repeat spacings were temperature-dependent but with dispersions in excess 40 wt.% DMSO, repeat spacings were less than that of the phospholipids in the dry state. One dimensional electron density profiles of the lipid bilayers were calculated and the thickness of the phosphatidylcholine bilayers were determined. It was inferred from the data that the thickness of liquid-crystal bilayers decreases in the presence of DMSO and that the DMSO molecules penetrate between the polar head groups of the phosphatidylcholines, resulting in an increase in area occupied by phospholipid at the bilayer surface.

1,2-Dipalmitoylphosphatidylcholine↗

Successful treatment with plasmapheresis, cyclophosphamide, and cyclosporin A in type B syndrome of insulin resistance. Case report.

CASE HISTORY: A woman born in 1949 was diagnosed in 1990 with systemic lupus erythematosus. She was treated with prednisolone, and < 1 year later she presented with marked hyperglycemia. Large doses of insulin were given four times per day. Even though the patient was thin (BMI 17.4 kg/m2), very little improvement was seen. INVESTIGATIONS AND TREATMENT: Serum insulin levels were high, and a euglycemic clamp investigation confirmed severe insulin resistance. The patient's serum contained insulin receptor antibodies inhibiting insulin binding, and thus the patient had a type B syndrome of insulin resistance. After diet and exercise, glycemic control stabilized and insulin treatment was withdrawn. However, in late 1993 she was in a catabolic and hyperglycemic state even though prednisolone doses were increased and azathioprin was added. In early 1994 she was treated with plasmapheresis and cyclophosphamide i.v. Subsequently, cyclosporin A was started as a maintenance therapy in addition to azathioprin. There was a rapid and sustained clinical improvement. Since late 1994 and onward, there is no sign of diabetes or glucose intolerance and there are no demonstrable insulin receptor antibodies in the patient's serum. DISCUSSION: Severe type B insulin resistance may respond favorably to treatment with plasmapheresis and cyclophosphamide followed by cyclosporin A in combination with azathioprin.

Adipocytes↗

Peroxovanadate and insulin action in adipocytes from NIDDM patients. Evidence against a primary defect in tyrosine phosphorylation.

We studied the effects of insulin and the stable peroxovanadate compound potassium bisperoxopicolinatooxovanadate (bpV(pic)), a potent inhibitor of phosphotyrosine phosphatases, on lipolysis and glucose uptake in subcutaneous adipocytes from 10 male patients with non-insulin-dependent diabetes mellitus (NIDDM) and 10 matched non-diabetic control subjects. Lipolysis stimulated by isoprenaline or the cAMP analogue, 8-bromo-cyclic AMP (8-br-cAMP), was reduced by approximately 40% in NIDDM compared to control subjects. In both groups bpV(pic) exerted an antilipolytic effect that was similar to insulin (approximately 50 % inhibition). 14C-U-glucose uptake was dose-dependently increased by bpV(pic) treatment, but this effect and also that of insulin were impaired in NIDDM compared to control (bpV(pic) 1.6-fold vs 2.4-fold and insulin 2.2-fold vs 3.4-fold). Furthermore, low concentrations of bpV(pic) did not affect insulin-stimulated glucose uptake, although tyrosine phosphorylation of the insulin receptor beta-subunit was clearly increased by bpV(pic). In conclusion, 1) beta-adrenergic stimulation of lipolysis in vitro is attenuated in NIDDM adipocytes due to post-receptor mechanisms. 2) Both insulin and bpV(pic) decrease lipolysis and enhance glucose uptake in control as well as NIDDM adipocytes. The effect on glucose uptake, but not that on lipolysis, is impaired in NIDDM cells. 3) Peroxovanadate does not improve sensitivity and responsiveness to insulin in NIDDM adipocytes, showing that insulin-resistant glucose uptake in NIDDM is not overcome by phosphotyrosine-phosphatase inhibition and, thus, probably is not caused by impaired tyrosine phosphorylation events alone.

8-Bromo Cyclic Adenosine Monophosphate↗

Thermotropic properties of dioleoylphosphatidylethanolamine in aqueous dimethyl sulfoxide solutions.

The effect of dimethyl sulfoxide (DMSO) on the phase behavior and structural parameters of aqueous dispersions of 1,2-dioleoylglycerophosphoethanolamine (DOPE) was characterized using differential scanning calorimetry and time-resolved synchrotron X-ray diffraction. In the range of concentrations of DMSO in water up to 50 wt%, there was a progressive increase in the temperature of the lamellar gel to lamellar liquid-crystal phase transition temperature and a marked decrease in temperature of the transition to the hexagonal II phase. At DMSO concentrations greater than about 25 wt% a direct transition from lamellar gel to hexagonal II structure was observed. A partial phase diagram of the phospholipid in DMSO has been constructed from thermal and X-ray diffraction data. Concentrations of DMSO greater than 50 wt% resulted in the formation of several lamellar crystal phases and interdigitated phases. Phase transition sequences measured on heating included low-temperature exothermic transitions between the interdigitated phase and lamellar crystal phases and an endothermic transition at about 30 degrees C between a lamellar crystal and an interdigitated phase. This latter phase converted directly into an hexagonal II phase at a temperature of about 50 degrees C. On cooling, a second nonbilayer phase (I) first appeared which subsequently reorganized to form a mixture of interdigitated and lamellar crystal phases. The significance of this data in terms of its possible relevance to the biological role of DMSO is discussed.

Biophysical Phenomena↗

Cryptic receptors for insulin-like growth factor II in the plasma membrane of rat adipocytes--a possible link to cellular insulin resistance.

To further elucidate the mechanisms for short-term regulation of the receptor for insulin-like growth factor II (IGF-II), we investigated effects of insulin, cAMP and phosphatase inhibitors on cell surface 125I-IGF-II binding in rat adipocytes. Preincubation with the serine/threonine phosphatase inhibitor okadaic acid (OA, 1 microM) or the non-hydrolysable cAMP analogue N6-mbcAMP (4 mM) markedly impaired insulin-stimulated 125I-IGF-II binding. Furthermore, addition of OA enhanced the inhibitory effect exerted by N6-mbcAMP. N6-mbcAMP also induced an insensitivity to insulin which was normalized by concomitant addition of the tyrosine phosphatase inhibitor vanadate (0.5 mM). In contrast, vanadate did not affect the impairment in maximal insulin-stimulated 125I-IGF-II binding produced by either OA or N6-mbcAMP. Phospholipase C (PLC), which cleaves phospholipids at the cell surface, markedly enhanced cell surface 125I-IGF-II binding in a concentration-dependent manner. Scatchard analysis demonstrated that the effect of PLC was due to an increased number of binding sites suggesting that "cryptic' IGF-II receptors are associated with the plasma membrane (PM). PLC (5 U/ml) also reversed the N6-mbcAMP-induced decrease of 125I-IGF-II binding at a low insulin concentration (10 microU/ml). Taken together, these data indicate that cAMP, similar to its effects on the glucose transporter GLUT 4 and the insulin receptor, may increase the proportion of functionally cryptic IGF-II receptors in the PM through mechanisms involving serine phosphorylation, possibly of a docking or coupling protein. Tyrosine phosphorylation appears to exert an opposite effect promoting the full cell surface expression of receptors.

Adipocytes↗

Effects of peroxovanadate and vanadate on insulin binding, degradation and sensitivity in rat adipocytes.

The effects of vanadate and the stable peroxovanadate compound bpV(pic) on insulin binding and degradation were investigated in rat adipocytes under conditions of ongoing receptor cycling. Both bpV(pic) and vanadate increased 125I-insulin binding to intact cells through an increase in apparent receptor affinity. The maximal effect of bpV(pic) was to increase binding approximately 4-fold (EC50 0.06 +/- 0.01 mM), whereas vanadate increased binding approximately 2-fold (EC50 1.4 +/- 0.2 mM). Removal of cell surface insulin-receptor complexes with trypsin showed that the effects on binding exerted by bpV(pic) and vanadate were due to a similar increase in both cell surface binding and intracellular accumulation of radioactivity. Both bpV(pic) and vanadate inhibited the degradation of 125I-insulin in medium containing 1% bovine serum albumin. The ratio of degraded/intact intracellular 125I-insulin was also markedly reduced by these agents, suggesting that they inhibit intracellular insulin-degrading proteases. Similar to previous findings with vanadate, bpV(pic) stimulated glucose transport and, at low concentrations, enhanced insulin sensitivity. Taken together, these data demonstrate that both bpV(pic) and vanadate inhibit insulin degradation. In addition, they significantly enhance cell surface insulin binding in rat fat cells and this is associated with an improved insulin sensitivity.

3-O-Methylglucose↗

Phase behaviour of distearoylphosphatidylethanolamine in glycerol--a thermal and X-ray diffraction study.

The phase behaviour of 1,2-distearoylphosphatidylethanolamine in glycerol has been examined using differential scanning calorimetry and real-time synchrotron X-ray diffraction methods. Dry phospholipid and phospholipid dispersed in glycerol over the concentration range 2.4%-90% (w/w) was equilibrated for 30 min at 20 degrees C and thermal and structural parameters on the temperature range 60 degrees C to 110 degrees C recorded during an initial heating and subsequent reheating. The characteristic feature of the initial heating scan was a direct lamellar crystalline to inverted hexagonal phase transition. In the subsequent cooling scan a lamellar gel structure was formed from the non-lamellar phase which transformed, on reheating, to a lamellar crystalline phase in which the acyl chain packing was titled with respect to the bilayer plane. The mechanism of the formation of the two crystalline phases was examined in the context of a relaxation model, where the liquid-crystal phase below the transition temperature from lamellar crystalline phase is metastable. A binary phase diagram over the temperature range 60 degrees C to 110 degrees C has been constructed.

Glycerol↗

Phase stability of phosphatidylcholines in dimethylsulfoxide solutions.

The temperature dependence of the phase stability of dispersions of dimyristoyl, dipalmitoyl, and distearoyl derivatives of phosphatidylcholines in excess aqueous dimethylsulfoxide has been examined by differential scanning calorimetry and synchrotron x-ray diffraction methods. There was a close correlation between the enthalpic transitions and the structural changes associated with the pre- and main transitions of the phospholipids in the range of concentrations up to mole fractions of dimethylsulfoxide in water of 0.1333. The temperature of the pre- and main transitions of the three phospholipids were found to increase linearly with increasing mole fraction of dimethylsulfoxide. The difference in phase stability between the lamellar gel and ripple phases induced by increasing dimethylsulfoxide concentration resulted in disappearance of the ripple phase and direct transition between lamellar gel and lamellar liquid-crystal phases. The effect of changing the properties of the solvent by the addition of dimethylsulfoxide on the dimensions of dipalmitoylphosphatidylcholine and solvent layers of the bilayer repeat structure has been determined from electron density distribution calculations. The lamellar repeat spacing recorded at 25 degrees C decreased from 6.36 nm in aqueous dispersion to 6.04 nm in a dispersion containing a mole fraction of 0.1105 dimethylsulfoxide. The results indicate that dipole interactions between solvent and phospholipid and dielectric properties of the solvent are important factors in the determination of the structure of saturated phosphatidylcholines.

1,2-Dipalmitoylphosphatidylcholine↗