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F Chen

Publications and source records attributed to F Chen.

At least 577 records · Page 32Linked to original sources
PubMed1992

Effect of ambasilide, a new class III agent, on plateau currents in isolated guinea pig ventricular myocytes: block of delayed outward potassium current.

The actions of ambasilide (LU-47110) on the action potential and membrane currents of isolated guinea pig ventricular myocytes were studied using voltage clamp techniques. Ambasilide (1 microM) prolonged the action potential (APD) at 20, 50 and 90% repolarization by 11.2 +/- 4.3, 13.8 +/- 3.9 and 13.6 +/- 3.7%, respectively, compared to control (n = 10). APD prolongation was attributed to the block of delayed rectifier outward current (Ik) in a concentration-dependent fashion (0.01-10 microM). The effects on the APD and Ik were both partially reversed after perfusion with drug-free Tyrode's solution. The block of Ik by ambasilide was compared to that by E-4031 (5 microM), a putative selective blocker of that fast, inwardly rectifying component of Ik identified in guinea pig ventricle. E-4031 produced about 65% block of Ik for pulse durations between 80 and 420 msec, but the block decreased as the pulse duration increased further, the block accounting for 34 +/- 5% of Ik at 6.3 sec. In contrast, the percentage of reduction of Ik by 10 microM ambasilide did not produce a consistent magnitude of block over a similar range of short depolarizations, but rather progressively decreased Ik as the pusle duration lengthened. Block at the end of a 2-sec pulse was about 48 +/- 8%, more block than could be attributed to an E-4031-sensitive current block alone. Whereas E-4031 (5 microM) shifted the activation curve of Ik 10 mV toward positive potentials and decreased the slope factor, k, by about 4 mV, ambasilide (5 microM) had no effect on these parameters.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1991

Contributions of arginines-43 and -94 of human choriogonadotropin beta to receptor binding and activation as determined by oligonucleotide-based mutagenesis.

Members of the glycoprotein hormone family contain a common alpha subunit and a hormone-specific beta subunit. Human choriogonadotropin (hCG) beta is a 145 amino acid residue protein glycosylated at 6 positions (2 N-linked and 4 O-linked oligosaccharides). In an effort to elucidate receptor determinants on hCG beta, we have used site-directed mutagenesis to prepare and express several mutant cDNAs with replacements at arginines-43 and -94. Arg-43 is invariant in all known mammalian CG/lutropin beta amino acid sequences, and Arg-94 is conserved in 10 of the 12 sequences. Moreover, various studies involving synthetic peptides and enzymatic digestions of intact beta chains suggest that these residues may be important in hCG receptor binding. Point mutants were made in which these two arginines were replaced with the corresponding residues in human follitropin beta, Leu-43 and Asp-94. The wild-type and mutant beta chains were expressed in CHO cells containing a stably integrated gene for bovine alpha, and heterodimer formation occurred. These heterologous gonadotropins were active in assays using transformed Leydig cells, competitive binding with standard 125I-hCG, and cAMP and progesterone production, but the potency was considerably less than that associated with the hCG beta wild-type-containing gonadotropin. The double-mutant protein Arg-43 to Leu/Arg-94 to Asp also associated with bovine alpha, but the resultant heterodimer exhibited only low activity. Replacement of each arginine with lysine yielded heterodimers that were at least as potent as bovine alpha-hCG beta wild type, but the Lys-43-containing beta chain appeared to exhibit a low degree of subunit association or reduced stability relative to the expressed hCG beta wild type. These results demonstrate that arginines-43 and -94 contribute to receptor binding through a positive charge.

Amino Acid Sequence↗
PubMed1991

Role of the invariant aspartic acid 99 of human choriogonadotropin beta in receptor binding and biological activity.

The four human glycoprotein hormones are heterodimers that contain a common alpha subunit and a hormone-specific beta subunit. Within this hormone family, 23 amino acid sequences from 11 mammalian species are available. There are 19 invariant amino acid residues in the beta subunits, 12 of which are Cys that form six disulfide bonds. Of the remaining seven conserved amino acid residues, we have investigated the role of an Asp which occurs at position 99 in human choriogonadotropin beta (hCG beta). Site-directed mutagenesis was used to replace hCG beta Asp99 with three residues, Glu, Asn, and Arg, and to prepare an inversion double mutant protein, Arg94----Asp and Asp99----Arg. The cDNAs were placed in a eukaryotic expression vector, and the plasmids were transiently transfected into Chinese hamster ovary cells containing a stably integrated gene for bovine alpha. Radioimmunoassays demonstrated that the mutant forms of hCG beta were capable of subunit assembly to the same extent as hCG beta wild type. The heterologous heterodimers were assayed in vitro using transformed mouse Leydig cells (MA-10) by competitive inhibition of 125I-hCG binding and stimulation of progesterone production. The gonadotropins containing Glu and Asn were active, although the potency was less than that associated with the hCG beta wild type-containing gonadotropin. In contrast, the Arg99-containing mutant protein and the inversion mutant protein Asp94/Arg99 were devoid of activity. Thus, in hCG beta Asp99 can be substituted with certain residues without total loss of function, although replacement with a positively charged residue leads to an inactive heterodimer. The primary role of Asp99 in hCG beta seems to involve, either directly or indirectly, receptor recognition.

Amino Acid Sequence↗
PubMed1991

Delineation via site-directed mutagenesis of the carboxyl-terminal region of human choriogonadotropin beta required for subunit assembly and biological activity.

The choriogonadotropin beta subunit is unique in the human glycoprotein hormone family in containing a carboxyl-terminal extension, with four sites of O-glycosylation, that is not present in the other beta subunits. We have used site-directed mutagenesis to define boundaries on the extent to which truncations can be made at the COOH terminus without abolishing subunit assembly and biological activity. Two COOH-terminal deletion mutant chains of human choriogonadotropin beta, des(93-145) and des(101-145), were prepared and expressed in Chinese hamster ovary cells containing a stably integrated gene for bovine alpha. The heterologous gonadotropin, bovine alpha-human choriogonadotropin des(101-145) beta, formed a heterodimer and, when assayed with transformed murine Leydig cells in vitro, competed with the binding of standard human choriogonadotropin and stimulated both cAMP and progesterone production, albeit with a reduced potency relative to bovine alpha-human choriogonadotropin beta wild type. In contrast, human choriogonadotropin des(93-145) beta, expressed under identical conditions in the presence of bovine alpha, failed to form heterodimer and thus exhibited no competitive binding and was without effect on cAMP and progesterone levels. Consequently, removal of the putative determinant loop region of the beta subunit (residues 93-100), which is believed to be important in determining receptor specificity, abolishes association with alpha. Hence, in addition to its possible role as a receptor determinant, this region of the molecule appears to be critical for proper folding or subunit interaction. The truncated form of human choriogonadotropin beta lacking residues 101-145 is the shortest form of the subunit yet described that retains biological activity. Moreover, these results demonstrate that the proposed disulfide between Cys-26 and Cys-110 is not required for subunit assembly or for receptor binding and subsequent intracellular signaling.

1-Methyl-3-isobutylxanthine↗
PubMed1991

Single-channel recording of inwardly rectifying potassium currents in developing myocardium.

Properties of the inwardly rectifying K+ channel, which contributes to the maintenance of the resting membrane potential, were studied in neonatal rabbit ventricular myocytes using the patch-clamp technique. Inward rectification was evident in single-channel current-voltage (I-V) relations at potentials positive to the potassium equilibrium potential (Ek = 0 mV with [K+]o = [K+]i = 150 mM, [Mg2+]i = 2 mM). The single-channel conductance was 3.2 +/- 0.1 pS in physiological (5.4 mM) [K+]o. The zero-current potential shifted 48.4 +/- 2.4 mV for a ten-fold change in [K+]o in neonatal cells. External Ba2+ blocked the current in a dose-dependent manner. The voltage dependence, open-state probability and channel density of this channel were compared between neonatal and adult ventricular myocytes isolated by similar techniques. The open-state probability of the channel was approximately the same in neonatal (0.39 +/- 0.06, n = 13) as in adult cells (0.4 +/- 0.05, n = 11). However, in symmetrical transmembrane K+ concentration [( K+]o = [K+]i = 150 mM), the single channel conductance was significantly smaller in neonatal (25 +/- 0.3 pS, n = 25) as compared with adult cells (31 +/- 0.4 pS, n = 12). In addition, the relationship between resting membrane potential and [K+]o was measured in neonatal and adult myocytes. The resting membrane potential in the neonate was less dependent on [K+]o than in the adult. These results are consistent with an age-related change in resting membrane K+ permeability which may result from a developmental change in the single-channel conductance properties of the inwardly rectifying K+ channel.

Action Potentials↗
PubMed1991

Genetic diversity among toxigenic and nontoxigenic Vibrio cholerae O1 isolated from the Western Hemisphere.

Multilocus enzyme electrophoresis was used to examine genetic relationships among and between toxigenic and non-toxigenic isolates of Vibrio cholerae O1 obtained from patients and the environment in the US Gulf Coast and surrounding areas. A total of 23 toxigenic and 23 non-toxigenic strains were examined. All the toxigenic and 7 of the non-toxigenic strains had the same alleles at 16 enzyme loci, whereas the balance of the nontoxigenic strains had 9 distinct combinations of alleles. This study suggests that all of the toxigenic strains belong to a single clone, and that while some of the non-toxigenic isolates were related, most were of diverse origin.

Alleles↗
PubMed1991

Isolation and sequencing of the Epstein-Barr virus BNLF-1 gene (LMP1) from a Chinese nasopharyngeal carcinoma.

The BamHI fragment containing the Epstein-Barr virus (EBV) LMP1 gene was cloned from a genomic library of the nude mouse-propagated Chinese nasopharyngeal carcinoma CAO. The sequence of the LMP1 gene and its promoter and enhancer was determined. The nucleotide sequence of the CAO isolate differed from those of the B95-8 and Raji isolates in the promoter/enhancer region; the amino acid sequence of the protein also differed. Structural differences in the protein were located mainly in the 20 N-terminal residues and the array of repeated amino acids in the C-terminal part of the protein, in which the CAO isolate displays a cluster of seven perfect repeats of 11 amino acids (aa). Three of these repeats have no counterpart in the other virus strains. This, together with two deletions of five and 10 aa in the C-terminal part, yields a protein of 404 aa, compared to 386 aa for B95-8 and Raji. The larger LMP1 protein was detected on immunoblots of tissue samples from the CAO nude mouse tumour, and was also present in EBV-negative B cell lines and immortalized keratinocytes transfected with the cloned gene. A XhoI restriction site in exon 1 of the B95-8 BNLF-1 gene was absent from the CAO EBV isolate, as well as from 36 of 37 Chinese NPC biopsies tested. In contrast, 17 of 19 NPC biopsies of African origin retained this XhoI site.

Amino Acid Sequence↗
PubMed1991

Nonneoplastic enhancing lesions mimicking intracanalicular acoustic neuroma on gadolinium-enhanced MR images.

The authors describe five patients with nonneoplastic lesions of the facial and/or vestibulocochlear nerves that demonstrated focal enhancement within the internal auditory canal on magnetic resonance (MR) images. MR and surgical findings for four patients with unilateral sensorineural hearing loss and one with Ramsay Hunt syndrome were reviewed. Three patients with hearing loss underwent surgical exploration and decompression of the internal auditory canal. The MR findings in all four patients with hearing loss were similar: Focal enhancement of the internal auditory canal was depicted on postcontrast T1-weighted images. Nonneoplastic lesions of the seventh and eighth cranial nerves may show focal enhancement on MR images, which mimics the appearance of a small intracanalicular neuroma. This potential for misdiagnosis may have important therapeutic implications.

Cranial Nerve Diseases↗
PubMed1991

Treatment of S antigen uveoretinitis with lipoxygenase and cyclo-oxygenase inhibitors.

The role of metabolites of arachidonic acid (AA) in experimental autoimmune uveoretinitis was studied using inhibitors of AA metabolism. Nordihydroguaiaretic acid (NDGA), which inhibits predominantly the lipoxygenase (LO) pathway, or indomethacin, a specific cyclo-oxygenase (CO) inhibitor, was administered to rats immunized with retinal S antigen. Levels of various AA metabolites were measured in the inflamed uvea, and the severity of intraocular inflammation was quantitated by morphometric analysis. Histopathologically, the uveoretinitis was significantly suppressed following treatment with NDGA, while indomethacin treatment resulted in augmentation of the disease (p less than 0.05). These results tend to indicate that the inhibition of the LO rather than the CO pathway may be more beneficial in the treatment of autoimmune uveitis.

Animals↗
PubMed1991

Calcium current measurements in acutely isolated neonatal cardiac myocytes.

Action potentials and voltage clamp-induced ionic currents were recorded in acutely isolated neonatal rabbit cardiac myocytes using the whole-cell voltage clamp technique. Time- and voltage-dependent Ca2+ currents in neonatal myocytes were elicited by depolarizations from a holding potential of -80 mV to various clamp potentials. The maximal measured inward Ca2+ current was 206 +/- 10 pA (mean +/- SEM, n = 51). The peak current occurred at a mean membrane potential of 7.8 +/- 1.3 mV (n = 51). The Ca2+ current voltage relation was shifted 26 mV in the positive direction when the external Ca2+ concentration was increased 10-fold. Ca2+ current rundown was observed with a half-time of approximately 20 min. Cells dialyzed with solution containing the Ca2+ chelating agent, EGTA (0.04 mM), had action potential durations similar to those previously reported in papillary muscle. In contrast, a higher concentration of EGTA (14 mM) prolonged the action potential duration. Control of the cell internal ionic composition was achieved by dialysis of the cell with a time constant for Na+ ions of 1.2 to 2.6 min. Tetrodotoxin (10 microM), included in some experiments to block Ca2+ entry via Na+ channels, was shown to be more than 98% effective. These results characterize the whole-cell voltage clamp technique as applied to immature heart cells.

Action Potentials↗
PubMed1991

L- and T-type calcium channels in acutely isolated neonatal and adult cardiac myocytes.

We have compared transsarcolemmal Ca2+ currents in acutely isolated neonatal (1- to 5-d-old) and adult rabbit cardiac myocytes prepared using similar enzymatic techniques. Time- and voltage-dependent inward Ca2+ currents were measured using the whole-cell voltage clamp technique. In neonatal myocytes, peak Ca2+ currents measured 114 +/- 10 pA (mean +/- SEM, n = 18) as compared with 2014 +/- 403 pA in adult myocytes (n = 5, p less than 0.001). Although adult myocytes had a larger surface area (estimated from cell capacitance) than neonatal cells (113 +/- 15 x 10(-6) versus 28 +/- 2 x 10(-6) cm2, p less than 0.001), the calculated peak current density was also significantly larger in adult cells (17.9 +/- 2.5 compared to 4.3 +/- 0.4 microA/cm2 for neonatal cells, p less than 0.001). The voltage dependence of the peak Ca2+ current was similar in neonatal and adult myocytes. Early transient (T-type) Ca2+ currents were also studied by comparing the current induced by depolarization to -20 mV from holding potentials of -40 and -80 mV. T-type Ca2+ channels were present in 91% of the adult cells but were evident in only 39% of the neonatal cells. In summary, voltage-gated Ca2+ current amplitude, current density, and T-type Ca2+ channel prevalence all increase with maturation. These data suggest that neonatal myocytes may be relatively deficient in Ca2+ channel activity when compared to adult myocytes.

Age Factors↗
PubMed1991

A field study of cold effects among cold store workers in China.

A field study was carried out among cold store workers in China. A self administrative questionnaire and health check-up were taken among 463 male workers from two different types of cold stores, all during their whole daily work. 296 men from Lower Temperature Stores (LTS), where the air temperature was between -10 to -25 degrees C, and 167 men from Ice Stores (IS), where the air temperature was between -5 and +5 degrees C participated. Another group of 152 men working in normal stores and exposed to an air temperature between 20 and 30 degrees C served as a control group. The study did not indicate any special disease attributable to the cold environment. The number of complaints of lower back pain and knee pain in the cold exposed group were significantly higher than that of the control group (in LTS: 42.3%, 46.6%; IS: 52.7%, 50.8%; control group: 9.2%, 14.5%; low back and knee pain, respectively). After 5 years of cold exposure work, the lower back and knee symptoms were very frequent. The point press pain on the knees of the cooler group (LTS) was higher than in the moderate cold group (IS). We suppose that the cold factor contributed to lower back and knee pain. 12.2% of 463 cold exposed workers had frostbite on the body extremities viz: hand, foot and ear. Self estimation of health indicated more problems among cold exposed workers (28.0%) than control group (2.7%).

Adult↗
PubMed1990

The distribution of S-100 protein in articular cartilage from osteoarthritic joints.

The distribution and localization of S-100 protein was compared with safranin-O staining and H and E morphology in relatively unaffected, degenerative, and osteophytic regions of human articular cartilage from 26 joints obtained at the time of total joint replacement for osteoarthritis. S-100 protein immunoreactivity in osteophyte cartilage was more intense than in unaffected regions and comparable to the staining intensity found in metaplastic cartilage tufts in degenerating cartilage. Immunoreactivity for S-100 protein appeared more prominent in the deep layers of osteophyte cartilage adjacent to areas of matrix calcification.

Adult↗
PubMed1990

[The anchoring of newly synthesized adenovirus DNA to the nuclear matrix].

After adenovirus infected HeLa cells were pulse labeled and pulse-chase labeled with 3H-thymidine, the nuclear matrix and DNA remaining tightly bound to the matrix were obtained by sequential cell fractionation. Measuring the radioactivity of labeled DNA indicated that newly synthesized viral DNA specifically attach to the nuclear matrix and the amount of binding DNA is in direct proportion to the viral DNA replication activity: then the DNA gradually detach from the matrix and are involved in viral assembly. Electron microscopic autoradiography of the extracted cells showed the virion and viral DNA associated with the nuclear matrix, and thus further confirmed the anchoring of newly synthesized viral DNA to the nuclear matrix.

Adenoviruses, Human↗
PubMed1990

Receptor binding of epidermal growth factor in cultured human choriocarcinoma cell lines: effects of actinomycin-D and methotrexate.

Binding of epidermal growth factor (EGF) to its receptor was evaluated in the four cultured choriocarcinoma cell lines BeWo, NaUCC-1, NaUCC-2, and NaUCC-3. Also, the effect of the anti-tumor drugs actinomycin-D (Act-D) and methotrexate (MTX) on the EGF receptor binding was investigated in these cell lines. Incubation of these cells with [125I]EGF at 37 degrees C resulted in a higher binding than that at 22 degrees C or at 4 degrees C. These bindings were saturable during 30- to 60-min incubation, and were specific and reversible. Scatchard analysis showed that the maximal number of receptor binding sites was 2.89 X 10(3)/cell in BeWo cells, 2.04 X 10(3)/cell in NaUCC-1 cells, 1.84 X 10(3)/cell in NaUCC-2 cells, and 1.01 X 10(3)/cell in NaUCC-3 cells. Preincubation with Act-D or MTX for 24 hr decreased the number of receptor binding sites (26%-53%) and slightly increased the receptor binding affinities. Combination of the two drugs resulted in a further diminution of EGF receptor binding sites in BeWo, NaUCC-1, and NaUCC-3 cells, respectively, but reversed the Act-D effect in NaUCC-2 cells. These results indicated that choriocarcinoma tissue is rich in EGF receptors, that the anti-tumor drugs Act-D and MTX diminish the receptor binding sites in the tissue, and suggest that MTX might induce a drug resistance to Act-D in some choriocarcinoma tissue.

Choriocarcinoma↗