Biomedical subjects

S Ding

Publications and source records attributed to S Ding.

At least 37 records · Page 2Linked to original sources

PubMed2000

A noninvasive transesophageal signal averaging technique for detection of sinus node electrogram.

We have developed a noninvasive transesophageal signal averaging technique for direct recording of sinus node electrogram. In this study, sinus node electrograms were recorded from 106 of 138 patients (77%), comparable to that (46%) recorded by conventional transesophageal technique, 59 were male and 47 were female ranging in age from 10-74 years (mean 44.2+/-12.4 years). The signals from lead I, surface averaged lead and esophagus averaged lead were amplified (up to 100 microV/cm), filtered (0.1-50 Hz), AD converted to 16-bit accuracy at a sampling rate of 2 KHz and averaged by using the three channel low-noise amplifier. The signal averaged esophageal sinus node potentials are deflections of low-amplitude and low-frequency preceding the P wave. Two morphologies, the domed wave (64 of 106 patients, 60%) and the smooth upstroke slope (42 of 106 patients, 40%), can be seen. The directly recorded sinoatrial conduction time was 82.3 +/-18.6 msec (mean+/-2 SD), ranged from 23-112 msec, amplitude was 3.8-27.7 microV and dv/dt was 0.42-1.92 mV/sec. The sinoatrial conduction time recorded by the transesophageal catheter technique was comparable to that (80.4+/-18. 1 msec) recorded by the transvenous catheter method perfectly. We think that signal averaged sinus node electrogram could be recorded in sinus rhythm in most patients with normal sinus node function and proper filter settings, high amplification and anti-drift technique are important in recording signal averaged esophageal sinus node electrogram.

Adolescent↗

PubMed2000

Immobilizing the moving parts of voltage-gated ion channels.

Voltage-gated ion channels have at least two classes of moving parts, voltage sensors that respond to changes in the transmembrane potential and gates that create or deny permeant ions access to the conduction pathway. To explore the coupling between voltage sensors and gates, we have systematically immobilized each using a bifunctional photoactivatable cross-linker, benzophenone-4-carboxamidocysteine methanethiosulfonate, that can be tethered to cysteines introduced into the channel protein by mutagenesis. To validate the method, we first tested it on the inactivation gate of the sodium channel. The benzophenone-labeled inactivation gate of the sodium channel can be trapped selectively either in an open or closed state by ultraviolet irradiation at either a hyperpolarized or depolarized voltage, respectively. To verify that ultraviolet light can immobilize S4 segments, we examined its relative effects on ionic and gating currents in Shaker potassium channels, labeled at residue 359 at the extracellular end of the S4 segment. As predicted by the tetrameric stoichiometry of these potassium channels, ultraviolet irradiation reduces ionic current by approximately the fourth power of the gating current reduction, suggesting little cooperativity between the movements of individual S4 segments. Photocross-linking occurs preferably at hyperpolarized voltages after labeling residue 359, suggesting that depolarization moves the benzophenone adduct out of a restricted environment. Immobilization of the S4 segment of the second domain of sodium channels prevents channels from opening. By contrast, photocross-linking the S4 segment of the fourth domain of the sodium channel has effects on both activation and inactivation. Our results indicate that specific voltage sensors of the sodium channel play unique roles in gating, and suggest that movement of one voltage sensor, the S4 segment of domain 4, is at least a two-step process, each step coupled to a different gate.

Base Sequence↗

PubMed2000

Detoxication of vinca alkaloids by human P450 CYP3A4-mediated metabolism: implications for the development of drug resistance.

Vinca alkaloids are important chemotherapeutic agents, and their pharmacokinetic properties display significant interindividual variations, possibly due to CYP3A4-mediated metabolism. We have evaluated the relevance of this metabolism for the chemotherapeutic and the toxicological properties of these drugs. Analysis was performed using Chinese hamster ovary cell lines that expressed either CYP2D6 or CYP3A4. The latter cells metabolized vinblastine with a turnover number of 0.4 min(-1), resulting in a decreased cytotoxicity of this compound. Whereas vincristine and vinblastine at a concentration of 100 nM killed more than 90% of the parental cells, more than 50 and 35%, respectively, of cells that coexpressed CYP3A4 and cytochrome P450 (P450) reductase survived these treatments. No additional increase in cytotoxicity was noted above 100 nM. Similarly, preincubation of vinblastine with bacterial membranes that contained recombinant CYP3A4 and P450 reductase decreased the cytotoxicity of vinblastine for parental Chinese hamster ovary cells. We also demonstrate that the presence of vinblastine in a coculture of cells that expressed beta-galactosidase together with cells that expressed CYP3A4 strongly selected for the latter cells, resulting in an increased level of CYP3A4 in the surviving cell population. Similarly, treatment of the human colon adenocarcinoma cell line LS174T with vinblastine selected for a cell population with higher levels of endogenous CYP3A4 as revealed by immunohistochemistry without simultaneous increase of multidrug resistance protein 1 (MDR1). This is the first evidence that tumor P450s have the potential to contribute to the development of drug resistance during chemotherapy.

Animals↗

PubMed2000

[The effect of intraperitoneal chemotherapy on retroperitoneal lymph node metastasis of ovarian cancer].

The objective of this study was to investigate the effect of intraperitoneal chemotherapy(repeated injections of cisplatin in small doses during a short time) on retroperitoneal lymph node metastasis of ovarian cancer. Thirty patients with ovarian cancer were selected and divided into two groups. The patients in the first group received intraperitoneal chemotherapy only once (48 hours before operation); the patients in the second group received intraperitoneal chemotherapy twice (96 hours and 48 hours before operation). After the intraperitoneal chemotherapy, according to the predicted time, the Pt concentrations of the tissues were measured by flameless atomic absorption spectrometry; the pathologic examinations of the lymph nodes were carried out. The results showed that the drug concentrations of the aortic, obturator and iliac lymph nodes of the first group were not significantly different, neither were the drug concentrations of the lymph nodes of the second group. The drug concentrations of the lymph nodes of the first and the second groups were 2.06 and 1.91 times that of the surrounding tissues respectively. Also, the drug concentrations of peritoneum of the first and the second groups were 4.14 and 2.50 times that of the lymph nodes. The drug concentrations of the iliac, obturator and aortic lymph nodes of the second group were 2.27, 2.75 and 2.54 times that of the first group (P < 0.005). Under light microscope, the metastatic cancer tissues in lymph nodes showed liquefaction necrosis in the second group. The results indicate that small doses of cisplatin given by repeated injections during a short time can lead to drug accumulation and consequently high efficacy with low toxicity, suggesting that this method may be used for the treatment of lymphatic metastasis of ovarian cancer.

Antineoplastic Agents↗

PubMed2000

[Jugulotympanic glomus tumors: a report of 11 cases].

OBJECTIVE: To evaluate the surgical technique in removal of jugulotympanic glomus tumor. METHODS: Retrospective analysis of surgical techniques and effects of the surgical treated 11 cases (1982-1998) of jugulotympanic glomus tumors (tympanic type I, 1, II, 1, IV, 3; jugular type I, 1, III, 5). RESULTS: The tumors of all 11 patients were removed completely and the wounds healed smoothly. Only one case had postoperative laryngeal nerve paralysis. There was no recurrence after 1-8 years follow-up. CONCLUSION: Suitable surgical techniques are available for removal of jugulotympanic glomus tumors and are chosen according to the size and location of the tumor. Postauricular incision is suitable for Glomous tympanicum Type II, III, IV. Since postauricular incision with superior and inferior extension provides a good exposure of the neck and temporal bone, it is suitable for surgical removal of Glomus jugulare type I. In case of Glomus jugulare type III, postauricular large "C" incision may be chosen for resection of the mid- and infra-temporal fosse, neck and skull base tumor.

Adult↗

PubMed2000

[Catalytic spectrophotometric determination of trace amounts of copper with the system of ascorbic acid and ammonium molybdate].

Ammonium molybdate was reduced to molybdenum blue by ascorbic acid in the medium of phosphorous acid in the presence of copper catalyst. A new method for catalytic spectrophotometric determination of trace copper has been developed. The linear range was 0-0.024 microgram.25 mL-1. The limit of detection was 1.54 x 10(-11) g.mL-1. The method has been used to determine trace copper in human hair with satisfactory results.

Ascorbic Acid↗

PubMed1999

Ion permeation and block of P2X(2) purinoceptors: single channel recordings.

P2X(2) purinoceptors are cation-selective channels activated by ATP and its analogues. Using single channel measurements we studied the channel's selectivity for the alkali metal ions and organic monovalent cations NMDG(+), Tris(+), TMA(+), and TEA(+). The selectivity sequence for currents carried by alkali metal ions is: K(+) > Rb(+) > Cs(+) > Na(+) > Li(+), which is Eisenman sequence IV. This is different from the mobility sequence of the ions in free solution suggesting there is weak interaction between the ions and the channel interior. The relative conductance for alkali ions increases linearly in relation to the Stokes radius. The organic ions NMDG(+), Tris(+), TMA(+) and TEA(+) were virtually impermeant. The divalent ions (Mn(2+), Mg(2+), Ca(2+) and Ba(2+)) induced a fast block visible as a reduction in amplitude of the unitary currents. Using a single-site binding model, the divalent ions exhibited an equilibrium affinity sequence of Mn(2+) > Mg(2+) > Ca(2+) > Ba(2+).

Adenosine Triphosphate↗

PubMed1999

Synthesis and activity of a new generation of ruthenium-based olefin metathesis catalysts coordinated with 1,3-dimesityl-4,5-dihydroimidazol-2-ylidene ligands.

[formula: see text] A new family of 1,3-dimesityl-4,5-dihydroimidazol-2-ylidene-substituted ruthenium-based complexes 9a-c has been prepared starting from RuCl2(=CHPh)(PCy3)2 2. These air- and water-tolerant complexes were shown to exhibit an increased ring-closing metathesis activity at elevated temperature when compared to that of the parent complex 2 and the previously developed complex 3. In many instances the activity of these complexes also rivaled or exceeded that of the alkoxy-imido molybdenum complex 1. Catalyst loadings of as low as 0.05 mol% could be used.

Alkenes↗

PubMed1999

DENSE: displacement encoding with stimulated echoes in cardiac functional MRI.

Displacement encoding with stimulated echoes (DENSE) was developed for high-resolution myocardial displacement mapping. Pixel phase is modulated by myocardial displacement and data spatial resolution is limited only by pixel size. 2D displacement vector maps were generated for the systolic action in canines with 0.94 x 1.9 mm nominal in-plane resolution and 2.3 mm/pi displacement encoding. A radial strain of 0.208 was measured across the free left ventricular wall over 105 ms during systole. DENSE displacement maps require small first-order gradient moments for encoding. DENSE magnitude images exhibit black-blood contrast which allows for better myocardial definition and reduced motion-related artifacts.

Animals↗

PubMed1999

Distribution of cyclosporin A in ocular tissues after topical administration to albino rabbits and beagle dogs.

PURPOSE: To determine the ocular pharmacokinetics of cyclosporin A after topical ophthalmic administration. METHODS: Radiolabled cyclosporin A in either a castor oil-in-water emulsion or a corn oil ointment was applied to the eyes of beagle dogs or albino rabbits using the following paradigms: (i) single doses of 0.2% emulsion to rabbits and dogs, (ii) single doses of 0.05%, 0.2%, or 0.4% emulsion to rabbits, (iii) multiple doses of 0.2% emulsion to dogs, (iv) single and multiple doses of 0.2% ointment to rabbits. The distribution of cyclosporin A was determined by measuring the distribution of radioactivity. RESULTS: After a single dose, cyclosporin A was rapidly absorbed into the conjunctiva (Cmax: dogs, 1490 ng/g; rabbits, 1340 ng/g) and cornea (Cmax: dogs, 311 ng/g; rabbits, 955 ng/g). High concentrations (>300 ng/g) could be detected in the cornea up to 96 hours post-dose. Lower concentrations were found in the intraocular tissues, and systemic absorption was minimal. After multiple doses, there was some accumulation in the cornea, lens, lacrimal gland, and iris-cilliary body, but limited accumulation in the conjunctiva and sclera. Ocular tissue concentrations of cyclosporin A increased with increasing dose concentration; proportionally in lacrimal gland and intraocular tissues; less than proportionally in conjunctiva and cornea. The pharmacokinetic profile of the cyclosporin A corn oil ointment was similar to that of the emulsion. CONCLUSIONS: Topical ophthalmic cyclosporin A penetrated into extraocular tissues at concentrations adequate for local immunomodulation while penetration into intraocular tissues was much less and absorption into the blood was minimal.

Absorption↗

PubMed1999

Merits and limitations of recombinant models for the study of human P450-mediated drug metabolism and toxicity: an intralaboratory comparison.

A wide variety of pharmacological and toxicological properties of drugs are determined by cytochrome P450-mediated metabolism. Characterization of these pathways and of the P450 isoenzymes involved constitutes an essential part of drug development. Similarly, because P450s are catalyzing the toxication and detoxication of environmental pollutants, an understanding of these reactions facilitates risk assessment in environmental toxicology. Recently, a variety of recombinant expression systems has been employed to study the role of human P450s in these reactions. These include insect, bacterial, yeast, and mammalian models. As these were developed and characterized by different laboratories, evaluation of their merits and limitations is inherently difficult. To resolve this problem, we have established and characterized the latter three systems and present the key results here. In general, the catalytic properties of P450 isozymes in the various models were rather similar. However, taking technical considerations into account as well as the high level of functional expression of P450s achieved in bacteria make this system ideally suited for drug metabolism research, including the generation of milligram quantities of metabolites for structural determinations. For toxicological studies, however, expression of P450s in mammalian cells was most appropriate. This is exemplified here by studies into the role of human P450s in the activation and inactivation of chemotherapeutic drugs.

Amino Acid Sequence↗

PubMed1999

Single channel properties of P2X2 purinoceptors.

The single channel properties of cloned P2X2 purinoceptors expressed in human embryonic kidney (HEK) 293 cells and Xenopus oocytes were studied in outside-out patches. The mean single channel current-voltage relationship exhibited inward rectification in symmetric solutions with a chord conductance of approximately 30 pS at -100 mV in 145 mM NaCl. The channel open state exhibited fast flickering with significant power beyond 10 kHz. Conformational changes, not ionic blockade, appeared responsible for the flickering. The equilibrium constant of Na+ binding in the pore was approximately 150 mM at 0 mV and voltage dependent. The binding site appeared to be approximately 0.2 of the electrical distance from the extracellular surface. The mean channel current and the excess noise had the selectivity: K+ > Rb+ > Cs+ > Na+ > Li+. ATP increased the probability of being open (Po) to a maximum of 0.6 with an EC50 of 11.2 microM and a Hill coefficient of 2.3. Lowering extracellular pH enhanced the apparent affinity of the channel for ATP with a pKa of approximately 7.9, but did not cause a proton block of the open channel. High pH slowed the rise time to steps of ATP without affecting the fall time. The mean single channel amplitude was independent of pH, but the excess noise increased with decreasing pH. Kinetic analysis showed that ATP shortened the mean closed time but did not affect the mean open time. Maximum likelihood kinetic fitting of idealized single channel currents at different ATP concentrations produced a model with four sequential closed states (three binding steps) branching to two open states that converged on a final closed state. The ATP association rates increased with the sequential binding of ATP showing that the binding sites are not independent, but positively cooperative. Partially liganded channels do not appear to open. The predicted Po vs. ATP concentration closely matches the single channel current dose-response curve.

Adenosine Triphosphate↗

PubMed1999

Instability of the EPM1 minisatellite.

Inherited mutations in the cystatin B gene ( CSTB ) are responsible for progressive myoclonus epilepsy type 1 (EPM1; MIM 254800). This autosomal recessive disease is characterized by variable progression to mental retardation, dementia and ataxia. The majority of EPM1 alleles identified to date contain expansions of a dodecamer repeat located upstream of the transcription start site of the CSTB gene. Normal alleles contain two or three copies of the repeat, whereas pathogenic alleles contain >40 repeats. We examined the meiotic stability of pathogenic, expanded EPM1 alleles from 17 EPM1 families by employing a fluorescence-based PCR-based genotyping assay capable of detecting single dodecamer repeat unit differences on an automated DNA sequencer. We followed 74 expanded allele transmissions to 30 affected individuals and 22 carriers. Thirty-five of 74 expanded allele transmissions demonstrated either contraction or expansion of the minisatellite, typically by a single repeat unit. Thus expanded alleles of the EPM1 minisatellite demonstrate a mutation rate of 47%, the highest yet observed for pathogenetic alleles of a human minisatellite.

Alleles↗

PubMed1999

Distinct mutation patterns of breast cancer-associated alleles of the HRAS1 minisatellite locus.

DNA sequence analysis of 130 alleles of the HRAS1 minisatellite has demonstrated that breast cancer-associated variants arise as a consequence of both replication errors and gene conversions. Unlike mutations at other variable number of tandem repeats (VNTRs), high-risk variants of the HRAS1 minisatellite do not demonstrate positional polarity. Instead, most mutations occur at three hotspots, with replication errors confined to one hotspot, gene conversions to a second and a mixed pattern of mutation at the third. DNA sequence analysis of 66 low-risk a1 alleles revealed no evidence for hypermutation. Therefore, while the HRAS1 minisatellite may serve as a reporter for a broad-based group of mutational mechanisms, these results are consistent with a direct pathogenetic contribution by high-risk alleles as the biological basis underlying cancer association of this VNTR.

Alleles↗

PubMed1999

Cloning and expression of the Ca2+ channel alpha1C and beta2a subunits from guinea pig heart.

Complimentary DNA clones encoding the alpha1C and beta2a subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2,169 amino acids for the alpha1C and 597 amino acids for the beta2a subunit. The proteins showed 94.2 and 94.8%, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig alpha1C and beta2a subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the alpha1C subunit is expressed exclusively in the heart, while the beta2a subunit is expressed in the heart, cerebellum, whole brain, and stomach. The alpha1C and beta2a subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing alpha1C alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of beta2a with alpha1C did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig alpha1C and rabbit beta1+alpha2/delta, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced 3-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.

Amino Acid Sequence↗

PubMed1999

[Biological properties of reconstructed mouse hematopoiesis with hematopoietic stem cells transfected by bcl-2 gene].

OBJECTIVE: To observe resistance to radiochemical agents in the mice reconstructed hematopoiesis with hematopoietic stem cells transfected by bcl-2 gene. METHODS: The plasmid containing bcl-2 cDNA retrovirus vector was transfected to PA317 cells with Lipofecaine kite. A PA317 cell clone(D3) with high virus titer was obtained through G418 selection. The virus supernatant of D3 clone cells was used to transfect mouse embryo-liver hematopoietic stem cells. Hematopoiesis in the mice treated with lethal dose irradiation was reconstructed by hematopoietic stem cells transfected with bcl-2 gene. The resistance to radiochemical agents in the mice with reconstructed hematopoiesis was evaluated according to the number of WBC of periphereal blood and mononuclear cells of bone marrow, CFU-S and CFU-S regeneration. RESULTS: The expression of bcl-2 protein was increased in the hematopoietic cells transfected by bcl-2 gene. The mice reconstructed hematopoiesis by stem cells transfected bcl-2 gene can tolerate high dose combined with chemotherapy and radiation of middle dose. The mice reconstructed hematopoiesis with stem cells transfected bcl-2 gene survived over one year, no tumors were found. CONCLUSION: The mice with reconstructed hematopoiesis by stem cells carried bcl-2 gene possess the ability of resistance to radiochemical agents, even only part of hematopoietic stem cells were transfected with bcl-2 gene.

Animals↗

PubMed1999

[Purification and properties of glutamate dehydrogenase from Pseudomonas pseudoalcaligenes].

Glutamate dehydrogenase was purified from the crude extract of Pseudomonas pseudoal-caligenes. The enzyme had a molecular weight of 290,000 and was composed of six subunits with identical molecular weight of 47,000. The enzyme was highly specific for NADP(H) and the substrates. The biochemical properties such as kinetic parameters and heat stability were also examined. The purified GDH showed considerable loss of activity upon freezing.

Glutamate Dehydrogenase↗

PubMed1998

[Heat strain for personnel wearing three levels of protective ensembles at moderate and heavy workloads].

The effects of individual protective ensembles and workloads on personnel's physiological heat strain were experienced by personnel in a hot environment (35 degrees C, 55% relative humidity). Six young males performed treadmill tests at both moderate (ML: 1.11 m/s, 5% grade) and heavy (HL: 1.34 m/s, 5% grade) workloads. They wore three levels of protective ensembles: protective mask only (PM), protective clothing only (PC), and full protective ensembles (FP). Tests lasted 60 min and 45 min, respectively, for ML and HL. Physiological measurements included heart rate (HR), rectal (Tre) and skin (Tsk) temperature, sweat rate (SR), and sweat evaporative rate (ER). Differences between the ensembles and the workloads were significant for HR responses. The increase rate in Tre and Tsk, and SR, ER were significantly reduced from PM and PC to FP condition during ML and Hl tests. The sweat evaporation restriction of ensembles and the increase of heat production from exercise are the main factors of enhancing the heat strain.

Adult↗