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Biomedical subjects

B L Ng

Publications and source records attributed to B L Ng.

At least 19 recordsLinked to original sources
PubMed2006

Ultra-high resolution array painting facilitates breakpoint sequencing.

OBJECTIVE: To describe a considerably advanced method of array painting, which allows the rapid, ultra-high resolution mapping of translocation breakpoints such that rearrangement junction fragments can be amplified directly and sequenced. METHOD: Ultra-high resolution array painting involves the hybridisation of probes generated by the amplification of small numbers of flow-sorted derivative chromosomes to oligonucleotide arrays designed to tile breakpoint regions at extremely high resolution. RESULTS AND DISCUSSION: How ultra-high resolution array painting of four balanced translocation cases rapidly and efficiently maps breakpoints to a point where junction fragments can be amplified easily and sequenced is demonstrated. With this new development, breakpoints can be mapped using just two array experiments: the first using whole-genome array painting to tiling resolution large insert clone arrays, the second using ultra-high-resolution oligonucleotide arrays targeted to the breakpoint regions. In this way, breakpoints can be mapped and then sequenced in a few weeks.

Adult↗
PubMed2005

Prenatal detection of unbalanced chromosomal rearrangements by array CGH.

BACKGROUND: Karyotype analysis has been the standard method for prenatal cytogenetic diagnosis since the 1970s. Although highly reliable, the major limitation remains the requirement for cell culture, resulting in a delay of as much as 14 days to obtaining test results. Fluorescent in situ hybridisation (FISH) and quantitative fluorescent PCR (QF-PCR) rapidly detect common chromosomal abnormalities but do not provide a genome wide screen for unexpected imbalances. Array comparative genomic hybridisation (CGH) has the potential to combine the speed of DNA analysis with a large capacity to scan for genomic abnormalities. We have developed a genomic microarray of approximately 600 large insert clones designed to detect aneuploidy, known microdeletion syndromes, and large unbalanced chromosomal rearrangements. METHODS: This array was tested alongside an array with an approximate resolution of 1 Mb in a blind study of 30 cultured prenatal and postnatal samples with microscopically confirmed unbalanced rearrangements. RESULTS: At 1 Mb resolution, 22/30 rearrangements were identified, whereas 29/30 aberrations were detected using the custom designed array, owing to the inclusion of specifically chosen clones to give increased resolution at genomic loci clinically implicated in known microdeletion syndromes. Both arrays failed to identify a triploid karyotype. Thirty normal control samples produced no false positive results. CONCLUSIONS: Analysis of 30 uncultured prenatal samples showed that array CGH is capable of detecting aneuploidy in DNA isolated from as little as 1 ml of uncultured amniotic fluid; 29/30 samples were correctly diagnosed, the exception being another case of triploidy. These studies demonstrate the potential for array CGH to replace conventional cytogenetics in the great majority of prenatal diagnosis cases.

Chromosome Aberrations↗
PubMed2005

The RASSF1A isoform of RASSF1 promotes microtubule stability and suppresses tumorigenesis.

The RASSF1A isoform of RASSF1 is frequently inactivated by epigenetic alterations in human cancers, but it remains unclear if and how it acts as a tumor suppressor. RASSF1A overexpression reduces in vitro colony formation and the tumorigenicity of cancer cell lines in vivo. Conversely, RASSF1A knockdown causes multiple mitotic defects that may promote genomic instability. Here, we have used a genetic approach to address the function of RASSF1A as a tumor suppressor in vivo by targeted deletion of Rassf1A in the mouse. Rassf1A null mice were viable and fertile and displayed no pathological abnormalities. Rassf1A null embryonic fibroblasts displayed an increased sensitivity to microtubule depolymerizing agents. No overtly altered cell cycle parameters or aberrations in centrosome number were detected in Rassf1A null fibroblasts. Rassf1A null fibroblasts did not show increased sensitivity to microtubule poisons or DNA-damaging agents and showed no evidence of gross genomic instability, suggesting that cellular responses to genotoxins were unaffected. Rassf1A null mice showed an increased incidence of spontaneous tumorigenesis and decreased survival rate compared with wild-type mice. Irradiated Rassf1A null mice also showed increased tumor susceptibility, particularly to tumors associated with the gastrointestinal tract, compared with wild-type mice. Thus, our results demonstrate that Rassf1A acts as a tumor suppressor gene.

Animals↗
PubMed2004

Applications of combined DNA microarray and chromosome sorting technologies.

The sequencing of the human genome has led to the availability of an extensive mapped clone resource that is ideal for the construction of DNA microarrays. These genomic clone microarrays have largely been used for comparative genomic hybridisation studies of tumours to enable accurate measurement of copy number changes (array-CGH) at increased resolution. We have utilised these microarrays as the target for chromosome painting and reverse chromosome painting to provide a similar improvement in analysis resolution for these studies in a process we have termed array painting. In array painting, chromosomes are flow sorted, fluorescently labelled and hybridised to the microarray. The complete composition and the breakpoints of aberrant chromosomes can be analysed at high resolution in this way with a considerable reduction in time, effort and cytogenetic expertise required for conventional analysis using fluorescence in situ hybridisation. In a similar way, the resolution of cross-species chromosome painting can be improved and we present preliminary observations of the organisation of homologous DNA blocks between the white cheeked gibbon chromosome 14 and human chromosomes 2 and 17.

Cell Line↗
PubMed2004

Corrosion behaviour of high copper dental amalgams.

This study evaluated the corrosion behaviour of two high copper dental amalgam alloys [Dispersalloy (Dentsply-Caulk) and Tytin (Kerr)] in different electrolytes. Amalgam specimens were prepared, coupled to a copper wire, cemented into glass tubes and polished to a 600-grit finish. A corrosion cell was prepared using a carbon counter-electrode, a standard calomel electrode as the reference and amalgam as the working electrode. The alloys were tested in the following mediums at 37 degrees C: (i) artificial saliva based on Fusayama's solution (FS), (ii) artificial saliva with citric acid adjusted to pH 4.0 (FC) and (iii) 1% sodium chloride solution (SC). Corrosion potentials (E(corr)) and corrosion rates (I(corr)) were determined using potentiostatic and impedance spectroscopy methods. Data was subjected to anova/Scheffe's post hoc test at 0.05 significance level. For both alloys, the corrosion potential in FS was significantly greater than in SC. Corrosion potential of Tytin in FS and SC was also significantly greater than in FC. The corrosion rate of Dispersalloy in FC was significantly greater than in FS and SC. For Tytin, corrosion rate in SC was significantly greater than in FS and FC. Although no significant difference in corrosion potential/rate was observed between the alloys when tested in FS, significant differences were observed when electrochemical testing was carried out in FC and SC. The corrosion behaviour of high copper amalgam alloys are both material and environment dependent. Certain food substances may increase the corrosion of high copper amalgams.

Citric Acid↗
PubMed2001

Helicobacter pylori in familial clusters based on antibody profile.

Studies have shown a high prevalence of Helicobacter pylori infection in close communities and that intrafamilial spread during early childhood may be a route of transmission. A total of 72 household members from 21 families were enrolled in this study. Sera from individuals showed 50/72 (69.4%) seropositive for IgG against H. pylori by ELISA. Western blots showed diversity in the protein profiles with molecular masses ranging from approximately 8 to 130 kDa. Cohen's kappa statistical analysis of the blot patterns showed that nine families demonstrated similar profiles (100%), while 4 other families showed varying similarities (17-50%). The results support the hypothesis of intrafamilial transmission of H. pylori. Furthermore, serological studies can be used as an effective approach to determine the familial status in relation to H. pylori infection.

Adolescent↗
PubMed2001

The Alvarado score and acute appendicitis.

INTRODUCTION: This is a retrospective study to assess the accuracy of the Alvarado score in predicting appendicitis for patients with right iliac fossa pain. MATERIALS AND METHODS: This is a retrospective study of consecutive patients with suspected appendicitis. The Alvarado score was computed from the admission notes and correlated with the final outcome of the patient. Patients discharged without surgery were reviewed in the outpatient's clinic to ascertain that they did not need surgery. RESULTS: There were 148 patients in the study and 63 had appendectomies with intention to treat appendicitis. The normal appendicectomy rate was 21%. The number of patients with score 1-4, 5-6, 7-8 and 9-10 were 46, 40, 44 and 18, respectively, while the number of appendicitis in each group were 0, 2 (5%), 30 (68%) and 18 (100%), respectively. The positive and negative predictive values of Alvarado's scores of 7 or more were 77% and 97.6%, respectively. CONCLUSION: Alvarado score is a useful tool in the diagnosis of appendicitis, especially at both ends of the scale.

Acute Disease↗
PubMed1997

Macrophages/microglia as 'sensors' of injury in the pineal gland of rats following a non-penetrative blast.

The pineal gland of adult rats was examined immunohistochemically and electron microscopically following exposure of the animals to a single blast equivalent to 110 kg TNT explosive. The most dramatic feature in rats killed at 7, 14 and 21 days after the blast was the upsurge of a large number of macrophages/microglia intensely immunostained with OX-42, OX-18, OX-6 and ED1 antibodies. These antibodies recognise the complement type three (CR3) receptors, major histocompatibility complex class I and class II (MHC I and MHC II) antigens and monocyte/macrophage antigens. Cell counts in OX-42 immunostained sections showed a two-fold increase at these intervals but returned to normal values at 28 days. The immunolabelled cells appeared extremely hypertrophic after the blast when compared with those in normal rats. In the latter and in rats killed at 28 days after the blast, immunoreactive cells were sparsely distributed. Ultrastructural study confirmed a wider occurrence of perivascular macrophages/microglia after the blast and the cells were laden with massive amounts of phagosomes resembling degenerating pinealocyte processes. It is concluded that the seemingly quiescent macrophages/microglia present normally in pineal gland were activated by the external blast force. The induced changes including the increase in cell numbers and endocytosis, however, were reversible in longer surviving animals.

Animals↗
PubMed1997

Ultrastructural changes of macroglial cells in the rat brain following an exposure to a non-penetrative blast.

The present study examined the ultrastructural changes in astrocytes and oligodendrocytes in rats following an exposure to a non-penetrative blast. At 1 and 7 days after the blast, the astrocytes in the cerebral and cerebellar cortex were hypertrophied; their end-feet associated with the blood vessels were also swollen, bearing sparsely distributed organelles. The above changes were not observed in experimental rats when the survival interval was prolonged. It is concluded from this study that the blast could have disrupted the integrity of the blood-brain barrier resulting in possible abnormal entry of serum-derived substances thereby leading to astrocytic hypertrophy. The reversible nature of the changes is evidenced by the seemingly normal appearance of astrocytes in rats killed at 14, 21 and 28 days after the blast. Oligodendrocytes remained unaffected at various time intervals after the blast.

Animals↗
PubMed1996

Studies of the choroid plexus and its associated epiplexus cells in the lateral ventricles of rats following an exposure to a single non-penetrative blast.

The choroid plexus in rats exhibited ultrastructural changes following a non-penetrative blast. The immunophenotypic features of epiplexus cells associated with the choroid plexus epithelium were also altered. In rats killed at 1 and 7 days after the blast, the intercellular spaces between the epithelial cells were greatly widened, coupled with the massive eruption and possible extrusion of the apical cytoplasm into the ventricular lumen. Associated with these changes was the passage of some monocytes/lymphocytes across the epithelium. The incidence of such a migratory phenomenon was more frequent in rats killed 7 days after the blast. In rats killed 14 days after the blast, the ultrastructural changes of the epithelial cells became less pronounced. At 21 and 28 days after the blast, the ultrastructure of the choroid plexus was comparable to that of normal specimens. The immunoreactivity of epiplexus cells in terms of their cell number and staining intensity with the monoclonal antibodies OX-42, OX-18, OX-6 and ED1 was noticeably augmented at 7 and 14 days after the blast; this, however subsided at 21 and 28 days. It is concluded that the choroid plexus is extremely sensitive to a blast wave as manifested by its structural alterations and the vigorous expression of CR3 receptors and MHC antigens by the epiplexus cells. It is suggested that a possible immune response might have been triggered in the cerebrospinal fluid ventricular system following the blast.

Animals↗
PubMed1995

The response of neurons and microglia to blast injury in the rat brain.

Rats subjected to a single non-penetrative blast were examined for possible neuronal damage and glial reaction by immunohistochemistry and electron microscopy. The most dramatic feature in rats killed between 1 and 14 days after the blast was the widespread response of microglial cells in various parts of the brain in which the cells were hypertrophied and their surface antigens, like complement type three receptors (CR3), were upregulated. The blast wave also induced the vigorous expression of major histocompatibility complex (MHC) class I and II (Ia) antigen. In rats killed 21 days after the blast, the elevated immunoreactivity of microglia had subsided and at 28 days both the microglial external morphology and immunoreactivity were comparable to those of normal animals. In rats killed 4-7 days after the blast, the neurons in the cerebral and cerebellar cortex appeared normal except for the occurrence of some 'darkened' dendrites. The incidence of 'darkened' dendrites was most common in rats killed at day 14 but they were absent at 21 and 28 days. Microglial cells were closely associated with some of the 'darkened' dendrites. Results in this study show that a non-penetrative blast in rats provokes a widespread microglial activation suggesting increased endocytosis and immunological responses. However, it remains uncertain whether such a drastic response was a direct activation of the cells by the blast wave or elicited indirectly by some chemical factors released from the damaged brain tissues.

Animals↗
PubMed1994

Medical selection of military pilots: a Republic of Singapore Air Force perspective.

A comprehensive selection examination in the Republic of Singapore Air Force (RSAF) aims to minimise medical wastage of military pilots who have to function safely in the unnatural environment. Of the 8642 applicants examined, 657 (7.6%) were rejected for non-medical reasons before completion of medical examination. Of the remaining 7778, 58.7% passed the selection examination while 41.3% failed. Ophthalmological (34.3%), anthropometry (23.7%), and ENT (13.7%) conditions were the three major causes for failure (71.7%). Myopia and astigmatism accounted for 57.6% while squints accounted for 22.1% of the ophthalmological conditions. Amongst ENT conditions, 70.8% were for marked vasomotor rhinitis, sinusitis and nasal septum deviation with marked narrowing of one or both nasal passages, while 22.4% were for permanent abnormal hearing threshold shifts above the minimum standards.

Aerospace Medicine↗
PubMed1992

Plasminogen activators and inhibitors in normal late pregnancy, postpartum and in the postnatal period.

Tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasminogen activator inhibitors (PAI) are elevated in late pregnancy with t-PA and u-PA remaining so at 6 weeks postnatal. PAI-2 remains at postpartum but was absent by 6 weeks postnatal unlike PAI activity which was absent at postpartum and returned to nonpregnant level at postnatal. The potential fibrinolytic response to stress is much reduced in pregnancy thus increasing the risk of thromboembolism.

Adult↗
PubMed1992

Concentration of plasminogen activators and inhibitor in the human endometrium at different phases of the menstrual cycle.

The concentrations of tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasminogen activator inhibitor (PAI-1) have been determined in endometrial curettings obtained from 46 subfertile women during proliferative, early or late secretory phases of the menstrual cycle. t-PA activity and antigen concentrations was significantly higher (P < 0.001) in late secretory endometrium than in proliferative or early secretory endometrium. Higher concentrations of PAI-1 antigen (P < 0.05) were also noted in late secretory phase than in proliferative and early secretory endometrium. However, u-PA concentration was not significantly different and no PAI activity could be demonstrated in the menstrual phases studied. Zymography studies confirmed the presence of both t-PA and u-PA in the endometrium. Ovarian hormonal patterns may therefore influence the activity of plasminogen activators especially of t-PA in the endometrium during various phases of the menstrual cycle.

Adult↗
PubMed1991

Plasminogen activators t-PA, u-PA and its inhibitor (PAI) in normal males and females.

We determined the plasma levels of tissue plasminogen activator (t-PA), plasminogen activator inhibitor (PAI) activity and their antigen levels including urokinase plasminogen activator (u-PA) in 33 male and 27 female normal subjects. Males had mean t-PA activity of 0.50 iu/ml which was significantly lower (p less than 0.01) than the females 0.64 iu/ml. Males had higher (p less than 0.001) mean PAI activity (15.5 AU/ml) as compared to females 10.3 AU/ml. The respective mean levels of t-PA and PAI antigen were significantly higher (p less than 0.01) in males (8.1 ng/ml and 17.6 ng/ml) than in females (6.2 ng/ml and 12.1 ng/ml). The mean u-PA level in males was 1.54 ng/ml which was significantly higher (p less than 0.01) than in females with 1.02 ng/ml. In post-venous occlusion studies, females had a greater mean response of 8.6 fold in t-PA activity as compared to males with a mean of 4.5 fold increase. The mean t-PA antigen response in males was 2.0 fold increase as compared to 2.6 fold increase in the females. No significant responses were seen in both sexes in either PAI activity or antigen levels when compared with the resting state. In zymography studies, free t-PA, its inhibitor complexes and u-PA were demonstrated in the euglobulin fractions of stored plasma. This study demonstrates that significant differences in t-PA, u-PA and PAI exist between male and female subjects which should be taken into account when determining their levels in clinical conditions.

Adult↗
PubMed1989

A plasmin generation method for the determination of tissue plasminogen activator (t-PA) activity in blood.

A plasmin generation method to determine tissue plasminogen activator (t-PA) activity in plasma is described. A protein solution of homogenized fibrin was used as a stimulator in the presence of plasminogen and the plasmin generated was measured by the release of para-Nitroanilide (p-NA) from the chromogenic substrate S-2251. Plasmin generation by 5 iu/mL t-PA in the presence of 1 CU/mL of plasminogen and 850 micrograms/mL of fibrin solution reaches a peak at about 5 h incubation whilst in plasma, plasmin generation peaks after about 16 h incubation. The highest t-PA activity in plasma was determined using an assay involving 18 h incubation. In the 21 subjects studied by this method the t-PA activity at rest ranged from 0.34 to 0.92 iu/mL with a mean of 0.57 +/- 0.15 iu/mL of plasma whilst in the post-occlusion state the activity ranged from 1.12 to 18.0 iu/mL, with a mean of 5.25 +/- 4.49 iu/mL of plasma. We also found that subjects who developed petechiae during occlusion had significantly higher t-PA activity both at pre- and post-occlusion when compared with those who did not develop petechiae. The t-PA activity of acid-treated plasma stored at -70 degrees C showed no significant changes in activity after 12 weeks of storage when compared with the t-PA activity of the same plasma tested prior to storage.

Adult↗