Timing of insulin therapy for diabetic recipients with islet transplantation.
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Animals↗
Biomedical subjects
C H Kuo
Publications and source records attributed to C H Kuo.
Regulation of GLUT4 protein and glycogen synthase during muscle glycogen synthesis after exercise.
The pattern of muscle glycogen synthesis following its depletion by exercise is biphasic. Initially, there is a rapid, insulin independent increase in the muscle glycogen stores. This is then followed by a slower insulin dependent rate of synthesis. Contributing to the rapid phase of glycogen synthesis is an increase in muscle cell membrane permeability to glucose, which serves to increase the intracellular concentration of glucose-6-phosphate (G6P) and activate glycogen synthase. Stimulation of glucose transport by muscle contraction as well as insulin is largely mediated by translocation of the glucose transporter isoform GLUT4 from intracellular sites to the plasma membrane. Thus, the increase in membrane permeability to glucose following exercise most likely reflects an increase in GLUT4 protein associated with the plasma membrane. This insulin-like effect on muscle glucose transport induced by muscle contraction, however, reverses rapidly after exercise is stopped. As this direct effect on transport is lost, it is replaced by a marked increase in the sensitivity of muscle glucose transport and glycogen synthesis to insulin. Thus, the second phase of glycogen synthesis appears to be related to an increased muscle insulin sensitivity. Although the cellular modifications responsible for the increase in insulin sensitivity are unknown, it apparently helps maintain an increased number of GLUT4 transporters associated with the plasma membrane once the contraction-stimulated effect on translocation has reversed. It is also possible that an increase in GLUT4 protein expression plays a role during the insulin dependent phase.
Animals↗
Vacuolating toxin gene polymorphism among Helicobacter pylori clinical isolates and its association with m1, m2, or chimeric vacA middle types.
BACKGROUND: Helicobacter pylori vacuolating cytotoxin encoded by vacA plays an essential role in H. pylori-related pathogenesis. Specific vacA alleles are believed to be associated with increased virulence. Association among vacA polymorphism, vacA middle genotypes, and various H. pylori-related diseases was thus investigated. METHODS: Eighty-nine isolates from patients with various gastrointestinal diseases were examined for restriction fragment length polymorphism (RFLP) of the 2.0-kb polymerase chain reaction-amplified vacA middle region. Further genetic heterogeneity was assessed with ureA-ureB RFLP. RESULTS: Twenty-eight distinct vacA RFLPs were seen among 89 isolates. Each pattern was associated with one specific vacA middle genotype. The association of specific RFLPs with certain clinical manifestations was noted among six common groups. Further RFLP analysis of the 2.4-kb ureA-ureB segment from isolates in four popular vacA RFLPs showed high genetic variation. CONCLUSIONS: The vacA genetic polymorphism may be associated with different gastrointestinal diseases.
Bacterial Proteins↗
Vacuolating toxin production in clinical isolates of Helicobacter pylori with different vacA genotypes.
A vacuolating cytotoxin encoded by vacA in Helicobacter pylori is known as a potential virulent determinant. The relationship between different vacA alleles, vacuolating ability, and H. pylori-related diseases was investigated. Genetic analysis of 119 isolates from Taiwanese patients revealed that 104 strains were s1a/m2, 13 strains were characterized as the s1a/m1T type, which was more homologous to the s1a/m1 strains, and 2 were characterized as the s1a/m1Tm2 chimeric type. Production of high-grade cytotoxin among 11 strains with s1a/m1T was higher (72.7%) than among 66 strains with s1a/m2 (21.2%) (P < .01). Peptic ulcer occurred in 76.9% of 13 patients with s1a/m1T strains compared with 46.2% of 104 patients with s1a/m2 strains (P < .05). These results suggest that s1a/m1T strains are associated with increased cytotoxic activity and higher ulcer prevalence than are s1a/m2 strains.
Amino Acid Sequence↗
Amylin influences insulin-stimulated glucose metabolism by two independent mechanisms.
The effects of amylin on fiber type-specific muscle glucose metabolism under hyperglycemic (10 mmol/l) and hyperinsulinemic (2.1 nmol/l) conditions were investigated using a rat hindlimb perfusion system. Amylin concentration ranged from 1 to 100 nM. Efficacy for inhibition of glucose uptake traced with 2-deoxyglucose by amylin was demonstrated in all three fiber types. The incorporation of 2-deoxy-[3H]glucose tracer decreased from control values by 41% in fast oxidative (FO), 36% in fast glycolytic (FG), and 37% in slow oxidative (SO) muscle with 100 nM amylin. Amylin increased intracellular glucose 6-phosphate (G-6-P), and G-6-P was negatively correlated with 2-deoxyglucose uptake in both FO (r = -0.65; P < 0.01) and FG (r = -0.53; P < 0.01) muscle. Muscle glycogen concentration increased under control conditions and decreased in the presence of 100 nM amylin. Lactate arteriovenous efflux across the hindlimb increased significantly above control with 100 nM amylin (5.03 +/- 0.81 to 11.28 +/- 0.94 mumol.g-1.h-1). Adenosine 3',5'-cyclic monophosphate (cAMP) increased in FO and FG muscle with amylin. Salmon calcitonin-(8-32), an amylin antagonist, ameliorated the effect of amylin on all responses other than 2-deoxyglucose uptake and G-6-P concentration. These results suggest that amylin may work through at least two independent mechanisms, a cAMP-mediated effect on glycogen metabolism and a non-cAMP-mediated inhibition of glycolysis.
Amyloid↗
Amylin-mediated inhibition of insulin-stimulated glucose transport in skeletal muscle.
We examined the effects of amylin on 3-O-methyl-D-glucose (3-O-MG) transport in perfused rat hindlimb muscle under hyperinsulinemic (350 microU/ml, 2,100 pmol/l) conditions. Amylin at 100 nmol/l concentration inhibited 3-O-MG transport relative to control in all three basic muscle fiber types. Transport decreased in slow-twitch oxidative (from 5.65 +/- 1.13 to 3.46 +/- 0.71 micromol . g-1 . h-1), fast-twitch oxidative (from 6.84 +/- 0.90 to 4.84 +/- 0.76 micromol . g-1 . h-1), and fast-twitch glycolytic (from 1.27 +/- 0.20 to 0.60 +/- 0.05 micromol . g-1 . h-1) muscle. Amylin inhibition of insulin-stimulated glucose transport in skeletal muscle was accompanied by a 433 +/- 72% increase in intracellular glucose 6-phosphate (G-6-P) despite the absence of extracellular glucose. The source of hexose units for the formation and maintenance of G-6-P was likely glycogen. Amylin increased glycogenolysis, increased lactate formation, and decreased glycogen synthase activity. Furthermore, the kinetics of glycogen synthase suggest that this enzyme may control intracellular G-6-P concentration. Despite the large increase in G-6-P, no detectable increase in uridine diphosphate-N-acetylhexosamines occurred, suggesting that the proposed glucosamine pathway may not be involved in transport inhibition. However, decreases in uridine diphosphate hexoses were detected. Therefore, uridine or hexosamine-based metabolites may be involved in amylin action.
3-O-Methylglucose↗
Modified Mitchell osteotomy for hallux valgus.
From 1988 to 1995, 96 patients (161 feet) underwent a modified Mitchell distal metatarsal osteotomy performed for mild-to-moderate hallux valgus. On AP x-rays of the standing foot, the average intermetatarsal angle was corrected from 15 degrees to 9 degrees, and the first metatarsophalangeal angles were corrected from an average of 41 degrees to 15 degrees. Criteria for evaluation of clinical results included relief of pain, appearance of foot, and shoe wear. After an average follow-up of 38 months, the overall satisfaction rate was 92.5%. Complications included 13 pin tract infections, two delayed unions, and two correction losses. The most common late sequela was transfer metatarsalgia of the lesser toes, which occurred in 20 feet (12.4%), leading to some dissatisfaction. The Mitchell osteotomy can be used on cases with less than 20 degrees of intermetatarsal angle, offering a stable construct with easy postoperative care.
Adolescent↗
A single stranded DNA-binding protein, ssCRE-BP/Pur alpha, in rat lung and its increase in allergic airway inflammation.
ssCRE-BP/Pur alpha is a single stranded DNA-binding protein and may be involved in gene replication and transcription and in the development of morphine dependence. We found a ssCRE-BP/Pur alpha (45 kDa) in rat lung that was larger than those (40 kDa) identified in rat and mouse brains and mouse lung. Immunohistochemistry showed that ssCRE-BP/Pur alpha is primarily distributed in the lung epithelium. As allergic inflammation induces various gene expressions, we investigated the changes of Pur alpha during airway inflammation. Ovalbumin-sensitized rats were used for inducing allergic airway inflammation. The expression and DNA-binding activity of 45-kDa ssCRE-BP/Pur alpha were significantly increased in the sensitized rat lungs 24 hr after antigen challenge, but not in those of rats nonsensitized or sensitized with ovalbumin and challenged with saline. Immunohistochemistry and in situ hybridization demonstrated that the vascular endothelial cells and numerous infiltrated eosinophils around the airways were stained with anti-Pur alpha antibody. These data suggest that rat lung and the eosinophils contain a 45-kDa ssCRE-BP/Pur alpha that is increased when airway inflammation occurs.
Animals↗
Survival of Taenia solium cysticerci in carcasses of pigs kept at 4 C.
In the present study, the survival of cysticerci of Taenia solium in carcasses of 1 Small-Ear-Miniature and 2 Landrace Small-Ear-Miniature pigs kept in a refrigerator at 4 C was determined. The viability of the cysticerci was determined by observing evagination and active movement of the scolex under a fluorescent light after digestion in pig bile. Although no viable cysticerci were found after day 30, 58.8% cysticerci survived between day 26 and day 30. Moreover, more than 80% of the cysticerci were viable between day 1 and day 25. These findings indicate that taeniasis solium can be transmitted through eating raw or undercooked pork or viscera of pigs following refrigeration at 4 C for less than 30 days.
Animals↗
Surgical treatment for pilon fracture of the ankle-open reduction and internal fixation.
From 1991 to 1994, 39 ankles of 38 patients underwent surgical open reduction and internal fixation for pilon fractures. These patients included 29 males and 9 females with an average age of 38.6 y/o (range 28 y/o-58 y/o). The follow up and evaluation period averaged 31.7 months (range 22Ms-44Ms), during which time a standing x-ray for arthrosis grading and functional scale was used for clinical evaluation. Complications included 1 case of infection, 1 case of loss reduction, 2 cases of partial skin necrosis and 2 cases of delayed union. Post-traumatic arthritis occurred in 23 ankles (59%) but only 4 ankles of grade 4 arthrosis resulted in poor functional scale and the overall satisfactory rate was 82%. It was found that anatomic reduction, rigid fixation and early motion exercise are important to successful treatment of ankle fractures. Regarding pilon fracture, specifically the severity of fracture pattern and delay of reduction are important problems to overcome to ensure successful results. Therefore, adequate surgical approach for entire view of ankle joint, reduction and fixation of fibula, sufficient bone graft for articular support, intraoperative x-ray check and postoperative immobilization are essential for the achievement of better clinical results.
Adult↗
Molecular nature of a sperm acrosomal antigen recognized by HS-13 monoclonal antibody.
Among the numerous monoclonal antibodies generated against human sperm antigens, HS-13 monoclonal antibody was shown to react with an intra-acrosomal antigen from human, mouse and rat. In this study, HS-13 was used as the affinity ligand for the purification of the cognate antigen from human sperm by immunoaffinity chromatography. The purified cognate antigen from human sperm, designated as HSAg-13, was found to be a protein with a molecular weight of approximately 80 kDa on SDS-PAGE in the presence of reducing reagents. This monoclonal antibody was used as the probe to study the tissue distributions and developmental expression of the cognate antigen from human, mouse and rat by immunohistochemical assays. It was concluded that the antigen recognized by HS-13 antibody is highly sperm specific and found only in sperm and mature testis, but not in any other somatic tissues examined in human and mouse. The antigen was shown to be expressed at the postmeiotic stages of spermatogenesis in mouse and rat. By using indirect immunofluorescent staining assay, HS-13 was shown to react only with the methanol-fixed acrosome-intact sperm but not with the live sperm. Following calcium ionophore A23187 treatment, acrosome-reacted sperm showed either negative staining or residual staining in the equatorial region, suggesting the intra-acrosomal location of HSAg-13. The spontaneous acrosome reaction following overnight incubation in BWW medium resulted in a statistically significant decrease of antibody-stained human sperm. In view of excellent correlations for the scoring of acrosome-intact sperm with that of fluorescence-labeled Pisum sativum agglutinin (PSA) probe, HS-13 monoclonal antibody can be routinely used for monitoring sperm capacitation and acrosome reaction.
Acrosome↗
A comparative study on egg hatching methods and oncosphere viability determination for Taenia solium eggs.
The efficiencies of an enzyme method and the sodium hypochlorite method in hatching eggs of Taenia solium (Henan strain) were compared. The viability of the hatched oncospheres was determined by staining with trypan blue. For the former method, the hatching rate and viability percentage were 85.7% and 79.7%, respectively. The corresponding figures for the latter method were 97.9% and 86.7%, respectively. These findings indicate that the sodium hypochlorite method is a simple, efficient and practical method for hatching eggs of Taenia solium.
Animals↗
Fate of a small number of islets transplanted into diabetic mice.
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Animals↗
Analysis of the T beta gamma-binding domain of MEKA/phosducin.
MEKA/phosducin, a 33 kDa phosphoprotein in the photoreceptor cell, associates with transducin beta gamma (T beta gamma) with its N-terminal domain (N-terminal 105 amino acids of MEKA), and translocates T beta gamma from the photoreceptor disc membrane to the soluble fraction. The present study further localized the T beta gamma-binding domain to aa 17-105 of MEKA, and showed that the activity of MEKA to translocate T beta gamma depends on the domain. A series of deletion mutant MEKA proteins were prepared to investigate the domain of MEKA which binds to and translocates T beta gamma. Both binding and translocation activities were not impaired by the deletion of the N-terminal 16 amino acids of MEKA, but completely abolished by further deletion to 42Val. Although anti-MEKA serum inhibited the T beta gamma-MEKA association, the antiserum absorbed with a recombinant peptide corresponding to aa 17-105 of MEKA did not, confirming that aa 17-105 of MEKA directly interacts with T beta gamma.
Amino Acid Sequence↗
Characterization of a nuclear factor that enhances DNA binding activity of SSCRE-BP/PUR alpha, a single-stranded DNA binding protein.
Pur alpha has been identified as a single-stranded DNA binding protein that specifically binds to the purine-rich strand present in the DNA replication initiation zone of the human c-myc gene. We have previously demonstrated that chronic morphine treatment decreases the DNA binding activity of ssCRE-BP (single-stranded cyclic AMP response element-binding protein), which has been shown to be identical to pur alpha by cDNA cloning, and is abundant in the brain. In this report we identified an activator of ssCRE-BP/pur alpha in the brain and characterized it. Although purified ssCRE-BP/pur alpha or its GST-fusion protein exhibited very low DNA binding activities, they were markedly enhanced by including nuclear extract in the binding assay. The enhanced binding activity is trypsin-sensitive, heat-stable and has a molecular weight of approximately 66 kDa. Casein could substitute for the activator and increased the DNA binding activity of ssCRE-BP/pur alpha by one order. A series of deletion mutants were prepared in order to determine the DNA binding and activator interacting domains, and both of them were found to reside in AA 50-215 of ssCRE-BP/pur alpha. These data suggest that the DNA binding activity of ssCRE-BP/pur alpha is augmented by a nuclear protein, which may modulate the ssCRE-BP/pur alpha activity to develop morphine dependence and tolerance.
Animals↗
Genetic analysis of the cytotoxin-associated gene and the vacuolating toxin gene in Helicobacter pylori strains isolated from Taiwanese patients.
OBJECTIVE: Two virulence factors encoded by the cytotoxin-associated (cagA) gene and the vacuolating toxin (vacA) gene of Helicobacter pylori are known to be associated with gastroduodenal pathologic conditions. In this report, nucleotide sequence of cagA and vacA and the serum antibody response to the CagA antigen in H. pylori isolates from Taiwanese patients were studied. METHOD: cagA and vacA were characterized in 173 H. pylori strains by polymerase chain reaction and DNA hybridization analyses. The presence of serum IgG antibodies against CagA was assessed by Helico-blot Western blot system. Sequence analysis of polymerase chain reaction-amplified vacA and cagA was performed for two strains. RESULTS: cagA was detected in all the isolates with peptic ulcers and gastric cancer. Of 54 strains with non-ulcer dyspepsia, 51 were found to be cagA+. Serum antibodies to CagA were detected for 99.2% of cagA+ strains compared with 0% of strains lacking cagA. vacA was detected in all isolates. Sequence analysis of vacA and cagA indicated that sequences of two Taiwanese strains were closely related to each other (95.9% and 97% nucleotide identity, respectively) but less homologous to the published strains (90.9% and 91.4% mean nucleotide sequence identity, respectively). CONCLUSIONS: The strong association (98%) of cagA+ strains with Taiwanese patients suggests that the cagA+ phenotype could not be used as a single marker of high-risk patients in Taiwan. Moreover, sequence analysis indicates that Taiwanese strains contain different genetic sequences from those in other geographic regions.
Adolescent↗
Involvement of a single-stranded DNA binding protein, ssCRE-BP/Pur alpha, in morphine dependence.
We have purified a nuclear protein from mouse cerebella that binds to single-stranded oligo-DNA of cAMP response element and is modulated by morphine treatment. Isolation of the cDNA clone showed that the nuclear protein (ssCRE-BP) was identical to Pur alpha, a DNA binding protein for single-stranded purine-rich sequences that was originally isolated as a replication factor. ssCRE-BP/Pur alpha and mRNA were abundant in the brain. The levels of ssCRE-BP/Pur alpha and the transcript were not changed by chronic morphine treatment, however, the levels of an activator of ssCRE-BP/Pur alpha, which is necessary for the DNA binding, may be modulated by the treatment.
3T3 Cells↗
MEKA/phosducin attenuates hydrophobicity of transducin beta gamma subunits without binding to farnesyl moiety.
Hydrophobic modifications of transducin (T) gamma, such as farnesyl-and carboxyl-methylation, are essential for the association of T beta gamma with the photoreceptor disc membrane, and MEKA/phosducin is known to inhibit the association. In this study, we examined the effect of MEKA on the hydrophobicity of T beta gamma. MEKA could bind to T beta gamma without farnesyl/carboxyl-methyl moieties as well as native T beta gamma. In the Triton X-114 phase separation assay, T beta gamma-MEKA complex was recovered in the aqueous phase, whereas T beta gamma was recover in the detergent phase. N-terminal portion of MEKA which includes T beta gamma-binding domain was not sufficient to reduce the hydrophobicity of T beta gamma or to dissociate T beta gamma from the membrane. The data suggest that MEKA attenuates the hydrophobicity of T beta gamma to result in the dissociation of T beta gamma from the membrane without directly binding to farnesyl/carboxyl-methyl moieties.
Animals↗