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Biomedical subjects

J Han

Publications and source records attributed to J Han.

At least 379 records · Page 21Linked to original sources
PubMed1993

Endotoxin induces rapid protein tyrosine phosphorylation in 70Z/3 cells expressing CD14.

CD14, a glycosylphosphatidylinositol-anchored glycoprotein of leukocytes, binds endotoxin (lipopolysaccharide (LPS)) with high affinity. After the murine pre-B cell line 70Z/3 is transfected with DNA encoding human CD14 (hCD14), the resultant stably transfected cell line, 70Z/3-hCD14, responds to 1000-fold lower LPS concentrations than the parental CD14-negative line. We have used 70Z/3-hCD14 cells, RAW264.7 cells, and elicited murine peritoneal exudate macrophages (PEM) to study LPS-induced protein tyrosine phosphorylation. LPS induces the rapid tyrosine phosphorylation of a 38-kDa protein (p38) in 70Z/3-hCD14 cells, PEM, and RAW264.7 cells and of two isoforms of mitogen-activated protein kinases (MAPK) in only RAW264.7 cells and PEM. p38 can be distinguished from the MAPK isoforms based on differences in mobilities on SDS-polyacrylamide gel electrophoresis and the lack of reactivity of p38 with anti-MAPK antibody even after dephosphorylation with potato acid phosphatase. Synthetic lipid A induces p38 phosphorylation in 70Z/3-hCD14 cells, whereas phorbol 12-myristate 13-acetate and interferon-gamma fail to induce tyrosine phosphorylation of p38. Pretreatment of 70Z/3-hCD14 cells with anti-hCD14 monoclonal antibody or the tyrosine kinase inhibitor herbimycin A inhibits LPS-induced tyrosine phosphorylation of p38. These results suggest that increased protein tyrosine phosphorylation occurs rapidly after LPS binds to CD14 and is likely to be an important event in mediating LPS-induced cell activation.

Animals↗
PubMed1993

Resonance Raman spectroscopy of the azurin His117Gly mutant. Interconversion of type 1 and type 2 copper sites through exogenous ligands.

The copper center of the Pseudomonas aeruginosa His117Gly azurin mutant is accessible to exogenous ligands through an aperture in its surface created by the removal of the endogenous imidazole ligand. Depending on the exogenous ligand, a surprising variety of type 1 and type 2 copper sites can be obtained that are readily distinguished by electronic, EPR, and resonance Raman (RR) spectroscopy. The RR spectrum of type 1 H117G with exogenous imidazole is nearly identical to that of wild-type azurin, indicating that the trigonal geometry and short Cu-S(Cys) bond of approximately 2.15 A have been maintained. With anionic ligands (e.g., Cl-, Br-, N3-), the RR spectra show increased intensity at 370 and 400 cm-1 and a corresponding decrease in intensity at 410 cm-1, suggesting a lengthening of the Cu-S(Cys) bond as the site achieves a more tetrahedral character. An extreme example is the hydroxide adduct of H117G which is green in color and has optical and RR spectra reminiscent of the tetrahedral type 1 site in Achromobacter cycloclastes nitrite reductase. The fact that the basic RR pattern is little changed in most of the type 1 adducts indicates that the RR spectrum is due primarily to vibrations of the Cu-cysteinate moiety and that its coplanar conformation is conserved. Type 2 H117G proteins are formed by the addition of bidentate exogenous ligands such as histidine and histamine. They have their absorption maxima blue-shifted to 400 nm and their EPR A parallel values increased to approximately 160 x 10(-4) cm-1, both of which are characteristic of tetragonal Cu sites with Cu-S(thiolate) bonds of > 2.25 A. The RR spectra of the type 2 H117G proteins are still dominated by multiple cysteinate-related vibrational modes. However, the vibrational modes with the greatest intensity and Cu-S(Cys) stretching character have shifted approximately 100 cm-1 to lower energy compared to the type 1 sites, consistent with a longer (Cys)S-Cu bond. It is proposed that the tetragonal type 2 character of the bidentate ligand complexes is due to the addition of a fourth strong ligand in the equatorial ligand plane.

Anions↗
PubMed1993

Glycosyl-phosphatidylinositol-anchored or integral membrane forms of CD14 mediate identical cellular responses to endotoxin.

Endotoxin stimulates leukocytes to release cytokines that initiate septic shock in humans and animals. CD14, a glycosyl-phosphatidylinositol-anchored membrane glycoprotein, is an endotoxin receptor on leukocytes, and endotoxin binding to CD14 induces cytokine production. Here we show that glycosyl-phosphatidylinositol-anchored or integral membrane CD14 mediates identical cellular responses to endotoxin, including NF-kappa B activation and protein tyrosine phosphorylation. We also show that an anti-CD14 monoclonal antibody that does not block endotoxin binding to CD14 nonetheless inhibits cell activation by endotoxin. These findings suggest that binding of endotoxin to cell-surface CD14 is followed by subsequent interactions of the endotoxin-CD14 complex with additional membrane component(s) that enable transmembrane signaling. This function of CD14 may be prototypic for other members of the glycosyl-phosphatidylinositol-anchored family of proteins that do not play a primary role in signal transduction but rather are the principal ligand-binding units of membrane-bound receptor complexes.

Amino Acid Sequence↗
PubMed1993

Glutathione ester delays the onset of scurvy in ascorbate-deficient guinea pigs.

Previous studies showed that administration of ascorbate to glutathione (GSH)-deficient newborn rats and guinea pigs prevented toxicity and mortality and led to increased tissue and mitochondrial GSH levels; ascorbate thus spares GSH. In the present work, we tried to answer the converse question: Does administration of GSH spare ascorbate? Because administered GSH is not well transported into most cells, we gave GSH monoethyl ester (which is readily transported and converted into GSH intracellularly) to guinea pigs fed an ascorbate-deficient diet. We found that treatment with GSH ester significantly delays appearance of the signs of scurvy and that this treatment spares ascorbate; thus, the decrease of tissue levels of ascorbate was delayed. The findings support the conclusions that (i) GSH is essential for the physiological function of ascorbate because it is required in vivo for reduction of dehydroascorbate and (ii) there is metabolic redundancy and overlap of the functions of these antioxidants. The sparing effect of GSH in scurvy may be mediated through an increase in the reduction of dehydroascorbate (which would otherwise be degraded) and to antioxidant effects of GSH that are also produced by ascorbate. Other studies indicate that GSH deficiency in adult mice stimulates ascorbate synthesis in liver. During this work we found that administration of GSH itself is highly toxic to ascorbate-deficient guinea pigs when given in divided i.p. doses totaling 3.75 mmol/kg daily.

Animals↗
PubMed1993

Highlights of the cancer chemoprevention studies in China.

New retinoids have been synthesized and screened in the search for chemopreventive agents of cancer. N-4-(Carboxyphenyl) retinamide showed a significant inhibitiory effect on carcinogenesis of cancers in the buccal pouch of hamsters and in the forestomach of mice. Clinical studies have demonstrated that N-4-(carboxyphenyl) retinamide is effective against oral leukoplakia, vulvar leukoplakia, and dysplasia of the uterine cervix and stomach. Field studies among a population at high risk for esophageal cancer in Linxian County, Henan Province, revealed that N-4-(ethoxycarbophenyl) retinamide decreased the incidence of this cancer. Qidong County is a high-risk area for hepatoma in China. This has been correlated to the low levels of selenium in the blood of the residents as well as in grain grown in the area. S. Y. Yu, W. G. Li, Y. J. Zhu, et al. (Biol. Trace Element Res. 1985; 7:22-26) reported that the administration of selenium inhibited the incidence of hepatoma induced by aflatoxin B in rats and in ducks. Experimental studies demonstrated that green tea extract inhibited 12-O-tetradecanoylphorbol-3-acetate-induced epidermal ornithine decarboxylase activity and counteracted 12-O-tetradecanoylphorbol-3-acetate-induced ear edema in mice. It is interesting that green tea extract inhibited the transformation of Balb/c 3T3 cells induced by methylcholanthrene and 12-O-tetradenanoylphorbol-3-acetate. Garlic has been used for thousands of years in Chinese cooking and folk medicine. Epidemiological studies show that the dietary intake of garlic is inversely related to gastric cancer incidence in Shandong Province.

Allyl Compounds↗
PubMed1993

ATP-sensitive potassium channels are modulated by intracellular lactate in rabbit ventricular myocytes.

During myocardial ischemia, increased anaerobic glycolysis results in the accumulation of large amount of intracellular lactate. Effects of lactate on the ATP-sensitive potassium (KATP) channels were examined in rabbit ventricular myocytes, using the inside-out patch-clamp technique. Millimolar concentrations of lactate, applied to the cytosolic side of the patch membrane, induced openings of the KATP channel. This effect was inhibited by 0.1 mM glybenclamide. Lactate-induced openings of the channel were increased in a dose-dependent fashion. In dose-response relation for lactate, Kd (the lactate concentration producing half-maximal activation) and n (Hill coefficient) were 20 mM and 1.3, respectively (n = 5). Activation of KATP channels by lactate occurred even in the presence of 2 mM ATP. Lactate also caused a significant increase in Ki, the ATP concentration causing half-maximal inhibition, from 70 microM in control (n = 7) to 232 microM (n = 5). From the above results it could be concluded that intracellular lactate modulate KATP channels directly and such modulation may resolve the discrepancy between the low Ki in excised membrane patches and high levels of intracellular ATP concentration during myocardial ischemia or hypoxia.

Adenosine Triphosphate↗
PubMed1993

Macromolecular diffusion in crowded solutions.

The effects of crowding on the self or tracer diffusion of macromolecules in concentrated solutions is an important but difficult problem, for which, so far, there has been no rigorous treatment. Muramatsu and Minton suggested a simple model to calculate the diffusion coefficient of a hard sphere among other hard spheres. In this treatment, scaled particle theory is used to evaluate the probability that the target volume for a step in a random walk is free of any macromolecules. We have improved this approach by using a more appropriate target volume which also allows the calculation to be extended to the diffusion of a hard sphere among hard spherocylinders. We conclude that, to the extent that proteins can be approximated as hard particles, the hindrance of globular proteins by other proteins is reduced when the background proteins aggregate (the more so the greater the decrease in particle surface area), the hindrance due to rod-shaped background particles is reduced slightly if the rod-like particles are aligned, and the anisotropy of the diffusion of soluble proteins among cytoskeletal proteins will normally be small.

Biophysical Phenomena↗
PubMed1993

Prenatal diagnosis and a donor splice site mutation in fibrillin in a family with Marfan syndrome.

The Marfan syndrome, an autosomal dominant connective tissue disorder, is manifested by abnormalities in the cardiovascular, skeletal, and ocular systems. Recently, fibrillin, an elastin-associated microfibrillar glycoprotein, has been linked to the Marfan syndrome, and fibrillin mutations in affected individuals have been documented. In this study, genetic linkage analysis with fibrillin specific markers was used to establish the prenatal diagnosis in an 11-wk-gestation fetus in a four-generation Marfan kindred. At birth, skeletal changes suggestive of the Marfan syndrome were observed. Reverse transcription-PCR amplification of the fibrillin gene mRNA detected a deletion of 123 bp in one allele in affected relatives. This deletion corresponds to an exon encoding an epidermal growth factor-like motif. Examination of genomic DNA showed a G-->C transversion at the +1 consensus donor splice site.

Adult↗
PubMed1993

Modification of catalytic properties of chicken liver fructose 1,6-bisphosphatase by allicin.

The activity of chicken liver fructose 1,6-bisphosphatase increases dramatically after incubation with allicin, a major biologically active compound produced by garlic. Activation is more pronounced when the enzyme is assayed with Mn2+ than Mg2+. Maximum activation is accompanied by the disappearance of 4 highly reactive sulfhydryl groups per molecule of enzyme. This modification also leads to loss of activation by K+, and reduced sensitivity to inhibition by AMP, fructose 2,6-bisphosphate, and high concentration of fructose 1,6-bisphosphate. All the altered properties induced by allicin can be reversed by dithiothreitol or tris(2-carboxyethyl)phosphine, the latter being much more effective.

Animals↗
PubMed1993

Modification of opioid receptors and uncoupling of receptors from G proteins as possible mechanisms underlying suppression of opioid binding by cholecystokinin octapeptide.

Previous studies have shown that central cholecystokinin (CCK) octapeptide (CCK-8) suppresses the binding of opioid receptors to the opioid agonists. In the present study, tritium-labelled etorphine (opioid agonist) and naloxone (opioid antagonist) as well as guanosine-5'-0-(3-thiotriphosphate) (GTPrS) were used to investigate the possible mechanisms underlying the suppression of opioid binding by CCK-8 in rat brain membranes. CCK-8 at a concentration range of 10 nmol/L to 1 mumol/L dose-dependently suppressed the binding of [3H]-naloxone to opioid receptors, with a decrease in Bmax and an increase in Kd. GTPrs was found to reduce the affinity of [3H]-etorphine binding in a dose-dependent manner. This effect was markedly diminished in the presence of 10 nmol/L of CCK-8. These results suggest that CCK-8 might suppress opioid binding at both the receptor and post-receptor levels; that is, 1) by reducing the number and affinity of opioid receptors, and 2) by uncoupling opioid receptors from their G proteins.

Animals↗
PubMed1992

Involvement of opioid receptors in nucleus tractus solitarii in modulating endotoxic hypotension in rats.

Opioid antagonists selective for the mu-receptor (naltrexone) and kappa-receptor (nalmefene), respectively, were unilaterally microinjected into the nucleus tractus solitarii (NTS) of the pentobarbital anaesthetized (50 mg/kg, i.p.) rat subjected to E. coli endotoxin (15 mg/kg, i.v.). Naltrexone (0.5 microgram) ameliorated or totally abolished the endotoxic hypotension and tachycardia, and nalmefene at a dose of 1/25 (0.02 microgram) that of naltrexone showed a similar effect. The results suggest that the kappa- and mu-opioid receptors in NTS are actively involved in mediating the hypotensive and tachycardiac effects induced by systemic endotoxin in rats.

Animals↗
PubMed1992

Differential expression of two sodium channel subtypes in human brain.

Two partial human brain sodium channel cDNA sequences (designated HBSC I and II) have been cloned and mapped to chromosome 2q23-2q24 by chromosome microdissection-PCR (CMPCR). The distribution of HBSC I and II mRNA in human brain was studied by means of a novel approach based on the ligase detection reaction. These studies demonstrate that HBSC I and II mRNA is heterogeneously distributed in brain, and that the relative ratio of the two forms can vary as much as 7-fold between different regions.

Amino Acid Sequence↗
PubMed1992

Isolation, characterization, and amino acid sequences of auracyanins, blue copper proteins from the green photosynthetic bacterium Chloroflexus aurantiacus.

Three small blue copper proteins designated auracyanin A, auracyanin B-1, and auracyanin B-2 have been isolated from the thermophilic green gliding photosynthetic bacterium Chloroflexus aurantiacus. All three auracyanins are peripheral membrane proteins. Auracyanin A was described previously (Trost, J. T., McManus, J. D., Freeman, J. C., Ramakrishna, B. L., and Blankenship, R. E. (1988) Biochemistry 27, 7858-7863) and is not glycosylated. The two B forms are glycoproteins and have almost identical properties to each other, but are distinct from the A form. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis apparent monomer molecular masses are 14 (A), 18 (B-2), and 22 (B-1) kDa. The amino acid sequences of the B forms are presented. All three proteins have similar absorbance, circular dichroism, and resonance Raman spectra, but the electron spin resonance signals are quite different. Laser flash photolysis kinetic analysis of the reactions of the three forms of auracyanin with lumiflavin and flavin mononucleotide semiquinones indicates that the site of electron transfer is negatively charged and has an accessibility similar to that found in other blue copper proteins. Copper analysis indicates that all three proteins contain 1 mol of copper per mol of protein. All three auracyanins exhibit a midpoint redox potential of +240 mV. Light-induced absorbance changes and electron spin resonance signals suggest that auracyanin A may play a role in photosynthetic electron transfer. Kinetic data indicate that all three proteins can donate electrons to cytochrome c-554, the electron donor to the photosynthetic reaction center.

Amino Acid Sequence↗
PubMed1992

A randomized comparative trial of the MLCu375, TCu220C, steel ring and uterine cavity-shaped IUDs.

The Stainless Steel Ring, Uterine Cavity-Shaped Device, TCu220C and MLCu375 IUDs were evaluated in a randomized trial of 1536 women in Guangdong Province, China. The first-year continuation and expulsion rates were significantly lower (p less than 0.05) for the latter 3 IUDs than for the Stainless Steel Ring. The first year pregnancy rates for the MLCu375 (0.76 per 100 women) and TCu220C (1.2 per 100 women) were significantly lower (p less than 0.05) than those of the other two devices (6.1 and 4.1 per 100 women). The results of the study show that the MLCu375 and TCu220C are the most suitable IUDs for women in Guangdong Province.

Adult↗
PubMed1992

The effects of two air-powder abrasive prophylaxis systems on the surface of machined titanium: a pilot study.

This in vitro pilot project compared the effect of two air-abrasive prophylaxis systems on the surface of machined titanium. Single Brånemark titanium abutment cylinders were exposed to the Prophy-Jet and Microprophy systems for 90 seconds each. Both of the test cylinders were compared with an untreated control cylinder by scanning electron microscopy. Machining marks were completely removed by the Prophy-Jet and only partially removed by the Microprophy. Both of the resultant surfaces appeared to be smoother and thus may be more resistant to plaque formation. A rationale for the removal of machining marks is presented, although the reason for the difference in removal by the two systems is unclear. The prophylaxis cleaning powders were also examined by scanning electron microscopy and exhibited similar particle dimensions and morphology. A noncrystalline deposit was observed on the surface of the abutment cylinder exposed to the Microprophy. Energy dispersive spectroscopy analysis revealed that the deposit consisted almost entirely of sodium. Further investigation of the deposit is needed.

Air Pressure↗
PubMed1992

Miniature urea sensor based on H(+)-ion sensitive field effect transistor and its application in clinical analysis.

An urea-ENFET (Enzyme field effect transistor) probe was made by covering one of the grids of the dual ISFET (Ion sensitive field effect transistor) with a membrane of cross-linked bovine serum albumin (BSA)-urease and the other with cross-linked BSA, and the response characteristics of the probe was then tested through differential measurements. In different concentrations of phosphate buffer, the sensor responded to various concentrations of urea solution within 10-60s. From the calibration curve plotted on logarithmic scales a linear concentration range of 1.0-8.0 mg/dl was acquired, and the correlation coefficient and response sensitivity were 0.997 and 50mV/dec. (mg/dl), respectively. However, in dilute urea solution, the sensor responded linearly to the contents of urea over the range of concentration of 0.1-1.0 mg/dl with a correlation coefficient of 0.998 and a response sensitivity of 12-15mV/mg/dl. The standard deviation and the variation coefficient for 20 performances responding to 100mg/dl urea in 0.01M pH7.0 phosphate buffer were found to be 1.39mV and 1.44%, respectively. The urea-ENFET was used for the determination of BUN (Blood urea nitrogen) and the BUN values were compared with those determined by enzymatic method, the repression equation and correlation coefficient for 50 assays were y = -0.1272 + 0.9695x and r = 0.9912, respectively. When the urea-ENFET was used for determining urea either in buffer solution or in serum for 250 runs over a period of 1.5 months (the enzyme FET was stored at 4 degrees C between measurements during this period), the observed decrease of response sensitivity was only about 10%.(ABSTRACT TRUNCATED AT 250 WORDS)

Biosensing Techniques↗