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Biomedical subjects

W Hua

Publications and source records attributed to W Hua.

At least 19 recordsLinked to original sources
PubMed2001

Evaluation of the Hexaplex assay for detection of respiratory viruses in children.

The Hexaplex assay (Prodesse, Inc., Milwaukee, Wis.) is a multiplex reverse transcriptase (RT)-PCR assay for the detection of parainfluenza virus types 1, 2, and 3, respiratory syncytial virus (RSV) types A and B, and influenza virus types A and B. We evaluated the Hexaplex assay in comparison with conventional viral cell cultures and rapid enzyme immunoassays (EIAs) for RSV (Directigen; Becton Dickinson Inc., Cockeysville, Md.) and influenza A virus (Abbott Test Pack; Abbott Laboratories, Abbott Park, Ill.) for the detection of respiratory viruses from pediatric respiratory specimens obtained from children seen at Children's Hospital of Wisconsin from December 1997 through May 1998. A total of 363 respiratory specimens were evaluated. The tissue culture prevalence of parainfluenza virus during this period of time was low (1.1%). The sensitivity, specificity, and positive and negative predictive value of Hexaplex compared to tissue culture for the detection of parainfluenza virus were 100, 95.8, 19.0, and 100%, respectively. Only one specimen was determined to contain influenza B virus by Hexaplex; it was tissue culture negative. A specimen was considered to contain RSV or influenza A virus when it was either culture positive or culture negative but Hexaplex and EIA positive. Prior to the analysis of discrepant results, the sensitivity, specificity, and positive and negative predictive value for the detection of RSV were 91.2, 100, 100, and 98.0%, respectively, for tissue culture; 84.5, 100, 100, and 96.6% for EIA; and 98.5, 91.5, 72.8, and 99.6% for Hexaplex, respectively. The sensitivity, specificity, and positive and negative predictive value for the detection of influenza A virus prior to the analysis of discrepant results were 100, 100, 100, and 100%, respectively, for culture, 78.0, 100, 100, and 89.4% for EIA, respectively, and 95.1, 94.1, 67.2, and 99.3% for Hexaplex, respectively. Culture- and/or EIA-negative, Hexaplex-positive specimens were analyzed by a second RT-PCR assay which used primers specific for a different genomic region than that used in the Hexaplex assay. After analysis of these discrepant results, the sensitivity, specificity, and positive and negative predictive value for the detection of RSV were 74.3, 100, 100, and 93.5%, respectively, for tissue culture; 70.3, 100, 100, and 92.5% for EIA; and 98.6, 97.4, 91.2, and 99.6% for Hexaplex. The sensitivity, specificity, and positive and negative predictive value for the detection of influenza A virus were 83.3, 100, 100, and 97.4%, respectively, for tissue culture; 69.4, 100, 100, and 83.3% for EIA; and 95.8, 98.7, 92.0, and 99.3% for Hexaplex. Hexaplex is a rapid, sensitive, and specific method for the detection of the seven most common respiratory viruses in children.

Child↗
PubMed2000

Computer-aided design, synthesis and biological assay of p-methylsulfonamido phenylethylamine analogues.

Class III antiarrhythmic agents selectively delay the effective refractory period (ERP) and increase the transmembrance action potential duration (APD). Based on our previous studies, a set of 17 methylsulfonamido phenylethylamine analogues were investigated by 3D-QSAR techniques of CoMFA and CoMSIA. The 3D-QSAR models proved a good predictive ability, and could describe the steric, electrostatic and hydrophobic requirements for recognition forces of the receptor site. According to the clues provided by this 3D-QSAR analysis, we designed and synthesized a series of new analogues of methanesulfonamido phenylethylamine (VIa-i). Pharmacological assay indicated that the effective concentrations of delaying the functional refractory period (FRP) 10ms of these new compounds have a good correlation with the 3D-QSAR predicted values. It is remarkable that the maximal percent change of delaying FRP in microM of compound VIc is much higher than that of dofetilide. The results showed that the 3D-QSAR models are reliable.

Animals↗
PubMed2000

Synthesis and characterization of pyridine-based polyamido-polyester optically active macrocycles and enantiomeric recognition for D- and L-amino acid methyl ester hydrochloride.

Five new chiral macrocycles, 3a-e, have been prepared by the acylation cyclization of chiral diamine dihydrobromide intermediates 2a-c with 2,6-pyridinedicarbonyl dichloride in highly diluted solution at room temperature. The chiral diesters 1a-c needed for the preparation of the macrocycles were obtained from condensation of corresponding N-(Z)-L-amino acids and 2,6-bishydroxymethyl pyridine in the presence of DCC and DMAP. The enantiomeric recognition of chiral macrocycles 3a-e for D- and L-amino acid methyl ester hydrochlorides has been characterized by fluorescence spectra, which indicate that some of them exhibited significant chiral recognition for the enantiomers of D- and L-amino acid methyl ester hydrochlorides. The stoichiometry and binding constants of 3a-L-Am(2) and 3c-L-Am(2) complexes have been determined. An X-ray analysis of the chiral macrocycle 3b show that the chiral ligand is rather rigid and strained.

Acylation↗
PubMed2000

Study of infrared and mass spectra of N-2-(3'-phthalide-yl)phthalazin-1-one.

The infrared and mass spectra of N-2-(3'-phthalide-yl)phthalazin-1-one were studied. The loss of one or two natural CO residues was observed from the title compound and its partial fragment ions. Using the MIKE spectrum of ion m/z 250, its most possible structure was confirmed. Two possible paths of the formation of the title compound were also proposed.

Mass Spectrometry↗
PubMed2000

Determination of edible bird's nest and its products by gas chromatography.

A specific gas chromatographic (GC) detection method for edible bird's nest (EBN) based on identifying the composition of the oligosaccharide chain combined with glycoprotein in EBN is developed. Five monoses (D-mannitose, D-galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and N-acetyl neuraminate) that constitute the oligosaccharide chain are detected using GC and GC-mass spectrometry techniques; their characteristic GC spectrum can reliably be regarded as EBN's fingerprint. The peak-area ratios in GC spectrum of those five monoses are found to be fixed; therefore, the GC technique developed in this work can conveniently be used to determine various raw EBNs and their products both qualitatively and quantitatively, distinguishing between fake and genuine EBN rapidly.

Animals↗
PubMed1999

An alternative HER-2/neu transcript of 8 kb has an extended 3'UTR and displays increased stability in SKOV-3 ovarian carcinoma cells.

UNLABELLED: HER-2/neu is a potent oncogene that predicts poor outcome when overexpressed in ovarian cancer. The SKOV-3 ovarian carcinoma cell line, one of the only models for HER2-driven ovarian cancer, expresses a major uncharacterized 8-kb alternative HER-2 transcript. OBJECTIVES: The aim of this study was to characterize the structure and determine the origin of the alternative sequence and examine the possible role of the 8-kb alternative transcript in overexpression of the HER-2 gene. METHODS: The structure of the 8-kb transcript was investigated using polymerase chain reaction (PCR) and nucleotide sequencing of cDNA clones. PCR analysis of genomic DNA was used to assess the origin of the 8-kb transcript. The stability of the 8-kb mRNA was assessed by Northern blot analysis of RNA extracted from cells treated with transcriptional inhibitors. RESULTS: Similar 5'UTR and coding sequence but an extended 3'UTR were contained in the 8-kb compared to the well-characterized 4.5-kb HER-2 transcript. Genomic DNA had continuity between the novel 3'UTR sequence from the 8-kb transcript and adjacent HER-2 terminal exon sequence. The 8-kb transcript had a half-life of 13 h compared to 5.5 h for the 4.5-kb transcript (P<0.01). CONCLUSIONS: The 8-kb transcript is generated from alternative polyadenylation site usage rather than gene rearrangement. Since the 8-kb transcript contains alternative sequence found at the 3' end of the normal HER-2 gene, it could be expressed in other cells. Increased stability of the 8-kb transcript may confer a selective advantage for SKOV-3 cells by providing enhanced HER-2 expression.

3' Untranslated Regions↗
PubMed1999

Nimodipine inhibits calcium-independent nitric oxide synthase activity in transient focal cerebral ischemia rats and cultured mouse astroglial cells.

The effect of nimodipine on nitric oxide synthase (NOS) activities in brains in transient focal cerebral ischemia rats, in cultured mouse neurons and astroglial cells and bovine brain capillary endothelial cells (BCECs) was investigated. The administration of nimodipine (3 mg.kg(-1), p.o., twice a day, for 3 days) before middle cerebral artery (MCA) occlusion significantly reduced infarct size, decreased nitrite/nitrate (NOx) content and inhibited Ca2+-independent NOS activity in the infarct area. Nimodipine inhibited the Ca2+-independent NOS activity induced by lipopolysaccharide (LPS) + tumor necrosis factor alpha (TNF alpha) in mouse astroglial cells with an IC50 value of 0.036+/-0.003 mM and Ca2+-dependent NOS activity in mouse neurons with an IC50 value of 0.047+/-0.003 mM, but did not affect Ca2+-dependent NOS activity in BCECs. The inhibition of Ca2+-independent NOS activity by nimodipine in astroglial cells was competitive with respect to L-arginine. Nimodipine also inhibited the induction of Ca2+-independent NOS activity in vitro. These results suggest that nimodipine in addition to its cerebral vasodilating effect may protect brain from ischemic neuronal damage through modifying NOS activity.

Animals↗
PubMed1999

Transvenous pectoral implantation of single lead implantable cardioverter defibrillator.

OBJECTIVE: To report our initial experience of transvenous pectoral implantation of single lead implantable cardioverter defibrillator (ICD) in fifteen patients. METHODS: Fifteen patients (male 13, female 2, mean age of 51.7 years), including dilated cardiomyopathy in 4, hypertrophic cardiomyopathy in 2, coronary artery disease in 5 and syncope of unknown etiology in 2, underwent ICD implantation. Indications for ICD implantation were ventricular fibrillation (8 patients) and refractory ventricular tachycardia (7 patients). All patients had syncope out of hospital. The ICD system were Medtronic Jewel active can in 5 patients (Model 7219C in 2, 7220C in 3) and CPI Ventak PRxIII in 9 patients. For active can, shocks were delivered through distal coil to ICD shell and for PRxIII through the endotak lead of proximal to distal coil. Successful implantation criteria was 2 consecutive defibrillation at less than 24 J. RESULTS: All 15 patients had successful implantation using single lead ICD system in catheter laboratory. The mean defibrillation threshold was 12.8 J (5-15 J) and R wave was 9.9 mV (3.7-14.6 mV). All devices were of pectoral implantation. No operative and perioperative complications occurred. In 7.6 months (1-18 months) of follow-up, 3 patients had VT terminated by antitachycardia pacing (ATP) and 1 patient VF terminated by shock. CONCLUSION: Transvenous pectoral implantation of single lead ICD is easy to perform and has high defibrillation efficacy and should be a preferred approach.

Defibrillators, Implantable↗
PubMed1998

Clinical use of nonthoracotomy cardioversion-defibrillation system.

OBJECTIVE: To summarize our experience in 22 patients who had attempted nonthoracotomy implantable cardioverter-defibrillators (ICD) for malignant ventricular arrhythmias (VA). METHODS: Indications for implantation were ventricular fibrillation (VF) in 17 patients and refractory ventricular tachycardia in 5. Thirteen patients of them had underlying ischaemic heart disease. Seven had dilated cardiomyopathy and two had arrhythmogenic right ventricular dysplasia. RESULTS: 20 out of 22 patients were successfully implanted nonthoracotomic ICD with defibrillating threshold of 16.7 J. The mean hospital stay was 7.8 days. The complications included chest wall haematoma and ventricular lead dislodgment requiring repositioning. In the follow-up period of 10.1 +/- 8 months, 6 patients with VA were treated successfully by ICD. CONCLUSION: Nonthoracotomy ICD is possible in a vast majority of patients (91%) and should be considered in all patients requiring ICD without concomitant surgery.

Adolescent↗
PubMed1997

Coupling of kinesin steps to ATP hydrolysis.

A key goal in the study of the function of ATP-driven motor enzymes is to quantify the movement produced from consumption of one ATP molecule. Discrete displacements of the processive motor kinesin along a microtubule have been reported as 5 and/or 8 nm. However, analysis of nanometre-scale movements is hindered by superimposed brownian motion. Moreover, because kinesin is processive and turns over stochastically, some observed displacements must arise from summation of smaller movements that are too closely spaced in time to be resolved. To address both of these problems, we used light microscopy instrumentation with low positional drift (< 39 pms[-1]) to observe single molecules of a kinesin derivative moving slowly (approximately 2.5nm s[-1]) at very low (150nM) ATP concentration, so that ATP-induced displacements were widely spaced in time. This allowed increased time-averaging to suppress brownian noise (without application of external force), permitting objective measurement of the distribution of all observed displacement sizes. The distribution was analysed with a statistics-based method which explicitly takes into account the occurrence of unresolved movements, and determines both the underlying step size and the coupling of steps to ATP hydrolytic events. Our data support a fundamental enzymatic cycle for kinesin in which hydrolysis of a single ATP molecule is coupled to a step distance of the microtubule protofilament lattice spacing of 8.12 nm. Step distances other than 8nm are excluded, as is the coupling of each step to two or more consecutive ATP hydrolysis reactions with similar rates, or the coupling of two 8-nm steps to a single hydrolysis. The measured ratio of ATP consumption rate to stepping rate is invariant over a wide range of ATP concentration, suggesting that the 1 ATP to 8nm coupling inferred from behaviour at low ATP can be generalized to high ATP.

Adenosine Triphosphatases↗
PubMed1997

Estrogen action in human ovarian cancer.

Evidence is accumulating for a facilitative role for estrogen in ovarian cancer. Although response to antiestrogen therapy has been poor, there is a distinct subset of patients that respond. Strategies for treatment of ovarian cancer would be improved by identification of patients likely to respond to hormonal therapy. Cell culture models that are responsive or resistant to estrogen and antiestrogen may be of value in finding markers that predict responsiveness to hormonal therapy. Several model cell lines have been generated that express ER and proliferate in response to estrogen in vitro. Further studies are needed to better characterize the response of these ER positive cells lines to estrogen in vivo in mouse xenograft models. Expression of many of the same genes are regulated by estrogen in breast and in ovarian cancer cell lines. One exception may be the HER-2/neu oncogene product, which is down-regulated by estrogen in responsive breast carcinoma cells but not in two ovarian carcinoma cell lines. Initial analyses of several estrogen responsive and one resistant cell model suggests the potential value of progesterone receptor presence and low levels of HER-2/neu expression for predicting responsiveness to hormonal therapy. Additional cell models need to be investigated to determine the frequency with which these markers are associated with antiestrogen resistance.

Animals↗
PubMed1997

Chronic steroid-eluting lead performance: a comparison of atrial and ventricular pacing.

It is generally believed that atrial pacing leads have higher stimulation thresholds and long-term complication rates than ventricular leads, and this is one of the factors limiting the use of dual chamber pacing. A study was undertaken to compare atrial and ventricular bipolar tined steroid-eluting leads in two designs: the Medtronic CapSure SP and the Telectronics Encor Dec. There were 123 pairs of leads: 81 CapSure SP and 42 Encor Dec. Bipolar atrial and ventricular stimulation thresholds, electrograms, and pacing impedance were measured using the Telectronics META DDDR pulse generator immediately postimplantation, and at 1, 3, and 6 months for all leads and at 12, 18, and 24 months for the CapSure SP. The only major lead complication was a 2% atrial lead dislodgment rate. All leads demonstrated low stimulation thresholds, with the CapSure SP leads having lower values than comparable Encor Dec leads. All leads had a mean range of 0.53-0.89 V at all testing periods with P < 0.05 for atrial leads only. There were no differences in electrogram size between manufacturers and no instances of atrial and ventricular undersensing. Pacing impedance was about 100 omega higher for the Encor Dec leads (P < 0.05, atrial leads only), suggesting that these leads will result in lower pacing energy losses provided the pulse generators are at identical settings. More than 90% of patients could be paced chronically in the atrium and ventricle at 2.5 V, but for chronic 1.6-V pacing, the CapSure SP leads were superior. In conclusion, atrial and ventricular steroid-eluting leads of both manufacturers gave excellent stimulation threshold results allowing low energy dual chamber pacing.

Aged↗
PubMed1997

Long-term performance of steroid-eluting lead in dual chamber pacing.

The atrial pacing lead is believed having higher stimulation thresholds and long-term complication rates than ventricular lead, this being one of the factors limiting the use of dual chamber pacing. A prospective study was undertaken to evaluate both atrial and ventricular bipolar tined steroid-eluting leads in long-term dual chamber pacing. There are 81 pairs of leads (Medtronic Capsure SP) used in 81 patients. Bipolar atrial and ventricular stimulation thresholds were measured immediately post implantation and 1, 3, 6, 12, 18 and 24 months. All leads demonstrated low mean stimulation thresholds during the follow-up and more than 94% of leads could be paced chronically in the atrium and ventricle at 2.5 volts. In conclusion, atrial and ventricular steroid-eluting leads gave excellent stimulation thresholds allowing low energy long-term dual chamber pacing.

Aged↗
PubMed1996

Radiofrequency ablation of idiopathic ventricular tachycardia.

BACKGROUND: Radiofrequency ablation (RFA) has been shown to be very effective in the treatment of supraventricular tachycardias and has replaced surgical ablation. Only a few reports of RFA for idiopathic ventricular tachycardia (VT) have appeared in the literature during the last two years. AIM: This paper presents our experience with RFA for idiopathic VT in 19 patients. MATERIAL: The age range of patients was 22-60, with a mean of 37.9 years. Twelve out of 19 were females, two patients had cardiac failure due to almost incessant VT while the rest had normal left ventricular function. Twelve patients had VT arising from the right ventricle (RV); of these, nine were from the outflow tract, two from the RV apex, and one from the mid-anterior RV. Seven patients had VT arising from the left ventricle (LV); of these, five were from the inferobasal portion of the septum and two were from the anterolateral area. METHODS: In all patients the diagnostic study and therapeutic RFA were combined in a single procedure. Pacemapping was used to guide the site of RFA in patients with VT arising from the RV. Local activation time (LAT), Purkinje potentials (PP) and pacemapping were used to guide RFA in those patients with LV septal tachycardias. RESULTS: A total of 21 RF procedures were performed in 19 patients and 15 out of 19 patients had successful VT ablation. Ten of the 12 patients with RV tachycardias and all five patients with LV septal (left axis, right bundle branch block) tachycardias were successfully ablated. One patient with mid anterior RV VT required two attempts for successful ablation. One patient with RV outflow tract (RVOT) VT could not be ablated despite two attempts. Two patients with LV tachycardias arising from the antero-lateral LV could also not be ablated. During a follow up period of two to 16 months none of the successful patients had recurrence of VT. The number of RF applications was one to 27, mean 10; fluoroscopy times were four to 75, mean 26.9 minutes. CONCLUSION: Idiopathic VT frequently arises from the RVOT and inferobasal portion of the LV septum. These tachycardias can be diagnosed on clinical and ECG grounds. RFA for idiopathic VT arising from these areas has a high success rate and this mode of treatment should be considered as a nonpharmacological curative treatment for symptomatic patients.

Adolescent↗
PubMed1995

Developmentally expressed Ca(2+)-sensitive adenylyl cyclase activity is disrupted in the brains of type I adenylyl cyclase mutant mice.

The type I Ca(2+)-sensitive adenylyl cyclase has been implicated in several forms of synaptic plasticity in vertebrates. Mutant mice in which this enzyme was inactivated by targeted mutagenesis show deficient spatial memory and altered long term potentiation (Wu, Z. L., Thomas, S. A., Villacres, E. C., Xia, Z., Simmons, M. L., Chavkin, C., Palmiter, R. D., and Storm, D. R. (1995) Proc. Natl Acad Sci. U. S. A. 92, 220-224). Long term potentiation in the CA1 region of the rat hippocampus develops during the first 2 weeks after birth and reaches maximal expression at postnatal day 15 with a gradual decline at later stages of development. Here we report that Ca(2+)-stimulated adenylyl cyclase activity in rat hippocampus, cerebellum, and cortex increases significantly between postnatal days 1-16. This increase appears to be due to enhanced expression of type I adenylyl cyclase rather than type VIII adenylyl cyclase, the other adenylyl cyclase that is directly stimulated by Ca2+ and calmodulin. Type I adenylyl cyclase mRNA in the hippocampus increased 7-fold during this developmental period. The developmental expression of Ca(2+)-stimulated adenylyl cyclase activity in mouse brain was attenuated in mutant mice lacking type I adenylyl cyclase. Changes in expression of the type I adenylyl cyclase during the period of long term potentiation development are consistent with the hypothesis that this enzyme is important for neuroplasticity and spatial memory in vertebrates.

Adenylyl Cyclases↗
PubMed1995

SKOV3 ovarian carcinoma cells have functional estrogen receptor but are growth-resistant to estrogen and antiestrogens.

Estrogen receptor positive ovarian cancer is often refractile to antiestrogen therapy. Here we describe the SKOV3 human ovarian carcinoma cell line as an in vitro model for estrogen and antiestrogen resistant ovarian cancer. While SKOV3 cells expressed estrogen receptor (ER) mRNA and protein at a similar level as the estrogen responsive T47D breast carcinoma cell line, their growth was not responsive to estradiol (E2) and was not inhibited by the antiestrogens OH-tamoxifen and ICI 164,384. The ER in SKOV3 cells was normal with respect to apparent Kd for binding with E2, E2 regulation of a transiently transfected ERE driven reporter gene, and E2 stimulation of expression of the early growth response genes c-myc and c-fos. However, the SKOV3 cells exhibited no expression of the progesterone receptor gene (PR) even after addition of E2, and the protein products of the estrogen responsive genes HER-2/neu and cathepsin D were expressed at constitutive levels that were not regulated by E2. Therefore, estrogen resistance in these cells may be a result of constitutive expression and loss of E2 regulation of selected growth regulatory gene products rather than a defect in estrogen activation of ER as a transcriptional regulator.

Blotting, Northern↗