Biomedical subjects

D S Hwang

Publications and source records attributed to D S Hwang.

At least 19 recordsLinked to original sources

PubMed2001

Expression of the seqA gene is negatively modulated by the HU protein in Escherichia coli.

The SeqA protein acts as a regulator of chromosomal replication initiation in Escherichia coli by sequestering hemi-methylated oriC, effectively blocking methylation and therefore preventing rapid re-initiation. The level of SeqA protein is maximal at mid-log phase and decreases when cells enter late-log phase. In hup mutants that lack the HU protein, the maximal seqA expression is also seen at mid-log phase, but seqA expression, as well as SeqA levels and activity, is increased by up to four fold relative to that in the wild type. These results suggest that the HU protein functions as a negative modulator of seqA expression.

Bacterial Outer Membrane Proteins↗

PubMed2000

The arc two-component signal transduction system inhibits in vitro Escherichia coli chromosomal initiation.

Under anaerobic growth conditions, Escherichia coli operates a two-component signal transduction system, termed Arc, that consists of ArcB protein, a transmembrane sensor kinase and ArcA protein, the cognate response regulator. In response to low oxygen levels, autophosphorylated ArcB phosphorylates ArcA, and the resulting phosphorylated ArcA (ArcA-P) functions as a transcriptional regulator of the genes necessary to maintain anaerobic growth. Under anaerobic conditions, cells maintain a slow growth rate, suggesting that the initiation of chromosomal replication is regulated to reduce the initiation frequency. DNase I footprinting experiments revealed that ArcA-P binds to the left region of the chromosomal origin, oriC. ArcA-P did not affect the in vitro replication of plasmid DNA containing the ColE1 origin nor the in vitro replication of viral DNAs; however, ArcA-P specifically inhibited in vitro E. coli chromosomal replication. This inhibition was caused by the prevention of open complex formation, a necessary step in the initiation of chromosomal replication. Our in vitro results suggest that the Arc two-component system participates in regulating chromosomal initiation under anaerobic growth conditions.

Bacterial Outer Membrane Proteins↗

PubMed2000

High temperature stress resistance of Escherichia coli induced by a tobacco class I low molecular weight heat-shock protein.

TLHS1 is a class I low molecular weight heat-shock protein (LMW HSP) of tobacco (Nicotiana tabacum). For a functional study of TLHS1, a recombinant DNA coding for TLHS1 with a hexahistidine tag at the amino-terminus was constructed and expressed in Escherichia coli. An expressed fusion protein, H6TLHS1, was purified using a Ni2+ affinity column and a Sephacryl S400 HR column. A polyclonal antibody against H6TLHS1 was produced to follow the fate of H6TLHS1 in E. coli. The fusion protein in E. coli maintained its solubility at a temperature of up to 90 degrees C and most of the proteins in the E. coli cell lysate with H6TLHS1 were prevented from thermally induced aggregation at up to 90 degrees C. We compared the viability of E. coli cells expressing H6TLHS1 to the E. coli cells without H6TLHS1 at a temperature of 50 degrees C. After 8 h of high temperature treatment, E. coli cells with H6TLHS1 survived about three thousand times more than the bacterial cells without H6TLHS1. These results showed that a plant class I LMW HSP, TLHS1, can protect proteins of E. coli from heat denaturation, which could lead to a higher survival rate of the bacterial cells at high temperature.

Cloning, Molecular↗

PubMed2000

Involvement of retinoblastoma protein in p27Kip1-induced apoptosis.

p27Kip1, a cyclin-dependent kinase (CDK) inhibitor, plays a critical role in cell cycle regulation. Expression of p27Kip1 is shown to increase during apoptosis in mammalian cells. Here, to directly address the role of p27Kip1 in apoptosis, p27Kip1 is overexpressed in human SK-Hep1 hepatoma cells. This leads to apoptotic cell death and this reduces protein, but not mRNA, levels of the retinoblastoma (Rb). Consistently, accumulation of Rb protein blocks p27Kip1-mediated apoptosis. These studies demonstrate an involvement of Rb in the apoptotic cell death which is induced by overexpression of p27Kip1.

Apoptosis↗

PubMed2000

Quantitative analysis of ultrahigh molecular weight polyethylene (UHMWPE) wear debris associated with total knee replacements.

The size and morphology of particulate wear debris retrieved from tissues around 18 failed total knee replacements (TKR) were characterized. Interfacial membranes from nine cemented and nine uncemented TKR were harvested from below the tibial components during revision surgery. Wear debris were extracted using papain and potassium hydroxide digestion. Ultrahigh molecular weight polyethylene (UHMWPE) particles from around cemented or uncemented TKR were similar in size and morphology. The mean size was 1.7 +/- 0. 7 microm with a range of 0.1-18 microm. Thirty-six percent of the particles were less than 1 microm and 90% were less than 3 microm. Morphologically the particles were predominantly spherical with occasional fibrillar attachments and flakes. Particles from TKR were greater than threefold larger than previously characterized particles from total hip replacements, which were 0.5 microm in mean size. Differences in joint conformity and wear patterns between the hip and knee articulations may explain the disparity in size of the wear debris. Since particle size represents an important variable influencing the magnitude of the biological response, it is possible that in vivo the larger TKR debris results in a diminished mediator release, which in turn may account for the lower incidence of osteolysis and aseptic loosening in some designs of TKR.

Aged↗

PubMed1999

Interaction of SeqA and Dam methylase on the hemimethylated origin of Escherichia coli chromosomal DNA replication.

Preferential binding of SeqA protein to hemimethylated oriC, the origin of Escherichia coli chromosomal replication, delays methylation by Dam methylase. Because the SeqA-oriC interaction appears to be essential in timing of chromosomal replication initiation, the biochemical functions of SeqA protein and Dam methylase at the 13-mer L, M, and R region containing 4 GATC sequences at the left end of oriC were examined. We found that SeqA protein preferentially bound hemimethylated 13-mers but not fully nor unmethylated 13-mers. Regardless of strand methylation, the binding of SeqA protein to the hemimethylated GATC sequence of 13-mer L was followed by additional binding to other hemimethylated GATC sequences of 13-mer M and R. On the other hand, Dam methylase did not discriminate binding of 13-mers in different methylation patterns and was not specific to GATC sequences. The binding specificity and higher affinity of SeqA protein over Dam methylase to the hemimethylated 13-mers along with the reported cellular abundance of this protein explains the dominant action of SeqA protein over Dam methylase to the newly replicated oriC for the sequestration of chromosomal replication. Furthermore, SeqA protein bound to hemimethylated 13-mers was not dissociated by Dam methylase, and most SeqA protein spontaneously dissociated 10 min after binding. Also, SeqA protein delayed the in vitro methylation of hemimethylated 13-mers by Dam methylase. These in vitro results suggest that the intrinsic binding instability of SeqA protein results in release of sequestrated hemimethylated oriC.

Bacterial Outer Membrane Proteins↗

PubMed1999

PhoB-dependent transcriptional activation of the iciA gene during starvation for phosphate in Escherichia coli.

The IciA protein from Escherichia coli has been shown specifically to inhibit the in vitro initiation of chromosomal DNA replication. However, the in vivo role of IciA has not yet been established. In order to investigate the in vivo function of this protein, expression of the iciA gene was studied by monitoring the beta-galactosidase activity specified by an iciA promoter-lacZ fusion inserted into the chromosome. Among the conditions tested (carbon starvation, the stringent response, phosphate starvation, and the SOS response), only phosphate depletion increased iciA expression. Supplementation of phosphate-depleted cultures with inorganic phosphate reduced the beta-galactosidase activity to basal levels. Enhanced expression of iciA-lacZ was dependent upon the PhoB protein. PhoB is known to be a transcriptional activator of the Pho regulon, expression of which is activated during phosphate starvation. It was also found that the iciA promoter contains a PhoB protein-binding sequence, termed the Pho box, which is necessary for the activation of genes of the Pho regulon. These results suggest that the iciA gene is a member of the Pho regulon.

Bacterial Proteins↗

PubMed1999

Evaluation of arterial compliance in patients with carotid arterial atherosclerosis.

BACKGROUND: High-resolution peripheral vascular sonography has the capability to determine vessel compliance. A number of factors affect compliance in humans, including age, hypertension and atherosclerosis. However, compliance in patients with coronary artery disease (CAD) combined with carotid artery lesions has not been well studied. The objectives of this study were: 1) to determine whether carotid artery compliance is reduced in patients with CAD and carotid artery lesions, and 2) to determine whether decreased arterial compliance is related to an abnormality in vascular wall structure. METHODS: The study participants included 12 patients with CAD and carotid artery disease (group III), 13 patients with CAD only (group II) and 13 age-matched normal subjects who served as controls (group I). High-resolution peripheral vascular ultrasonography was performed to directly visualize the common carotid artery and to measure its diameter and wall thickness. Carotid artery elastic properties were determined relative to arterial diameter and pressure generated within the heart. RESULTS: Carotid artery walls were thicker in Group II and III patients than in normal subjects (1.49 +/- 0.03 mm, 1.98 +/- 0.04 mm, vs 1.22 +/- 0.03 mm, p < 0.05 and p < 0.01). There were significant differences in wall thickness between subgroups of CAD patients (p < 0.01). Carotid distensibility was lower (21.8 +/- 1.2 x 10(-6).N-1.m2, 14.8 +/- 1.7 x 10(-6).N-1.m2, vs 25.6 +/- 1.5 x 10(-6).N-1.m2; p < 0.05 and p < 0.01) and Young's modulus of elasticity was higher (3.12 +/- 0.43 x 10(5).Nm-2, 4.18 +/- 0.30 x 10(5).Nm-2, vs 2.34 +/- 0.23 x 10(5).Nm-2; p < 0.05 and p < 0.01) in group II and III patients than in normal subjects. These two indices of carotid artery compliance also differed between subgroups of CAD (p < 0.01). Increased wall thickness may occur secondary to changes in the architectural structure of the vessel wall, and these atherosclerotic changes lead to decreased compliance of arteries. CONCLUSION: The mechanical properties of the carotid arteries provide reliable information regarding changes caused by atherosclerosis.

Arteriosclerosis↗

PubMed1998

Effect of IciA protein on the expression of the nrd gene encoding ribonucleoside diphosphate reductase in E. coli.

The E. coli nrd operon contains the genes encoding the two subunits of ribonucleoside diphosphate reductase. We found that the IciA protein binds specifically to the AT-rich upstream region of nrd promoter. In vivo overexpression of IciA increases the expression of nrd gene by four- to five-fold, suggesting that IciA functions as a transcriptional activator for the nrd gene.

Bacterial Proteins↗

PubMed1998

Prediction of right ventricular infarction from standard surface ECG in patients with inferior myocardial infarction.

BACKGROUND: Patients with inferior myocardial infarction (MI) have a 45% chance of having concurrent right ventricular infarction (RVI); of these, 5-10% suffer hemodynamic collapse. Immediate correct diagnosis and appropriate management of such patients is vital. ST-segment elevation in the right precordial V4 lead (V4R) has a high diagnostic value in identifying RVI, but this determination requires additional time and cost. An attempt was made to use a collection of patients' standard surface electrocardiograms (ECG) to find any available data to detect RVI and to lead to a new way to diagnose RVI. METHODS: Fifty patients (males/females, 44/6; mean age, 64.3 +/- 6.9 years) with acute inferior myocardial infarction were enrolled in a first group to develop new diagnostic criteria for RVI. As a first step, the ST-segment change in every standard surface ECG lead was analyzed and compared with corresponding changes in V4R. RVI was diagnosed by typical clinical symptoms (chest pain for more than 30 minutes, ST elevation > 0.1 mV and enzyme changes) accompanying ST elevation of more than 0.1 mV in V4R (by Lopez-Sendon criteria) and echocardiographic findings. RVI was diagnosed in 24 (48%) patients using ECG. The new criteria were then tested in a secondary group of 48 patients (males/females, 43/5; mean age, 65.5 +/- 7.9 years) with inferior MI. RESULTS: Analysis of these patients found that ST depression in lead I and aVL was a specific characteristic of RVI (I + aVL > 0.2 mV). This criterion was applied to another group of patients with acute inferior MI to check the predictive value (sensitivity, 94.7%; specificity, 89.7%; positive predictive value, 85.7%; negative predictive value, 96.3%). CONCLUSIONS: In patients with evolving inferior MI, standard surface ECG analyzed for this criterion could aid clinical recognition of concomitant RVI.

Aged↗

PubMed1997

Safety and tolerability of cyclosporine microemulsion versus cyclosporine: two-year data in primary renal allograft recipients: a report of the Neoral Study Group.

BACKGROUND: The new microemulsion formulation of cyclosporine (CsA-ME) is more bioavailable than cyclosporine (CsA) in de novo renal transplant patients. Therefore, it was of interest to compare the safety profile of each formulation in such patients. METHODS: In a multicenter, double-blind, parallel-group study, 101 renal transplant recipients were randomized after transplantation to receive either CsA (n=50) or CsA-ME (n=51) capsules twice daily for 2 years. Of these patients, 54 (CsA, n=26; CsA-ME, n=28) completed 1 year of the study and entered the second-year, double-blind extension. Initial dose at the time of transplantation was 5 mg/kg b.i.d.; doses were titrated to target trough levels. METHODS: The mean (+/- SD) doses at the end of 2 years were 4.6 +/- 1.8 and 3.8 +/- 1.1 mg/kg per day for CsA- and CsA-ME-treated patients, respectively. The mean (+/- SD) CsA trough levels at end point were 187 +/- 63 and 210 +/- 95 ng/ml for CsA- and CsA-ME-treated patients, respectively. At least one adverse event was reported by 25/26 (96%) of CsA- and 27/28 (96%) of CsA-ME-treated patients. No patient discontinued the study because of adverse events. No deaths occurred during the study. Renal function, as measured by serum creatinine levels, and blood pressure were comparable over time in both treatment groups. CONCLUSIONS: There was no significant difference in safety and tolerability between CsA- and CsA-ME-treated kidney recipients for 2 years after transplantation.

Adolescent↗

PubMed1997

Occlusion of RNA polymerase by oligomerization of DnaA protein over the dnaA promoter of Escherichia coli.

DnaA protein, the initiator protein for initiation of Escherichia coli chromosomal replication, has been shown to repress its own expression from two dnaA promoters, 1P and 2P. The sequence-specific binding of DnaA protein to the DnaA box, located between the two promoters, results in subsequent oligomerization of DnaA protein. Upon increasing the concentration of DnaA protein, the oligomerization proceeds to both dnaA promoters from the DnaA box and inhibits RNA polymerase binding to both promoters. This results in the repression of transcription, suggesting that the extent of oligomerization of DnaA proteins over two dnaA promoters contributes to the autoregulation of expression of the dnaA gene. When the two dnaA promoters were bound and repressed by DnaA protein, the interaction of RNA polymerase with IciA protein, which is a specific inhibitor of initiation of in vitro E. coli chromosomal replication, appeared to dissociate the oligomerized DnaA proteins from the 1P promoter and allowed RNA polymerase to be loaded for its transcription.

Bacterial Proteins↗

PubMed1997

Description of blood pressure changes in patients beginning cyclosporin A therapy.

Cyclosporin A (CyA) is the primary immunosuppressive agent for the prophylaxis of rejection episodes in renal, cardiac, liver, and other transplants. Recently, its use in autoimmune diseases has been investigated as well. Although several studies have produced promising results, nephrotoxicity and hypertension can result from CyA treatment, and their development must be understood in order to facilitate patient management. This article describes the diastolic blood pressure (DBP) responses in two populations of patients during three months of CyA therapy. Study A involved psoriasis patients and Study B involved postoperative renal transplant patients. The relationship between blood pressure and systemic CyA exposure and other covariates was evaluated using linear mixed effects modeling. Temporal patterns of blood pressure changes with varying duration of CyA exposure were investigated. In Study A, the psoriasis patients showed transient exposure-related increases in DBP on CyA. These elevations, while statistically significant, were clinically insignificant. In Study B, the renal transplant patients showed no CyA-related rises in DBP. In neither study was there evidence for a difference in effect on DBP between Sandimmune and Neoral, the two formulations of CyA presently approved for marketing by the Food and Drug Administration, after differences in CyA exposure were taken into account.

Adult↗

PubMed1997

Histamine 2 receptor blocker-ranitidine and sinus node dysfunction.

BACKGROUND: Histamine 2 (H2) receptors were observed to be present in the sinus node. H2 receptor blockers could attenuate the histamine-induced increase in sinus rate and could thereby worsen sinus node dysfunction. However, the data pertaining to electrophysiological effects of H2 receptor blocker ranitidine on sinus node function are not available as yet, although bradycardia has been occasionally reported along with ranitidine. The present study evaluates the electrophysiological effects of ranitidine on sinus node function, and also determines the safety of administering ranitidine to patients suffering from sinus node dysfunction. METHODS: Thirty-nine patients with clinical diagnosis of sinus node dysfunction were selected for this study. Electrophysiological effects of intravenous injection of 150 mg ranitidine on sinus node function was evaluated. Tests were performed for 21 patients in the basal state (Group 1) and for 18 patients (Group 2) after autonomic blockade (propranolol 0.2 mg/kg, and atropine 0.04 mg/kg). The sinus cycle length (SCL), sinus node recovery time (SNRT), corrected sinus node recovery time (CSNRT), sinoatrial conduction time (SACT), atrio-His (AH) and His-ventricle (HV) intervals and blood pressure (BP) before and after ranitidine were analyzed. RESULTS: In Group 1, changes in SCL, SNRT, CSNRT, SACT, AH, HV and BP were not found to be significant. However, in Group 2, wherein the patients received ranitidine after autonomic blockade, it was observed that ranitidine significantly prolonged SCL (905.0 +/- 31.0 to 1000.0 +/- 40.0 msec, p = 0.001), decreased systolic BP (134.7 +/- 4.7 to 124.8 +/- 4.8 mmHg, p < 0.001), and diastolic BP (71.9 +/- 2.0 to 67.9 +/- 1.9 mmHg, p = 0.001), while the changes in SNRT, CSNRT, SACT, AH and HV were not significant. CONCLUSIONS: A clinical dose of 150 mg ranitidine must be administered by a slow intravenous infusion for at least 5 minutes in case of patients suffering from sinus node dysfunction. Continuous monitoring of BP and heart rate are necessary, especially in cases of autonomic denervation.

Adult↗

PubMed1996

Purification and characterization of mitochondrial ribonuclease P from Aspergillus nidulans.

Mitochondrial ribonuclease (RNase) P from Aspergillus nidulans was purified to near homogeneity using whole-cell extract as the starting material. A 4400-fold purification with a yield of 5.2% was achieved by ammonium sulfate fractionation, heat treatment, and five types of column chromatography, including tRNA-affinity column chromatography. This enzyme, which has a molecular mass of 232 kDa determined by glycerol gradient sedimentation analysis, appears to be composed of seven polypeptides and an RNA moiety. These seven polypeptides consistently copurified with the RNase P activity through two ion-exchange chromatography columns and in a glycerol gradient. As judged by nuclease sensitivity, the enzyme requires an RNA component for its activity. The 3'-end-labeled RNAs that copurified with the enzyme displayed identical sequences but had variable lengths for the 5' end, indicating that they originated from a common RNA molecule, the putative RNA component of RNase P. The purified enzyme cleaved mitochondrial precursor tRNAHis, resulting in an 8-bp acceptor stem. This implies that the purified RNase P is a mitochondrial enzyme and that an additional guanylate residue (at position -1) of tRNAHis in A. nidulans mitochondria is generated by a mode that is analogous to the generation of their counterparts in prokaryotes and chloroplasts.

Aspergillus nidulans↗

PubMed1996

Transcriptional activation of the dnaA gene encoding the initiator for oriC replication by IciA protein, an inhibitor of in vitro oriC replication in Escherichia coli.

Transcription of the Escherichia coli dnaA gene, encoding DnaA protein required for initiation of chromosomal DNA replication at oriC in E. coli, starts from two promoters, 1P and 2P. Gel-shift and DNase I-protection assays revealed that IciA protein, an inhibitor of initiation of in vitro E. coli chromosomal DNA replication at oriC, bound to two sites in the dnaA promoter region. One site is located upstream of promoter 1P, and the second is located downstream of promoter 2P. Whereas IciA protein did not affect transcription from the promoter 2P, transcription from the promoter 1P was specifically enhanced by IciA protein in vivo and in vitro. DnaA protein bound to the DnaA box between the two promoters 1P and 2P, acts as an transcriptional repressor. Under this condition, IciA protein counteracted the repressive effect of DnaA protein on the promoter 1P. These findings suggest that IciA protein may regulate the initiation of chromosomal DNA replication at oriC by controlling expression of the dnaA gene, as well as by inhibiting the initiation of chromosomal DNA replication at oriC.

Bacterial Proteins↗

PubMed1996

Evidence of genetic heterogeneity of hypertrophic cardiomyopathy in eight Chinese patients.

BACKGROUND: The genetic basis causing hypertrophic cardiomyopathy (HCM) was found due to missense mutations in cardiac beta-myosin heavy chain (beta-MHC), cardiac troponin T and alpha-tropomyosin genes in certain affected families. However, most mutations and majority of the affected families were reported to be related to beta-MHC gene. Till now, 20 different missense mutations of beta-MHC gene identified in more than 40 independent families were distributed in exons 8, 9, 13, 14, 15, 16, 19, 20, 21 and 23. Therefore, we chose these 10 exons for screening. METHODS: Eight probands with HCM and 1 normal control were included for screening. 32P-labeled PCR products of these 10 exons of beta-MHC gene were amplified from genomic DNA obtained from peripheral lymphocytes. PCR-DNA single strand conformation polymorphism (PCR-SSCP) analysis was performed using electrophoresis with polyacrylamide gels with and without 10% glycerol. Large amount copies of these 10 exons were also made from genomic DNA with PCR. Detection of sequencing variation of these exons was determined by the direct sequencing method with dideoxy chain termination method and 35S. RESULTS: No abnormal extra bands were noted on PCR-SSCP analysis. Sequencing analysis showed no missense mutation in these probands. CONCLUSIONS: Genetic heterogeneity of HCM is evident in Chinese patients with HCM.

Aged↗

PubMed1995

Stereospecific blocking effects of naloxone against hemodynamic compromise and ventricular dysfunction due to myocardial ischemia and reperfusion.

Endogenous opioid peptides subserve regulatory roles in cardiovascular function and are released upon myocardial ischemia contributing to the development of ischemic arrhythmias and cardiogenic shock, which are reversed by the opioid antagonist naloxone. Since the hallmark of myocardial infarction is the impairment of hemodynamics and ventricular function, we evaluated further if blockade of opioids reverses the ischemia induced hemodynamic compromise, and if the effects are mediated by opioid receptors. Thirty-two mongrel dogs were anesthetized and artificially ventilated. Median thoracotomy was performed, the heart exposed, and the left anterior descending coronary artery isolated for subsequent occlusion and reperfusion. All cardiac parameters were recorded on an Electronics for Medicine recorder through the intracardiac catheters advanced from femoral vessels. Results indicate that naloxone significantly reversed the ischemic and reperfusion induced reduction in aortic, left ventricular and pulmonary arterial pressures, and left ventricular dp/dt. The inactive (+) stereoisomer of naloxone was without effect. These data demonstrate that opioids may have a role in the pathophysiology of myocardial infarction, mediated by opioid receptors, and provide new insight and strategies for the understanding and treatment of ischemic heart disease.

Animals↗