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Biomedical subjects

M L Ng

Publications and source records attributed to M L Ng.

At least 73 records · Page 4Linked to original sources

A supersensitive in-house enzyme-linked immunosorbent assay (ELISA) for measurement of thyroid-stimulating hormone (TSH) and its clinical applications.

A supersensitive ELISA was developed for measurement of thyroid-stimulating hormone (TSH) concentrations in serum using in-house rabbit polyclonal antisera and a commercial monoclonal antibody. The assay was optimised and validated by recovery, linearity and cross-reactivity experiments and further compared to other available assays and EQAS samples. Good precision was obtained with a working assay range of 0.2 to 100 mIU/L with < 10% coefficient of variation (CV) for both intra and interassay. The assay is highly sensitive and specific with a minimum detectable limit of 0.07 mIU/L and negligible cross-reactivities against LH, FSH, HCG and other pituitary peptides. Good correlations were obtained when compared to Abbott hTSH EIA (r = 0.993; p < 0.001; n = 85) and NETRIA IRMA (r + 0.995; p < 0.001; n = 76). The normal reference range established was 0.4 to 4.0 mIU/L (n = 76). TSH levels in serum of thyrotoxic patients (n = 83) were significantly lower (0.07 to 0.20 mIU/L, p < 0.0001) and completely distinct from normal values thereby obviating the requirement of a TRH-stimulation test. Stability studies showed that coated wells can be stored at 4 degrees C for at least 2 months. This highly sensitive in-house hTSH ELISA which is cheap, stable and readily available is useful for diagnosis and management of patients with various thyroid disorders.

Enzyme-Linked Immunosorbent Assay↗

Evaluation of suppression of growth hormone levels following a 75g oral glucose tolerance test.

Growth hormone (GH) levels were measured after a 75g oral glucose load (OGTT) in normal adults, patients with impaired glucose tolerance (IGT), insulin-dependent diabetes mellitus (IDDM) and acromegaly. Nadir GH levels at 2-hour post-OGTT in normal subjects ranged from 0.4 to 8.4 mIU/L, the 95% confidence interval being 0.4-4.4 mIU/L. In IGT and IDDM subjects basal fasting GH levels were not significantly different from normal and did not alter during OGTT. The high fasting GH level measured in one each of the IGT and IDDM patients was suppressible at 1-hour after glucose intake. In contrast, acromegalic patients had elevated fasting GH levels (11.8-178 mIU/L) although in 3 patients, the levels were mildly elevated and overlapped with normal. OGTT failed or only partially suppressed GH secretion in all acromegalics. Therefore, elevated fasting GH levels are not diagnostic and OGTT is required for accurate diagnosis and assessment of treatment of acromegalic patients.

Acromegaly↗

Clinical applications of salivary cortisol measurements.

The clinical applications of salivary cortisol measurements were evaluated by radioimmunoassay of time-matched saliva and plasma samples. Salivary cortisol levels of normal subjects exhibited a significant (p less than 0.001) diurnal variation with a mean (+/- SD) concentration of 8.7 +/- 4.8 nmol/L at 0800-1000 h and 2.4 +/- 1.1 nmol/l at 1500-1700 h. After an overnight dexamethasone suppression test, morning salivary cortisol levels decrease to 2.7 +/- 0.7 nmol/L (p less than 0.001 vs normal). An excellent correlation (r = 0.805) of cortisol measurements with time-matched saliva and plasma samples was obtained (y = 0.03x + 0.88, p less than 0.001, n = 91). Hypercortisolism was confirmed by raised salivary cortisols in only half of patients with elevated total plasma levels, thereby indicating that salivary cortisol measurements is a better index of adrenal status.

Adrenal Cortex Function Tests↗

Collection and storage of capillary blood in glass fibre filters for glycated haemoglobin measurement by a microcolorimetric method.

A filter method for collection and storage of capillary blood spots for glycated haemoglobin (gHb) has been developed. Glass fibre filters (GFB) impregnated with 0.8 M boric acid were used to collect and store capillary blood. Haemoglobin from the dried blood spots was eluted into water and determined by Drabkin's method, while gHb in the eluates was determined by the microcolorimetric method. The intraassay coefficients of variation (CVs) were 4.5, 4.5 and 3.1% at 882, 1101 and 1704 pmol HMF/mg Hb, respectively. The corresponding inter-assay CVs were 8.6, 8.6 and 6.3%, respectively. A total of 63 paired capillary and venous blood samples were measured by both the direct and GFB method. The GFB method showed excellent correlation with the direct method (r = 0.948 and r = 0.994) after 7 and 14 days' storage at room temperature. The GFB method will enable prior collection and postage of blood samples by patients.

Blood Specimen Collection↗

Thyroid eye disease--medical or surgical therapy?

Thyroid eye disease is autoimmune in nature and associated with Graves' Disease. Autoantibodies to the 64 kDa antigen in thyroid membranes cross-react to the 64 kDa proteins in human eye muscle membranes. Antibody dependent cell mediated cytotoxicity against eye muscle cells are also found in patients with thyroid eye disease. The purpose of this paper is to review the treatment available and to share the authors' experience using cyclosporin A. In the majority of cases, thyroid eye disease is mild, manifest only as bilateral or unilateral proptosis, with/without grittiness of the eyes. This is usually treated conservatively with eye drops. If proptosis is more severe and there is incomplete closure of eyelids, epiphora and conjunctival injection, then lateral tarrsorrhaphy is usually effective, combined with use of eye pads and eye drops. The problem of diplopia can be treated conservatively with special lenses, or with surgical correction of tethered muscles. However when proptosis is severe, with raised intraocular pressure, severe chemosis and danger of blindness, then the choice of therapy is controversial: rapid decompression by surgical means or use of high doses of prednisolone. Most prefer prednisolone therapy initially, surgical decompression if it fails. Various other methods have been tried, aimed at the immunological nature of the disease, namely plasmapheresis, radiotherapy and immunosuppressive drugs such as cyclosporin, with variable success. Our experience with cyclosporin had been mixed and inconclusive.

Cyclosporins↗

Ultrastructural study of Helicobacter pylori-associated gastritis.

Endoscopic biopsies of antral mucosa from 26 patients with Helicobacter pylori-associated gastritis were studied by electron microscopy (EM). Scanning electron microscopy (SEM) showed clustering of H. pylori in the intercellular areas, being entrapped by the microvilli which were decreased at the sites where the bacilli were seen. The observations of SEM were confirmed by transmission electron microscopy (TEM), which showed adherence of the bacilli to the cell surface, producing cup-shaped depressions in the epithelial cells, and occasionally intracellular infiltration by H. pylori. There were also depletion of mucus granules, degenerative changes, and disruption of intercellular junction complexes of the epithelial cells. Post-treatment biopsies showed complete disappearance of the bacilli, and ultrastructural changes associated with H. pylori infection were resolved.

Campylobacter↗

Sexual attitudes in the Chinese.

The sexual attitudes in the Chinese have been described by scholars as suppressive or nonsuppressive, based on observations made on various aspects of the Chinese culture. Many characteristics of the Chinese history and society are responsible for this controversy and confusion. The overall picture may be better understood using the developmental model propounded by some modern sociologists in the study of social units and collective behaviors. It is time to proceed from static to dynamic to try to apply this model in comprehending the sexual attitude of the Chinese and those of other cultures.

Attitude↗

Prevalence of diabetes, hypertension and renal disease amongst railway workers in Malaysia.

A survey was done to determine the prevalence of diabetes mellitus, hypertension and renal disease, as well as extent of diabetic control, amongst the workers of Malaysian Railways. The prevalence of diabetes was high at 6.6%, with 3.8% of these being insulin dependent diabetes. The highest prevalence was in Indians (16.0%) followed by Chinese (4.9%) and Malays (3.0%). Using HbA1 measurements, diabetic control was poor in 70.6% of the diabetics. Hypertension was found in 37% and proteinuria in 35%. Renal impairment was present in 30% of the diabetics. This survey shows that diabetes, hypertension and renal disease are high amongst the railway workers in Malaysia.

Adult↗

Cytostatic effect of antiestrogens in lymphoid cells: relationship to high affinity antiestrogen-binding sites and cholesterol.

The antiproliferative effect of 10(-6) M antiestrogens in an estrogen receptor-negative lymphoid cell line (K36) was enhanced in lipoprotein-poor growth medium. The enhancement was not due to increased bioavailability because cellular uptake of [3H]tamoxifen was not increased and the lipoprotein fraction of serum had negligible [3H]tamoxifen-binding capacity. Cholesterol and lipoproteins, but not mevalonate, reversed the cytostatic effect of antiestrogens. Reversal by cholesterol was dose-related (10(-7) M to 10(-5) M), while that by lipoproteins could also be demonstrated in medium undepleted of lipoproteins. The cytostatic efficacy of a series of ten compounds correlated well with their relative binding affinities for solubilized antiestrogen-binding sites from K36 cells when log IC50 values (concentration required to reduce [3H]thymidine incorporation by 50%) were plotted against log RBA50 values (concentration required to reduce [3H]tamoxifen binding by 50%) (correlation coefficient 0.94). Transmission electron microscopy of antiestrogen-treated cells showed evidence of disordered cytokinesis which was partially reversed by cholesterol. These observations implicate the antiestrogen-binding protein in the antiproliferative effect of antiestrogens in nonestrogen target cells.

Cell Division↗

Regulation of epidermal growth factor receptor by progestins and glucocorticoids in human breast cancer cell lines.

Human breast cancer cells secrete a number of autocrine peptides which modulate their proliferation rates. The known effects of steroid hormones on breast cancer cell proliferation may be mediated in part by altering the production of these growth factors and/or their interactions with cellular receptor sites. Receptors for epidermal growth factor (EGF), which also bind the autocrine growth factor, alpha-transforming growth factor, are present on a number of breast cancer cell lines and it has previously been shown that T-47D and MCF-7 cells respond to progestins with an increase in the concentration of EGF receptors (EGF-R). In the present study we examined the effects of both progestins and glucocorticoids on EGF binding in 10 human breast cell lines. Five of these lines were progesterone receptor positive and all lines expressed the glucocorticoid receptor (GR). All cell lines were initially incubated for 24 hr with increasing concentrations of the synthetic progestin, medroxyprogesterone acetate (MPA), and the level of specifically bound EGF was determined. An increase in specific binding of EGF was confirmed in two PR-positive lines but, in addition, increases in EGF binding were observed in 4 PR-negative cell lines. In these last lines the synthetic glucocorticoid, dexamethasone, was a more potent inducer of EGF binding than MPA, a known glucocorticoid agonist, while the high-affinity PR ligand, ORG 2058, was without effect. Furthermore, MPA competed with dexamethasone for binding to GR in these cell lines, supporting the view that the induction of EGF binding by MPA in these cells was mediated via the GR. This conclusion was further supported by studies in which addition of the glucocorticoid and progestin antagonist, RU 486, inhibited the effect of ORG 2058 in two cell lines and completely abrogated the effect of dexamethasone in two other lines. Detailed binding studies revealed that the increase in EGF binding was accompanied by an increase in the concentration of EGF-R. This effect was observed when EGF binding was assayed at either 0 degree or 37 degrees C. Further studies demonstrated that the increases in EGF binding following ORG 2058 and dexamethasone treatment were accompanied by increases in EGF-R mRNA levels. Our data illustrate that the binding of EGF by some human breast cancer cells can be regulated by both progestins and glucocorticoids acting via their respective receptors and inducing increases in EGF-R mRNA levels.

Breast Neoplasms↗

Flavivirus infection: essential ultrastructural changes and association of Kunjin virus NS3 protein with microtubules.

Virus-induced vesicles evolved early in the Kunjin virus replication cycle around 9 to 10 h p.i. just before the end of the latent period in infected Vero cells. About 2 h following the appearance of the vesicles, microtubule paracrystals were also formed. These two virus-induced structures seemed interlinked and have essential roles in Kunjin virus replication. A viral protein NS3 was found to be associated with the microtubule component of the cells. When vinblastine sulphate was added to the cells immediately after infection, formation of the paracrystals was delayed by two hours, and the affiliation of NS3 protein was also observed to be rearranged.

Animals↗

Detection of some dengue-2 virus antigens in infected cells using immuno-microscopy.

Immunoelectron microscopy was used to detect the distribution of some dengue-2 virus proteins in infected Vero and Aedes albopictus (C6/36) cells. It was found that the envelope protein (GP 60) was located in clumps on the surface of plasma membrane, and accumulated very little in the infected cytoplasm. However no envelopment of dengue-2 virus nucleocapsids through the plasma membranes was observed. In contrast, the NS 3 (P 67) protein was distributed throughout the whole cytoplasm. No specific association of this protein with the proliferated virus-induced structures was seen. The NS 1 (GP 46) protein showed almost similar distribution as the NS 3 but its quantity appeared to be lower. The NS 3 and NS 1 distribution patterns observed supported the results of immunofluorescent staining but results on E protein were not consistent in both methods.

Aedes↗

Definition of the carboxy termini of the three glycoproteins specified by dengue virus type 2.

The carboxy termini of the three glycoproteins (prM, E, and NS1) specified by dengue virus type 2 (DEN-2) were determined. The glycoproteins were radiolabeled with selected amino acids chosen following analysis of the deduced amino acid sequence of the polyprotein and then digested with carboxypeptidase A. The pattern of release of radioactive amino acids enabled definition of the carboxy termini. In addition, the amino terminus of NS2A was determined by Edman degradation of the radiolabeled protein. The results showed that no amino acids were lost at the carboxy termini of prM, E, and NS1 during their cleavage from the DEN-2 polyprotein. For each glycoprotein, the carboxy terminal amino acid immediately preceded the amino terminal acid of the following polypeptide.

Amino Acid Sequence↗

Effects of Lugol's solution on thyroid function in normals and patients with untreated thyrotoxicosis.

Thirty-eight normal volunteers and 10 patients with untreated thyrotoxicosis were each given 0.5 ml of Lugol's solution daily for 10 days. On days 0, 5, 10, 15 and 20, serum levels of T4, free T4, T3 and TSH (by sensitive immunoradiometric assay) were measured. In normal subjects, the serum concentrations of free T4 declined significantly at day 10 while TSH levels were significantly increased at days 5, 10 and 15. Serum levels of T4 and T3 did not change significantly. All the observed changes took place within the limits of normal ranges for the hormones mentioned. In contrast, in the thyrotoxic subjects, both T4 and T3 were significantly decreased at days 5 and 10, while serum TSH remained below detection limit (0.14 mU/l) throughout the study. Short exposure to excessive iodide in normal subjects affects T4 and T3 release and this effect could be partially overcome by compensatory increase in TSH. In thyrotoxicosis, lack of compensatory increase in TSH results in rapid decreases in T4 and T3 levels. The integrity of the hypothalamo-pituitary-thyroidal axis may be effectively assessed by measuring TSH response to iodide suppression, using a highly sensitive immunoradiometric assay.

Adult↗

Semantic facilitation and translation priming effects in Chinese-English bilinguals.

In two experiments, semantic facilitation and translation priming effects in Chinese-English bilingual speakers were demonstrated with a lexical decision task. A 300-msec stimulus-onset asynchrony (SOA) was used between display of the prime and the target item. Experiment 1 showed that subjects' lexical decision responses were facilitated to a greater extent when primed by a translation equivalent than a semantically related between-language word. In Experiment 2, we found that pictorial, between-language, and within-language primes produced comparable effects of semantic facilitation. These results are in line with the hypothesis that lexical items in different languages and pictures are processed by means of an amodal conceptual system.

Adult↗

Hyperthyroid graves disease--a 5 year retrospective study.

The clinical, biochemical and immunological features of 180 patients with hyperthyroid Graves' disease managed at the Universiti Kebangsaan Malaysia (UKM) Endocrine Clinic from 1983 to 1987 were examined. The prevalence of the disease is highest in Chinese and lowest in Indians. The female: male ratio is 2.8:1. Hypokalaemic periodic paralysis and hypercalcaemia were present in 5.0% and 1.7% of the cases respectively. Pretibial myxoedema was extremely rare. Thyrotropin - binding inhibitory immunoglobulins, anti-thyroglobulin and anti-microsome antibodies were positive in 61.5%, 25.8% and 42.3% of the patients respectively. A eumetabolic state could be achieved in the majority of patients with antithyroid drugs alone. Definitive therapy with subtotal thyroidectomy or radioiodine were needed in 31.3% of cases.

Adolescent↗

Possible involvement of receptors in the entry of Kunjin virus into Vero cells.

The results obtained from electron microscopy, adsorbed and internalised virus assays and immunofluorescence studies supported that the most likely mode of entry of Kunjin virus into Vero cells was by receptor-mediated endocytosis. This was deduced indirectly from the time sequence of events that occurred. Electron microscopy revealed that endocytosis of the virus through coated vesicles had occurred. The adsorbed and internalised virus assay and immunofluorescence studies showed that there were two factors being recycled during endocytosis: the receptor for the virus and clathrin, the protein found on coated pits and vesicles. The study showed that clathrin was recycled first, followed by the receptor.

Adsorption↗