PubMed Health⌕ Search

Biomedical subjects

C Tu

Publications and source records attributed to C Tu.

At least 19 recordsLinked to original sources
PubMed2006

Circulating endothelial progenitor cell deficiency contributes to impaired arterial elasticity in persons of advancing age.

Reduced arterial elasticity is a hallmark of ageing in healthy humans and appears to occur independently of coexisting disease processes. Endothelial-cell injury and dysfunction may be responsible for this fall in arterial elasticity. We hypothesized that circulating endothelial progenitor cells (EPCs) are involved in endothelial repair and that lack of EPCs contributes to impaired arterial elasticity. A total of 56 healthy male volunteers were divided into young (n=26) and elderly (n=30) groups. Large and small artery elasticity indices were noninvasively assessed using pulse wave analysis. The number of circulating EPCs was measured by using flow cytometry. Cells demonstrating DiI-acLDL and FITC-ulex lectin double-positive fluorescence were identified as EPCs. C1 large artery elasticity and C2 small artery elasticity indices were significantly reduced in the elderly group compared with the young group (11.73+/-1.45 vs 16.88+/-1.69 ml/mm Hg x 10, P<0.001; 8.40+/-1.45 vs 10.58+/-1.18 ml/mm Hg x 100, P<0.001, respectively). In parallel, the number of circulating EPCs was significantly reduced in the elderly group compared with the young group (0.13+/-0.02 vs 0.17+/-0.04%, P<0.05). The number of circulating EPCs correlated with C1 large and C2 small artery elasticity indices (r=0.47, P<0.01; r=0.4, P<0.01). The present findings suggest that the fall in circulating EPCs with subsequently impaired endothelial-cell repair and function contributes to reduced arterial elasticity in humans with ageing. The decrease in circulating EPCs may serve as a surrogate biologic measure of vascular function and human age.

Adult↗
PubMed2006

In situ hybridization and RT-PCR detection of Macrobrachium rosenbergii nodavirus in giant freshwater prawn, Macrobrachium rosenbergii (de Man), in Taiwan.

Between January and March 2006, suspected outbreaks of white tail disease were observed in post-larvae and juveniles of Macrobrachium rosenbergii in hatcheries and nursery ponds at Kaohsiung and Pingtung Counties in southern Taiwan. Pathognomonic lesions showed the presence of large oval and/or irregular basophilic, cytoplasmic inclusion bodies in the infected muscles and hepatopancreas. Reverse transcriptase polymerase chain reaction (RT-PCR) assay and in situ hybridization (ISH) revealed evidence of M. rosenbergii nodavirus (MrNV) infection but did not detect extra small virus. Phylogenetic analysis indicated that there were very high identities between nucleotide sequences among six strains obtained in this study (99.5-100%), moderate identities with Caribbean and Indian strains (98.2-98.6%), but slightly lower identity with a Chinese strain (95.2%). This is the first confirmation of MrNV in giant freshwater prawns, using an RT-PCR and ISH, in Taiwan.

Abdominal Muscles↗
PubMed2003

Root-parasitic nematodes enhance soil microbial activities and nitrogen mineralization.

Obligate root-parasitic nematodes can affect soil microbes positively by enhancing C and nutrient leakage from roots but negatively by restricting total root growth. However, it is unclear how the resulting changes in C availability affect soil microbial activities and N cycling. In a microplot experiment, effects of root-parasitic reniform nematodes ( Rotylenchulus reniformis) on soil microbial biomass and activities were examined in six different soils planted with cotton. Rotylenchulus reniformis was introduced at 900 nematodes kg(-1) soil in May 2000 prior to seeding cotton. In 2001, soil samples were collected in May before cotton was seeded and in November at the final harvest. Extractable C and N were consistently higher in the R. reniformis treatments than in the non-nematode controls across the six different soils. Nematode inoculation significantly reduced microbial biomass C, but increased microbial biomass N, leading to marked decreases in microbial biomass C:N ratios. Soil microbial respiration and net N mineralization rates were also consistently higher in the nematode treatments than in the controls. However, soil types did not have a significant impact on the effects of nematodes on these microbial parameters. These findings indicate that nematode infection of plant roots may enhance microbial activities and the turnover of soil microbial biomass, facilitating soil N cycling. The present study provides the first evidence about the direct role of root-feeding nematodes in enhancing soil N mineralization.

Animals↗
PubMed2003

Impact of endoscopic ultrasound combined with fine-needle aspiration biopsy in the management of esophageal cancer.

BACKGROUND AND STUDY AIMS: Endoscopic ultrasound (EUS) in combination with fine-needle aspiration biopsy (FNA) is a highly accurate method for the preoperative staging of esophageal cancer. Its impact on medical decision-making and the cost of care is unknown. This prospective case series was undertaken in order to determine the impact of EUS in combination with FNA on patients' choice of therapy and on the cost of care. PATIENTS AND METHODS: Sixty consecutive patients with esophageal cancer, referred for preoperative EUS staging in a large tertiary-care academic medical center, were enrolled. The accuracy of EUS, the impact of EUS-based staging on the patients' choice of therapy, and costs were studied. RESULTS: The accuracy rates for EUS combined with FNA in tumor and lymph-node staging were 83 % and 89 %, respectively. Twenty-five patients (42 %) had EUS stage I and II and were candidates for curative surgery. Twenty-eight patients (47 %) had stage III, and seven (12 %) had stage IV. All patients with stage I had surgery, while all patients with stage IV had medical therapy. The majority (62 %) of patients with stage II had surgery, while only a minority (25 %) of patients with stage III had surgery. Thirty-six patients (60 %) underwent medical therapy. Patients' medical decisions in favor of surgical or medical therapy correlated strongly with the results of their EUS staging ( P = 0.005), but not with age, sex, or referring physicians (surgeons vs. nonsurgeons). EUS-guided therapy potentially decreased the cost of care by $ 740 424 ($ 12 340/patient) by reducing the number of thoracotomies. CONCLUSIONS: Patients' decisions regarding therapy correlated with their overall tumor staging, suggesting that the information provided by EUS played a significant role in patients' decision-making. The use of EUS in combination with FNA reduces the cost of managing patients with esophageal cancer.

Adenocarcinoma↗
PubMed2001

Phylogenetic comparison of classical swine fever virus in China.

An N-terminal fragment of the E2 gene of classical swine fever (CSF) virus encoding major immunogenic sites was amplified by RT-PCR directly from 110 clinical specimens representing 109 epizootic sites during the last decade in China. Phylogenetic relationships between these viruses as well as 20 reference strains were determined by comparison of their nucleotide sequences. A phylogenetic tree showed that 103 of the 110 field viruses (93.6%) were clustered within group 2 and subdivided into three subgroups, while the remaining seven viruses (6.4%), along with two Chinese reference strains, Shimen and HCLV (attenuated vaccine strain), were clustered into subgroup 1.1 within group 1. However, none of the Chinese CSF viruses were members of subgroup 1.2 (represented by reference strain Brescia). This is the first report on the distribution of CSF virus genotypes in China. Results indicated that CSF viruses predominating in recent epizootics within China are genetically divergent from the reference strain Shimen and the vaccine strain HCLV.

Animals↗
PubMed2001

Liquid-phase microextraction of phenolic compounds combined with on-line preconcentration by field-amplified sample injection at low pH in micellar electrokinetic chromatography.

This paper describes a novel method that applies field-amplified sample injection (FASI) in micellar electrokinetic chromatography (MEKC) with a low pH background electrolyte (BGE). Six phenolic compounds prepared in water or NaOH solution were used as the test analytes. Sample was injected electrokinetically after the introduction of a plug of water. During the injection, the water plug was pumped out of the capillary inlet by the electroosmotic flow, and the phenolic anions migrated very quickly in the direction of the outlet. When the anions reached the boundary between the water plug and BGE, they were neutralized and ceased moving. Thereafter, MEKC was initiated for the separation. This on-line preconcentration method could be conveniently coupled with a liquid-liquid-liquid microextraction procedure, in which a hollow fiber was used as an extraction solvent support to extract the analytes from the water sample. The acceptor phase consisted of 8 mM NaOH. After extraction, the extract was analyzed directly by MEKC, as described.

Journal Article↗
PubMed2001

The catalytic properties of human carbonic anhydrase IX.

Human carbonic anhydrase IX (CA IX) is an integral membrane protein and a member of the alpha class of carbonic anhydrases that includes the human and animal enzymes. We have prepared a truncated, recombinant form of human CA IX of 255 residues consistent with full-length human CA II, among the most efficient of the carbonic anhydrases. Catalysis by and inhibition of this form of human CA IX has been investigated using stopped-flow spectrophotometry and 18O exchange measured by mass spectrometry. In kinetic constants for the hydration of CO2, CA IX closely resembled CA II with maximal proton transfer-dependent 18O exchange near 1 micros(-1) and kcat/Km near 55 microM(-1) x s(-1). Human CA IX was very strongly inhibited by three classic sulfonamides and cyanate, with inhibition constants that are close to those for CA II.

Amino Acid Sequence↗
PubMed2001

Bicarbonate as a proton donor in catalysis by Zn(II)- and Co(II)-containing carbonic anhydrases.

Catalysis of (18)O exchange between CO(2) and water catalyzed by a Co(II)-substituted mutant of human carbonic anhydrase II is analyzed to show the rate of release of H(2)(18)O from the active site. This rate, measured by mass spectrometry, is dependent on proton transfer to the metal-bound (18)O-labeled hydroxide, and was observed in a site-specific mutant of carbonic anhydrase II in which a prominent proton shuttle residue His64 was replaced by alanine, which does not support proton transport. Upon increasing the concentration of bicarbonate, the rate of release of H(2)(18)O increased in a saturable manner to a maximum of 4 x 10(5) s(-)(1), consistent with proton transfer from bicarbonate to the Co(II)-bound hydroxide. The same mutant of carbonic anhydrase containing Zn(II) had the rate of release of H(2)(18)O smaller by 10-fold, but rate of interconversion of CO(2) and HCO(3)(-) about the same as the Co(II)-containing enzyme. These data as well as solvent hydrogen isotope effects suggest that the bicarbonate transferring the proton is bound to the cobalt in the enzyme. The enhancement of (18)O exchange caused by increasing bicarbonate concentration during catalysis by the Zn(II)-containing carbonic anhydrase from the archaeon Methanosarcina thermophila suggests that a very similar mechanism for proton donation by bicarbonate occurs with this wild-type enzyme.

Amino Acid Substitution↗
PubMed2001

Structural and kinetic analysis of the chemical rescue of the proton transfer function of carbonic anhydrase II.

Histidine 64 in human carbonic anhydrase II (HCA II) functions in the catalytic pathway of CO(2) hydration as a shuttle to transfer protons between the zinc-bound water and bulk water. Catalysis of the exchange of (18)O between CO(2) and water, measured by mass spectrometry, is dependent on this proton transfer and was decreased more than 10-fold for H64A HCA II compared with wild-type HCA II. The loss of catalytic activity of H64A HCA II could be rescued by 4-methylimidazole (4-MI), an exogenous proton donor, in a saturable process with a maximum activity of 40% of wild-type HCA II. The crystal structure of the rescued complex at 1.6 A resolution shows 4-MI bound in the active-site cavity of H64A HCA II, through pi stacking interactions with Trp 5 and H-bonding interactions with water molecules. In this location, 4-MI is about 12 A from the zinc and approximates the observed "out" position of His 64 in the structure of the wild-type enzyme. 4-MI appears to compensate for the absence of His 64 and rescues the catalytic activity of the H64A HCA II mutant. This result strongly suggests that the out conformation of His 64 is effective in the transfer of protons between the zinc-bound solvent molecule and solution.

Alanine↗
PubMed2001

DNA-mediated protection against classical swine fever virus.

Four eukaryotic expression plasmids containing the entire E2 gene sequence of classical swine fever virus (CSFV) were constructed: (a) pcDST, with 5' signal and 3' transmembrane sequences; (b) pcDSW, with 5' signal sequence only; (c) pcDWT, with transmembrane sequences only; and (d) pcDWW, containing the E2 gene alone. All four plasmids were readily transfected into BHK-21 cells, with pcDST and pcDSW resulting in secretion of E2 antigen. The latter two plasmids were also shown to induce a humoral immune response against CSFV in mice when administered intramuscularly, but no immune responses were detected with either pcDWT or pcDWW. The antibody level elicited by pcDSW was higher than that induced by pcDST. When pcDSW was used to immunize rabbits and pigs, both species were shown to be protected from challenge with virulent CSFV (hog cholera lapinized virus for rabbits and Shimen strain for pigs).

Animals↗
PubMed2001

Catalytic effect of soil colloids on the reaction between CrVI and p-methoxyphenol.

Adsorption of CrVI and p-methoxyphenol (PMP) on soil colloids at different pH media was studied. The resulting k1 and n of 1.89 x 10(2) and 0.53 (r2 = 0.99) and k2 and b of 0.13 and 1.25 x 10(3) (r2 = 0.96) were obtained from Freundlich (Q = k1Caqn) and Langmuir [Q = k2bCaq/(1 + k2Caq)] simulation equations, respectively, for CrVI adsorption on soil colloids (pH 4.20). The adsorption of PMP on soil colloids in pH 5.72 media was simulated by five different equations and the results indicated that the Fritz-Schluender one (r2 = 1.00) was the most suitable among them. Adsorption quantity of CrVI and PMP on colloids increased with increasing acidity in the pH range of 3.5-9.0. Study of CrVI adsorption kinetics indicated that the adsorption equilibrium of CrVI was reached rapidly within 2 h. In pure aqueous solution, CrVI reduction by PMP was observed only when the media's pH was lower than 4.0. Oxidation and reduction reaction between CrVI and p-methoxyphenol obviously occurred when soil colloids were involved in this system, even at pH > or = 7.0, which strongly suggested that minerals in soil colloids acted as catalysts to speed the reaction of CrVI and PMP. The oxidized product of PMP by CrVI, extracted by chloroform in acid media and analyzed by gas chromatography-mass spectrometry, was identified as benzoquinone. The reaction included two steps of one electron process.

Adsorption↗
PubMed2001

Levels of neuropeptide-Y in the plasma and skin tissue fluids of patients with vitiligo.

In order to study the possible role of neuropeptide-Y (NPY) in the pathogenesis of vitiligo, the authors measured the levels of NPY in the plasma from 47 patients with vitiligo compared with 25 healthy volunteers, and the tissue fluids of skin lesions and uninvolved skin from 32 patients, employing a NPY 125I RIA Kit. The results showed that the levels of NPY in the patients with vitiligo of all of the generalized, local and segmental types were significantly higher than the normal controls. In both local and segmental type, the levels in progressive stage were significantly higher than those in stable stage, while in generalized type, there was no significant difference between those in progressive stage and stable stage. The levels of NPY in the tissue fluids from skin lesions were significantly higher than those from uninvolved skin in both the local type and segmental type, while in the generalized type, there was no significant difference between the NPY level in the tissue fluid from skin lesion and that from uninvolved skin. It is speculated that NPY may play certain roles in the pathogenesis of vitiligo, via neuro-immunity mechanism or neuronal affection on the melanocytes.

Adolescent↗
PubMed2001

Biochemical, histological, and inhibitor studies of membrane carbonic anhydrase in frog gastric acid secretion.

Gastric acid secretion is dependent on carbonic anhydrase (CA). To define the role of membrane-bound CA, we used biochemical, histochemical, and pharmacological approaches in the frog (Rana pipiens). CA activity and inhibition by membrane-permeant and -impermeant agents were studied in stomach homogenates and microsomal fractions. H(+) secretion in the histamine-stimulated isolated mucosa was measured before and after mucosal addition of a permeant CA inhibitor (methazolamide) and before and after mucosal or serosal addition of two impermeant CA inhibitors of differing molecular mass: a 3,500-kDa polymer linked to aminobenzolamide and p-fluorobenzyl-aminobenzolamide (molecular mass, 454 kDa). Total CA activity of frog gastric mucosa is 2,280 U/g, of which 10% is due to membrane-bound CA. Membrane-bound CA retains detectable activity below pH 4. Histochemically, there is membrane-associated CA in surface epithelial, oxynticopeptic, and capillary endothelial cells. Methazolamide reduced H(+) secretion by 100%, whereas the two impermeant inhibitors equally blocked secretion by 40% when applied to the mucosal side and by 55% when applied to the serosal side. The presence of membrane-bound CA in frog oxynticopeptic cells and its relative resistance to acid inactivation and inhibition by impermeant inhibitors demonstrate that it subserves acid secretion at both the apical and basolateral sides.

4-Aminobenzoic Acid↗
PubMed2001

Characterization of carbonic anhydrase from Neisseria gonorrhoeae.

We have investigated the steady state and equilibrium kinetic properties of carbonic anhydrase from Neisseria gonorrhoeae (NGCA). Qualitatively, the enzyme shows the same kinetic behaviour as the well studied human carbonic anhydrase II (HCA II). This is reflected in the similar pH dependencies of the kinetic parameters for CO(2) hydration and the similar behaviour of the kinetics of (18)O exchange between CO(2) and water at chemical equilibrium. The pH profile of the turnover number, k(cat), can be described as a titration curve with an exceptionally high maximal value of 1.7 x 10(6) s(-1) at alkaline pH and a pK(a) of 7.2. At pH 9, k(cat) is buffer dependent in a saturable manner, suggesting a ping-pong mechanism with buffer as the second substrate. The ratio k(cat)/K(m) is dependent on two ionizations with pK(a) values of 6.4 and 8.2. However, an (18)O-exchange assay identified only one ionizable group in the pH profile of k(cat)/K(m) with an apparent pK(a) of 6.5. The results of a kinetic analysis of a His66-->Ala variant of the bacterial enzyme suggest that His66 in NGCA has the same function as a proton shuttle as His64 in HCA II. The kinetic defect in the mutant can partially be overcome by certain buffers, such as imidazole and 1,2-dimethylimidazole. The bacterial enzyme shows similar K(i) values for the inhibitors NCO(-), SCN(-) and N(3)(-) as HCA II, while CN(-) and the sulfonamide ethoxzolamide are considerably weaker inhibitors of the bacterial enzyme than of HCA II. The absorption spectra of the adducts of Co(II)-substituted NGCA with acetazolamide, NCO(-), SCN(-), CN(-) and N(3)(-) resemble the corresponding spectra obtained with human Co(II)-isozymes I and II. Measurements of guanidine hydrochloride (GdnHCl)-induced denaturation reveal a sensitivity of the CO(2) hydration activity to the reducing agent tris(2-carboxyethyl)phosphine (TCEP). However, the A(292)/A(260) ratio was not affected by the presence of TCEP, and a structural transition at 2.8--2.9 M GdnHCl was observed.

Carbonic Anhydrase Inhibitors↗
PubMed2001

[Study on the interaction between Pd(II) and Rhus vernicifera laccase].

The influence of the mixing time of Pd(II) and Rhus vernicifera laccase on the oxidation of 5,6-dibromo-2,3-dicyanohydroquinone (DDBQH2) catalyzed by laccase has been studied through spectrophotometer at pH 4.5 and (30 +/- 0.1 degree C). The activation of Pd(II) on the laccase activity is gradually converted into inhibition and ultimately inactivation with the extension of the mixing time of Pd(II) and laccase, and the rate of such conversion goes up with the increase of Pd(II) concentration. The influence of Pd(II) on the absorption spectrum of laccase suggests that it is the type I site in laccase with which Pd(II) interacts and Pd(II) may gradually and at last substitute Cu(II) from the site. The substitution may be the cause of decrease and disappearance of laccase activity.

Binding Sites↗
PubMed2000

Purification and kinetic analysis of recombinant CA XII, a membrane carbonic anhydrase overexpressed in certain cancers.

Carbonic anhydrase XII (CA XII) is a transmembrane glycoprotein with an active extracellular CA domain that is overexpressed on cell surfaces of certain cancers. Its expression has been linked to tumor invasiveness. To characterize its catalytic properties, we purified recombinant secretory forms of wild-type and mutant CA XIIs. The catalytic properties of these enzymes in the hydration of CO(2) were measured at steady state by stopped-flow spectrophotometry and at chemical equilibrium by the exchange of (18)O between CO(2) and water determined by mass spectrometry. The catalysis of CO(2) hydration by soluble CA XII has a maximal value of k(cat)/K(m) at 34 microM(-1) small middle dots(-1), which is similar to those of the membrane-associated CA IV and to soluble CA I. The pH profiles of this catalysis and the catalyzed hydrolysis of 4-nitrophenylacetate indicate that the pK(a) of the zinc-bound water in CA XII is 7.1. His64 in CA XII acts as a proton shuttle residue, as evidenced by the reduced rate constant for proton transfer in the mutants containing the replacements His64 --> Ala and His64 --> Arg, as well as by the selective inhibition of the proton transfer step by cupric ions in wild-type CA XII. The catalytic rate of CO(2) hydration by the soluble form of CA XII is identical with that of the membrane-bound enzyme. These observations suggest a role for CA XII in CO(2)/HCO(3)(-) homeostasis in cells in which it is normally expressed. They are also compatible with a role for CA XII in acidifying the microenvironment of cancer cells in which CA XII is overexpressed, providing a mechanism for CA XII to augment tumor invasiveness and suggesting CA XII as a potential target for chemotherapeutic agents.

Animals↗