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Biomedical subjects

W C Chen

Publications and source records attributed to W C Chen.

At least 91 records · Page 5Linked to original sources
PubMed1999

Role of the cyclic AMP-protein kinase A pathway in lipopolysaccharide-induced nitric oxide synthase expression in RAW 264.7 macrophages. Involvement of cyclooxygenase-2.

The signaling pathway for lipopolysaccharide (LPS)-induced nitric oxide (NO) release in RAW 264.7 macrophages involves the protein kinase C and p38 activation pathways (Chen, C. C., Wang, J. K., and Lin, S. B. (1998) J. Immunol. 161, 6206-6214; Chen, C. C., and Wang, J. K. (1999) Mol. Pharmacol. 55, 481-488). In this study, the role of the cAMP-dependent protein kinase A (PKA) pathway was investigated. The PKA inhibitors, KT-5720 and H8, reduced LPS-induced NO release and inducible nitric oxide synthase (iNOS) expression. The direct PKA activator, Bt(2)cAMP, caused concentration-dependent NO release and iNOS expression, as confirmed by immunofluorescence studies. The intracellular cAMP concentration did not increase until after 6 h of LPS treatment. Two cAMP-elevating agents, forskolin and cholera toxin, potentiated the LPS-induced NO release and iNOS expression. Stimulation of cells with LPS or Bt(2)cAMP for periods of 10 min to 24 h caused nuclear factor-kappaB (NF-kappaB) activation in the nuclei, as shown by detection of NF-kappaB-specific DNA-protein binding. The PKA inhibitor, H8, inhibited the NF-kappaB activation induced by 6- or 12-h treatment with LPS but not that induced after 1, 3, or 24 h. The cyclooxygenase-2 (COX-2) inhibitors, NS-398 and indomethacin, attenuated LPS-induced NO release, iNOS expression, and NF-kappaB DNA-protein complex formation. LPS induced COX-2 expression in a time-dependent manner, and prostaglandin E(2) production was induced in parallel. These results suggest that 6 h of treatment with LPS increases intracellular cAMP levels via COX-2 induction and prostaglandin E(2) production, resulting in PKA activation, NF-kappaB activation, iNOS expression, and NO production.

Animals↗
PubMed1999

Relations of diastolic left ventricular filling to systolic chamber and myocardial contractility in hypertensive patients with left ventricular hypertrophy (The PRESERVE Study).

Abnormalities of left ventricular (LV) diastolic filling and stress-corrected midwall shortening (MWS) have been described in hypertensive patients with normal ejection fraction (EF). However, whether stress-corrected MWS parallels LV diastolic filling better than EF does remains uncertain. Blood pressure, body mass index, echocardiographic LV mass and LV geometry, EF and stress-corrected MWS, LV diastolic filling (peak E- and A-wave velocities, E-wave deceleration time, and atrial filling fraction) were evaluated in 212 hypertensive patients with LV hypertrophy enrolled in the Prospective Randomized Enalapril Study Evaluating Regression of Ventricular Enlargement study. LV structure, geometry, as well as LV diastolic filling, were compared between patients with reduced EF (<55%, n = 39, 18%) and those with normal EF (>55%) as well as between patients with reduced stress-corrected MWS (<89.2%, n = 31, 15%) and those with normal stress-corrected MWS (>89.2%). Patients with reduced EF had higher LV mass, eccentric LV geometry, and higher heart rate than those with normal EF, although they did not differ in age, blood pressure, or body mass index. LV filling pattern was also similar in those 2 groups. Patients with reduced stress-corrected MWS had higher atrial filling fraction, body mass index, heart rate, LV mass, and concentric geometry than those with normal stress-corrected MWS. Atrial filling fraction was negatively associated with stress-corrected MWS, but not with EF in multivariate models, independently of age, gender, heart rate, and body mass index. Thus, in hypertensive patients with LV hypertrophy, abnormal LV diastolic filling is more closely related to impaired myocardial contractility than to LV chamber EF.

Aged↗
PubMed1999

Expression of the focal adhesion protein paxillin in lung cancer and its relation to cell motility.

Lung cancer can lead to abnormalities of the actin cytoskeleton structure which may be important in transformation. In this study, we have investigated the expression of the cytoskeletal associated protein paxillin in lung cancer. Paxillin is a 68 kDa focal adhesion protein, with four tandem LIM domains at the C-terminus, involved in growth factor receptor, integrin and oncogenic signaling such as v-src, BCR/ABL, and E6 of the papilloma virus. In non-small cell lung cancer (NSCLC) cell lines, paxillin localized to the focal adhesions. The possible role of paxillin in lung cancer cells was assessed by overexpressing green fluorescence protein (GFP)-paxillin construct in two separate NSCLC cell lines (Calu-1 and H661). Over the course of 48 h, GFP-paxillin consistently caused the cells to become round and to decrease cell motility as compared to normal controls, GFP-N-terminus paxillin, or GFP-LIM transfected cells. Because some lung cancers may be quite aggressive and metastasize quickly, which may be related to the cytoskeleton, we determined the expression of paxillin in NSCLC and small cell lung cancer (SCLC) cell lines and patient tumor tissues. Expression of paxillin in NSCLC and SCLC cell lines were determined by Northern blot and Western blot analysis. The expression of paxillin was consistently low in SCLC cell lines, whereas there was paxillin expression in NSCLC cell lines. There was a variability of expression of paxillin in NSCLC tumor tissue as compared to normal lung tissue. In contrast, by immunohistochemistry, we show that there was no detectable expression of paxillin in 5/5 SCLC patients. This data suggests that absence or low level of paxillin protein expression may cause certain lung cancers, such as SCLC, to be more motile and possibly more aggressive.

3T3 Cells↗
PubMed1999

MRI in recurrent nasopharyngeal carcinoma.

In this study, we retrospectively reviewed the MRI features of recurrent nasopharyngeal carcinoma (NPC) in 72 patients who underwent MRI before and after gadolinium injection. Recurrent NPC exhibited a high degree of regional spread and a variety of signal intensities and contours. MRI showed a nasopharyngeal mass in 50 patients (69.4 %); other sites of involvement included the parapharyngeal space (44.4 %), nasal cavity (12.5 %), paranasal sinuses (27.8 %), oropharynx (4.2 %), orbit (8.3 %), infratemporal fossa (18.1 %), skull base (59.8 %), intracranial area (51.4 %) and regional lymph nodes (15.3 %). On T2-weighted images, the nasopharyngeal mass gave high signal in 9 of 50 cases (18 %), intermediate in 27 (54 %), mixed in 8 (16 %) and low signal in 6 (12 %). Contrast enhancement was strong in 12 cases (24 %), moderate in 29 (58 %) and heterogeneous in 9 (18 %). The lesion was convex in 31 cases (62 %) and concave or straight in 19 (38 %). Recognition of the distribution and the appearance of recurrent NPC on MRI is essential for timely diagnosis and appropriate treatment.

Adult↗
PubMed1999

Phosphorylation of transcription factor cyclic-AMP response element binding protein mediates c-fos induction elicited by sustained hypertension in rat nucleus tractus solitarii.

We investigated the role of cyclic-AMP response element binding protein signaling in the induction of the immediate-early gene c-fos by baroreceptor activation in neurons of the nucleus tractus solitarii of anesthetized rats. Activation of the arterial baroreceptors with sustained hypertension significantly increased the number of neurons in the caudal nucleus tractus solitarii that were immunoreactive to an antiserum that detects Ser133-phosphorylated cyclic-AMP response element binding protein. This implied increase in phosphorylation of cyclic-AMP response element binding protein was subsequently followed by an elevation in the expression of Fos protein in neurons of the nucleus tractus solitarii. Microinjection bilaterally into the nucleus tractus solitarii of a phosphorothioated antisense oligonucleotide directed against the initiation site of cyclic-AMP response element binding protein messenger RNA discernibly reduced the manifested immunoreactivity of phosphorylated cyclic-AMP response element binding protein in response to baroreceptor activation. This was accompanied by a decline in the transcription of c-fos messenger RNA and the expression of Fos protein, along with an appreciable potentiation of the baroreceptor reflex response. Control injections of the sense oligonucleotide or artificial cerebrospinal fluid were ineffective. These findings suggest that phosphorylation of cyclic-AMP response element binding protein is crucial to Fos expression in the nucleus tractus solitarii elicited by sustained hypertension. As such, phosphorylation of cyclic-AMP response element binding protein may be an important early nuclear event that mediates the long-term inhibitory modulation of the baroreceptor reflex response by Fos protein at the nucleus tractus solitarii.

Animals↗
PubMed1999

The influence of delay and the effect of fibrin sealant on the cut surface of the peripheral nerve. An experimental study in the rat.

Thirty-two sciatic nerves in 16 rats were divided to investigate the effect of delay in fixation and the use of fibrin sealant at the site of the division on the nerve end. Specimens were assessed by morphological and morphometric criteria using scanning electron microscopy and longitudinal sections. All specimens showed a protruded nerve end. Wrapping the nerve with fibrin sealant before division and immediate fixation of the specimen resulted in less protrusion.

Animals↗
PubMed1999

Radiation-induced hearing impairment in patients treated for malignant parotid tumor.

Radiation-induced hearing loss was evaluated in 21 patients with unilateral malignant parotid tumors treated with surgery and radiotherapy. The contralateral ear was used as a control. Eight patients (38%) were found to have a reduction in static compliance of the tympanic membrane (type B tympanogram) in the irradiated ear. By audiometry, significant hearing loss was found in 9 patients (43%). These hearing losses were mainly sensorineural, as shown by a similar reduction in both air and bone conduction, although mixed-type hearing loss existed in some patients. A statistically significant difference in incidence of 67% versus 0% (p = .0085) was noted for patients with a cochlear dose of greater than or equal to 60 Gy, in comparison to those receiving doses of less than 60 Gy. A type B tympanogram was also found to be a prognostic factor for significant sensorineural hearing loss. Patients with type B tympanograms had a much higher incidence of significant sensorineural hearing loss than those with type A tympanograms (88% versus 15%, p = .02). This study clearly shows that radiotherapy can induce significant hearing impairment, especially when the cochlear doses are higher than 60 Gy.

Acoustic Impedance Tests↗
PubMed1999

Signal transduction of arginine vasopressin-induced arachidonic acid release in H9c2 cardiac myoblasts: role of Ca2+ and the protein kinase C-dependent activation of p42 mitogen-activated protein kinase.

The mechanism of arginine vasopressin (AVP)-induced arachidonic acid (AA) release was examined in the cardiac myoblast cell line, H9c2. Stimulation of cells with AVP induced dose-dependent AA release, and this effect was completely inhibited by the V1 receptor antagonist, d(CH)5[Tyr(Me)2]AVP. AVP also produced dose-dependent stimulation of inositol phosphate formation; this was not affected by pertussis toxin, indicating the presence of the V1 receptor/Gq protein/PLCbeta pathway in H9c2 cells. The concentration-response curves for these two effects of AVP overlapped. AVP induced a rapid increase in [Ca2+]i, followed by a sustained increase. The Ca2+ ionophore, A23187 or ionomycin, mimicked the effect of AVP, whereas the protein kinase C (PKC) activator, TPA, only induced a slight increase in AA release. Both the AVP- or A23187-stimulated AA release and the AVP-induced sustained [Ca2+]i increase were completely blocked in the absence of external Ca2+. The receptor-operated Ca2+ channel blocker, SKF 96365, and the inorganic Ca2+ channel blockers, Ca2+ and Ni2+, also inhibited the AVP-induced AA release. Western blots demonstrated expression of PKCalpha, betaI, epsilon, delta, and zeta in H9c2 cells; PKC inhibitors (staurosporine or Ro 31-8220) or down-regulation of PKCalpha, betaI, epsilon, and delta by long-term (24 h) TPA treatment caused a partial blockade of the AVP-induced response, whereas the A23187-induced AA release was unaffected by down-regulation of these isoforms. AVP-induced, but not A23187-induced, AA release was partially blocked by the p42 MAPK cascade inhibitor, PD 98059. AVP and TPA, but not A23187, induced an increase in activity and tyrosine phosphorylation of p42 MAPK, together with a molecular weight shift, consistent with phosphorylation, of cytosolic PLA2. AVP- or TPA-induced activation and tyrosine phosphorylation of p42 MAPK were completely blocked by down-regulation of PKCalpha, betaI, epsilon, and delta, but still occurred, together with the cytosolic PLA2 mobility shift, in the absence of external Ca2+. These results show that AVP-induced AA release in H9c2 cells is secondary to activation of the V1 receptor/Gq protein/PLCP pathway, leading to an influx of extracellular Ca2+ and activation of PKCalpha, betaI, epsilon, and delta. The influx of extracellular Ca2- and DAG act, respectively, through PKC-/MAPK-independent or PKC-dependent MAPK pathways to mediate AA release.

Arachidonic Acid↗
PubMed1999

Production of cholesterol oxidase by Rhodococcus equi No. 23 in a jar fermenter.

Rhodococcus equi No. 23 was grown in a batch fermenter. The effects of cultivation temperature, pH of the culture medium, aeration rate and agitation speed on the production of cholesterol oxidase (CholOx) by the test organism were examined. Results revealed that the cultivation temperature, the pH of the medium, the aeration rate and the agitation speed all affected the production of CholOx by R. equi No. 23. Adjusting the operation variables during the cultivation period increased the production of CholOx effectively and prevented the occurrence of overflow of foam during the fermentation period. A maximum CholOx activity of 0.34 unit/ml with a volumetric production rate of 0.011 unit/h per ml could be achieved in 30 h of cultivation at an aeration rate of 5.0 l/min, if the pH of the culture medium, the cultivation temperature and the agitation speed were controlled at 6.5, 39 degrees C and 200 rev. /min respectively during the first 24 h of cultivation, then shifted to 7.5, 37 degrees C and 300 rev./min respectively.

Bioreactors↗
PubMed1999

[Different age of asthmatic patients affected by different aeroallergens].

A total of 1070 cases of bronchial asthma, aged from 3 to 70 years, were analyzed for their hypersensitivity to the four most common aeroallergens, Dermatophagoides pteronyssinus, German cockroaches, Penicillium notatum and Candida albicans. The severity of hypersensitivity was classified as mild, moderate and severe based on the concentration of allergen specific IgE in the sera. Results showed that there were 77.9% patients allergic to D. pteronyssinus, 40.0% to G. cockroaches, 9.6% to P. notatum and 10.4% to C. albicans. The incidence and severity varied with age. For D. pteronyssinus, the highest incidence was 96.8% with the age between 11 and 20 years. Seventy percent of them were severe hypersensitivity. For P. notatum, the highest incidence was 18.9% with the age above 61 years. None of them were severe hypersensitivity and 11.9% of them were moderate hypersensitivity. For C. albicans, the highest incidence was 27.3% with the age between 51 and 60 years. All of them were mild hypersensitivity. For G. cockroaches, the highest incidence was 55.4% with the age between 21 and 30 years. 76.8% of them were mild hypersensitivity and 23.2% were moderate hypersensitivity. We concluded that different age of asthmatic patients affected by different aeroallergens. D. pteronyssinus was the most potent aeroallergen. Most asthmatic patients sensitized by mite were severe, particularly those patients with the age below 20 years of age.

Adolescent↗
PubMed1999

[Relation of different food allergens to different age of asthmatic patients].

A total of 1,070 cases of bronchial asthma, age from 3 to 70 years, were analyzed for their hypersensitivity to the four most common food allergens, egg white, milk, shrimp and peanuts. The severity of hypersensitivity was classified as mild, moderate and severe based on the concentration of allergen specific IgE in the sera. Results showed that there were 16.2% patients allergic to egg white, 22.3% allergic to milk, 12.6% allergic to peanuts and 34.9% allergic to shrimp. The incidence and severity varied with age. For milk and egg white allergy, the age with highest incidence rate appeared to be under 10 years old. In this age group, there were 38.1% patients allergic to milk, 8.9% of them were moderate hypersensitivity and the rest were mild hypersensitivity. There were 27.3% patients allergic to egg white, 17.3% of them were moderate hypersensitivity and the rest were mild hypersensitivity. For shrimp and peanuts allergy, the age with highest incidence rate appeared to be 11 to 20 years old. In this age group, there were 51.9% patients allergic to shrimp, 42.9% of them were moderate hypersensitivity and the rest were mild hypersensitivity. There were 18.2% patients allergic to peanuts, 25.0% of them were moderate hypersensitivity and the rest were mild hypersensitivity. We concluded that different age of asthmatic patients were affected by different food allergen. Shrimp and peanuts allergy occurred later and more severe than milk and egg white allergy, however, most of them were mild hypersensitivity and might be only partially contributed to their asthmatic symptoms.

Adolescent↗
PubMed1998

Nifedipine, verapamil and diltiazem block shock-wave-induced rises in cytosolic calcium in MDCK cells.

Nifedipine and verapamil have been shown previously to protect against renal function alterations induced by shock wave lithotripsy (SWL) in humans and rats; however, the mechanism is unclear. This study was aimed to examine whether these drugs could protect cultured kidney cells following shock wave exposure (SWE). The effect of nifedipine, verapamil and diltiazem on Madin Darby canine kidney (MDCK) cells following SWE was examined by determining the release of glutamate oxalactate transferase (GOT) and lactate dehydrogenase (LDH) in cell suspensions; and also cytosolic Ca2+ concentration ([Ca2+]i). Immediately after SWE, there was a transient release of GOT and LDH (16% and 4 fold, respectively). In contrast, [Ca2+]i measured within 1-6 hr after SWE gradually increased by 15-156%. The Ca2+ entry blockers (1 or 10 microM) failed to inhibit the enzyme release; however, they abolished the progressive rises in [Ca2+]i. The Ca2+ entry blockers may protect the cells from damage of SWE via maintaining a low resting [Ca2+]i.

Adenosine Triphosphate↗
PubMed1998

Synthesis and biological activity of [Tic5] didemnin B.

A didemnin B analog containing a Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) as a conformationally restrained replacement for tyrosine has been synthesized and shown to have comparable potency as a protein biosynthesis inhibitor. Synthetic highlights include an oxidation of an alcohol to an acid in the presence of the sensitive Tic heterocycle and a modified Schmidt-type one-pot macrocyclization.

Animals↗
PubMed1998

Protein kinase C eta mediates lipopolysaccharide-induced nitric-oxide synthase expression in primary astrocytes.

The signaling pathway involved in protein kinase C (PKC) activation and role of PKC isoforms in lipopolysaccharide (LPS)-induced nitric oxide (NO) release were studied in primary cerebellar astrocytes. LPS caused a dose- and time-dependent increase in NO release and inducible NO synthase (iNOS) expression. The tyrosine kinase inhibitor, genestein, the phosphatidylcholine-phospholipase C inhibitor, D609, and the phosphatidate phosphodrolase inhibitor, propranolol, attenuated the LPS effects, whereas the PI-PLC inhibitor, U73122, had no effect. The PKC inhibitors (staurosporine, Ro 31-8220, Go 6976, and calphostin C) also inhibited LPS-induced NO release and iNOS expression. However, long term (24 h) pretreatment of cells with 12-O-tetradecanoyl phorbol-13-acetate (TPA) did not affect the LPS response. Previous results have shown that TPA-induced translocation, but not down-regulation, of PKCeta occurs in astrocytes (Chen, C. C., and Chen, W. C. (1996) Glia 17, 63-71), suggesting possible involvement of PKCeta in LPS-mediated effects. Treatment with antisense oligonucleotides for PKCeta or delta, another isoform abundantly expressed in astrocytes, demonstrated the involvement of PKCeta, but not delta, in LPS-mediated effects. Stimulation of cells for 1 h with LPS caused activation of nuclear factor (NF)-kB in the nuclei as detected by the formation of a NF-kB-specific DNA-protein complex; this effect was inhibited by genestein, D609, propranolol, or Ro 31-8220 or by PKCeta antisense oligonucleotides, but not by long term TPA treatment. These data suggest that in astrocytes, LPS might activate phosphatidylcholine-phospholipase C and phosphatidylcholine-phospholipase D through an upstream protein tyrosine kinase to induce PKC activation. Of the PKC isoforms present in these cells, only activation of PKCeta by LPS resulted in the stimulation of NF-kB-specific DNA-protein binding and then initiated the iNOS expression and NO release. This is further evidence demonstrating that different members of the PKC family within a single cell are involved in specific physiological responses.

Animals↗
PubMed1998

ATP-induced arachidonic acid release in cultured astrocytes is mediated by Gi protein coupled P2Y1 and P2Y2 receptors.

ATP-induced arachidonic acid (AA) release was studied in [3H]AA-prelabeled cultured astrocytes. To characterize the P2 purinoceptor-mediated effect of ATP, the subtype-specific agonists 2-methylthio ATP (2-MeSATP) and UTP were compared. ATP, UTP, or 2-MeSATP induced a dose-dependent increase of [3H]AA release, with EC50 values of 22.7 microM, 29.4 microM, and 1.68 microM, respectively; alpha,beta-methyleneATP and adenosine had no effect. The order of potency was ATP = UTP > or = 2-MeSATP, indicating that ATP interacted with both P2Y1 and P2Y2 receptors to mediate AA release in astrocytes. The effect of ATP, UTP, or 2-MeSATP was markedly inhibited by pretreatment of cells with pertussis toxin. Ca2+ ionophore-A23187 and PKC activator-TPA mimicked the effects of these three agonists to stimulate AA release. ATP, UTP, and 2-MeSATP induced a rapidly initial rise of [Ca2+]i and a sustained [Ca2+]i increase. The AA release was blocked in the external Ca2+ free in condition the sustained [Ca2+]i increase was abolished. Both A23187- and TPA-induced AA release were also blocked in this condition. Furthermore, inorganic Ca2+ channel blocker Co2+ inhibited ATP, UTP, or 2-MeSATP induced AA release as well. Long-term (24 h) treatment of cells with TPA resulted in an attenuation of three agonists, TPA or A23187 response. Similarly, ATP or TPA promoted AA release was inhibited by the mitogen-activated protein kinase (MAPK) cascade inhibitor PD 98059. ATP, TPA, or A23187 induced an increase in the activity and tyrosine phosphorylation of p42 MAPK, as well as a molecular weight shift, consistent with phosphorylation, of cytosolic phospholipase A2 (cPLA2). ATP- and TPA-stimulated activation of p42 MAPK activity and tyrosine phosphorylation were inhibited by long-term TPA treatment, while A23187-stimulated effects were completely blocked. Furthermore, tyrosine phosphorylation and activation of p42 MAPK and mobility shift of cPLA2 induced by A23187 were reversed in the absence of external Ca2+, suggesting the involvement of PKCalpha in MAPK activation and mobility shift of cPLA2. Taken together, ATP-stimulated AA release was secondary to the activation of P2Y1 and P2Y2 receptors/PLC pathway. Ca2+ and PKC interact to regulate this response. Elevation of intracellular Ca2+, the mechanism involving extracellular Ca2+ influx, might act partly through PKCalpha activation and in turn MAPK might be activated, leading to cPLA2 phosphorylation and AA release.

Adenosine Triphosphate↗
PubMed1998

Comparison of the immunogenicity of wasp venom peptides with or without carbohydrate moieties.

Three synthetic vespulakinin analogues either with or without carbohydrate moieties and mastoparan B isolated from Vespa basalis venom were investigated for their immunogenic activity and solution conformation. Mice immunized with these wasp venom peptides, with the exception of (Gal alpha)Thr3, (Gal alpha)Thr4-vespulakinin 1, showed positive antibody responses. However, the response elicited by mastoparan B was much higher than those induced by vespulakinin analogues. The class of antibody induced by these peptides was identified as an IgG1 isotype with kappa-light chain, suggesting stimulation of a T-cell-dependent immune response by these peptides. According to the circular dichroism spectra of these peptides, the structures of the vespulakinin analogues in solution were largely unordered, while mastoparan B exhibited a conformation rich in alpha-helices. The presence of carbohydrate moieties and the rather random structure in vespulakinins may interfere with T-cell recognition of the peptides, leading to lower immune responses.

Allergens↗