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J L Lin

Publications and source records attributed to J L Lin.

At least 199 records · Page 11Linked to original sources
PubMed1988

Motility-dependence of the heterogenous staining of culture cells by a monoclonal anti-tropomyosin antibody.

A monoclonal antibody (CG1) which recognizes tropomyosin isoforms 1 and 3 of chicken embryo fibroblasts was used to detect what is a motility-dependent change in the availability of the antigenic determinant in tropomyosin molecules along microfilaments. Immunofluorescence microscopy with this antibody revealed a heterogenous staining pattern among chicken embryo fibroblasts cells such that a population (17%) of cells showed only background staining. Stress fibers in about half the population of the cells stained weakly with this antibody, while the stress fibers in another population of cells (35%) showed very strong staining. After glycerination or cytochalasin B treatment, all of the cells became positive in reaction to CG1 antibody, suggesting that the antigenic determinant was present in every cell. On the other hand, all of the cells after brief nonionic detergent treatment became negative to CG1 antibody. The CG1 staining pattern was not significantly changed in cells at different stages after release from colcemid blockage, nor was a brief treatment of cells with buffer containing 2 M urea, mild trypsin, chymotrypsin, or V.8 protease effective in changing the reactivity. However, most of the cells with a morphology typical of movement, and all of the contracted, glycerinated cells were strongly positive to CG1 antibody. These results suggest that the unmasking of the CG1 determinant may be motility-dependent. Immunoblot analysis showed that forced modification on the cysteine residue of tropomyosin molecules, caused either by performic acid oxidation or by disulfide cross-linking with the chemical 5,5'-dithiobis (2-nitrobenzoate), results in drastic changes in the reactivity of the different isoforms to CG1 antibody. These results indicate that the cysteine residue is involved in the CG1 determinant. The motility-dependent unmasking of this determinant may suggest an important role for nonmuscle tropomyosin in regulating cell motility.

Actin Cytoskeleton↗
PubMed1988

Differential localization of tropomyosin isoforms in cultured nonmuscle cells.

We have previously shown that chicken embryo fibroblast (CEF) cells and human bladder carcinoma (EJ) cells contain multiple isoforms of tropomyosin, identified as a, b, 1, 2, and 3 in CEF cells and 1, 2, 3, 4, and 5 in human EJ cells by one-dimensional SDS-PAGE (Lin, J. J.-C., D. M. Helfman, S. H. Hughes, and C.-S. Chou. 1985. J. Cell Biol. 100: 692-703; and Lin, J. J.-C., S. Yamashiro-Matsumura, and F. Matsumura. 1984. Cancer Cells 1:57-65). Both isoform 3 (TM-3) of CEF and isoforms 4,5 (TM-4,-5) of human EJ cells are the minor isoforms found respectively in normal chicken and human cells. They have a lower apparent molecular mass and show a weaker affinity to actin filaments when compared to the higher molecular mass isoforms. Using individual tropomyosin isoforms immobilized on nitrocellulose papers and sequential absorption of polyclonal antiserum on these papers, we have prepared antibodies specific to CEF TM-3 and to CEF TM-1,-2. In addition, two of our antitropomyosin mAbs, CG beta 6 and CG3, have now been demonstrated by Western blots, immunoprecipitation, and two-dimensional gel analysis to have specificities to human EJ TM-3 and TM-5, respectively. By using these isoform-specific reagents, we are able to compare the intracellular localizations of the lower and higher molecular mass isoforms in both CEF and human EJ cells. We have found that both lower and higher molecular mass isoforms of tropomyosin are localized along stress fibers of cells, as one would expect. However, the lower molecular mass isoforms are also distributed in regions near ruffling membranes. Further evidence for this different localization of different tropomyosin isoforms comes from double-label immunofluorescence microscopy on the same CEF cells with affinity-purified antibody against TM-3, and monoclonal CG beta 6 antibody against TM-a, -b, -1, and -2 of CEF tropomyosin. The presence of the lower molecular mass isoform of tropomyosin in ruffling membranes may indicate a novel way for the nonmuscle cell to control the stability and organization of microfilaments, and to regulate the cell motility.

Animals↗
PubMed1988

Monoclonal antibodies against caldesmon, a Ca++/calmodulin- and actin-binding protein of smooth muscle and nonmuscle cells.

Monoclonal antibodies, C2, C9, C18, C21 and C23, against chicken gizzard caldesmon have been prepared and characterized. These antibodies reacted with gizzard caldesmon (150 KDa) by enzyme-linked immunosorbent assay and protein immunoblotting. Immunofluorescence microscopy with these antibodies on cultured gizzard cells showed strong stress fiber and membrane ruffle stainings. Surprisingly, in addition to these cytoplasmic staining patterns, the C23 antibody also stained nuclei in these cells. Preabsorption of C23 with purified caldesmon abolished the staining of stress fibers and membrane ruffles as well as the staining of nuclei, suggesting that a common epitope existed in both gizzard caldesmon and the nuclear protein. Western blot analysis on the cell extract of chicken embryo fibroblast (CEF) showed that antibodies C2, C9, C18 and C21 recognized a nonmuscle caldesmon (66 KDa), whereas C23 reacted with a protein (60 KDa) different from nonmuscle caldesmon. Antibody C21 also crossreacted with a nonmuscle caldesmon (80 KDa) in normal rat kidney (NRK) cells, with a nonmuscle caldesmon (78 KDa) in human cells, and with a nonmuscle caldesmon (72 KDa) in gerbil fibroma cells. This antibody had broad-species specificity. Immunofluorescent staining of CEF cells with antibodies C2, C9, C18 and C21 showed some stress fibers and ruffles, but mostly diffuse staining. Antibody C23 crossreacted with 62 KDa and 55 KDa proteins in NRK cells, 63 KDa and 55 KDa proteins in gerbil fibroma cells and 66 KDa and 56 KDa proteins in human bladder carcinoma cells. These polypeptides were identified as nuclear lamins A and C by an anti-lamin antibody in immunoblots and two-dimensional gel analysis. Like the nuclear lamins, the C23 antigens also underwent a reversible disassembly during mitosis, as detected by double-label immunofluorescence with C23 antibody and a polyclonal anti-tubulin antibody. Tropomyosin-enriched microfilaments isolated from fibroblastic and epithelial types of NRK cells by monoclonal anti-tropomyosin antibody contained an 80 KDa protein, which had the heat-resistant property of caldesmon. The polyclonal antiserum generated against this 80 KDa protein showed a crossreactivity with purified gizzard caldesmon and vice versa. The amount of this nonmuscle caldesmon associated with the microfilaments of Kirstein virus-transformed NRK cells was greatly decreased.

Actin Cytoskeleton↗
PubMed1988

Reappraisal of atrioventricular junctional pacemaker automaticity in the sick sinus syndrome. Clinical significance and the role of autonomic chronotropic influences.

Postpacing impulse recovery times of the junctional tissue (junctional automaticity) were determined by atrial or ventricular overdrive pacing in 27 patients with dysfunction of the sinus node. The maximum junctional recovery time (MJRT) could be measured in 22 patients and ranged from 1,630 to 9,730 ms (mean 3,860 +/- 2,077); the maximum corrected junctional recovery time (MJRTc) could be measured in 18 patients and ranged from 140 to 5,986 ms (mean 2,089 +/- 1,529). Autonomic influence on the JRTs was evaluated by intravenous administration of atropine (1.5 mg) alone or in combination with propranolol (5 to 6 mg). Of the seven patients in whom MJRTc and/or MJRT could be measured before and after drug intervention, the JRTs shortened in four and prolonged in three after combination of atropine and propranolol. Atropine alone shortened MJRT in all eight patients studied. Our data reveal that both vagal and catecholamine-dependent factors (especially vagal over-activity) are operative in the escape mechanism of the junctional tissue.

Adult↗
PubMed1986

Terminal cardiac electrical activity in adults who die without apparent cardiac disease.

Prolonged electrocardiographic (Holter) recording was performed to analyze terminal electrical events in 23 hospitalized adults who died without apparent cardiac disease. Most patients showed a gradual slowing of heart rate with shifting of cardiac pacemaker downward from the sinus node or atria to the atrioventricular junction and ventricles, resulting in cardiac asystole. Dominant bradyarrhythmia was more common than ventricular tachyarrhythmia (83 vs 17%). Agonal ST-segment elevation was not uncommon (26%). These terminal electrical events became manifest from 1 to 450 minutes (mean 62) before cessation of cardiac electrical activity. Forty-eight percent of the patients continued to show deteriorating sinus or atrial activity up to the last moment. The mechanism of bradycardiac asystole in patients with no apparent cardiac disease may be attributed to generalized anoxic and toxic depression of the sinus node and subsidiary pacemakers, together with neurogenic suppression of these structures.

Adult↗
PubMed1986

Frequency of various congenital heart diseases in Chinese adults: analysis of 926 consecutive patients over 13 years of age.

A retrospective analysis was performed in 926 consecutive Chinese persons, aged 14 to 68 years, in whom congenital heart disease had been diagnosed and catheterization had been performed over the past 24 years. The incidence of various congenital cardiac malformations was compared with that in other series reported world-wide. About 60% of the 926 patients underwent surgery, and anatomic details of the malformations were examined. Congenital aortic valvular stenosis and coarctation of the aorta were uncommon (2.4% and 1.2%, respectively) among the Chinese adults, and the ethnic differences with Western countries in the incidence of these 2 defects were apparent and real. In contrast, aneurysm of the sinus of Valsalva, either ruptured or nonruptured, was more common (4.3%) and the incidence of outlet muscular type (or type 1) of ventricular septal defect was 4 to 9 times as common as has been reported in the West. Genetic differences rather than environmental factors or dietary habits may explain these special features.

Adolescent↗
PubMed1986

Assembly of different isoforms of actin and tropomyosin into the skeletal tropomyosin-enriched microfilaments during differentiation of muscle cells in vitro.

We have used a monoclonal antibody (CL2) directed against striated muscle isoforms of tropomyosin to selectively isolate a class of microfilaments (skeletal tropomyosin-enriched microfilaments) from differentiating muscle cells. This class of microfilaments differed from the one (tropomyosin-enriched microfilaments) isolated from the same cells by a monoclonal antibody (LCK16) recognizing all isoforms of muscle and nonmuscle tropomyosin. In myoblasts, the skeletal tropomyosin-enriched microfilaments had a higher content of alpha-actin and phosphorylated isoforms of tropomyosin as compared with the tropomyosin-enriched microfilaments. Moreover, besides muscle isoforms of actin and tropomyosin, significant amounts of nonmuscle isoforms of actin and tropomyosin were found in the skeletal tropomyosin-enriched microfilaments of myoblasts and myotubes. These results suggest that different isoforms of actin and tropomyosin can assemble into the same set of microfilaments, presumably pre-existing microfilaments, to form the skeletal tropomyosin-enriched microfilaments, which will eventually become the thin filaments of myofibrils. Therefore, the skeletal tropomyosin-enriched microfilaments detected here may represent an intermediate class of microfilaments formed during thin filament maturation. Electron microscopic studies of the isolated microfilaments from myoblasts and myotubes showed periodic localization of tropomyosin molecules along the microfilaments. The tropomyosin periodicity in the microfilaments of myoblasts and myotubes was 35 and 37 nm, respectively, whereas the nonmuscle tropomyosin along chicken embryo fibroblast microfilaments had a 34-nm repeat.

Actin Cytoskeleton↗
PubMed1985

Monoclonal antibodies against chicken tropomyosin isoforms: production, characterization, and application.

Eight mouse monoclonal antibodies, CH1, CH106, CH291, CL2, CG1, CG3, CG beta 2 and CG beta 6, against chicken tropomyosin isoforms have been prepared and characterized. The antigens recognized by these isoform-specific monoclonal antibodies were identified by both solid-phase radioimmunoassay and protein immunoblotting. To some extent, most antibodies showed isoform-specific, but one (CG3) recognized all isoforms of tropomyosin from chicken materials. The effects of monoclonal antibodies on the binding of cardiac tropomyosin to F-actin were investigated. Antibodies CH1, CH106, and CH291 had the ability to interfere with the binding of tropomyosin to F-actin, whereas others appeared to have no effect. Monoclonal antibody CL2 was able to distinguish the skeletal muscle tropomyosin-enriched microfilaments from the fibroblastic tropomyosin-enriched microfilaments of differentiating muscle cells. This antibody will be most useful for studying the compartmentalization of microfilaments and microfilament-associated proteins, particularly actin and tropomyosin isoforms during muscle differentiation. Immunofluorescence microscopy with CG1 antibody which recognized CEF tropomyosin isoforms 1 and 3 revealed the continuous staining of stress fibers in some populations of CEF cells. On the other hand, both periodic fluorescent staining and continuous staining of stress fibers were observed with CG3 antibody in all CEF cells.

Actin Cytoskeleton↗
PubMedDate not supplied

Recovery of respiratory function in survivors with paraquat intoxication.

The purpose of this study was to investigate chest radiograms and respiratory function changes, including pulmonary function tests and alveolar-arterial oxygen difference, in survivors with paraquat intoxication. Chest radiograms and pulmonary function tests for 21 paraquat-poisoned patients were performed 10 d after paraquat intoxication; 3 mo later, the tests were repeated in 16 patients who survived. Forced vital capacity, forced expiratory volume in 1 s, diffusing capacity of the lung, and alveolar-arterial oxygen difference were compromised after paraquat intoxication. Forced expiratory volume in 1 s and forced vital capacity correlated significantly with initial platelet counts (r = .453 and .443, respectively) 10 d after intoxication. The alveolar-arterial oxygen difference also correlated significantly with peak serum total bilirubin concentrations (r = .443) and initial platelet counts (r = .469). The follow-up data for respiratory functions forced expiratory volume in 1 s; 74.33 +/- 27.1% versus 97.89 +/- 16.39%; forced vital capacity: 71.44 +/- 26.03% versus 93.22 +/- 13.92%; diffusing capacity of lung: 60.11 +/- 27.61% versus 81.67 +/- 24.56%; alveolar-arterial oxygen difference: 37.95 +/- 24.32 mm Hg versus 7.75 +/- 9.94 mm Hg) and chest radiograms of survivors with moderate to severe paraquat poisoning showed significant improvements 3 mo after intoxication. The results demonstrated that paraquat-induced respiratory function impairments could recover significantly, at least partially, with time. In addition, pulmonary structure damage improved, as shown in the follow-up chest radiographs.

Adolescent↗
PubMedDate not supplied

Short-term small-dose intravenous iron trial to detect functional iron deficiency in dialysis patients.

BACKGROUND/AIM: Management of renal anemia in end-stage renal disease requires careful evaluation of the iron status before and in particular during erythropoietin treatment. However, there is no simple and practical iron index accurately predictive of functional iron deficiency in these patients till now. The purpose of this prospective study, therefore, is to evaluate whether a short course of low-dose intravenous iron challenge can detect functional iron deficiency in hemodialysis patients. METHODS: Twenty-four patients with baseline serum ferritin levels between 100 and 500 ng/ml were treated with intravenous saccharated ferric oxide, 960 mg over 24 hemodialysis treatments, and the hemoglobin level was checked every week. RESULTS: Patients whose hemoglobin value increased at least by 1 g/dl within the 8-week period were classified as having functional iron deficiency or as responders (n = 26; 81.2%). All other subjects were classified as having adequate iron levels or as nonresponders (n = 6; 18.8%). There were no significant differences in age, sex, dialysis years, Kt/V, dialyzers, hemoglobin, and basal and final transferrin saturation and ferritin between responders and nonresponders. In addition, there were no iron indices with acceptable levels of sensitivity and specificity. On the contrary, the cutoff value of increments of hemoglobin of at least 0.2 g/dl after a 2-week intravenous iron trial had a sensitivity of 96.2% and a specificity of 100% in all patients (n = 32) and a sensitivity of 100% and a specificity of 100% after patients with transferrin saturation <20% were excluded (n = 24). These values had the greatest utility of the tests studied in this work. CONCLUSION: A 240-mg intravenous iron challenge during a 2-week period may be a simple, accurate, and straightforward method to detect a functional iron deficiency status in hemodialysis patients undergoing erythropoietin therapy.

Adult↗
PubMedDate not supplied

Central diabetes insipidus following carbon monoxide poisoning.

Diabetes insipidus is a rare complication after carbon monoxide poisoning. We report on a woman and her daughter who were noted to have polyuria and hypernatremia after exposure to carbon monoxide. Both patients were responsive to desmopressin therapy, and the diagnosis of central diabetes insipidus was made. The development and correction of hypernatremia further complicated the neurological damages caused by carbon monoxide poisoning. These cases illustrate the possibility of central diabetes insipidus and hypernatremia after carbon monoxide poisoning and the importance of immediate antidiuretic hormone treatment and careful fluid therapy.

Adult↗
PubMedDate not supplied

Left atrial appendage blood flow determined by transesophageal echocardiography in patients with complete atrioventricular block.

Function of the left atrial appendage (LAA) represented by LAA outflow is an important predictor for thrombus formation in patients with nonrheumatic atrial fibrillation, but the pattern of LAA flow in patients with complete atrioventricular (AV) block has rarely been studied. Twenty-one patients with complete AV block and an implanted VVI or VVIR pacemaker were studied with transesophageal echocardiography. The LAA outflow velocity obtained during the ventricular diastolic phase was significantly higher than that obtained during the ventricular systolic phase (41.5 +/- 6.0 vs. 34.9 +/- 9.7 cm/s; p < 0.001). The LAA inflow velocity obtained during the ventricular diastolic phase was also significantly higher that that obtained during the ventricular systolic phase (29.9 +/- 7.8 vs. 26.4 +/- 5.3 cm/s; p < 0.01). In addition, the LAA outflow and inflow velocity time integrals during the ventricular diastolic phase were significantly higher than those during the ventricular systolic phase (4.66 +/- 0.96 vs. 4.08 +/- 1.05 cm, p < 0.01, and 2.81 +/- 0.77 vs. 2.56 +/- 0.65 cm, p < 0.05, respectively). Thus, due to both diastolic augmentation of the LAA flow related to active atrial contraction and minor early diastolic LAA flow formation, left ventricle diastolic function might have some influence on LAA flow. Thus may have implications for the pathogenesis of LAA thrombi in left ventricular dysfunction.

Adult↗
PubMedDate not supplied

Use of cardiac troponin T, creatine kinase and its isoform to monitor myocardial injury during radiofrequency ablation for supraventricular tachycardia.

To determine whether radiofrequency ablation for supraventricular tachycardia causes significant minor myocardial injury, 16 patients with supraventricular tachycardia undergoing radiofrequency ablation were studied. Cardiac troponin T, creatine kinase and its MB form (CKMB) were measured before, immediately after ablation and every 6 h thereafter for 24 h to detect myocardial injury. Elevation of creatine kinase, CKMB and cardiac troponin T was observed in 6, 4 and 5 patients, respectively. The peak mean creatine kinase concentration was 167 +/- 152 IU/l and that of CKMB was 9 +/- 6 IU/l. The peak mean cardiac troponin T level was 0.44 +/- 0.47 ng/ml. The frequency of elevated measurements was not statistically different among creatine kinase CKMB and cardiac troponin T. The mean pulse numbers of ablation, mean duration of ablation, radiofrequency current and mean total energy did not differ statistically between those with or without elevated cardiac troponin T. It was concluded that radiofrequency ablation for supraventricular tachycardia indeed caused some minor myocardial injury and the frequency of elevated cardiac troponin T was comparable to that of CKMB.

Adolescent↗