Biomedical subjects

W K Wang

Publications and source records attributed to W K Wang.

At least 19 recordsLinked to original sources

PubMed2006

Evaluating microcirculation by pulsatile laser Doppler signal.

Laser Doppler flowmetry (LDF) is a popular method for monitoring the microcirculation, but it does not provide absolute measurements. Instead, the mean flux response or energy distribution in the frequency domain is generally compared before and after stimulus. Using the heartbeat as a trigger, we investigated whether the relation between pressure and flux can be used to discriminate different microcirculatory conditions. We propose the following three pulsatile indices for evaluating the microcirculation condition from the normalized pressure and flux segment with a synchronized-averaging method: peak delay time (PDT), pressure rise time and flux rise time (FRT). The abdominal aortic blood pressure and renal cortex flux (RCF) signals were measured in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The mean value of the RCF did not differ between SHR and WKY. However, the PDT was longer in SHR (87.14 +/- 5.54 ms, mean +/- SD) than in WKY (76.92 +/- 2.62 ms; p < 0.001). The FRT was also longer in SHR (66.56 +/- 1.98 ms) than in WKY (58.02 +/- 1.77 ms; p < 0.001). We propose that a new dimension for comparing the LDF signals, which the results from the present study show, can be used to discriminate RCF signals that cannot be discriminated using traditional methods.

Animals↗

PubMed2006

Effects of whole-body mechanical stimulation at double the heart rate on the blood pressure waveform in rats.

The effects of mechanical stimulation on hemodynamics, such as due to mechanotransduction in vascular endothelial cells, have been widely discussed recently. We previously proposed a resonance model in which the arterial system is treated as a pressure-transmitting system, and suggested that the application of external mechanical stimulation with frequencies near the heart rate (HR) or harmonics thereof can be sensed by the arterial system and induce hemodynamic changes. In this study, we monitored the effects of external mechanical stimulation at a frequency of double the HR on BPW (blood pressure waveform), HRV (HR variability) and BPHV (blood-pressure-harmonics variability) in rats. A motor beating a waterbed mattress was used to generate pressure variations of 0.5 mmHg to apply onto the rats. The experiments were performed on three groups of rats with different beating frequencies: (A) double the HR, (B) 5% deviation from double the HR and (C) 1.5 times the HR. The experimental procedure was a 15 min control period followed by application of the mechanical stimulation for 15 min and further recording for 15 min (OFF period). During the OFF period, the amplitude of the second harmonic in the BPW significantly increased by >5% in group A with decreased HRV and BPHV. The second harmonic increased less in group B, and decreased in group C. The increase in the second-harmonic amplitude in group A may be due to the filtering properties of the renal arterial structure. This mechanism could be used to improve the local blood supply into the kidneys, and hence provide a new treatment modality for some important diseases, such as renal hypertension or nephrosis.

Animals↗

PubMed2005

Population structure of wild bananas, Musa balbisiana, in China determined by SSR fingerprinting and cpDNA PCR-RFLP.

Both demographic history and dispersal mechanisms influence the apportionment of genetic diversity among plant populations across geographical regions. In this study, phylogeography and population structure of wild banana, Musa balbisiana, one of the progenitors of cultivated bananas and plantains in China were investigated by an analysis of genetic diversity of simple sequence repeat (SSR) fingerprint markers and cpDNA PCR-RFLP. A chloroplast DNA (cpDNA) genealogy of 21 haplotypes identified two major clades, which correspond to two geographical regions separated by the Beijiang and Xijiang rivers, suggesting a history of vicariance. Significant genetic differentiation was detected among populations with cpDNA markers, a result consistent with limited seed dispersal in wild banana mediated by foraging of rodents. Nuclear SSR data also revealed significant geographical structuring in banana populations. In western China, however, there was no detected phylogeograpahical pattern, possibly due to frequent pollen flow via fruit bats. In contrast, populations east of the Beijiang River and the population of Hainan Island, where long-range soaring pollinators are absent, are genetically distinct. Colonization-extinction processes may have influenced the evolution of Musa populations, which have a metapopulation structure and are connected by migrating individuals. Effective gene flow via pollen, estimated from the nuclear SSR data, is 3.65 times greater than gene flow via seed, estimated from cpDNA data. Chloroplast and nuclear DNAs provide different insights into phylogeographical patterns of wild banana populations and, taken together, can inform conservation practices.

China↗

PubMed2005

Raising harmonic variation of arterial pulse in dying rats.

Our previous study revealed that the coefficient of variation of harmonic magnitude (HCV) of radial arterial pulse was significantly raised before the death of cancer patients. In this study, we recorded the caudate arterial pulse of 24 Sprague-Dawley rats that had a fatal dose of urethane injected into their abdomens. Twenty rats were dead within 3 hours after the injection and four survived. We defined the last 100 minutes of each rat's life as the dying process. During the dying process, we found that both the systolic blood pressure and diastolic blood pressure dropped steeply during the last 5 minutes. However, all HCVs, except HCV1, climbed steeply before the last 5 minutes. The HCV1 of the dying rats was significantly higher than that of rats that survived, starting from the first minute (P < 0.01). The HCV2 of the dying rats was significantly higher than that of the survived rats starting from the 52nd minute (P < 0.05). The HCV3 and HCV4 of the dying rats were significantly higher than those of the survived rats until the 70th minute and the 80th minute, respectively (P < 0.05). Furthermore, HCV2-HCV4 proceeded with the dying process and increased gradually. We concluded that HCVs, which failed first in the high-frequency components and then in the low-frequency components, could provide physicians with earlier information to prevent the coming failure of circulatory system, and could reflect quantitatively pathological severity and predict patient outcome. The specific Fourier components in the pulse provide more physiological information than systolic and diastolic blood pressures.

Acute Disease↗

PubMed2003

Influencing the heart rate of rats with weak external mechanical stimulation.

The ventricular-arterial coupling is assumed to minimize the expenditure of cardiac energy. From the conjecture of the resonance theory, the arterial system transmits pressure waves and resonates with the heartbeat, therefore, the arterial system is similar to a mechanical resonator. Theoretically, the heart rate can be paced with weak external mechanical stimulation and corresponding blood pressure changes can be observed. A waterbed was activated to generate 0.5-mmHg pressure vibrations as a stimulus and the rate was set to deviate 5% from the control heart rate. Among 13 studies on seven rats, the linear regression between X (stimulation frequency--control heart rate) and Y (actual changes of the heart rate) is Y = 0.992X = 0.062 (Hz) with a correlation coefficient of 0.97 (Y = X implies complete steering). The intercorrelation coefficient between the change in mean blood pressure and the heart rate was 0.79. The study showed that this weak mechanical stimulation influences the heart rate, and the blood pressure changes according to the heart rate. Cardiovascular optimization and the resonance theory may explain the way one may regulate the heart rate and the blood pressure of humans noninvasively in the future.

Animals↗

PubMed2003

Validation of fermentation processes.

The ability to prepare consistent biopharmaceutical products depends extensively on possession of banked and characterized cell substrates and on development of production processes which can be validated. While the attributes that define cell characterization have been extensively detailed by ICH and the regulatory agencies in the past decade, little has been specified regarding process validation for biological processes. The extent to which validation concepts can be applied to biological processes varies depending on the nature of the process, the nature of the product, and the level of knowledge regarding the relationship between process parameters and product quality. Expectations concerning the rigour of the validation programme should be adjusted accordingly. There is no single approach that is appropriate for all processes and products. At a minimum, there should be an attempt to define which process parameters are critical, and to focus the attention of validation efforts on these parameters.

Biotechnology↗

PubMed2001

Crystallization and preliminary crystallographic studies of a ribosomal protein L30e from the hyperthermophilic archaeon Thermococcus celer.

Ribosomal protein L30e from the hyperthermophilic archaeon Thermococcus celer is a good model for the study of the thermostability of proteins. It has been overexpressed, purified and crystallized using the hanging-drop vapour-diffusion method using PEG 8000 as precipitant at 290 K. The crystal belongs to the hexagonal space group P6(1)/P6(5), with unit-cell parameters a = b = 48.32, c = 86.42 A. The asymmetric unit contains a single molecule of L30e, with a corresponding crystal volume per protein mass (V(M)) of 2.68 A(3) Da(-1) and a solvent content of 54%. A complete data set diffracting to 1.96 A resolution was collected from a single crystal at 100 K.

Archaeal Proteins↗

PubMed2001

Structure of the C-terminal sterile alpha-motif (SAM) domain of human p73 alpha.

p73 is a homologue of the tumour suppressor p53 and contains all three functional domains of p53. The alpha-splice variant of p73 (p73 alpha) contains near its C-terminus an additional structural domain known as the sterile alpha-motif (SAM) that is probably responsible for regulating p53-like functions of p73. Here, the 2.54 A resolution crystal structure of this protein domain is reported. The crystal structure and the published solution structure have the same five-helix bundle fold that is characteristic of all SAM-domain structures, with an overall r.m.s.d. of 1.5 A for main-chain atoms. The hydrophobic core residues are well conserved, yet some large local differences are observed. The crystal structure reveals a dimeric organization, with the interface residues forming a mini four-helix bundle. However, analysis of solvation free energies and the surface area buried upon dimer formation indicated that this arrangement is more likely to be an effect of crystal packing rather than reflecting a physiological state. This is consistent with the solution structure being a monomer. The p73 alpha SAM domain also contains several interesting structural features: a Cys-X-X-Cys motif, a 3(10)-helix and a loop that have elevated B factors, and short tight inter-helical loops including two beta-turns; these elements are probably important in the normal function of this domain.

Amino Acid Motifs↗

PubMed2001

[Studies of FISH and karyotype of Gossypium barbadense].

Based on the fluorescent in situ hybridization (FISH) of somatic chromosome of Gossypium barbadense with the probe of genomic DNA (gDNA) of Gossypium arboreum, two sets of chromosomes were easily distinguished by signals hybridized or not. The FISH directly proved that G. barbadense originated from two different diploid species, but was not in concordance with the former point that every chromosome of A sub-genome of tetraploid species was bigger than that of other sub-genome (D genome). The karyotype formula of G. barbadense based on its FISH was 2n = 4x = 52 = 38 m + 14sm(6sat). There were three pairs of satellite chromosomes which were all sm types. Their satellites located in short arms but originated differently from their chromosomes of sub-genome. Fragment translocations occured in the long arms of homologous chromosomes of number 5, 6 and 9 of A sub-genome. It was suggested that the translated fragments come from D sub-genome. The fragments are fairly large with the relative lengths of 19.21%, 17.69% and 12.88% of their whole chromosomes, respectively. At least five pairs of chromosomes in D sub-genome show some hybridized signals of gDNA probe of G. arboreum in their centromere regions, which indicated that there would be chromatin introgressions from A sub-genome.

Gossypium↗

PubMed2000

Interaction between HIV type 1 glycoprotein 120 and CXCR4 coreceptor involves a highly conserved arginine residue in hypervariable region 3.

Several seven-transmembrane chemokine receptors are known to function as entry coreceptors for human immunodeficiency virus type 1. CCR5 and CXCR4 are the major coreceptors for non-syncytium-inducing (NSI) and syncytium-inducing (SI) viruses, respectively. During the natural course of infection, the emergence of variants with a phenotypic transition from NSI to SI and rapid disease progression is associated with expanded coreceptor usage to CXCR4. Characteristic amino acids at several positions in the hypervariable region 3 (V3) of gp120 have been linked to CXCR4 utilization. Previously, we reported that a highly conserved arginine residue of V3 played an important role in CCR5 utilization. In this study, the possible involvement of the same arginine residue in CXCR4 utilization was investigated. Amino acid substitutions introduced to this arginine on R5X4 viruses were found to have a significant effect on their utilization of CXCR4. These results, taken together with those reported previously, suggest that this highly conserved arginine may contribute to the functional convergence of chemokine coreceptor utilization by human immunodeficiency viruses and may represent a unique target for future antiviral design.

Amino Acid Sequence↗

PubMed2000

Crystallization and preliminary crystallographic studies of a SAM domain at the C-terminus of human p73alpha.

p73 is a recently discovered homologue of the tumour suppressor p53 and contains all three functional domains of p53. The alpha-splice variant of p73 (p73alpha) contains an additional structural domain near its C--terminus that has sequence homology with the sterile alpha-motif (SAM) domain. This domain is considered to be responsible for mediating protein-protein interactions. Pyramidal crystals of human p73alpha SAM domain were obtained by the hanging-drop vapour-diffusion method with ammonium dihydrogen orthophosphate as the precipitant. The crystals diffract to 2.54 A resolution and belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 32.02, c = 133.84 A. The structure was solved by molecular replacement using the NMR structure of the same protein as the search model.

Amino Acid Motifs↗

PubMed2000

Effect of length on the fundamental resonance frequency of arterial models having radial dilatation.

The pressure wave moving along an elastic artery filled with blood was examined as a moving Windkessel having a natural oscillation angular frequency nu 0 and a damping coefficient b. The radial directional motion for an element of the wall segment and the adherent fluid was considered. This equation was solved with conditions at both ends of an artery of length L. An external impulse force was applied at one end and a static pressure Po at the other. Analytic solution allowed only certain oscillation modes of resonance frequencies fn, where fn2 = a + cnL-2 with [formula: see text] and V infinity is the high frequency phase velocity. The relationship between f0 and L was examined experimentally for tubes constructed of latex, rubber, or dissected aorta. The effect of raising the static pressure P0 or increasing the tension in the tube was consistent with the prediction. The hypertension that accompanies an augmentation in arterial wall and the association between the heart rate and the mean blood pressure were discussed.

Animals↗

PubMed2000

Determination of human immunodeficiency virus type 1 subtypes in Taiwan by vpu gene analysis.

The genetic diversity of human immunodeficiency virus (HIV) type 1 (HIV-1) has been characterized mainly by analysis of the env and gag genes. Information on the vpu genes in the HIV sequence database is very limited. In the present study, the nucleotide sequences of the vpu genes were analyzed, and the genetic subtypes determined by analysis of the vpu gene were compared with those previously determined by analysis of the gag and env genes. The vpu genes were amplified by nested PCR of proviral DNA extracted from 363 HIV-1-infected individuals and were sequenced directly by use of the PCR products. HIV-1 subtypes were determined by sequence alignment and phylogenetic analysis with reference strains. The strains in all except one of the samples analyzed could be classified as subtype A, B, C, E, or G. The vpu subtype of one strain could not be determined. Of the strains analyzed, genetic subtypes of 247 (68.0%) were also determined by analysis of the env or gag gene. The genetic subtypes determined by vpu gene analysis were, in general, consistent with those determined by gag and/or env gene analysis except for those for two AG recombinant strains. All the strains that clustered with a Thailand subtype E strain in the vpu phylogenetic analyses were subtype E by env gene analysis and subtype A by gag gene analysis. In summary, our genetic typing revealed that subtype B strains, which constituted 73.8% of all strains analyzed, were most prevalent in Taiwan. While subtype E strains constituted about one-quarter of the viruses, they were prevalent at a higher proportion in the group infected by heterosexual transmission. Genetic analysis of vpu may provide an alternate method for determination of HIV-1 subtypes for most of the strains, excluding those in which intersubtype recombination has occurred.

Amino Acid Sequence↗

PubMed2000

Quantitative competitive reverse transcription-PCR for quantification of dengue virus RNA.

A quantitative competitive reverse transcription-PCR assay was developed to quantify dengue virus RNA in this study. The main features include a primer pair targeting a highly conserved region in the capsid and the addition of competing RNA that contains an internal deletion to provide a stringent internal control for quantification. It can be utilized to quantify RNA isolated from the four dengue virus serotypes but not RNA isolated from other flaviviruses, including Japanese encephalitis virus and hepatitis C virus, both prevalent in Asia. It can also be used to quantify dengue virus RNA isolated from the plasma of infected individuals. The sensitivity of the assay was estimated to be 10 to 50 copies of RNA per reaction, and twofold differences in virus titer are distinguishable. This assay is a convenient, sensitive, and accurate method for quantification and can be used to further understanding of the pathogenesis of dengue virus infection.

Culture Media↗

PubMed2000

Influence of spleen meridian herbs on the harmonic spectrum of the arterial pulse.

Pulse analysis is a powerful method in Chinese medicine. We suggest that the effect of herbal medicine is to redistribute the blood to organs and meridians. In this report, by injecting extracts into rats and then analyzing the blood pressure wave measured at the caudate arteries, we studied eight important spleen meridian related herbs: They were Semen Lablab, Fructus Amomi Globosi, Rhizoma Atractylodis Macrocephalae, Rhizoma Atractylodis, Tuber Pinelliae, Radix Codonopsitis, Pericarpium Aurantii and Rhizoma Polygonati. All eight herbs increased the intensity of the 3rd harmonic (C3) of the pressure pulse which is correlated to the spleen and spleen meridian, as described in traditional Chinese medical literature. All of them also increased the 2nd harmonic (which is correlated to the kidney and the kidney meridian) as well as decreased the heart load (DC term of pressure wave, C0). Tuber Pinelliae, Radix Codonopsitis, Pericarpium Aurantii and Rhizoma Polygonati decreased the 1st harmonic (correlated to the liver meridian) significantly, while Rhizoma Atractylodis Macrocephalae only decreased C1 slightly. Except for Semen Lablab, all the others decreased the intensity of the 5th (stomach meridian) and the 7th harmonics. The effects of kidney herbs: Cortex Eucommiae and Radix Achyranthis were also shown for comparison.

Animals↗

PubMed2000

Molecular biology of human immunodeficiency virus type 1.

Since the discovery of human immunodeficiency virus (HIV) as the etiologic agent of acquired immune deficiency syndrome (AIDS) more than a decade ago, tremendous progress has been made in various aspects of this virus and its interplay with the host immune system. The advent of potent combination therapy has made it possible to achieve effective and durable control of HIV-1 replication in vivo, yet the persistence of the latent reservoirs pose a new challenge. The recent identifications of several cellular proteins interacting with different viral gene products have not only shed new insights into our understanding of the HIV-1 and the host cell biology, but also provided the bases for developing novel strategies to block HIV-1 replication. It is from this perspective that we review the current understanding of the molecular mechanisms governing the HIV-1 life cycle.

Base Sequence↗

PubMed1999

Hypervariable region 3 residues of HIV type 1 gp120 involved in CCR5 coreceptor utilization: therapeutic and prophylactic implications.

Crystallographic characterization of a ternary complex containing a monomeric gp120 core, parts of CD4, and a mAb, revealed a region that bridges the inner and outer domains of gp120. In a related genetic study, several residues conserved among primate lentiviruses were found to play important roles in CC-chemokine receptor 5 (CCR5) coreceptor utilization, and all but one were mapped to the bridging domain. To reconcile this finding with previous reports that the hypervariable region 3 (V3) of gp120 plays an important role in chemokine coreceptor utilization, elucidating the roles of various V3 residues in this critical part of the HIV type 1 (HIV-1) life cycle is essential. Alanine-scanning mutagenesis was carried out to identify V3 residues critical for CCR5 utilization. Our findings demonstrated that several residues in V3 were critical to CCR5 utilization. Furthermore, these residues included not only those conserved across HIV-1 subtypes, but also those that varied among HIV-1 subtypes. Although the highly conserved V3 residues may represent unique targets for antiviral designs, the involvement of variable residues raises the possibility that antigenic variation in the coreceptor binding domain could further complicate HIV-1 vaccine design.

Alanine↗

PubMed1999

Stability and folding of the tumour suppressor protein p16.

The tumour suppressor p16 is a member of the INK4 family of inhibi tors of the cyclin D-dependent kinases, CDK4 and CDK6, that are involved in the key growth control pathway of the eukaryotic cell cycle. The 156 amino acid residue protein is composed of four ankyrin repeats (a helix-turn-helix motif) that stack linearly as two four-helix bundles resulting in a non-globular, elongated molecule. The thermodynamic and kinetic properties of the folding of p16 are unusual. The protein has a very low free energy of unfolding, Delta GH-2O/D-N, of 3.1 kcal mol-1 at 25 degreesC. The rate-determining transition state of folding/unfolding is very compact (89% as compact as the native state). The other unusual feature is the very rapid rate of unfolding in the absence of denaturant of 0.8 s-1 at 25 degreesC. Thus, p16 has both thermodynamic and kinetic instability. These features may be essential for the regulatory function of the INK4 proteins and of other ankyrin-repeat-containing proteins that mediate a wide range of protein-protein interactions. The mechanisms of inactivation of p16 by eight cancer-associated mutations were dissected using a systematic method designed to probe the integrity of the secondary structure and the global fold. The structure and folding of p16 appear to be highly vulnerable to single point mutations, probably as a result of the protein's low stability. This vulnerability provides one explanation for the striking frequency of p16 mutations in tumours and in immortalised cell lines.

Circular Dichroism↗