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Biomedical subjects

E Chan

Publications and source records attributed to E Chan.

At least 73 records · Page 4Linked to original sources
PubMed1998

Comparison of the combined cortical thickness of the second metacarpal with Sharp's method for scoring hand microradiographs in rheumatoid arthritis.

OBJECTIVE: To compare the correlation and specificity of the combined cortical thickness of the second metacarpal (CCT-MC) with Sharp's method (SM) for scoring joint erosions and joint space narrowing in rheumatoid arthritis (RA) and to compare the degree of interobserver agreement between the 2 methods. METHODS: Hand microradiographs of 22 women with RA, functional classes III and IV, were scored independently by 3 rheumatologists using the CCT-MC and the CCT of the middle phalanx and SM. RESULTS: (1) There was a highly significant correlation between the total SM score and the CCT-MC for the 3 observers (r = 0.61, p = 0.0026), but not between the CCT of the middle phalanx and SM (r = 0.15, p = 0.53). There was a lower degree of agreement between the observers for SM erosion scores compared to the CCT-MC (intraclass correlation 0.88 for the CCT-MC and 0.63 for the SM); (2) Both joint space narrowing and erosion scores correlated highly with the CCT-MC (r = -0.60, p = 0.004; and r = -0.51, p = 0.014, respectively); (3) CCT-MC measurements are more closely related to the inner (d) as opposed to the outer (D) diameter of the 2nd metacarpal; (4) The mean time to obtain the CCT-MC score was 3.43 min (SD = 1.38) versus 9.83 min (SD = 3.20) for SM (p = 0.0001); (5) the derivative, (D2-d2)/D2, was significantly correlated with SM (r = -0.72, p = 0.0002) and its erosion and joint space narrowing components (r = -0.63, p = 0.0019; and r = -0.71, p = 0.0002, respectively). CONCLUSION: The CCT-MC is a rapid, practical method with higher agreement among observers compared to SM and correlates highly with SM scores for joint damage in RA. CCT-MC appears to have a higher degree of specificity than other sites for CCT measurement. The CCT-MC is more closely related to the inner diameter than the outer diameter, which supports the notion that the principal site of accelerated bone loss due to RA in the hand occurs at the endosteal surface. The CCT-MC should be further assessed with respect to monitoring radiological progression in RA.

Arthritis, Rheumatoid↗
PubMed1997

Effects of ingredients on stability of captopril in extemporaneously prepared oral liquids.

The stability of captopril in several extemporaneously prepared oral liquid formulations was studied. Captopril 1-mg/mL oral liquid formulations were prepared from either powder or tablets in two grades of water, syrup, methylcellulose, and edetate disodium. The liquids were stored at 5 degrees C in amber glass containers, and samples were removed at intervals up to 30 days for assay of captopril concentration by stability-indicating high-performance liquid chromatography. The pH of the formulations remained fairly stable for 30 days. In general, captopril was more stable in formulations containing captopril from tablets than from powder. Captopril in formulations in which the only vehicle was highly purified water was slightly but not significantly more stable than in formulations made with sterile water for irrigation. Formulations made with undiluted syrup were more stable than formulations in which water was used to dilute syrup or formulations containing methylcellulose. The formulations containing edetate disodium were much more stable than those that lacked this component. The stability of captopril 1 mg/mL in oral liquid formulations was influenced by the captopril source (tablets versus powder) and by the presence of syrup, methylcellulose, and edetate disodium. Captopril in a preparation made with tablets and undiluted syrup was stable for 30 days at 5 degrees C, and the formulation should be palatable.

Captopril↗
PubMed1997

Propagation of fluorescent light.

BACKGROUND AND OBJECTIVE: In general, the remitted fluorescence spectrum is affected by the scattering and absorption properties of tissue. Other important factors are boundary conditions, geometry of the tissue sample, and the quantum yield of tissue fluorophores. Each of these factors is examined through a series of Monte Carlo simulations. STUDY DESIGN/MATERIALS AND METHODS: Monte Carlo modeling is used to simulate the propagation of excitation light and the resulting fluorescence. Remitted fluorescence is determined for semi-infinite single and multiple layer geometries and for cubic geometries representing small tissue samples. Monte Carlo results are compared to approximations obtained with a heuristic model. RESULTS: Remitted fluorescence as a function of (1) the depth of fluorescence generation and (2) radial escape position is presented for semi-infinite single and multiple layer geometries. Fluorescence from a small tissue sample is simulated in terms of a cubic geometry, and losses from the sides and bottom are presented as a function of cube dimensions in terms of optical depth of the excitation wavelength. Monte Carlo results for a homogeneous semi-infinite layer are compared to a simple, fast heuristic model. CONCLUSION: Both Monte Carlo simulations and the heuristic model clearly detail the volume of tissue interrogated by fluorescence. Since approximately 35-40% of the remitted fluorescence is due to photons originally directed away from the surface, distal layers affect the remitted fluorescence. Fluorescence spectra from small biopsy samples may not produce the correct line shape owing to wavelength dependent losses.

Aorta↗
PubMed1997

Minimum length of sequence homology required for in vivo cloning by homologous recombination in yeast.

With efficient homologous recombination in Saccharomyces cerevisiae, a rapid in vivo cloning technique has been available. Here we demonstrated that 30 bp of a homologous sequence at each end of a DNA fragment is sufficient to integrate the fragment into a linearized plasmid in yeast. To obtain a high yield of recombination transformants, however, more than 60 bp are desirable. Interestingly, we observed that 20 bp of homology at one end of a DNA fragment is sufficient for efficient recombination provided that the other end contains 80 bp of homologous sequence. Some applications, including high-throughput transferring of EST inserts to the yeast expression systems for the Human Genome Project, are discussed.

Animals↗
PubMed1997

Relationship between stiffness, internal cell pressure and shape of outer hair cells isolated from the guinea-pig hearing organ.

The mechanical properties of outer hair cells are of importance for normal hearing, and it has been shown that damage of the cells can lead to a reduction in the hearing sensitivity. In this study, we measured the stiffness of isolated outer hair cells in hyper- and hypotonic conditions, and examined the change in stiffness in relation to the corresponding changes in internal cell pressure and cell shape. The results showed that the axial stiffness of isolated outer hair cells (30-90 microns in length, 8-12 microns in diameter), ranging from 0.13-5.39 mN m-1, was inversely related to cell length. Exposure to hyper- and hypotonic external media with a small percentage change in osmolality caused a similar magnitude of change in cell length and cell diameter, but an average 60% change in cell stiffness. Therefore, a moderate osmotic change in the external medium can lead to a significant alteration in cell stiffness. The findings thus indicate an important contribution of internal cell pressure to cell stiffness.

Animals↗
PubMed1997

Single-point phenytoin dosage predictions in Singapore Chinese.

Phenytoin dosing is often difficult in a clinical situation because of the non-linear nature of phenytoin metabolism at therapeutic plasma concentrations. This study was performed to examine retrospectively five pharmacokinetic methods of adjusting phenytoin dosage based on a single steady-state plasma phenytoin concentration-dose pair in 66 epileptic Chinese children and adults. The methods compared included four fixed-parameter(s) methods (1 = the fixed Km (Michaelis-Menten constant) method; 2 = the fixed Vm (maximum rate of elimination) method; 3 = the fixed Vmax/Km method; 4 = the fixed S (slope of logarithmic growth model) method) and a Bayesian feedback method (method 5). Measures of bias or accuracy (mean error, percentage dose) and precision (root mean squared error, percentage dose) were 20.1/88.0, -1.85/21.5, 2.14/21.9, -1.07/21.1 and -1.98/22.2, respectively. Method 1 was significantly inferior to methods 2-5 with respect to accuracy and precision. The correlation between the predicted and observed doses was higher with methods 2-5 (r = 0.862, 0.855, 0.866 and 0.841, respectively) when compared to method 1 (r = 0.539). All methods had a sizeable number of poor predictions (range: 21.2% for method 4 to 37.9% for method 1), i.e. predictions with an error of greater than 20% of the dose. With respect to the frequency of poor versus good predictions, comparisons of all methods showed no significant differences.

Adolescent↗
PubMed1997

Lithium pharmacokinetics in Chinese manic-depressive patients.

The pharmacokinetics of lithium was studied in 16 manic-depressive Chinese adults who were on lithium therapy in Hong Kong. A two-compartment open model was used to describe the plasma data obtained. The steady-state volume of distribution and total plasma clearance were estimated to be 63.4 +/- 38.3 L and 1.47 +/- 0.49 L/hr, respectively, which were similar to those reported in studies of Caucasian subjects. The lithium clearance was found to be significantly related to lean body mass (r = 0.646, p < 0.01) but not to either total body weight, age, or creatinine clearance. Interindividual variability in lithium clearance was substantially reduced to a coefficient of variation of 24.6% from 33.3% when lean body mass was considered. The combined plasma and urine data indicate that the rate and extent of lithium absorption were similar to those of earlier studies in Caucasian subjects on a standard lithium dosage.

Adult↗
PubMed1997

The changing preference of T and B cells for partners as T-dependent antibody responses develop.

Recirculating virgin CD4+ T cells spend their life migrating between the T zones of secondary lymphoid tissues where they screen the surface of interdigitating dendritic cells. T-cell priming starts when processed peptides or superantigen associated with class II MHC molecules are recognised. Those primed T cells that remain within the lymphoid tissue move to the outer T zone, where they interact with B cells that have taken up and processed antigen. Cognate interaction between these cells initiates immunoglobulin (Ig) class switch-recombination and proliferation of both B and T cells; much of this growth occurs outside the T zones B cells migrate to follicles, where they form germinal centres, and to extrafollicular sites of B-cell growth, where they differentiate into mainly short-lived plasma cells. T cells do not move to the extrafollicular foci, but to the follicles; there they proliferate and are subsequently involved in the selection of B cells that have mutated their Ig variable-region genes. During primary antibody responses T-cell proliferation in follicles produces many times the peak number of T cells found in that site: a substantial proportion of the CD4+ memory T-cell pool may originate from growth in follicles.

Animals↗
PubMed1996

An isolated rat vibrissal preparation with stable responses of slowly adapting mechanoreceptors.

Sinus hairs were isolated from rats and examined in an isolated organ bath while superfused with oxygenated synthetic interstitial fluid. The distal end of the deep vibrissal nerve was teased for single unit recordings of responses from slowly adapting mechanoreceptors to standard bending of the hair. Sinus hair type I and type II receptors could be clearly identified by their respective characteristic firing pattern. Their responses were stable for at least 5 h even if the sinus hair had been stored at 4 degrees C for 24 h beforehand. Electron microscopic examination of these hairs at the end of experiments showed well preserved ultrastructure without abnormalities. The short diffusion distances in this preparation make it well suited for studying drug effects with the aim of investigating the mechanoelectric transduction process in these receptors.

Animals↗
PubMed1996

Cytoplasmic Ca2+ concentrations in intact Merkel cells of an isolated, functioning rat sinus hair preparation.

An isolated, functioning sinus hair preparation was developed to investigate cytoplasmic Ca2+ concentrations in intact Merkel cells using microfluorimetric techniques. Intracellular Ca2+ levels were monitored by means of photon counters in small groups of Merkel cells loaded with the calcium fluorescent indicators fura-2 or fluo-3. Mechanical stimulation of Merkel cells with fine glass rods resulted in small transient increases in intracellular Ca2+ levels (by about 20%) in the group of Merkel cells around the stimulating probe. A rise in Ca2+ is presumed to be essential for the postulated synaptic transmission to the afferent nerve terminal. Depolarization with a high concentration of potassium chloride (100 mM) caused increases in intracellular Ca2+ concentrations in Merkel cells (by about 70%) only in the presence of extracellular Ca2+, indicating an influx of Ca2+ through voltage-gated channels. The Ca2+ response was abolished neither by (+)-BayK8644 nor omega-conotoxin, suggesting that the Ca2+ channels are different from the classical L- or N-type channels. Extracellular application of ATP (10 microM to 5 mM) caused dose-dependent increases in intracellular Ca2+ levels in Merkel cells of up to sevenfold from the basal level of about 100 nM. Similar responses to ATP were also measured during superfusion with Ca(2+)-free medium, suggesting intracellular stores as the main Ca2+ source. Pre-incubation of Merkel cells with the purinoceptor antagonist suramin (100 microM) for 30 min reduced the Ca2+ responses to ATP by about 50% compared with control conditions. In conclusion, the results have demonstrated that a rise in intracellular Ca2+ in Merkel cells can be evoked by mechanical stimulation, membrane depolarization and chemical stimulation by ATP. These observations strongly suggest a possible contribution of Ca2+ to the normal responsiveness of Merkel cell mechanoreceptors, in turn supporting the hypothesis that Merkel cells are involved in the mechano-electric transduction process in sinus hair type I mechanoreceptors.

Adenosine Triphosphate↗
PubMed1996

Complement C6 and C7 DNA polymorphisms analysed by PCR in seven ethnic groups and characterisation of the C6 MspI RFLP.

Five polymorphisms in the C6 and C7 genes have been investigated in seven ethnic groups. The allele frequencies are broadly similar in most groups except C7 M/N which is monomorphic in our group of Africans, and C6 MspI and C7 S367T where the allele frequencies in African and Cape Coloured subjects are very different from the other ethnic groups. There is very little allelic association except between C6 A/B and C6 MspI. Seventeen of the 32 possible haplotypes have been observed, suggesting that much recombination has taken place. We describe a new method for the investigation of the MspI RFLP located in intron 3 of C6 (approximately 3 kbp 3' from exon 3 and 1.5 kbp 5' from exon 4) and its molecular basis, together with an improved method for the isolation of DNA from stored serum.

Alleles↗
PubMed1995

Mycobacterium tuberculosis chaperonin 10 forms stable tetrameric and heptameric structures. Implications for its diverse biological activities.

The chaperonin activity of sequence-related chaperonin 10 proteins requires their aggregation into heptameric structures. We describe size-exclusion chromatography and ultracentrifugation studies that reveal that while Escherichia coli chaperonin 10 exists as a heptamer, the Mycobacterium tuberculosis chaperonin 10 is tetrameric in dilute solutions and in whole M. tuberculosis lysate. At high protein concentration and in the presence of saturating amounts of divalent ions, the protein is heptameric. Human chaperonin 10 is predominantly heptameric, although smaller oligomers were detected. These differences in structural assembly between species may explain differences in biological activity such as antigenicity. Using C-terminal and N-terminal fragments, sequence 1-25 was identified as indispensable for aggregation. CD spectroscopy studies revealed that (i) a minimum at 202-204 nm correlates with aggregation and characterizes not only the spectrum of the mycobacterial protein, but also those of E. coli and human chaperonin 10 proteins; (ii) the interactions between subunits are of the hydrophobic type; and (iii) the anti-parallel beta-pleated sheet is the main secondary structure element of subunits in both tetrameric and heptameric proteins.

Amino Acid Sequence↗
PubMed1995

Sequence and structural homologies between M. tuberculosis chaperonin 10 and the MHC class I/II peptide binding cleft.

The peptide corresponding to the C-terminal half of M.tuberculosis hsp10 was synthesised based on the prediction that it might represent an independent structural region of the protein. This hypothesis was confirmed by aggregation and CD studies using this peptide and longer sequences of the protein. The peptide shares about 40-50% sequence homology with alpha 2 and beta 1 chains of MHC class I and II antigens. This and the CD results which indicated that the peptide at acidic pHs folds into an anti-parallel beta-sheet were used to generate a 3D model which has the same "W" fold contained in the MHC peptide binding groove. These data suggest that the hypothesis of molecular mimicry proposed to be one of the mechanisms which triggers autoimmune diseases may be extended to hsp10 proteins. Furthermore the suggested evolutionary relationship between hsp's and MHC antigens may find support from these data.

Amino Acid Sequence↗
PubMed1995

Corneal photocoagulation with continuous wave and pulsed holmium: YAG radiation.

In this study, the effectiveness of pulsed and continuous wave (CW) holmium: YAG lasers in coagulating in vitro pig corneas was analyzed. With the CW laser, irradiance and exposure time were varied; irradiance, from 162 to 324 W/cm2 and exposure time, from 200 to 800 ms. With the pulsed laser, number of pulses and radiant exposure were varied; number of pulses per lesion, from 4 to 30 and radiant exposure, from 10 to 25 J/cm2. Laser-induced corneal damage was determined by analyzing histological cross sections of each lesion. Depth and diameter of the lesions were plotted against the varying laser parameters. Light and birefringent photomicrographs of typical lesion histology show that the pulsed laser significantly damaged superficial layers of the cornea and could not achieve the coagulation depths produced by the CW laser. Additional histology demonstrates that minimal surface damage (intrastromal coagulation) occurred when the CW laser beam was delivered with a sapphire-tipped contact probe. The results provide empirical data on the sensitivity of each parameter in producing a range of coagulation end points. In addition, the experimental results describe trends between the parameters of either laser and the extent of coagulation.

Animals↗
PubMed1995

Transurethral resection of the prostate versus open prostatectomy: long-term mortality comparison.

To determine whether transurethral prostatectomy results in higher long-term mortality rates than open prostatectomy, we reviewed retrospectively 1,125 patients treated by transurethral and 190 treated by nonperineal open prostatectomy for benign disease at 1 institution from 1978 through 1987. Patients in whom prostatic cancer was found were excluded. We identified age, preoperative medical illnesses and urinary retention, American Society of Anesthesiologists category, type of anesthesia, length of followup, health status and cause of death. For statistical analysis the study cohort consisted of 527 patients in whom the charts were complete and followup was adequate (421 in the transurethral prostatectomy and 106 in the open prostatectomy groups). Mean age for the 2 groups was 66.3 and 67.5 years, respectively. With an average followup of 70.7 months 77% of the transurethral prostatectomy group were alive, compared to 78% of the open prostatectomy group at an average followup of 71.4 months. We found no supportive evidence that transurethral prostatectomy results in higher long-term mortality rates than does an open operation (log-rank test p = 0.74). Also, there was no significant survival difference in patients who required a preoperative Foley catheter. We also examined a subset of patients with adequate followup who had no significant medical history (for example hypertension, diabetes, heart disease and so forth) and compared them to patients with medical illnesses at prostatectomy. There was a significant survival difference between those with and without preoperative medical conditions (Wilcoxon test p = 0.047) in the transurethral prostatectomy group but not in the open group (p = 0.58). However, there was no significant survival difference between procedures among the healthiest subset of patients (p = 0.16).

Aged↗
PubMed1995

The lurcher gene induces apoptotic death in cerebellar Purkinje cells.

In the neurologically mutant mouse strain lurcher (Lc), heterozygous animals display cell autonomous degeneration of cerebellar Purkinje cells beginning in the second postnatal week. During the course of our studies to identify the genetic lesion responsible for this disease (Norman et al., 1991), we have formulated an hypothesis suggesting that in Lc Purkinje cells homeostasis is sufficiently perturbed to lead to the activation of programmed cell death, thus resulting in neuronal loss and the consequent neurologic disease (Heintz, 1993). To address this possibility, we have examined the properties of Lc Purkinje cells as they die during the second postnatal week. Our light and electron microscopic studies demonstrate that dying Lc Purkinje cells exhibit the characteristic morphologic features of apoptosis, including nuclear condensation, axon beading and membrane blebbing. Using an in situ end-labeling method, we have also detected nicked nuclear DNA in these cells. Furthermore, we have examined the expression of the sulfated glycoprotein 2 (SGP2), whose mRNA is induced in both T-cells and prostate epithelial cells undergoing apoptotic death. We show by in situ hybridization that SGP2 is not expressed at detectable levels in normal Purkinje cells, but that its mRNA is present in Lc Purkinje cells prior to their death. Also expression of the Kv3.3b potassium channel, which marks the terminal phase of Purkinje cell differentiation, is evident in Lc Purkinje cells prior to their death. These data demonstrate that the Lc mutation induces apoptosis in cerebellar Purkinje cells following their maturation in postnatal cerebellum. Isolation of the Lc mutation and further analysis of its action in eliciting apoptosis can provide an important opportunity for understanding the etiology of neurodegenerative disease.

Animals↗