Search PubMed⌕ Search

Biomedical subjects

J Cheng

Publications and source records attributed to J Cheng.

At least 343 records · Page 19Linked to original sources
PubMed1996

Microchip-based Devices for Molecular Diagnosis of Genetic Diseases.

Microchips, constructed with a variety of microfabrication technologies (photolithography, micropatterning, microjet printing, light-directed chemical synthesis, laser stereochemical etching, and microcontact printing) are being applied to molecular biology. The new microchip-based analytical devices promise to solve the analytical problems faced by many molecular biologists (eg, contamination, low throughput, and high cost). They may revolutionize molecular biology and its application in clinical medicine, forensic science, and environmental monitoring. A typical biochemical analysis involves three main steps: (1) sample preparation, (2) biochemical reaction, and (3) detection (either separation or hybridization may be involved) accompanied by data acquisition and interpretation. The construction of a miniturized analyzer will therefore necessarily entail the miniaturization and integration of all three of these processes. The literature related to the miniaturization of these three processes indicates that the greatest emphasis so far is on the investigation and development of methods for the detection of nucleic acid, followed by the optimization of a biochemical reaction, such as the polymerase chain reaction. The first step involving sample preparation has received little attention. In this review the state of the art of, microchip-based, miniaturized analytical processes (eg, sample preparation, biochemical reaction, and detection of products) are outlined and the applications of microchip-based devices in the molecular diagnosis of genetic diseases are discussed.

Journal Article↗
PubMed1996

The inversion of the exponential Radon transform for quantitative brain SPECT.

The mathematical derivation for the inversion of the exponential Radon transform is presented and the implementation of the inversion is detailed. The inversion can be verified and implemented by the readers for practical applications, such as for quantitative reconstruction of brain SPECT (single-photon emission computed tomography).

Brain↗
PubMed1996

Evaluation of the Technicon Immuno 1 free thyroxine assay.

The Technicon Immuno 1 free thyroxine (fT4) assay, a modified two-step procedure that is fully automated on a random access analyzer, was evaluated-at two clinical sites. The method had excellent precision and correlated well overall with three other estimates of free thyroxine: free thyroxine index (FTI) measured on the Immuno 1; the Abbott IMx fT4 assay (Abbott Park, IL); and the Clinical Assays two-step manual fT4 assay. Using a combination of thyrotropin and FTI assay results as a "gold standard" for defining thyroid status, the Immuno 1 fT4 method had a sensitivity of 100% and specificity of 98.3% for hyperthyroidism, versus 93.8% and 99.3%, respectively, for hypothyroidism. In conclusion, the Immuno 1 fT4 assay is useful in screening for thyroid disease.

Autoanalysis↗
PubMed1996

Genetic analysis of a severe case of Dowling-Meara epidermolysis bullosa simplex.

The epidermis serves an important protective function, which it manifests by producing an extensive cytoskeletal architecture, the unique feature of which are keratin filaments. Through studies that began with epidermolysis bullosa simplex (EBS) and now extend to a group of autosomal dominant human blistering skin disorders it was discovered that defects in the keratin genes lead to cell fragility and degeneration upon mechanical trauma. In most cases of EBS, point mutations occur in the keratin 5 (K5) and K14 genes expressed in the basal layer of the epidermis. The precise location of the mutation and the degree to which it causes perturbations in filament assembly correlate with disease severity. In the present study, we examine a case of EBS, which clinically lies at the severe end of the spectrum of Dowling-Meara EBS and which shows keratin filament clumping in suprabasal as well as basal cells. We show that one of the two K14 alleles has a single point substitution, giving rise to a Y129D mutation. This mutation resides 4 residues internal to the R125C/H hotspot known to account for the majority of Dowling-Meara cases. We provide functional and structural evidence to suggest why the Y129D mutation may be capable of creating such a severe form of EBS.

Adult↗
PubMed1996

Pseudocyst formation by adenoid cystic carcinoma cells in collagen gel culture and in SCID mice.

In order to reconstruct the characteristic three-dimensional architecture of adenoid cystic carcinoma, we cultured ACC2 cells, a cell system established from a human adenoid cystic carcinoma of the palate, in collagen gel matrix and transplanted them in SCID mice. In the collagen gel culture, the cells formed spherical colonies measuring 75.6 +/- 14.6 microns in diameter by 6 days after seeding. The tumor cell nests contained vacuolar structures that were immunopositive for heparan sulfate proteoglycan, type III collagen, type IV collagen, and fibronectin. The rim of the nests was argyrophilic and immunopositive for type I collagen, type IV collagen, laminin, and fibronectin. Transplants of ACC2 cells in SCID mice grew to form tumor masses in which pseudocysts were formed. The results indicate that our collagen gel culture system provides physiological conditions for ACC2 cells to secrete particular extracellular matrix molecules and form pseudocystic spaces.

Animals↗
PubMed1996

Na+/H+ antiport activity conferred by Bacillus subtilis tetA(L), a 5' truncation product of tetA(L), and related plasmid genes upon Escherichia coli.

An Escherichia coli transformant expressing the Bacillus subtilis tetA(L) gene from a weak promoter was challenged by growth on medium with low, increasing tetracycline concentrations. Changes in the substrate preference ratios of the TetA(L)-mediated resistances and antiports were examined in view of recent findings suggesting that TetA(L) catalyzes efflux of Na+ in exchange for protons in addition to having the ability to catalyze metal-tetracycline/H+ antiport. After growth of the transformant on 1 microgram or more of tetracycline per ml for 12 to 15 h, the tetA(L) gene in the plasmid was found to be disrupted by an IS10 element 50 bp from the 5' end of the coding sequence. This disrupted recombinant plasmid, pKB1, conferred greater tetracycline resistance and higher levels of membrane metal-tetracycline/proton antiport than the original plasmid, pJTA1, but conferred lower NA+ resistance and Na+/H+ antiport levels than the original plasmid. The results indicate that the 5' end of the gene is necessary for optimal Na+/H+ antiport but that some such activity as well as robust tetracycline/H+ antiport persists in its absence. Two plasmid genes, tet(K) and qacA, were compared with tetA(L) vis-à-vis their abilities to enhance the Na+/H+ antiporter activity of everted vesicles from E. coli transformants. tet(K), which is more closely related to tetA(L), catalyzed 22Na+ uptake by energized vesicles, whereas the less closely related qacA gene did not.

Bacillus subtilis↗
PubMed1996

Chromosomal tetA(L) gene of Bacillus subtilis: regulation of expression and physiology of a tetA(L) deletion strain.

Deletion of the tetA(L) chromosomal region of Bacillus subtilis in a strain designated JC112 increased the strain's sensitivity to low tetracycline concentrations. It also resulted in phenotypic changes that correlate with the previously found role of TetA(L) in mediating electrogenic NA+/H+ antiport. Growth of JC112 was impaired relative to that of the wild type at both pH 7.0 and 8.3; Na(+)- and K(+)-dependent pH homeostases were impaired at alkaline pH. The phenotype of JC112 was complemented by plasmid-borne tetA(L) and related tet(K) genes; the antiport activity conferred by the tet(K) gene had an apparently higher preference for K+ over Na+ than that conferred by tetA(L). The data were consistent with TetA(L) being the major Na+(K+)/H+ antiporter involved in pH homeostasis in B. subtilis as well as a significant Na+ extrusion system. The phenotype of JC112 was much more pronounced than that of an earlier transposition mutant, JC111, with a disruption in the putative tetA(L) promoter region. Northern (RNA) blot analysis of tetA(L) RNA from wild-type and JC111 strains revealed the same patterns. That JC111 nevertheless exhibited some Na+ and alkali sensitivity may be accounted for by disruption of regulatory features that, in the wild type, allow increased tetA(L) expression under specific conditions of pH and monovalent cation concentration. Evidence for several different regulatory effects emerged from studies of lacZ expression from the transposon of JC111 and from a tetA(L)-lacZ translational fusion introduced into the amyE locus of wild-type and JC112 strains.

Anti-Bacterial Agents↗
PubMed1996

Common core sequences are found in skeletal muscle slow- and fast-fiber-type-specific regulatory elements.

The molecular mechanisms generating muscle diversity during development are unknown. The phenotypic properties of slow- and fast-twitch myofibers are determined by the selective transcription of genes coding for contractile proteins and metabolic enzymes in these muscles, properties that fail to develop in cultured muscle. Using transgenic mice, we have identified regulatory elements in the evolutionarily related troponin slow (TnIs) and fast (TnIf) genes that confer specific transcription in either slow or fast muscles. Analysis of serial deletions of the rat TnIs upstream region revealed that sequences between kb -0.95 and -0.5 are necessary to confer slow-fiber-specific transcription; the -0.5-kb fragment containing the basal promoter was inactive in five transgenic mouse lines tested. We identified a 128-bp regulatory element residing at kb -0.8 that, when linked to the -0.5-kb TnIs promoter, specifically confers transcription to slow-twitch muscles. To identify sequences directing fast-fiber-specific transcription, we generated transgenic mice harboring a construct containing the TnIs kb -0.5 promoter fused to a 144-bp enhancer derived from the quail TnIf gene. Mice harboring the TnIf/TnIs chimera construct expressed the transgene in fast but not in slow muscles, indicating that these regulatory elements are sufficient to confer fiber-type-specific transcription. Alignment of rat TnIs and quail TnIf regulatory sequences indicates that there is a conserved spatial organization of core elements, namely, an E box, a CCAC box, a MEF-2-like sequence, and a previously uncharacterized motif. The core elements were shown to bind their cognate factors by electrophoretic mobility shift assays, and their mutation demonstrated that the TnIs CCAC and E boxes are necessary for transgene expression. Our results suggest that the interaction of closely related transcriptional protein-DNA complexes is utilized to specify fiber type diversity.

Animals↗
PubMed1996

Changes in action potentials and ion currents in long-term cultured neonatal rat ventricular cells.

A primary culture of neonatal ventricular myocytes isolated from day-old rats was established for investigating the changes in action potentials and ion currents over long periods. Cells at days 5 and 15 in culture were studied. These changes in vitro were compared with those in situ derived from the age-matched freshly isolated cells. During primary culture, quiescent cells demonstrated shortening of action potential durations (APD) resembling the developmental changes observed in situ. The beating cultured cells were not associated with APD shortening. Despite constant current amplitudes, the densities of Ca2+ currents (ICa) decreased in the quiescent cultures at later ages as a result of cell enlargement. ICa densities were maintained in the beating cultured and freshly isolated cells. Acceleration in the inactivation of ICa was observed during developments both in vitro and in situ. In addition, the densities of transient outward currents (Ito) tripled and doubled in the quiescent and beating cells during 15-day cultures. However, Ito in beating cultured cells made less contribution to APD in contrast to the quiescent cultured and freshly isolated myocytes. These findings demonstrate that electrophysiological properties differ between two types of long-term cultured cells. ICa densities remained constant in the beating cultures, suggesting that cell beating may be required for the maintenance of ICa density in developing cardiomyocytes.

Action Potentials↗
PubMed1996

Prostaglandin F2 alpha induces cardiac myocyte hypertrophy in vitro and cardiac growth in vivo.

Several prostaglandins [prostaglandin (PG) A2, -B2, -D2, -E2, -F2 alpha, and -I2 and carbaprostacyclin] and the thromboxane analogue U-46619 were analyzed for the ability to induce hypertrophy of rat neonatal cardiac ventricular myocytes. Myocyte hypertrophy was induced specifically by PGF2 alpha. Myocytes exposed to this prostanoid in culture increased in size and protein content. The contractile fibrils within the cells became organized into parallel arrays, and the cells tended to cluster and beat spontaneously. PGF2 alpha also induced the expression of c-fos, atrial natriuretic factor (ANF), and alpha-skeletal actin in these cells. The effects of PGF2 alpha were compared with several known cardiac myocyte hypertrophy factors (phenylephrine, endothelin-1, leukemia inhibitory factor, cardiotrophin-1, and angiotensin II). PGF2 alpha was found to be intermediate in potency among the factors but induced a level of ANF production that was approximately 10-fold higher than any of the other effectors. Responsiveness to PGF2 alpha was not limited to neonatal cardiocytes. Ventricular myocytes isolated from adult rats also responded specifically to PGF2 alpha with a morphological change similar to that observed with phenylephrine and by producing ANF. In rats, chronic administration of fluprostenol, a potent agonist analogue of PGF2 alpha, resulted in a dose-dependent increase in heart weight- and ventricular weight-to-body weight ratios. The amount of PGF2 alpha extractable from the hearts of rats with cardiac hypertrophy induced by myocardial infarction was also found to be greater than that in sham-operated control rats. These results indicate that PGF2 alpha may play an important role in inducing cardiac hypertrophy.

Aging↗
PubMed1996

Dynamics and energetics of scallop locomotion

A dynamic model for a swimming scallop was developed which integrates the mechanical properties of the hinge ligaments, valve inertia, the external fluid-flow reaction, the fluid pressure in the mantle cavity and the muscle contraction. Kinematic data were recorded for a swimming Placopecten magellanicus from high-speed film analysis. Dynamic loading experiments were performed to provide the required mechanical properties of the hinge for the same species. The swimming dynamics and energetics based on data from a 0.065 m long Placopecten magellanicus at 10 °C were analyzed. The main conclusions are as follows. 1. The mean period of a clapping cycle during swimming is about 0.28 s, which can be roughly divided into three equal intervals: closing, gliding and opening. The maximum angular velocity and acceleration of the valve movements are about 182 degrees s-1 and 1370 degrees s-2, respectively. 2. The hysteresis loop of the hinge was found to be close to an ellipse. This may be represented as a simple Voigt body consisting of a spring and dashpot in parallel, with a rotational stiffness of 0.0497 N m and viscosity coefficient of 0.00109 kg m2 s-1 for the 0.065 m long Placopecten magellanicus. 3. The external fluid reaction has three components, of which the added mass is about 10 times higher than the mass of a single valve, and the flow-induced pseudo-viscosity compensates for nearly half of the hinge viscosity for the 0.065 m long Placopecten magellanicus. 4. The locomotor system powered by the muscle can be divided into two subsystems: a pressure pump for jet production and a shell-hinge/outer-fluid oscillator which drives the pumping cycle. The dynamics of the oscillator is determined predominantly by the interaction of the external fluid reaction and the hinge properties, and its resonant frequency was found to be close to the swimming frequencies. 5. The momentum and energy required to run the oscillator are negligibly small (about 1 % for the 0.065 m long Placopecten magellanicus) compared with that for the jet. Almost all the mechanical energy from muscle contraction is used to perform hydrodynamic work for jet production. Thus, the Froude efficiency of propulsion in scallops is nearly the same as the entire mechanical efficiency of the locomotor system. This could be a fundamental advantage of jet propulsion, at least for a scallop. 6. The estimated maximum muscle stress is about 1.06x10(5) N m-2, the cyclic work is 0.065 J and power output is 1.3 W. Using an estimate of the mass of an adductor muscle, the work done by the muscle per unit mass is 9.0 J kg-1 and the peak power per unit mass is 185 W kg-1. 7. The time course of the force generation of the contracting adductor muscle is basically the same as that of the hydrodynamic propulsive force.

Journal Article↗
PubMed1996

Differential effects of MS-551 and E-4031 on action potentials and the delayed rectifier K+ current in rabbit ventricular myocytes.

OBJECTIVE: The frequency-dependent effects of MS-551 on the action potential duration (APD) and the underlying ionic mechanisms were investigated in comparison with those of E-4031. METHODS: Whole-cell clamp techniques were used to study action potentials and ionic currents in enzymatically isolated rabbit ventricular myocytes. RESULTS: The frequency-response obtained within the range of 0.1 to 3.3 Hz was different for MS-551 and E-4031. The APD prolongation by MS-551 (10 microM) was significant at 0.5-3.3 Hz, whereas that by E-4031 (1 microM) was significant at 0.1-1.0 Hz. The prolongation by MS-551 (10 microM) of APD of a test action potential, which was preceded by a train of 1.0 Hz stimulation, decreased progressively as the rest duration increased, whereas that by E-4031 (1 microM) remained at the same level. Both MS-551 (10 microM) and E-4031 (1 microM) significantly decreased IK, but showed no effects on the transient outward current (Ito) and the inward rectifier K+ current (IKl). The development of the block on IK and the recovery from the block by MS-551 were voltage dependent. At a holding potential of -50 mV, MS-551 reduced the tail current to a similar extent (21-34%, n = 6) across all the tested durations of the depolarizing pulses to +10 mV, whereas at -75 mV, the intensity of the block progressively increased as the durations of depolarizing pulses were prolonged. The recovery from the block by MS-551 was absent at -50 mV, but occurred at -75 mV with a time constant of 577 +/- 179 ms (n = 6). The development of the block on IK by E-4031 was voltage and time independent. No recovery from the block was observed for E-4031 at either -50 or -75 mV. CONCLUSIONS: These findings suggest that MS-551 produces frequency-dependent class III action, presumably due to the voltage-dependent binding and unbinding to the IK channels. The reverse frequency dependence of class III action by E-4031 cannot be explained by the effects on IK.

Action Potentials↗
PubMed1996

Autoimmune disease results from multiple interactive defects in apoptosis induction molecules and signaling pathways.

Activation induced cell death (AICD) plays a critical role in eliminating autoimmune cells and limiting inflammation after activation. The two major signaling molecules for AICD are the Fas and TNF-R pathways of apoptosis. Defective Fas apoptosis in lpr/lpr mice results in a compensatory increase in TNF-R/TNF-mediated apoptosis. TNF/TNF-R has been shown to be a compensatory pathway of apoptosis in T cells and macrophages of lpr/lpr mice. Therefore, early production of TNF/TNF-R limit an immune response by inducing AICD in the absence of an intact Fas/Fas ligand apoptosis pathway. However, increased TNF production in lpr mice also lead to increased susceptibility to septic shock and autoimmune disease such as arthritis. Therefore TNF production during an inflammatory response can downmodulate this response, but this also results in the failure to downmodulate TNF production leading to septic shock and arthritis. A second pathway of AICD is mediated by Nur77 after T cell stimulation through the CD3 molecule. Mice with defective Nur77 signaling undergo AICD using the Fas-Fas ligand pathway to eliminate autoreactive T cells. A third defect of AICD is observed in HCP-mutant me/me (motheaten) mice which develop autoimmune disease related to defective Fas apoptosis signaling. Therefore, multiple interactive pathways play a role in limiting development of autoimmunity.

Animals↗
PubMed1996

Endoscopic stenting in the management of biliary stones.

INTRODUCTION: Endoscopic sphincterotomy and stenting are established modes of treatment in the management of bile duct stones and cholangitis. It is the first choice of treatment when the patients are elderly and have other medical conditions rendering them unfit for surgery. AIMS: We studied the immediate and long-term outcome of endoscopic stenting in patients who underwent Endoscopic Retrograde Cholangiopancreatography (ERCP) for ductal stones. PATIENTS AND METHODS: From January 1990 to December 1992, 366 patients had ERCP done for biliary stone disease. Fifty-five of these patients received a 10 French biliary stent for the treatment of bile duct stones. Most of these patients (60%) had an advanced age of more than 70 years and 45% had co-existing medical conditions rendering them high risk for surgery. Ninety-one percent had stones greater than 1.2 cm in diameter and 82% had multiple stones. RESULTS: Eighty-nine percent of the patients had relief of jaundice and cholangitis resolved in 96% of the patients after stenting. None required immediate surgery after ERCP. Temporary bile drainage was achieved in 19 patients who went for surgery at a later date when they became more stable. Twenty-nine patients were on long-term follow-up for a mean period of 13 months. Nineteen patients still have their stents in place and remained well. The stents were repeatedly changed in 15 patients after an average duration of 5.5 months. Late complications were cholangitis in 2 patients and stent migration in one patient. There was no related mortality. In 5 patients, the stones have either disappeared spontaneously or become smaller and have been removed at subsequent ERCP. CONCLUSION: Endoscopic stenting is a valuable alternative to surgical bile duct exploration. It can be the definitive mode of treatment for large or multiple bile duct stones in the elderly and patients with multiple medical problems who are unfit for surgery.

Adult↗
PubMed1996

[Effect of electroacupuncture on cholecystokinin gene expression in rat hippocampus during penicillin-induced epileptic seizure].

Northern Blot and hybrization in situ techniques were used to investigate the effect of electroacupuncture (EA) on the changes of cholecystokinin (CCK) mRAN levels of the hippocampus in rat penicillin-induced epilepsy model. Epilepsy can significantly increase CCK mRNA levels in dentate gyrus and CA3 areas of hippocampus in diencephalic sections after penicillin-induced seizure, whereas EA not only can attenuate the seizure behaviors and EEG changes, but also can decrease the increase of CCK mRNA contents induced by the seizure. However, in the subiculum, dentate gyrus and CA3 areas of mesencephalic sections of rat hippocampus, EA can further increase the enhancement of CCK mRNA concentration induced by penicillin-induced seizure. The results suggest that EA inhibitory effects on the seizure's behaviors and epileptiform activities may be related to the alteration of CCK gene expression in the different area of hippocampus.

Animals↗
PubMed1996

[The research of radioimmunoassay using double Abs for bradykinin].

Bradykinin (BK) is a potent vasodilative substance, and plays great physiological and pathological roles in animals and human beings. To measure the quantity of BK, the radioimmunoassay (RIA) has been devised, but the traditional RIA method has certain defects, such as presence of numerows interfering factors and errors and time consuming. Now, we produce anti-BK serum in rabbits by using BK-ovalbumin conjugate as an immunogen, and the 125I labeled Tyr8-BK by using a modified chloramine-T method. High specific activity has been obtained after purification with DEAE-Sephadex A-25 column chromatography. We use the donkey anti-rabbit Ab and PEG 6000 to separate the bound from the free 125I-Tyr8-BK. The limitation range of standard curve is from 25 to 1600 pg, NSB is 3.1% affinity constant (K) is 0.8 x 10(10) L/mol, and there is no significant interference with other biological BK analogues. The blood samples are treated by adding Polybrene (inhibitor) and PEG 6000 to deposit the big serum proteins in order to reduce the disturbing substances. This method has been shown to be a sensitive, specific, reliable, simple and convenient measure of the serum BK level. By this method, the serum BK quantities in men, women and rats are respectively 1584 +/- 347 pg/ml, 1642 +/- 302 pg/ml and 1805 +/- 225 pg/ml, the recycling rate is 95%, the intergroup CV is 5.0% and outergroup CV = 9.2%.

Animals↗
PubMed1996

[Effects of gossypol acetic acid on human ejaculated sperm Ca2+ influx and rat vas deferens contraction action].

It has been reported that gossypol acetic acid (GAA) inhibits sperm motility and calcium plays an important role in regulating the function of the sperm flagella. In order to explore the mechanism by which GAA inhibits sperm motility, the effects of GAA on human ejaculated sperm Ca2+ influx and isolated rat vas deferens contraction action were studied. The results showed that GAA inhibited sperm Ca2+ influx in a dose-dependent manner, and breaked the dynamic equilibrium of sperm internal and external Ca2+ gradient concentrations. It may be one of important mechanism by which GAA inhibits sperm motility. But GAA exerts no effect on the high K+ induced contraction of smooth muscle of isolated rat vas deferens. It is likely that GAA has selective action on the sperm membrane of the rats.

Adult↗