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Biomedical subjects

C C Yang

Publications and source records attributed to C C Yang.

At least 19 recordsLinked to original sources

On-line concentration of s-triazine herbicides in micellar electrokinetic chromatography using a cationic surfactant.

On-line concentration of neutral species of s-triazine herbicides in micellar electrokinetic chromatography using tetradecylammonium bromide (TTAB) as a cationic surfactant was investigated. Factors affecting the stacking of analytes were examined. The results indicate that the stacking efficiency is markedly improved with addition of phosphate buffer in the sample matrix. It was found that, depending on the nature of the analytes, the most effective stacking of these analytes occurs when the ratio of the conductivity of buffer electrolyte to that of sample matrix is in the range 1.4-1.2, with sample matrix containing phosphate buffer. Micelle concentration in the separation buffer is also a crucial factor to enhance the stacking efficiency and detection sensitivity of analytes. Moreover, the stacking efficiency of each individual analyte depends on its binding constant to TTAB micelles. The concentration effect is primarily based on sweeping mechanism which is operated in a normal stacking mode with reversed electrode polarity in the presence of reversed electroosmotic flow. As a result of concentration enhancement, the detection limits of these herbicides can reach about 9-15 ng/ml with UV detection.

Chromatography, Micellar Electrokinetic Capillary↗

Refolding of Taiwan cobra neurotoxin: intramolecular cross-link affects its refolding reaction.

In order to explore the effect of intramolecular cross-linking in the folding reaction of cobrotoxin from Naja naja atra (Taiwan cobra) venom, the toxin molecule was modified with glutaraldehyde (GA). The monomeric GA-modified cobrotoxin (mGA-cobrotoxin) was separated from the dimeric and trimeric derivatives using gel filtration. The results of electrophoretic and chromatographic analyses revealed that mGA-cobrotoxin comprised two modified derivatives, which contained modified Lys residues at positions 26 and 27 and at positions 26, 27, and 47, respectively. Moreover, an intramolecular cross-linking of loops II and III by Lys residues was noted with the monomeric derivative containing three modified Lys residues. In sharp contrast to cobrotoxin observations, the folding rate of mGA-cobrotoxin decreased in the presence of GSH/ GSSG, but notably increased in the absence of thiol compounds. Particularly, the accelerated effect of GSH/GSSG on the refolding reaction was affected by the presence of the intramolecular cross-link. Comparative analyses on cobrotoxin and mGA-cobrotoxin CD spectra revealed that modification with the GA reagent caused a change in the gross conformation of cobrotoxin. Fluorescence measurement revealed that the stability of the microenvironment around the single Trp-29 in mGA-cobrotoxin and unfolded mGA-cobrotoxin was appreciably higher than in cobrotoxin and unfolded toxin. Moreover, the ordered structure formation around Trp-29 in refolded mGA-cobrotoxin was faster than in refolded cobrotoxin as evidenced by fluorescence quenching studies. Taken together, these results suggest that the structural flexibility of unfolded cobrotoxin should be favorable for the thiol catalyst to exert its action in the refolding reaction after modification with GA.

Amino Acids↗

Suppression of proline-directed protein kinase F(A) systemically inhibits the growth of human acute leukemia cells.

Initial studies revealed that proline-directed protein kinase F(A) (PDPK F(A)) was overexpressed in various cancerous tissues relative to normal controls. However, the functional role of overexpressed PDPK F(A) in cancer remains to be established. In this report, we explore the potential role of PDPK F(A) in leukemia cell growth by investigating the effects of partial inhibition of this kinase on human acute promyelocytic leukemia (HL-60) and acute lymphoblastic leukemia (Jurkat) cells. Cloning of PDPK F(A) cDNA and its recombinant antisense expression vector and antibody were successfully developed. Several stable antisense clones of HL-60 and Jurkat cells were subcloned, which expressed a low level of PDPK F(A) when compared with the control-transfected clone in immunoblot analysis. Moreover, these antisense clones potently inhibited cell growth, clonogenic growth in soft agar and serum-independent growth. The results taken together demonstrate that suppression of PDPK F(A) is able to interfere with the growth of HL-60 and Jurkat cells, suggesting an essential role of this PDPK in human acute leukemia cell growth.

Cell Division↗

Smad proteins function as co-modulators for MEF2 transcriptional regulatory proteins.

An emerging theme in transforming growth factor-ss (TGF-ss) signalling is the association of the Smad proteins with diverse groups of transcriptional regulatory proteins. Several Smad cofactors have been identified to date but the diversity of TGF-ss effects on gene transcription suggests that interactions with other co-regulators must occur. In these studies we addressed the possible interaction of Smad proteins with the myocyte enhancer-binding factor 2 (MEF2) transcriptional regulators. Our studies indicate that Smad2 and 4 (Smad2/4) complexes cooperate with MEF2 regulatory proteins in a GAL4-based one-hybrid reporter gene assay. We have also observed in vivo interactions between Smad2 and MEF2A using co-immunoprecipitation assays. This interaction is confirmed by glutathione S:-transferase pull-down analysis. Immunofluorescence studies in C2C12 myotubes show that Smad2 and MEF2A co-localise in the nucleus of multinuclear myotubes during differentiation. Interestingly, phospho-acceptor site mutations of MEF2 that render it unresponsive to p38 MAP kinase signalling abrogate the cooperativity with the Smads suggesting that p38 MAP Kinase-catalysed phosphorylation of MEF2 is a prerequisite for the Smad-MEF2 interaction. Thus, the association between Smad2 and MEF2A may subserve a physical link between TGF-ss signalling and a diverse array of genes controlled by the MEF2 cis element.

Animals↗

Evidence for alterations in circulating low-molecular-weight antioxidants and increased lipid peroxidation in smokers on hemodialysis.

BACKGROUND/AIM: Cardiovascular disease is the major cause of mortality in dialysis patients, accounting for about 40% of deaths in most large registries. Oxidative stress has been strongly implicated in the pathogenesis of these events. As end-stage renal disease is a state of elevated free radical activity, the aim of the present study was to investigate the negative impact of smoking in 57 male hemodialysis patients. METHODS: The patients, who were 20-85 years of age (mean age 51.0 +/- 14 years), had been on hemodialysis for at least 6 months before participating in this study. Fasting blood sampling for serum lipid, albumin, urate and lipophilic antioxidants such as tocopherols, carotenes, ascorbate and lipid peroxides was performed. RESULTS: The plasma malondialdehyde (MDA) concentration was significantly higher in hemodialysis patients who smoked compared to hemodialysis patients who were nonsmokers (1.92 +/- 0.52 vs. 1.59 +/- 0.42 nmol/ml, p = 0.006). No association was found between levels of MDA in smokers and parameters such as body mass index, serum cholesterol, serum triglycerides and smoking index. There were no significant differences in the plasma levels of uric acid, alpha-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-carotene, beta-carotene and retinol between the two groups. A significantly lower level of plasma ascorbate was observed in hemodialysis patients who smoked compared to the nonsmoking hemodialysis patients or healthy controls (4.59 +/- 4.0 vs. 9.57 +/- 4.0 and 10.16 +/- 4.6 microg/ml, p < 0.05). Moreover, in smokers, the plasma levels of ascorbate were negatively correlated with the levels of plasma MDA (r = -0.43, p < 0.001) of each patient. Partial correlation analysis of the plasma levels of the measured antioxidants and the smoking index revealed a negative correlation between the plasma levels of lipid-normalized lycopene and the smoking index (r = -0.53, p < 0.05). CONCLUSION: Our data suggest that cigarette smoking further increases plasma-circulating products of lipid peroxidation, which are already increased in nonsmoking hemodialysis patients as compared to matched healthy controls. The lower plasma levels of ascorbate in hemodialysis patients who smoke suggest that these patients may be more susceptible to oxidative tissue damage caused by smoking.

Adult↗

Tracheal length of infants under three months old.

The measurement of tracheal length in infants is difficult to perform in vivo. In adults, tracheal length may be consistent with age, but in infants, tracheal length may vary much more with age and other factors. This study used video rigid ventilation bronchoscopy to evaluate the length of the airway, concentrating on the population younger than 3 months old. There were 34 infants in this study: 14 boys and 20 girls. The mean length from the superior border of the vocal fold to the carina was 5.04 cm, and the mean tracheal length (from the ridge of the first tracheal ring to the carina) was 4.12 cm. There was no significant difference between boys and girls in the length from the vocal fold to the carina or in the tracheal length. The length from the vocal fold to the carina is best correlated to body weight, followed by body height and age.

Anthropometry↗

Suppression of proline-directed protein kinase F(A) expression potentiates erythroid differentiation of human myeloid leukemia cells.

BACKGROUND: Initial clinic studies revealed that the overexpression of proline-directed protein kinase F(A) (PDPK F(A)) is associated conversely with various stages of tumor tissue differentiation. However, the role of overexpressed PDPK F(A) in tumor cell differentiation remains unknown and needs to be established. In this report, the authors explore the potential role of PDPK F(A) in cellular differentiation by investigating the effects of partial inhibition of this kinase on erythroid differentiation of chronic myeloid leukemia cells (K562). METHODS: PDPK F(A) antisense expression vector and its specific antibody were developed successfully. Two stable, transfected antisense clones of human myeloid leukemia cells were subcloned that expressed approximately 80% and approximately 50% of the total PDPK F(A) existing in control-transfected clones, as determined by both immunoprecipitate activity assay and immunoblot analysis. In sharp contrast, the PDPK F(A) antisense clones expressed no significant suppression of any other related PDPK members' expression, demonstrating the specificity of these two antisense clones. RESULTS: The antisense clones proportionally induced spontaneous erythroid differentiation up to approximately 30% of the total K562 cells. Moreover, antisense suppression of PDPK F(A) expression appeared to potentiate sodium butyrate/hemin-induced erythroid differentiation of K562 cells to a more complete stage compared with the control. CONCLUSIONS: The results demonstrate that specific antisense suppression of overexpressed PDPK F(A) in human myeloid leukemia cells is sufficient to potentiate both spontaneous and drug-induced erythroid differentiation, indicating that PDPK F(A) is an important negative regulator in controlling the erythroid differentiation of human myeloid leukemia cells.

Antisense Elements (Genetics)↗

Opposition of rapid baroreceptor resetting by prostanoids in rabbits.

Arterial baroreceptors reset rapidly within minutes during acute hypertension; baroreceptor pressure threshold (Pth) is increased and the pressure-baroreceptor activity relation is shifted to the right. The purpose of the present study was to determine if prostacyclin (PGI2) or other prostanoids, released during acute hypertension modulate the magnitude of baroreceptor resetting. Baroreceptor activity was recorded from the vascularly-isolated carotid sinus during distension of the sinus with slow pressure ramp in rabbits anesthetized with chloralose. Pressure-activity curves were generated after holding carotid sinus pressure for 10-15 min from 30 to 100 mmHg. In control, the elevation of holding pressure increased Pth from 44+/- to 65+/-5 mmHg (p < 0.05, n = 12). In the presence of PGI2 (20 microM), Pth averaged 43+/-4 and 45+/-3 mmHg (n = 12) after holding pressure at 30 and 100 mmHg, respectively. In the control group before exposing the carotid sinus to indomethacin, an elevation of holding pressure increased Pth from 49+/-2 to 71+/-3 mmHg (p < 0.05, n = 12). After inhibition of the endogenous formation of prostanoids with indomethacin (20 microM), Pth increased by a significantly greater extent from 61+/-2 to 90+/-3 mmHg (p < 0.05, n = 12) with the increase in holding pressure. The slope of the pressure-activity curve (baroreceptor gain) was not influenced by the change in holding pressure. It was increased significantly by PGI2, while decreased by indomethacin. Neither the change in holding pressure nor PGI2 affected the circumferential wall strain of carotid sinus over a wide range of pressure alteration. The results suggest that PGI2 or other prostanoids released during acute hypertension sensitizes baroreceptors and provides a negative feedback mechanism that opposes and limits the magnitude of rapid baroreceptor resetting.

Animals↗

Resectional treatment for thyroid cancer with tracheal invasion: a long-term follow-up study.

HYPOTHESIS: Transmural invasion of the trachea by well-differentiated thyroid carcinomas is a locally advanced disease condition. It frequently causes deaths owing to airway obstruction. We hypothesized that resection of the invaded trachea followed by primary anastomosis provides the opportunity for cure. DESIGN: A retrospective review study of medical records. SETTING: The surgical department of a tertiary referral center. PATIENTS: Eight patients with well-differentiated thyroid carcinomas, complicated with tracheal invasion resulting in bleeding and airway obstruction, operated on by tracheal resection and immediate anastomosis, were included. INTERVENTIONS: All patients received total thyroidectomy and neck lymph node dissection as well as segmental tracheal resection followed by primary reconstruction. Postoperative radioactive sodium iodine I 131 treatment and suppression therapy with thyroxine were applied to all of them. MAIN OUTCOME MEASURE: Postoperative follow-up of serum levels of human thyroglobulin and abnormal radioactive iodine 131I from whole body scan. RESULTS: Seven patients are alive. Of these patients, 5 had no evidence of disease: (3 had no evidence of cancer for more than 10 years' follow-up), 2 had regional lymph node metastasis, and 1 had lung metastases. The remaining patient had anastomotic site recurrence with airway obstruction and needed tracheostomy to relieve stridor. She was lost to follow-up 39 months after undergoing the initial operation. CONCLUSIONS: Patients with mucosal invasion of the trachea by well-differentiated thyroid carcinomas should be treated by surgical resection followed by primary reconstruction when technically feasible. This facilitates postoperative care, and it is possible to achieve long-term survival with improvement of the quality of life and possible cures.

Adult↗

Impact of pulse pressure on the respiratory-related arterial pressure variability and its autonomic control in the rat.

The autonomic control of respiratory-related arterial pressure variability (RAPV) remains uncharacterised. We set out to test the hypothesis that the autonomic nervous system may participate in RAPV via its effect on pulse pressure (PP). Experiments were performed on adult male Sprague-Dawley rats anaesthetised with ketamine and paralysed with pancuronium under positive-pressure ventilation. Respiratory variability in each parameter was quantified by auto-spectral analysis and the relationship between variabilities in two signals assessed by cross-spectral analysis. PP and systolic pressure (SP) exhibited similar powers of respiratory variability that were far more pronounced than for diastolic pressure (DP) or mean pressure (MP). The variability in PP preceded that in SP, MP or DP. The largest phase lag occurred between PP and DP and was equivalent to the time delay of one pulse interval. Coherence between respiratory variabilities in PP and MP was nearly perfect. Propranolol dose dependently suppressed respiratory variability in each pressure parameter and eventually disrupted the PP-MP relationship in respiratory variability. However, the influence of lung volume on MP persisted with a near-zero phase lag. Combined administration of phentolamine and atropine facilitated respiratory variability in PP and SP and failed to block the effect of propranolol. However, the combined administration moved the dose/response curve of propranolol and respiratory variability in each pressure parameter to the right. We conclude that, in addition to non-autonomic mechanisms, RAPV may be facilitated by cardiac sympathetic function via the effect on PP variability. The autonomic mechanism can still be evoked during combined alpha-adrenoceptor and muscarinic blockade, possibly via baroreflex mechanisms.

Adrenergic alpha-Agonists↗

Antegrade continence enema for the treatment of neurogenic constipation and fecal incontinence after spinal cord injury.

OBJECTIVE: To describe the effects of an antegrade continence enema stoma formed in a paraplegic man with intractable constipation and fecal incontinence. DESIGN: Case report. SETTING: Spinal cord injury unit, Veterans Affairs hospital. PARTICIPANTS: Spinal cord injury (SCI) patient with T12 paraplegia. INTERVENTION: Surgical formation of antegrade continence enema stoma. MAIN OUTCOME MEASURES: Time of bowel program care, ease of fecal elimination, safety of procedure. RESULTS: Bowel care time was decreased from 2 hours to 50 minutes daily; 6 bowel medications were discontinued; fecal incontinence was eliminated; and no surgical or medical side effects noted after the procedure. CONCLUSION: The antegrade continence enema procedure is a safe and effective means of treating intractable constipation and fecal incontinence in the adult SCI patient. This option should be considered for those persons in whom medical management of bowel care has been unsuccessful.

Cecostomy↗

Rapid separation of lysozyme from chicken egg white by reductants and thermal treatment.

Reductants (0.1-2.0% ascorbic acid, cysteine, or cystine and 0.04-1. 0% beta-mercaptoethanol) were added to 5-fold diluted, salted duck egg whites (commercially and laboratory prepared) and fresh egg whites (chicken and duck), and subsequently the mixtures were heated at 70 degrees C for 1-10 min. The maximal recovery and purification fold of lysozyme obtained from fresh chicken egg whites added with 1. 0% ascorbic acid were 78% and 2.4, respectively. Storage tests showed that the obtained lyophilized lysozyme powder after dialysis was stable when refrigerated at 4 degrees C for 3 months.

Animals↗

Repair of CFTR mRNA by spliceosome-mediated RNA trans-splicing.

Most messenger RNA precursors (pre-mRNA) undergo cis-splicing in which introns are excised and the adjoining exons from a single pre-mRNA are ligated together to form mature messenger RNA. This reaction is driven by a complex known as the spliceosome. Spliceosomes can also combine sequences from two independently transcribed pre-mRNAs in a process known as trans-splicing. Spliceosome-mediated RNA trans-splicing (SMaRT) is an emerging technology in which RNA pre-therapeutic molecules (PTMs) are designed to recode a specific pre-mRNA by suppressing cis-splicing while enhancing trans-splicing between the PTM and its pre-mRNA target. This study examined the feasibility of SMaRT as a potential therapy for genetic diseases to correct mutations using cystic fibrosis (CF) as an example. We used several versions of a cystic fibrosis transmembrane conductance regulator (CFTR) mini-gene expressing mutant (deltaF508) pre-mRNA targets and tested this against a number of PTMs capable of binding to the CFTR target intron 9 and trans-splicing in the normal coding sequences for exons 10-24 (containing F508). When 293T cells were cotransfected with both constructs, they produced a trans-spliced mRNA in which normal exon 10-24 replaced mutant exon 10. To test whether SMaRT produced mature CFTR protein, proteins were immunoprecipitated from lysates of cotransfected cells and detected by Western blotting and PKA-phosphorylation. Tryptic phosphopeptide mapping confirmed the identity of CFTR. This proof-of-concept study demonstrates that exon replacement by SMaRT can repair an abnormal pre-mRNA associated with a genetic disease.

Blotting, Western↗

Reflex innervation of the bulbocavernosus muscle.

OBJECTIVE: To determine the somatic reflex innervation of the bulbocavernosus muscle (BCM) and to illustrate the role of the perineal nerve in BCM contraction, using electrophysiological techniques. Subjects and methods Ten healthy, potent men underwent genital electrophysiological testing. Stimuli were applied to the dorsal nerve of the penis at the base of the penis, glans and anterior urethra, and the perineal nerve branch to the BCM. The electromyographic (EMG) responses of the BCM were recorded and onset latencies measured at baseline, both after anaesthetising the perineal nerve and after recovery from the anaesthetic. RESULTS: A reflex BCM contraction was elicited from all stimulation sites. Anaesthetizing the perineal nerve resulted in alterations of the EMG response. After the anaesthetic effect wore off, the responses returned to baseline values. CONCLUSIONS: Three distinct somatic bulbocavernosus reflexes were detected which are components of normal ejaculation. BCM contraction can be elicited after stimulating the dorsal penile nerve at the glans and the anterior urethra; these are the flexor responses of the bulbocavernosus reflex. BCM contraction can also be induced on stimulating the perineal nerve, which is the pathway through which the stretch and the tendon organ reflexes are carried to mediate muscle tone. The motor innervation of the BCM for all reflexes is carried through a branch of the perineal nerve. The findings may be relevant to the evaluation of ejaculatory disorders.

Adult↗

Suppression of proline-directed protein kinase F(A) expression inhibits the growth of human chronic myeloid leukaemia cells.

Initial studies revealed that proline-directed protein kinase F(A) (PDPK F(A)) was overexpressed in various cancerous tissues relative to normal controls. However, the functional role of overexpressed PDPK F(A) in cancer remains to be established. In this report, we explore the potential role of PDPK F(A) in leukaemia cell growth by investigating the effects of partial inhibition of this kinase on the malignant phenotype of human chronic myeloid leukaemia cells (K562). Cloning of PDPK F(A) cDNA and its recombinant antisense expression vector and PDPK F(A)-specific antibody were successfully developed. Two stable antisense clones of K562 cells were subcloned which expressed 70% and 45% of PDPK F(A) respectively, compared with control-transfected clone in both immunoprecipitate activity assay and immunoblot analysis. In sharp contrast, these two antisense clones expressed no significant suppression of any other related PDPK family members, indicating the specificity of these two antisense clones. Moreover, these antisense clones proportionally and potentially exhibited cell growth retardation, poor clonogenic growth in soft agar and loss of serum independence. The results demonstrate that specific antisense suppression of PDPK F(A) is sufficient to interfere with the growth of K562 cells, indicating that PDPK F(A) is essential for human chronic myeloid leukaemia cell growth.

Amino Acid Sequence↗

Inadequate stocking of antidotes in Taiwan: is it a serious problem?

OBJECTIVE: Insufficient hospital stock of a variety of poisoning antidotes is a worldwide problem. In an attempt to establish an antidote storage and distribution system for the response of the various poisoning accidents, we conducted a nationwide survey to characterize the current availability of selected antidotes and their anticipated need in Taiwan. MATERIALS AND METHODS: A questionnaire was mailed to 834 hospitals to gather information on the availability, anticipated need, and preferred purchase policy of 20 selected antidotes. A survey on the availability of cyanide antidote in 523 cyanide-handling facilities and their neighboring hospitals was also conducted. RESULTS: Hospitals of different size and service levels had a statistically significant difference in response rates. Except for pyridoxine, the availability and anticipated need for antidotes also varied significantly among different hospital groups. We found that physostigmine, cyanide antidote kit, BAL, EDTA, methylene blue, Vipera Russell formosensis antivenin, and botulism antitoxin were not available in most (>90%) hospitals. Interestingly, these antidotes are also among the most needed antidotes. Most hospitals preferred a government-ordered purchase of antidotes. In the survey of cyanide-processing facilities, a response rate of 24.1% was obtained and only 9.3% of these 107 facilities that both replied to the questionnaire and continued handling cyanide products had stocked cyanide antidote. It is noteworthy that cyanide antidote was also frequently lacking in the neighboring hospitals. CONCLUSIONS: The appropriate storage of antidotes in hospitals or workplaces in rural areas is instrumental in the timely treatment of certain poisonings, while nationwide unavailability is the critical problem. Raising awareness of the importance of antidotes by education, regular review of antidote storage, distribution plans, and appropriate legislation might provide solutions.

Antidotes↗

Altered frequency characteristic of central vasomotor control in SHR.

Previous work from our laboratory has demonstrated that the very low-frequency (VLF: 0-0.25 Hz) and low-frequency (LF: 0.25-0.8 Hz) power of arterial pressure variability (APV) are related to vasomotor reactivity in response to control signals from the rostral ventrolateral medulla (RVLM) via the sympathetic system in the rat. The present study evaluated the differences in the dynamic property of central vasomotor control between spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Experiments were carried out in 10- to 12-wk-old rats that were anesthetized with continuous infusion of pentobarbital sodium, paralyzed with pancuronium, and maintained on mechanical ventilation. We found that SHR exhibited significantly higher arterial pressure (AP), heart rate (HR), and VLF, LF, and high-frequency (0.8-2.4 Hz) power of APV than WKY under resting state. Broad-band electrical stimulation of the RVLM elicited parallel APV in the VLF and LF ranges in both rat strains. The evoked APV and transfer magnitude of the APV to stimulus spike rate variability (RVLM-AP magnitude) were significantly higher in SHR, especially in the LF range. The response frequency of central vasomotor control, represented by the high-cut frequency of RVLM-AP magnitude, was also extended in SHR. The disparity in RVLM-AP transfer magnitude between SHR and WKY became virtually absent after combined alpha- and beta-adrenoceptor blockade by phentolamine and propranolol. These results suggest that the dynamic control of RVLM on AP reactivity is enhanced in SHR, in which the adrenergic system may play a major role.

Action Potentials↗