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Biomedical subjects

R Ling

Publications and source records attributed to R Ling.

At least 37 records · Page 2Linked to original sources
PubMed2000

Local anaesthetic techniques and pulsatile ocular blood flow.

AIM: To compare pulsatile ocular blood flow (POBF) and intraocular pressure (IOP) between eyes of patients receiving either peribulbar (with and without balloon compression) or subconjunctival local anaesthesia (LA). METHODS: 30 eyes of 30 patients undergoing cataract surgery by phacoemulsification were investigated in a study of parallel group design. Ten patients had peribulbar LA and 10 minutes compression with a Honan's balloon (group A). A further 10 patients who received peribulbar LA alone (group B) acted as controls for the effects of balloon compression. Ten other patients were given subconjunctival LA (group C). POBF and IOP were measured using a modified Langham pneumatonometer. Three measurements were made in each eye, the first recording immediately before LA, the second 1 minute after, and the third 10 minutes after LA. RESULTS: No significant change in POBF or IOP was recorded in eyes receiving subconjunctival LA. In the peribulbar groups (A and B), there was a drop in median POBF of 252 and 138 microl/min respectively 1 minute after LA, which was statistically significant in both groups (p<0. 01). By 10 minutes, POBF tended to return to baseline levels, but remained significantly reduced in group B (p<0.05). In addition, there was a significant (p<0.05) reduction in IOP (mean drop of 4.82 mm Hg) in group A following peribulbar LA with balloon compression. CONCLUSIONS: POBF was significantly reduced after peribulbar LA but was unchanged after subconjunctival LA. Balloon compression reduced IOP and improved POBF following peribulbar LA. The findings may have clinical implications in patients with compromised ocular circulation or significant glaucomatous optic neuropathy.

Aged↗
PubMed2000

Cytotoxic T lymphocyte responses and CTL epitope escape mutation in HBsAg, anti-HBe positive individuals.

BACKGROUND/AIMS: Clearance of hepatitis B virus (HBV) is characterised by a strong cytotoxic T cell response. Persistence of HBV in chronic hepatitis B carriers may be related to failure of this response. The aim of this study was to determine whether HLA class I restricted cytotoxic T lymphocyte (CTL) responses persist in anti-hepatitis B e (HBe) positive / HBV DNA negative individuals, and to correlate the presence of viral CTL epitope mutation with clinical outcome. METHODS: An HLA/HBV dual transfectant model was used to demonstrate these CTL responses in individuals chronically infected with HBV. Subsequently, a known hepatitis B core (HBc) CTL epitope was sequenced in a family of five chronically infected individuals all sharing a HLA allele (HLA-A68.1). RESULTS: Low level HLA class I restricted cytotoxic T cell responses were detected in the peripheral blood of five of eight anti-HBe positive individuals. In the family of HLA-A68.1 positive chronically infected individuals, mutation of the HLA-A68.1 restricted hepatitis B core antigen (HBcAg) CTL epitope STLPETTVVRR was found in all four anti-HBe positive individuals but not in the sole hepatitis B e antigen (HBeAg) positive patient. CONCLUSION: These data are consistent with a continued immune selection pressure on HBV in anti-HBe positive chronically infected individuals with low replicating HBV infection and suggest that mutation of a CTL epitope may be a consequence of the immune response, as opposed to the cause of viral persistence.

Adult↗
PubMed1999

A divergent genotype of hepatitis E virus in Chinese patients with acute hepatitis.

Recent studies have reported and provided nucleotide sequence data from divergent isolates of hepatitis E virus (HEV), including isolates from North America and Africa. Sera were investigated from 29 Chinese patients with a diagnosis of acute hepatitis and who were negative for hepatitis viruses A-E by serology (HEV was excluded by testing for IgG antibody only). To determine whether some patients were infected with HEV but had yet to seroconvert to antibody positivity, RT-PCR was carried out with primers designed within conserved sequences of the HEV open reading frame (ORF) 1 and ORF2 regions. Fifteen patients were found to harbour sequences related to HEV. Analysis of the HEV products revealed that nucleotide sequences from nine of the sera closely matched Burmese-like HEV sequences (more than 92% nucleotide identity across ORF1 and 88% in ORF2). The remaining six HEV isolates were similar to each other but divergent from all other known HEV sequences (74 to 83% nucleotide identity in ORF1 or ORF2). Phylogenetic analysis suggests that the six divergent isolates represent a fourth genotype of HEV, distinct from the previously described Burmese, Mexican and United States variants (genotypes 1, 2 and 3). This novel variant, referred to here as the Chinese genotype (genotype 4), may be responsible for a significant proportion of cases of acute hepatitis in China, as seen by the fact that 40% of the HEV-infected patients in this study were genotype 4 positive.

Acute Disease↗
PubMed1999

Functional analysis of mutations conferring lamivudine resistance on hepatitis B virus.

Two patterns of mutation are commonly observed in the polymerase gene of lamivudine [(-)2'-deoxy-3'-thiacytidine]-resistant hepatitis B virus (HBV). The M539I substitution in the conserved YMDD motif occurs independently of other changes, whereas the M539V substitution is associated with an additional upstream change (L515M). These mutations were introduced into a common background and their effects on HBV DNA replication and lamivudine resistance studied. The L515M and M539V mutations provided only partial resistance while the M539I mutation conferred a high degree of lamivudine resistance. The combination of the L515M and M539V mutations gave an intermediate level of replication competence, compared with either mutation alone, and increased resistance to lamivudine. This probably accounts for these two mutations always being observed together. The M539I mutation reduced replication competence.

Amino Acid Sequence↗
PubMed1999

Incomplete cement mantles in the sagittal femoral plane: an anatomical explanation.

The influence of operative technique on the formation of incomplete cement mantles in the sagittal plane has been rarely considered in the literature. In this article, we discuss the influence of the anatomy of the proximal femur on the formation of incomplete cement mantles and discuss how their incidence can be reduced by correct component positioning.

Cementation↗
PubMed1998

HBV core promoter mutations prevail in patients with hepatocellular carcinoma from Guangxi, China.

The development of primary liver cancer frequently is associated with persistent HBV infection, and tumours may arise in individuals who are anti-HBe positive. However, it is unclear whether viruses with an HBeAg-negative phenotype are associated with tumour development or are selected, during seroconversion, after chromosomal integration of wild-type viral DNA. In order to investigate the temporal evolution of the HBV genome in such individuals, the polymerase chain reaction was used to amplify HBV DNA from tumour tissue and serum of 14 patients from Guangxi, China with hepatocellular carcinoma. Comparison of nucleotide and amino acid sequences of the precore and proximal core region of HBV from the two sites in each patient produced evidence of divergence following integration in the tumour, but in most cases, HBeAg-negativity could not be explained by precore mutations. Sequences from the core promoter region were therefore examined and mutations were found in the majority, which are believed to upregulate transcription of the core (and pregenomic) RNA but to downregulate precore mRNA. To determine whether this finding merely reflected sequence variation among geographical isolates of HBV, the same region of HBV DNA from asymptomatic controls was sequenced and these mutations were found to be rare. We hypothesise that HBV with the core promoter mutations replicates at higher levels than the wild type, with the consequence that more integrations occur into the hepatocyte chromosomes during the early stages of infection. These hepatocytes may expand clonally and be targets for further mutagenic events leading to tumour development.

Adult↗
PubMed1998

Engineering in vivo instability of firefly luciferase and Escherichia coli beta-glucuronidase in higher plants using recognition elements from the ubiquitin pathway.

The ubiquitin pathway targets proteins for degradation through the post-translational covalent attachment of the 76 amino acid protein ubiquitin to epsilon-amino lysyl groups on substrate proteins. Two instability determinants recognized by the ubiquitin pathway in Saccharomyces cerevisiae have been identified. One is described by the N-end rule and requires specific destabilizing residues at the substrate protein N-termini along with a proximal lysyl residue for ubiquitin conjugation. The second is a linear uncleavable N-terminal ubiquitin moiety. The ability of these two determinants to function in higher plants was investigated in tobacco protoplast transient transfection assays using DNA encoding variants of well characterized reporter enzymes as substrates: firefly luciferase that is localized to peroxisomes (pxLUC), a cytosolic version of LUC (cLUC), and Escherichia coli beta-glucuronidase (GUS). cLUC with phenylalanine encoded at its mature N-terminus was 10-fold less abundant than cLUC with methionine at its mature N-terminus. GUS with phenylalanine encoded at its mature N-terminus was 3-fold less abundant than GUS with methionine at its mature N-terminus. The presence of a uncleavable N-terminal ubiquitin fusion resulted in 50-fold lower protein accumulation of cLUC, but had no effect on GUS. Both instability determinants had a much larger effect on cLUC than on pxLUC, suggesting that these degradation signals are either unrecognized or poorly recognized in the peroxisomes.

Amino Acid Sequence↗
PubMed1998

Energy reflectance in the ear canal can exceed unity near spontaneous otoacoustic emission frequencies.

There is some controversy in the literature over whether the so-called "active mechanism" or "cochlear amplifier" is actually a power amplifier that can produce an output signal with more power than its input, or whether it simply minimizes dissipative losses within the cochlea without providing an actual power gain greater than unity. A corollary of this controversy is whether spontaneous otoacoustic emissions (SOAEs) represent the output of a nonlinear oscillator mechanism, i.e., a power amplifier which can produce an oscillatory output signal in the absence of an input oscillatory signal, or whether they represent the output of a noise-driven, passive, nonlinear system. This paper describes measurements of energy reflectance, and acoustic impedance in the ear canals of human subjects with strong SOAEs. The reflectance, and the resistive and reactive parts of the acoustic impedance, all show a frequency fine structure which correlates with SOAE frequencies, and which becomes more pronounced at low stimulus levels. In some ears at some SOAE frequencies, energy reflectance exceeds unity, and correspondingly, acoustic resistance is negative. This result demonstrates that there is a power gain at these frequencies: The power reflected from the cochlea to the ear canal exceeds the power incident. It is also consistent with the theory that these SOAEs are produced by a nonlinear oscillator mechanism in the cochlea.

Ear Canal↗
PubMed1998

Double-evoked otoacoustic emissions. II. Intermittent noise rejection, calibration and ear-canal measurements.

Measurements of double-click-evoked otoacoustics emissions (2CEOAEs) and double-chirp distortion products (2ChDPs) are reported for normal-hearing adults based upon theory presented in an earlier report [Keefe, J. Acoust, Soc. Am. 103, 3489-3498 (1998)]. The nonlinear acoustic response of a probe assembly used in ear-canal measurements in tested in a calibration cavity to compare the double-evoked (2E) technique with existing OAE techniques. The 2E technique reduces the peak distortion by approximately 30 dB relative to existing click-evoked techniques. The 2E subtraction of click responses is partially analogous to current techniques in that the linear response is eliminated, but differs in that high-frequency measurements are improved by eliminating time gating of the cochlear response, and low-frequency measurements are improved by reducing probe distortion, especially when two acoustic sources are used. Because time gating is eliminated, it is straightforward to measure the onset of a click-evoked OAE. The nonlinear coherence function is used to measure the nonlinear distortion signal-to-noise ratio (DNR) for the 2ChDPs and 2CEOAEs. The DNR is typically 20-30 dB. An intermittent noise rejection technique is implemented in real time that compares a currently acquired ear-canal response with a stored response. Dissimilar responses indicate the presence of intermittent noise, and the noise-contaminated responses are thereby discarded before ensemble averaging.

Acoustic Stimulation↗
PubMed1998

White-centred retinal haemorrhages (Roth spots).

Roth spots (white-centred retinal haemorrhages) were classically described as septic emboli lodged in the retina of patients with subacute bacterial endocarditis. Indeed many have considered Roth spots pathognomonic for this condition. More recent histological evidence suggests, however, that they are not foci of bacterial abscess. Instead, they are nonspecific and may be found in many other diseases. A review of the histology and the pathogenesis of these white-centred haemorrhages will be provided, along with the work-up of the differential diagnosis.

Endocarditis, Subacute Bacterial↗
PubMed1998

A retrospective epidemiology study of contact eczema among the elderly attending a tertiary dermatology referral centre in Singapore.

AIM: The aim of this study was to examine the epidemiology of contact eczema in the elderly patients attending a skin clinic in Singapore. METHODOLOGY: This is a retrospective study on patients attending our skin clinic at the National Skin Centre. All patients older than 49 years old attending our Contact and Occupational Dermatoses Clinic between 1990 and 1993 were included in the study. The prevalence of contact eczema among different age groups (50-59 years, 60-69 years, and 70-79 years) were compared. Diagnosis of contact dermatitis was based on the clinical history, physical examination and relevant patch test reactions. Demographic data of the patients were collected and analysed. Chi-square test was used for statistical analysis. RESULTS: Two hundred and one patients who were older than 49 years old, were included in the study. There was no significant difference in the prevalence of personal history of atopy among the 3 age groups. There was no significant difference in the racial distribution of the 3 age groups. The proportion of patients who were occupationally active decreased with age (50% vs 23% vs 0% respectively). There was no significant difference in the proportion of patients with positive patch test reactions in the 3 age groups although the proportion appears to increase with age (68% vs 75% vs 75% respectively). Similarly, the proportion of patients with allergic contact dermatitis was slightly higher in the older age group although the difference was not statistically significant among the 3 age groups (32% vs 45% vs 39% respectively). The face (54%) and hands (31%) were the commonest sites of eczema in the younger age group (50-59 years age group) whereas the lower limbs (41%) and the upper limbs (33.3%) were the commonest sites of eczema in the older patients (70-79 years age group). Nickel allergy was significantly more common in the younger age group (23% vs 17% vs 14% respectively). Medicament allergy (clioquinol-mix) and medicament related allergens e.g. balsam of Peru and fragrance-mix and para-phenylenediamine were slightly more prevalent in the older patients. CONCLUSIONS: The findings appear to indicate that the prevalence of allergic contact dermatitis is higher in older patients and that contact dermatitis tends to occur on the extremities in older patients than younger patients.

Aged↗
PubMed1997

Production of hepatitis B virus covalently closed circular DNA in transfected cells is independent of surface antigen synthesis.

Covalently closed circular DNA (cccDNA) is the first hepatitis B virus (HBV) DNA replicative intermediate formed from the genomic DNA and serves as the template for synthesis of viral mRNA and pregenomic RNA. It also appears to be produced by intracellular recycling of relaxed circular DNA intermediates. Here, we report that none of the forms of HBV surface antigen affect this intracellular recycling of HBV DNA. Elimination of the initiation codons for the large and middle surface proteins did not affect the detection of surface antigen in culture supernatants. In contrast, detection of surface antigen was eliminated by the removal of, or the introduction of two stop codons downstream of, the initiation codon of the small (major) surface antigen. None of the mutations affected the production, in transfected HepG2 cells, of HBV DNA replicative intermediates, including cccDNA.

Cell Line↗
PubMed1996

Selection of mutations in the hepatitis B virus polymerase during therapy of transplant recipients with lamivudine.

We describe mutations in the hepatitis B virus (HBV) polymerase gene in viruses which reactivated in two patients during therapy with -2'-deoxy-3'-thiacytidine, or lamivudine (3TC), and following orthotopic liver transplantation for chronic hepatitis B. Virus resistance to 3TC is associated with mutations which lead to amino acid substitutions in the highly conserved tyr-met-asp-asp (YMDD) motif, part of the active site of the polymerase, and which parallel those seen in resistant human immunodeficiency virus (HIV). Substitutions of valine and isoleucine for methionine were found in the two cases. The significance of single secondary mutations, which differ between viruses from the two patients, remains to be determined. Thus, viral resistance to lamivudine of hepatitis B virus mimics that of HIV and can occur in the setting of immunosuppression after liver transplantations.

Adult↗
PubMed1996

Sequence of the nucleocapsid protein gene of subgroup A and B avian pneumoviruses.

The nucleocapsid protein (N) gene of two subgroup A and one subgroup B strains of avian pneumovirus has been cloned and sequenced. The gene of all three isolates comprised 1197 nucleotides (nt), which formed a single major open reading frame, potentially encoding a protein of 391 amino acid residues. The N gene of the two subgroup A isolates differed by only 1 nt but differed by 282 (24%) nt and 35 (11%) amino acids from the B isolate. The predicted protein was identical in length to that of human, bovine and ovine respiratory syncytial viruses, the amino acid identity being approximately 41% overall but with some regions of identity > 90%.

Amino Acid Sequence↗
PubMed1995

Sequence and in vitro expression of the phosphoprotein gene of avian pneumovirus.

The phosphoprotein (P) gene of two subgroup A strains of avian pneumovirus comprised 855 nucleotides containing only one substantial open reading frame encoding a protein of 278 amino acids, with a predicted M(r) of 30,323. In vitro translation of P mRNA in a wheat germ system resulted in the synthesis of two polypeptides of M(r) 35,000. Comparison of the deduced P protein sequence with that of the known mammalian pneumoviruses revealed overall amino acid identities ranging from 31 to 34.5%, suggesting a distant relationship. However, there was a much higher identity (63.2-68.4%) in a region of 57 residues, which included a heptad repeat sequence.

Amino Acid Sequence↗
PubMed1995

Case report: malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia.

Malignant fibrous histiocytoma commonly occurs spontaneously. In some cases it follows previous therapeutic or incidental irradiation, or miscellaneous pre-existing osseous conditions. Recently, it has been associated with total hip arthroplasty. We report a case of malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia and review the relevant literature.

Bone Neoplasms↗
PubMed1994

Use of single-cysteine mutants to probe the location of the disulfide bond in LamB protein from Escherichia coli.

The two cysteine residues of the LamB protein of Escherichia coli outer membrane have been shown to form an intrasubunit disulfide whose location differs greatly in the two current topology models of the LamB protein. This study probes the location of the disulfide by examining conditions for intersubunit disulfide formation in single-cysteine mutants of LamB protein. Formation of an intersubunit bond in the purified mutant proteins, which resulted in a disulfide-linked dimer, only occurred after heat treatment, suggesting the disulfide is not exposed on the surface in the native protein.

Bacterial Outer Membrane Proteins↗