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Biomedical subjects

W Y Cheung

Publications and source records attributed to W Y Cheung.

At least 37 records · Page 2Linked to original sources

Induction training, career counselling, and performance review: views of junior medical staff.

Surveys of senior house officers and registrars were undertaken by postal questionnaire to ascertain views on the need for and content of induction training, career counselling, and performance review. The questionnaire was sent out in May 1990 and repeated in May 1996, after measures had been taken to improve induction training, and assessment and appraisal of trainees. In 1990 there was a clear wish to receive information on career prospects, research and education opportunities, and clinical audit, but more ambivalence regarding information or training in communication, discharge policies, standards, and encoding procedures. There was also a firm view that career counselling could be improved and formal goal setting and performance appraisal interviews would be welcomed. In 1996 there was disappointingly little change in the views expressed by the junior medical staff, though there was a significant increase in confidence in the role of the consultant in career counselling.

Career Choice↗

Microvascular free flap reconstruction: the Kwong Wah Hospital experience.

The early experience of free flap reconstruction in a regional plastic surgery unit was reviewed. Forty-seven free flap surgical procedures were performed for 46 patients during a 2-year period from 1 January 1995 to 31 December 1996 in a regional plastic surgery centre that specialises in head and neck, and breast reconstruction. Twenty-six free transverse rectus abdominus myocutaneous flaps were performed for 25 breast cancer patients. Head and neck reconstructions were mostly performed after surgical ablation of oral and pharyngeal cancers. A variety of free flaps were used, including fibular osteocutaneous flaps, radial forearm fasciocutaneous flaps, jejunal grafts, and rectus abdominus musculocutaneous flaps. Lateral arm fasciocutaneous and scapular fasciocutaneous flaps were used for three patients who had foot ulceration. The success rate of free flap transfer was 94%. Flap loss was found in three patients who had pedicle complications that were due to traction, kinking, and thrombosis. There was no mortality in the series. The most common morbidity was wound infection. Two thirds of the free flap transfers were uneventful. Seven (15%) patients had major complications that required re-operation. Institutional support was essential for the development of microvascular surgery.

Journal Article↗

Effect of anastomosis and geometry of vessel curvature on blood flow velocity and patency in microvessels.

The effect of the geometry of the vessel and the number of anastomoses on the blood flow was studied. Four different shapes of the vessel were constructed by using a 6-cm-long double vein graft model with three anastomoses: (1) an alpha loop, (2) an omega loop, (3) a sigmoid curve, and (4) straight. Blood flow was measured by an ultrasound Doppler flowmeter. The result showed no alternation in blood flow across different geometry and through three patent microanastomoses. However, six out of seven vein grafts were thrombosed at 24 hr postoperative due to vascular kinks. This model demonstrates potential sites of kinking at the dissection end of the femoral artery, the microanastomoses, the side branches of the vein graft, and the adventitial adhesions. This model is recommended to microvascular trainees for the study of kinking and the management of redundant pedicles and vein grafting.

Anastomosis, Surgical↗

Brain L-glutamate decarboxylase. Inhibition by phosphorylation and activation by dephosphorylation.

Previously we showed that the activity of the gamma-aminobutyric acid-synthesizing enzyme L-glutamate decarboxylase (GAD) in crude brain extract is inhibited by ATP and protein phosphatase inhibitors. We suggested that GAD activity is regulated by protein phosphorylation. In this paper we further present evidence to support our hypothesis that protein kinase A and calcineurin may be involved in regulation of GAD activity through phosphorylation and dephosphorylation fo GAD, respectively. In addition, the effect of neuronal stimulation on GAD activity in cultured neurons is also included. A model to link neuronal excitation and activation of GAD by Ca(2+)-dependent phosphatase is proposed.

Animals↗

Stimulation of myosin light-chain kinase by Cd2+ and Pb2+.

The effect of Cd2+ on myosin light chain kinase (MLCK) reported in the literature is controversial, apparently because the level of Ca2+ contaminating the reaction mixture could not be accurately controlled by the addition of a metal chelator when Cd2+ was also present. In the present study, we have reduced the contaminating Ca2+ to a trace level that did not interfere with the enzyme activity; thus the use of a metal chelator was not necessary. We showed that Cd2+, or Pb2+ had a biphasic effect on MLCK isolated from chicken gizzard: stimulation at low and inhibition at high concentrations. (The stimulatory effect of on the enzyme activity isolated from chicken gizzard: stimulation at low and inhibition at high concentrations). The stimulatory effect of Cd2+ or Pb2+ on MLCK activity was not seen in the absence of calmodulin, and was abolished by trifluoperazine, a calmodulin antagonist, indicating that the heavy metals exert their activation via calmodulin. The inhibition of the enzyme activity by Cd2+ or Pb2+ at higher concentrations was also seen with the calmodulin-independent catalytic fragment of MLCK, suggesting that the inhibition is probably through their binding to sulfhydryl groups that are essential for catalytic activity. Pb2+ was more effective than Cd2+ in stimulating the enzyme activity, but less potent in inhibition. The extent of stimulation by heavy metals most likely resulted from a combination of the biphasic effects. Dithiothreitol and N,N,N',N'-tetrakis (2-pyridylmethyl) ethylenediamine selectively chelated Cd2+ and Pb2+ over Ca2+, and reversed their stimulatory or inhibitory effect on MLCK.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

A family of related sequences associated with (TTTAGGG)n repeats are located in the interstitial regions of wheat chromosomes.

A family of related sequences associated with (TTTAGGG)n repeats has been cloned from the wheat cultivar Chinese Spring. These sequences reveal a high level of polymorphism between wheat varieties when used as restriction fragment length polymorphism (RFLP) probes. Although this family of sequences contains motifs homologous to the repeats in the telomeres of wheat, they are located at interstitial sites on wheat chromosomes.

Blotting, Southern↗

H+ is involved in the activation of calcineurin by calmodulin.

We have studied recently the surface hydrophobic properties of several calmodulin (CaM) target enzymes and found that a certain amphipathic domain(s) of the enzyme was exposed in response to a H+ increase within the neutral pH range. The exposed domain appeared to be related to their CaM-binding domains and associated with their activation by CaM (Huang, S., Carlson, G. M., and Cheung, W. Y. (1994) J. Biol. Chem. 269, 7631-7638). In this paper, the involvement of H+ in the activation of calcineurin (CaN), one of the CaM target enzymes, was further studied. Using dansylated CaM to monitor its conformational change, we found that the binding of Ca2+ to CaM occurred at a lower range of Ca2+ concentrations (pCa from 7 to 6) than the formation of CaM.CaN complex (pCa from 6 to 5). However, addition of H+ within the neutral pH range shifted the formation of CaM.CaN complex to the lower range of Ca2+ concentrations. Similarly, the addition of H+ shifted the Ca2+ requirement for enzyme activation to a lower Ca2+ range. These results show that the interaction of CaM with CaN could be initiated either by Ca2+ or by H+ in the presence of suboptimal Ca2+, suggesting a positive cooperative effect between Ca2+ and H+. The CaM-stimulated activity of CaN was accompanied by a time-dependent autoinactivation. Kinetic analysis showed that the autoinactivation initiated by Ca2+ or by H+ was biphasic and that the decay rate constants were comparable. In heavy water (D2O), the Ca(2+)-induced formation of CaM.CaN complex as well as the activity of CaN were decreased, indicating a solvent deuterium isotope effect. The same solvent isotope effect was observed in the H(+)-induced formation of CaM.CaN complex and in the H(+)-induced enzyme activity. When the enzyme was rendered CaM-independent by limited proteolysis, the isotope effect was abolished, suggesting that H+ was involved in the CaM-dependent and not the CaM-independent activity. Collectively, these data suggest that the interaction of CaM with CaN depends on the two cations: Ca2+ exposes an amphipathic domain on CaM, and H+ exposes the CaM-binding domain on the target enzyme.

Animals↗

Calmodulin-dependent enzymes undergo a protein-induced conformational change that is associated with their interactions with calmodulin.

The anionic hydrophobic (amphipathic) fluorescent probe 2-(p-toluidinyl)-naphthalene-6-sulfonate was used to investigate the surface hydrophobic properties of calmodulin (CaM)-dependent enzymes as follows: calcineurin, myosin light chain kinase, cyclic nucleotide phosphodiesterase, CaM-dependent protein kinase II, and the gamma-subunit of phosphorylase kinase. We found that certain domains of these enzymes that interacted with 2-(p-toluidinyl)-naphthalene-6-sulfonate were exposed by a transient proton (H+) increase within the neutral pH range. This H(+)-induced exposure, which could be caused either by direct addition of H+ or by the release of H+ from metal chelators upon their binding of Ca2+, seemed to be more closely linked with a change in pH value (i.e. transient H+ increase) than with the actual equilibrium pH value of the system. Unlike the case with CaM-dependent enzymes, the H(+)-induced conformational change was uncommon in CaM-independent enzymes. When CaM-binding domains were removed from calcineurin and smooth muscle myosin light chain kinase, the resultant enzymes no longer exposed new domains in response to H+ increase. Using dansylated CaM to monitor the formation of CaM-enzyme complexes, we found that complex formation occurred with an uptake of H+ from solution. When CaM-dependent enzymes were evaluated at suboptimal concentrations of Ca2+, addition of H+ enhanced both the formation of CaM-enzyme complexes and the CaM-dependent catalytic activities, but this synergistic H+ effect occurred within only a narrow range of Ca2+ concentrations. These findings suggest that the H(+)-exposed domains in CaM-dependent enzymes are involved in the binding of CaM and that both conformational changes in CaM and its enzyme targets are necessary for complex formation. Further, the findings are consistent with the notion that CaM-binding domains are masked in the nonactivated (uncomplexed) conformations of CaM-dependent enzymes. The interplay between H+ and Ca2+ is discussed in relation to other systems that display interdependent effects of these two ions.

Animals↗

Practice characteristics associated with audit activity: a medical audit advisory group survey.

AIM: A survey of general practices in West Glamorgan was carried out to assess the development of record systems and practice organization and to relate this to audit activity. METHOD: Following mutual agreement practices were visited by two assessors. Visits were conducted using an agreed protocol and practice data were recorded on a semi-structured questionnaire, which had been sent to the practices in advance of the visits. Practices varied from single handed practices to group practices with six partners. RESULTS: Fifty seven of the 63 practices in the county suitable for inclusion were visited (90%). Data were analysed from 54 practices (three visits were regarded as a pilot). The practices more likely to be involved in audit were those with three or more partners and which had modern medical record systems. Audit activity had occurred in 87% of practices who kept clinical summaries in the notes (compared with 38% who did not), in 87% of practices with long-term medication summaries in the notes (compared with 40% with no summaries) and in 85% of practices with a computerized age-sex register (compared with 50% with no register). All training practices had undertaken audit compared with 63% of non-training practices. CONCLUSION: The survey enabled the medical audit advisory group to identify the type and degree of audit undertaken locally and highlighted the characteristics which encourage this activity. As a consequence the group is able to target practices who have limited or no involvement in audit and to offer assistance and advice on record modification likely to enhance audit activity.

England↗

Latent sites of Epstein-Barr virus infection.

Epstein-Barr virus (EBV) has been detected in a wide spectrum of tumors. This study investigates the detection rate of EBV-DNA by Southern blot hybridization analysis (SOBH) and polymerase chain reaction (PCR) in different tissues from persons without apparent EBV-related diseases. Of 20 tonsillectomy specimens studied, SOBH indicated positivity for EBV-DNA in 1 case, and PCR indicated positivity in 10. In autopsies performed on patients with no apparent evidence of EBV-related diseases, the viral DNA was only detected by PCR in the following: parotid gland (7/15), submandibular gland (8/20), nasopharynx (8/10), tonsil (8/10), larynx (5/6), lung (5/9), cervical lymph node (7/10), mediastinal lymph node (7/10), abdominal lymph node (4/10), spleen (6/10), thyroid (5/10), liver (1/10), pancreas (1/4), kidney (4/10), uterine cervix (1/4), ovary (1/5) and testis (1/3). These results provide a baseline for interpreting the role of EBV in carcinogenesis.

Adolescent↗

Long-range physical mapping of the alpha-amylase-1 (alpha-Amy-1) loci on homoeologous group 6 chromosomes of wheat.

Long-range physical maps of the small multigene family of the malt alpha-amylase genes (alpha-Amy-1) located on the long arms of wheat chromosomes 6A (the alpha-Amy-A1 locus) and 6B (alpha-Amy-B1) were generated by pulsed-field gel electrophoresis analysis. By using three methylation-sensitive rare-cutter restriction endonucleases, NotI, NruI and MluI, and an alpha-Amy-1 cDNA probe and four gene-specific genomic probes from the alpha-Amy-B1 locus, the size of the alpha-Amy-A1 locus was estimated to be about 700 kb and of the alpha-Amy-B1 locus to be about approximately 4300 kb. These two maps indicate clustering of GC-rich and C-methylation-sensitive restriction enzyme recognition sites. At least five regions reminiscent of 'CpG islands' are apparent in alpha-Amy-B1, and three in alpha-Amy-A1. Correlation between recombination frequency and physical distance within the alpha-Amy-B1 locus suggests that 1 cM approximates to 1 Mb in physical distance.

Bacterial Proteins↗

The isolation of high molecular weight DNA from wheat, barley and rye for analysis by pulse-field gel electrophoresis.

A method is presented for the preparation of large DNA molecules from protoplasts embedded in agarose blocks of three different cereals--hexaploid bread wheat (Triticum aestivum), barley (Hordeum vulgare) and rye (Secale cereale). Pulse-field gel electrophoresis (PFGE) analysis of these DNA preparations using a contour-clamped homogeneous field (CHEF) apparatus indicated that the size of the DNA molecules was greater than 6 Mb. DNA samples prepared by this method were shown to be useful for restriction analysis using both frequent and rare cutting enzymes.

DNA↗

Activation of troponin C by Cd2+ and Pb2+.

Certain heavy metal actions such as Cd2+ and Pb2+ mimic Ca2+ effectively in stimulating calmodulin (CaM). We now show that these cations also activate skeletal muscle troponin C (TnC), a Ca2(+)-binding protein highly homologous to CaM. Like Ca2+, these cations allow TnC to alter its electrophoretic mobility on polyacrylamide gels, and to bind to phenyl-Sepharose. Moreover, they activate TnC to stimulate myofibrillar ATPase. When TnC was removed from the skeletal myofibrils by treatment with trans-1,2-cyclohexanediamine-N,N,N',N'-tetraacetic acid (CDTA), the ATPase activity was no longer stimulated by the cations. However, after reconstitution of CDTA-treated skeletal myofibril with TnC, the response of ATPase to Ca2+, Cd2+ or Pb2+ was restored. These findings suggest that the activation of myofibrillar ATPase by Cd2+ and Pb2+ is mediated through TnC. The ability of the heavy metals to stimulate TnC-supported ATPase activity correlated quite well with the ability to increase the extent of the myofibrillar superprecipitation. The activation of TnC by Cd2+ or Pb2+ could constitute a possible molecular basis for their toxicity.

Adenosine Triphosphatases↗

Phosphatidylinositol modulates the response of calmodulin-dependent phosphatase to calmodulin.

Phosphatidylinositol (PtdIns) and many other phospholipids activated calmodulin (CaM)-dependent phosphatase in the presence or absence of Ca2+, and the stimulation was more pronounced in the presence of Ca2+. In addition, PtdIns modulated the response of phosphatase to CaM: at low and nonstimulatory concentrations (less than 70 microM), PtdIns augmented the activity of phosphatase by a submaximum concentration of CaM, giving a synergistic effect; and at high concentrations (greater than 100 microM), PtdIns suppressed the synergistic effect. Kinetic experiments indicated that PtdIns (both nonstimulatory and stimulatory concentrations) increased the affinity of phosphatase for CaM. In addition to the CaM regulatory site, phosphatase appears to have two PtdIns regulatory sites: a high-affinity site the occupation of which does not stimulate enzyme activity, and a low-affinity site the occupation of which stimulates enzyme activity in the absence of CaM and inhibits it in the presence of CaM. Modulating the response of phosphatase to CaM is not unique to PtdIns, and was observed with other phospholipids, including some that did not stimulate the enzyme. This raises the possibility that certain phospholipids may regulate phosphatase in two ways: (i) direct activation of the enzyme and (ii) modulation of its response to CaM.

Animals↗

Calmodulin-dependent phosphatase preferentially dephosphorylates a 28 kDa protein in human platelets.

1. Human platelets contain a calmodulin-dependent phosphatase (calcineurin) that has many properties similar to those of bovine brain calmodulin-dependent phosphatase. 2. The activity of calcineurin phosphatase accounts for a small fraction of the total phosphatase activity in human platelets. 3. Labeling of human platelets with 32P yielded many phosphoproteins. 4. Incubation of a lysate of the 32P-labeled platelets with bovine brain calmodulin-dependent phosphatase led to preferential dephosphorylation of a 28 kDa protein (P28), a minor component of platelet proteins. 5. P28 is one of several proteins that were rapidly labeled upon stimulation of platelets with thrombin. 6. Even though the enzyme is known to catalyze the dephosphorylation of many substrates in vitro, its apparent preference for P28 suggests that its activity is highly selective.

Animals↗