The role of insulin-like growth factor-1 (IGF-1) in follicular thyroid cancer cell line CGTH W-1.
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Aged↗
Biomedical subjects
H Y Chang
Publications and source records attributed to H Y Chang.
Effects of intravenously administered vitamin B12 on sleep in the rat.
Vitamin B12 (VB12) has been reported to normalize the entrainment of circadian rhythms in the non-24-h sleep wake cycle and delayed sleep phase insomnia in humans. The purpose of this work was to clarify whether the peripheral administration of VB12 has any sleep-promoting effect on the sleep-wake rhythm in freely moving rats. After a baseline day of saline infusion. VB12 (500 micrograms/kg/day) was administered continuously for 4 days via the jugular vein. Polysomnographic recordings were carried out concurrently. In both the light and the 24-h periods, the amount of non-rapid eye movement (NREM) sleep increased significantly on VB12-days 2 and 3, while the amount of REM sleep increased significantly on VB12-day 2. In the light period, the increase in NREM sleep was due to increased duration of the episode, while the tendency to an increase in REM sleep was due to an increased number of episodes. Changes in the diurnal sleep-wake rhythm tended to appear in the earlier light period. The serum VB12 concentrations in the VB12 group were 40 times higher than in controls. These findings suggest that peripherally infused VB12 has promoting effects on the rat's sleep, especially in the light period.
Animals↗
Heart rate variability during sleep in Down's syndrome.
We observed heart rate (HR) variability in the sleep-wake cycle of young adults with Down's syndrome (DS) and their age-matched normal controls. Spontaneous HR fluctuations for 10 min during each sleep stage were analyzed by spectral analysis. Only during rapid eye movement (REM) sleep, a middle-frequency (0.05-0.15 cycles/beat) component of the HR fluctuations jointly mediated by the sympathetic and parasympathetic nerves, was significantly lower in the DS than in the control subjects. As the high-frequency (0.15-0.40 cycles/beat) component mediated by the parasympathetic nerve was not different in the two groups, it is suggested that the sympathetic activity during REM sleep is lower in DS than in controls.
Adult↗
Fatty acyl-CoA inhibits 1-alkyl-sn-glycero-3-phosphate acetyltransferase in microsomes of immature rabbit cerebral cortex: control of the first committed step in the de novo pathway of platelet-activating factor synthesis.
Microsomal fractions of cerebral cortices of 15-day-old rabbits were used to study the 1-alkyl-sn-glycero-3-phosphate (AGP) acetyltransferase that generates 1-alkyl-2-acetyl-sn-glycero-3-phosphate in the de novo path of platelet-activating factor synthesis. The AGP acetyltransferase activity was inhibited by small concentrations of medium-long chain fatty acyl-CoA thioesters. In contrast, the AGP acyltransferase used oleoyl-CoA as substrate and was not inhibited by the presence of acetyl-CoA in high molar excess. The inhibition of AGP acetyltransferase was seen at concentrations of oleoyl-CoA as low as 0.5 microM using 12.5 microM AGP and 200 microM acetyl-CoA. The inhibition by oleoyl-CoA was noncompetitive for the acetyl-CoA substrate. However, there was evidence that the oleoyl-CoA was competing with AGP in the acetyltransferase reaction, as the inhibition was lessened by increasing the AGP substrate concentration. Several acyl-CoA thioesters were effective as inhibitors of the AGP acetyltransferase, including oleoyl-, palmitoyl-, lauroyl-, and octanoyl-CoA. Propionyl- and butyryl-CoA were less effective as inhibitors, and propionyl-CoA was found to be a competitive inhibitor for acetyl-CoA. We have noted earlier that MgATP is an effective inhibitor of the AGP acetyltransferase and here we show that the inhibition by oleoyl-CoA can be increased by the presence of 0.1 mM MgATP. In brain ischemia, a decline in ATP levels would likely lead to a corresponding fall in acyl-CoA concentrations, thereby relieving the inhibition of AGP acetyltransferase and permitting the flow of AGP into the de novo pathway of platelet-activating factor synthesis.
1-Acylglycerol-3-Phosphate O-Acyltransferase↗
Diaphragmatic microcirculation measured by laser-Doppler flowmetry in the rat.
The applicability of laser-Doppler flowmetry (LDF) in measuring regional blood flow of the diaphragm (Qdi) was evaluated in anesthetized and mechanically ventilated rats. Specifically, we assessed the issues of the degree of tissue perfusion, spatial variations of Qdi signals, and the correlation between these signals and total Qdi. Qdi was detected from an area 1 mm in diameter and 0.5 mm in depth. With Ringer solution continuously suffusing the abdominal surface of the left hemidiaphragm, drugs were applied to the surface of the diaphragm at 1/100 of the flow rate of the suffusing fluid. In 13 rats, resting Qdi and the response of Qdi to a single concentration of adenosine (10(-4) M) remained unchanged over a 2-h period. Increasing adenosine concentrations (10(-9) to 10(-3) M) in eight rats elicited a progressive rise in Qdi. In six rats, the variability of Qdi in a given rat was assessed over five different sites at 2-mm intervals in a line on the diaphragm. A wide range of variability in Qdi was noticed among different sites. The depth of tissue penetration of LDF was little affected when a piece of freshly diaphragm was interposed between the probe and the perfused diaphragm, suggesting that LDF signals arise from complete penetration of the perfused diaphragm. Finally, significant linear relationships between mean Qdi and total Qdi were observed in all animals; however, the slopes of these relationships varied significantly between animals. In conclusion, LDF provides a stable and continuous method for monitoring regional microvascular diaphragmatic perfusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine↗
Rapidly fatal outcome of bacteremic Klebsiella pneumoniae pneumonia in alcoholics.
Twenty-eight alcoholic patients with community-acquired pneumonia who were admitted to a referral medical center during a 3-year period were reviewed. All were men and with few exceptions were heavy smokers. The overall mortality was 64.3%. The most dramatic group was that of 11 patients (39.3%) with bacteremic Klebsiella pneumoniae pneumonia (BKPP), which had high mortality rate (100%), short onset of illness before hospital admission (42.6 +/- 8.2 h, mean +/- SD), and short survival time after the admission (24.6 +/- 7.9 h). All these 11 patients needed intensive care unit (ICU) management and ventilatory support. ARterial blood gas values showed marked hypoxemia and metabolic acidosis in most of these 11 patients, and presence of shock at arrival in the hospital was noted in 8 patients. Acute renal failure and disseminated intravascular coagulation developed in six patients. Chest radiographs showed pleural effusion and radiographic spread in nearly 50% of patients. Combination chemotherapy consisting of one aminoglycoside and one second- or third-generation cephalosporin was initiated in all patients. The rapidly fatal outcome of these 11 patients with BKPP despite management with adequate antibiotics and intensive care indicated the fulminant nature of this disease. High virulence of the microorganism, altered immune response, and increased susceptibility to infection may all have contributed to the fulminancy in this group of patients. The disease continues to present many difficulties in management.
Adult↗
Effect of prostaglandins and nitric oxide on basal blood flow and acetylcholine-induced vasodilation in rat diaphragmatic microcirculation.
To assess the effect of prostaglandins and endothelium-derived relaxing factor (EDRF) on diaphragmatic microcirculation under basal conditions and after acetylcholine (ACh) stimulation, we studied a diaphragmatic preparation in anesthetized rats. With bicarbonate-buffered Ringer's solution suffusing the abdominal surface of the left costal diaphragm, laser-Doppler flowmetry was used to record microvascular blood flow (QLDF). Microvascular conductance (CLDF) was derived from QLDF by dividing by the systemic blood pressure. Drugs were applied to the surface of the diaphragm. Four series of experiments were performed. In Series 1 (n = 9), ACh (3 x 10(-5)-10(-3) mol/L) elicited a concentration-dependent increase in QLDF and CLDF. In Series 2 (n = 11), ACh-induced QLDF and CLDF changes were significantly attenuated after 30 minutes suffusion of indomethacin (10(-5) mol/L), although baseline QLDF and CLDF were little affected. In Series 3 (n = 7), following suffusion of N omega-nitro-L-arginine methyl ester (L-NAME) (10(-4) mol/L) for 30 minutes, there was no change in baseline QLDF and CLDF. The ACh-induced QLDF change was abolished, while there was still a slight increase in CLDF (172 +/- 26%) at high concentrations of ACh (10(-3) mol/L). In Series 4 (n = 5), co-administration of indomethacin (10(-5) mol/L) and L-NAME (10(-4) mol/L) for 30 minutes did not completely prevent the increase in CLDF (143 +/- 13%) induced by high concentrations of ACh (10(-3) mol/L). The data suggest low basal activities of both vasodilatory prostaglandins and EDRF in diaphragmatic microvascular beds of the anesthetized rat, while both mediators independently modulate microvascular responses to ACh.(ABSTRACT TRUNCATED AT 250 WORDS)
Acetylcholine↗
MgATP inhibits the synthesis of 1-alkyl-2-acetyl-sn-glycero-3-phosphate by microsomal acetyltransferase of immature rabbit cerebral cortex.
The activity of 1-alkyl-sn-glycero-3-phosphate (AGP) acetyltransferase was studied using microsomal fractions isolated from cerebral cortices of 15-day-old rabbits. Fraction P3A was isolated using buffered 0.32 M sucrose containing mercaptoethanol, EDTA and NaF. This fraction had specific AGP acetyltransferase activities which were 4.9-times those of microsomal fraction P3B isolated in 0.32 M sucrose alone. This P3B activity was increased 2.4-times after a preincubation in the presence of ATP, MgCl2 and a high-speed supernatant fraction from cerebral cortex. Further, the activities of both P3A and P3B were almost completely eliminated by preincubation in the presence of alkaline phosphatase. Thus an activation of the AGP acetyltransferase by phosphorylation was indicated. While there was little inhibition of the P3A AGP acetyltransferase in the presence of added ATP, the magnesium salt form of ATP (1 mM) was severely inhibitory, bringing about 86% inhibition for P3A and 91% for P3B. The inhibitory effects of MgADP and MgAMP were smaller, and MgATP was a much more effective inhibitor than MgCTP, MgGTP and MgUTP which brought about 20-38% inhibitions of P3A activity at 1 mM concentrations. The effect of MgATP may be of particular relevance to the synthesis of platelet activating factor (PAF) following a period of ischemia in brain. Falling MgATP levels during energy failure could relieve the inhibition of AGP acetyltransferase seen in healthy cells and allow the formation of 1-alkyl-2-acetyl-sn-glycero-3-phosphate, which is the first committed intermediate in the de novo pathway of PAF synthesis.
Acetyltransferases↗
Starch-binding domain of Aspergillus glucoamylase-I. Interaction with beta-cyclodextrin and maltoheptaose.
The characterization is reported of two peptide fragments (SBD106 and SBD122) containing the starch-binding domain (SBD) of Aspergillus sp. glucoamylase I. The starch-binding peptides were produced in Escherichia coli as fusion proteins of the maltose-binding protein (MBP). SBD106 (11.9 kDa) and SBD122 (13.8 kDa) were purified from the factor Xa digest of MBP fusion proteins. The amino acid compositions were similar to those deduced from their amino acid sequences. The interactions of beta-cyclodextrin and maltoheptaose with purified SBD peptides were investigated by UV difference spectroscopy. SBD106 and SBD122 bound specifically beta-cyclodextrin with a dissociation constant (Kd) of 34 microM and 23.5 microM, respectively. Maltoheptaose binding to SBD106 and SBD122 was weaker than that of beta-cyclodextrin; dissociation constants were 0.57 and 0.50 mM, respectively. The results indicate that the intramolecular disulfide bonding is not required for the domain functioning and that O-glycosylation is not critical for the functioning of the starch-binding domain, but may affect its conformation and dynamics.
Amino Acid Sequence↗
Nucleotide sequence and expression in Escherichia coli of the Klebsiella pneumoniae deaD gene.
The deaD gene of Klebsiella pneumoniae was isolated and its nucleotide sequence determined. The K. pneumoniae gene is highly homologous with the Escherichia coli analog throughout most of the coding region. The deduced primary sequence of the K. pneumoniae deaD gene product is 659 amino acids in length, in contrast with the 571 amino acids of the E. coli deaD product published previously. Sequence comparison revealed several differences near the 3' end of the deaD genes which result in the frame-shift effect. The 3' end sequence of the E. coli deaD gene was therefore analyzed to verify the discrepancy. Our result indicates that the E. coli deaD gene encodes a product of comparable size to the K. pneumoniae DeaD protein, and the carboxyl terminal sequences of the two proteins are highly homologous. In vivo expression of the K. pneumoniae deaD gene in E. coli yielded a 65-kDa protein. Primer extension analysis of the mRNA from K. pneumoniae identified a major transcription start site at an A residue 44 nt upstream of the first in-frame ATG codon.
Amino Acid Sequence↗
Population structure of the Japanese eel, Anguilla japonica.
Mitochondrial DNA (mtDNA) sequences that include (a) a part of the cytochrome b gene, (b) two tRNA genes, and (c) a part of the noncoding D-loop region of 31 Anguilla japonica (Japanese eel) and 1 A. marmorata collected from Taiwan, Japan, and mainland China were determined to evaluate the population structure of Japanese eel. Among 30 genotypes identified from the 31 Japanese eel mtDNAs sequenced, there are 58 variable sites, predominantly clustered at the D-loop region. The phylogenetic tree constructed by the unweighted pair-group method with arithmetic mean shows neither significant genealogical branches nor geographic clusters. Furthermore, the sequence-statistics test reveals little, if any, significant genetic differentiation. These results indicate that the 31 Japanese eels might come from a single population. Analysis of sequence variation in mtDNA by using the relationship between the number of segregating sites and the average number of nucleotide differences under the neutral mutation hypothesis reveals that neutral mutation acts as a major factor influencing the evolutionary divergence of the Japanese eel mitochondrial genome sequenced, especially in the noncoding region.
Animals↗
Acetoin catabolic system of Klebsiella pneumoniae CG43: sequence, expression, and organization of the aco operon.
A cosmid clone which was capable of depleting acetoin in vivo was isolated from a library of Klebsiella pneumoniae CG43 cosmids. The smallest functional subclone contained a 3.9-kb DNA fragment of the cosmid clone. Sequencing of the DNA fragment revealed three open reading frames (ORFs A, B, and C) encoding polypeptides of 34, 36, and 52 kDa, respectively. The presence of these proteins was demonstrated by expression of the recombinant DNA clone in Escherichia coli. Considerable similarities between the deduced amino acid sequences of the ORFs and those of the following enzymes were found: acetoin dissimilation enzymes, pyruvate dehydrogenase complex, 2-oxoglutarate dehydrogenase complex, and branched-chain 2-oxo acid dehydrogenase complex of various origins. Activities of these enzymes, including acetoin-dependent dichlorophenolin-dohenol oxidoreductase and dihydrolipoamide acetyltransferase, were detected in the extracts of E. coli harboring the genes encoding products of the three ORFs. Although not required for acetoin depletion in vivo, a possible fourth ORF (ORF D), located 39 nucleotides downstream of ORF C, was also identified. The deduced N-terminal sequence of the ORF D product was highly homologous to the dihydrolipoamide dehydrogenases of several organisms. Primer extension analysis identified the transcriptional start of the operon as an A residue 72 nucleotides upstream of ORF A.
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
Contribution of potassium channels to active hyperemia of the canine diaphragm.
Glibenclamide, iberiotoxin, and apamin (blockers of ATP-sensitive, large-conductance, and small-conductance Ca(2+)-activated K+ channels, respectively) were infused into the diaphragmatic vasculature of anesthetized indomethacin-treated dogs to assess the contribution of K+ channels to active hyperemia. Diaphragmatic blood flow (Qphr) and O2 uptake (VO2di) were measured at rest and during 2 min of continuous left phrenic nerve stimulation at 0.5, 1, 2, and 4 Hz. These measurements were repeated before (control) and after the infusion of a selective K+ channel blocker in three groups of animals. Glibenclamide at 10(-5) M significantly attenuated Qphr at rest and in response to all stimulation frequencies. Whereas resting VO2di remained unchanged, glibenclamide infusion significantly reduced VO2di in response to all stimulation frequencies. The slope of the linear relationship between Qphr and VO2di, however, was not affected by glibenclamide. By comparison, infusion of iberiotoxin (10(-7) M) in a second group reduced Qphr at rest and in response to 0.5- and 1-Hz stimulation, whereas Qphr measured in response to 2- and 4-Hz stimulation remained similar to control values. Apamin (10(-6) M) infusion in a third group reduced only resting Qphr with no effect on active hyperemia during phrenic nerve stimulation. Neither iberiotoxin nor apamin influenced resting or stimulated VO2di. In all groups diaphragmatic tension measured after the infusion of K+ channel blockers remained similar to control values. These results indicate that K+ channels, especially those sensitive to glibenclamide, modulate the increase in Qphr and VO2di in response to moderate augmentation of metabolic demands.
1-Methyl-3-isobutylxanthine↗
Response of the sleep-wake rhythm to an 8-hour advance of the light-dark cycle in the rat.
We have studied the effects of an 8-h advance of the environmental light-dark (LD) cycle on the sleep-wake rhythm in the rat. Electroencephalograms and electromyograms were recorded simultaneously on chart paper through a two-channel telemetry system for 3 days before phase shift (baseline) and 8 days during and after phase shift. Phase advance of the LD cycle led to an increase in both non-rapid eye movement (NREM) and REM sleep. The amount of NREM sleep in the light period correlated positively with that in the preceding dark period for 4 days after phase advance. The duration of REM sleep in the light period correlated negatively with that in the preceding dark period. The results suggest that homeostatic control of the amount of NREM sleep between the preceding dark period and the following light period is disturbed by phase advance of the LD cycle.
Activity Cycles↗
Differential effects of nitric oxide synthesis inhibitor on rat diaphragmatic microcirculation under basal conditions and after vasodilator stimulation.
To assess the effect of endothelium-derived relaxing factor (EDRF) on the diaphragmatic microcirculation under basal conditions and after vasodilator stimulation, prepared diaphragms from anesthetized rats were studied. With bicarbonate-buffered Ringer's solution suffusing the diaphragm, the microcirculatory blood flow was recorded by laser-Doppler flowmetry (QLDF). Drugs were applied to the surface of the diaphragm. Five series of experiments were performed. In Series 1, the effects of acetylcholine (ACh, 10(-4) M) and sodium nitroprusside (SNP, 3 x 10(-6) M) were measured before and after 15 minutes suffusion of N omega-nitro-L-arginine methyl ester (L-NAME, 10(-4) M). Trend analyses revealed that the vasodilator effect of SNP was not affected, while that of ACh was abolished by L-NAME. In Series 2, both ACh and SNP elicited a concentration-dependent increase in QLDF. In Series 3, following 15 minutes suffusion with L-NAME (10(-4) M), there was no change in baseline QLDF and the ACh-induced QLDF change was abolished, while the effect of SNP was maintained. In Series 4, five minutes of pretreatment with L-arginine (L-arg, 10(-2) M), followed by co-administration of L-arg (10(-2) M) and L-NAME (10(-4) M) for another 15 minutes, did not prevent the inhibitory effect on ACh-induced QLDF change by L-NAME. In Series 5, 20 minutes application of L-arg (10(-2) M) alone did not affect baseline QLDF. Also, L-arg did not influence the ACh-induced vasodilating response, whereas it slightly increased the SNP-induced vasodilating response.(ABSTRACT TRUNCATED AT 250 WORDS)
Acetylcholine↗
Insulinoma--clinical experience in ten cases.
During the period between January 1981 and December 1992, a total of ten patients with pathologically confirmed insulinoma were studied. All patients presented with variable degrees of neuroglycopenic symptoms and Whipple's triad. The ratio of insulin to glucose after an overnight fast was greater than 0.3 in 90% (9/10) of patients. Eight patients who received supervised fasting developed neuroglycopenia with relative hyperinsulinemia within 48 hours of fasting. Four of 8 patients developed hypoglycemia during a 5-hour oral glucose tolerance test (OGTT). Calcium infusion test was more sensitive in patients with an initially higher plasma glucose. The detection rates of various localization studies were 12.5% (1/8) by abdominal ultrasonography, 37.5% (3/8) by abdominal CT scan, 50% (5/10) by selective superior mesenteric and celiac arteriography. Transhepatic portal venous sampling (THPVS) detected insulinomas in 4 of 4 cases. Endoscopic ultrasonography and intraoperative ultrasonography were performed on 1 and 2 cases respectively, and were able to localize the lesions successfully. All patients received surgical treatment including enucleation (n = 2), subtotal pancreatectomy (n = 3) and distal pancreatectomy (n = 5). All patients had single tumors which were all benign islet cell adenomas. The mean size of the tumors was 15.5 +/- 2.0 mm in diameter (range: 8 to 30 mm) and mainly located in the body (50%) and tail (40%), only 1 in the pancreatic head. All symptoms of hypoglycemia subsided after operation. Hyperglycemia was observed in all patients immediately after operation, and most of them resumed normoglycemia within 8 days. However, the 2 patients who had impaired OGTT preoperatively had persistent hyperglycemia after operation and therefore were diagnosed as having diabetes (NIDDM).
Adult↗
The potential for platelet-activating factor synthesis in brain: properties of cholinephosphotransferase and 1-alkyl-sn-glycero-3-phosphate acetyltransferase in microsomal fractions of immature rabbit cerebral cortex.
The synthesis of platelet-activating factor (PAF) was studied in microsomal fractions of cerebral cortices of 15-day-old rabbits. These included: a total microsomal fraction P3, rough and smooth microsomes, R and S, and microsomal fraction P derived from isolated nerve cell bodies. Cholinephosphotransferase (CPT) generating PAF from alkylacetylglycerol had the highest specific activities in fractions R and P (24 and 6 times the homogenate values, based on membrane phospholipid content). This CPT activity differed from that which synthesized phosphatidylcholine as the latter was sensitive to dithiothreitol inhibition and was more readily inhibited by Triton X-100. As the CPT activity for PAF synthesis relies on the production of alkylacetylglycerol we studied the acetyltransferase which forms 1-alkyl-2-acetyl-sn-glycero-3-phosphate (AAGP). This enzyme had the highest specific activity in fraction R, followed by fractions P3 and P. There was evidence that the acetyltransferase was more active in a phosphorylated form. NaF maximized the recovery of AAGP products in the assays. The pH optimum for acetylation was in a range of 8.0-9.0. Lyso PAF did not inhibit the formation of AAGP and the rates of formation of PAF by acetylation were less than 5% of values for AAGP synthesis. During AAGP formation there was no evidence for subsequent alkylacetylglycerol formation in the absence of NaF, but a small formation of radioactive PAF could be demonstrated from AAGP under the CPT assay conditions.(ABSTRACT TRUNCATED AT 250 WORDS)
Acetyltransferases↗
Cloning of a human liver UDP-glucose pyrophosphorylase cDNA by complementation of the bacterial galU mutation.
A human liver cDNA clone which encodes the UDP-glucose pyrophosphorylase was isolated by complementation of a bacterial galU mutant. The deduced amino acid sequence of the human enzyme comprised 508 amino acids with a calculated molecular mass of 56,950. The human enzyme significantly resembles those of potato tuber and slime mold with a homology of 46.6% and 43.2%, respectively, in amino acid sequence. No homology was found between the eukaryotic and the prokaryotic enzymes. Northern blotting analysis revealed that the gene was expressed at the highest level in skeletal muscle, followed by liver, heart and kidney.
Amino Acid Sequence↗