Biomedical subjects

Lynette Lim

Publications and source records attributed to Lynette Lim.

6 recordsLinked to original sources

PubMed2004

A method for analysis of gene expression in isolated mouse photoreceptor and Müller cells.

PURPOSE: Molecular analysis of complex phenomena, such as selective death of photoreceptors and their rescue by neuro-protective agents, has been hindered by limitations of techniques for investigating gene expression in individual cells within a heterogeneous tissue such as the retina. The purpose of this study was to develop methods to assess gene expression in single retinal cells. METHODS: Individual cells from papain-dissociated mouse retinae were captured with micropipettes and identified by morphology and by immunocytochemistry. Single cell cDNA libraries were generated by poly-d(T)-primed reverse transcription, poly-d(A) tailing of first strand cDNA, and en masse PCR-amplification using a custom made oligo-d(T). PCR was used to investigate gene expression in cDNAs from individual cells. RESULTS: Dissociated rod and Müller glia cells maintained their morphology, which correlated with their immunocytochemical properties. RPE cells were recognized by their pigmentation. With the exception of bipolar cells, non-photoreceptor neurons were only identifiable by immunocytochemistry. Abundant cDNA could be synthesized from each individual cell. Cell-specific "markers" were detected by PCR almost exclusively in the predicted cell types. The expression of neurotrophic factor receptors was consistent with previous biological studies. CONCLUSIONS: These studies establish a method to compare, investigate, and analyze gene expression in individual cells of the retina.

Animals↗

PubMed2004

The MacNew Heart Disease health-related quality of life instrument: a summary.

BACKGROUND: The measurement of health, the effects of disease, and the impact of health care include not only an indication of changes in disease frequency and severity but also an estimate of patients' perception of health status before and after treatment. One of the more important developments in health care in the past decade may be the recognition that the patient's perspective is as legitimate and valid as the clinician's in monitoring health care outcomes. This has lead to the development of instruments to quantify the patients' perception of their health status before and after treatment. METHODS: We review evidence supporting the measurement properties of the MacNew Heart Disease Health-related Quality of Life [MacNew] Questionnaire which was designed to evaluate how daily activities and physical, emotional, and social functioning are affected by coronary heart disease and its treatment. RESULTS: Reliability was demonstrated by using internal consistency and the intraclass correlation coefficients for the three domains in the Dutch, English, Farsi, German, and Spanish versions of the MacNew. With internal consistency and intraclass correlation coefficients =>0.73, reliability is high. Validity of the MacNew was examined with factor analysis and three core underlying factors, physical, emotional, and social, were identified, explaining 63.0 - 66.5% of the observed variance and replicated in the translations with psychometric data. Construct validity of the MacNew was further demonstrated by extensive substantiation of the logical relationships, defined a priori, between items and other comparison tools. The MacNew is responsive and sensitive to changes in HRQL following various interventions for patients with heart disease with 11 of 13 effect size statistics >0.80. Taking an average of 10 minutes or less to complete, the respondent-burden for the MacNew is low and its acceptability is demonstrated by response rates of over 90%. Normative data are available for patients with myocardial infarction, angina, and heart failure in the English version. CONCLUSION: The MacNew may be a valuable tool for assessing and evaluating health related quality of life in patients with heart disease.

Attitude to Health↗

PubMed2004

Frames of reference in mobile augmented reality displays.

In 3 experiments, the authors investigated spatial updating in augmented reality environments. Participants learned locations of virtual objects on the physical floor. They were turned to appropriate facing directions while blindfolded before making pointing judgments (e.g., "Imagine you are facing X. Point to Y"). Experiments manipulated the angular difference between the learning heading and the imagined heading and between the actual heading and the imagined heading. The effect of actual-imagined on pointing latency was observed for naive users but not for users with brief training or instructions concerning the fact that objects can move with body movements. The results indicated that naive users used an environment-stabilized reference frame to access information arrays, but with experience and instruction the nature of the representation changed from an environment stabilized to a body stabilized reference frame.

Environment↗

PubMed2003

The use of mutually exclusive categories for atopic sensitization: a contrasting effect for family size on house dust mite sensitization compared with ryegrass sensitization.

Our aim was to examine the relative importance of family size on sensitization to two different allergens: ryegrass and house dust mite (HDM), using a mutually exclusive classification for allergen-specific sensitization. An 8-year follow-up birth cohort study of children born between 1988-89 was conducted. The follow-up sample consisted of 498 children residing in Northern Tasmania in 1997 (84% of eligible). Outcome measures included skin prick test (SPT) reaction to nine aeroallergens and parental questionnaire. Family size was defined as sibling number in 1997. Children with a positive SPT to either Der p or Der f house dust mite but not ryegrass were classified as HDM-exclusive (n = 84). Children with a positive SPT to ryegrass but not HDM were classified as ryegrass-exclusive (n = 43). Family size was associated with reduced ryegrass-exclusive sensitization [AOR 0.57 (0.39, 0.84) per increase in sibling number] but not HDM-exclusive sensitization [AOR 0.97 (0.77,1.23)]. The difference in the family size effect on these sensitization outcomes was significant (p = 0.02). Similarly, family size tended to be associated with reduced asthma among ryegrass-exclusive sensitized children [AOR 0.45 (0.18,1.12)] but not HDM-exclusive sensitized children [(AOR 1.46(0.80-2.65)]. Large family size was strongly associated with reduced sensitization for ryegrass allergens but not HDM allergens using mutually exclusive sensitization categories. If this difference is confirmed in other studies, the contrasting effect of family size may reflect differences between these allergens with regard to level or timing of early life exposure, differences in allergen -specific potentiation for sensitization or unidentified confounding. The use of mutually exclusive categories for allergen sensitization will assist future work on child atopic disease.

Allergens↗

PubMed2002

Adenylate kinase AK1 knockout heart: energetics and functional performance under ischemia-reperfusion.

Deletion of the major adenylate kinase AK1 isoform, which catalyzes adenine nucleotide exchange, disrupts cellular energetic economy and compromises metabolic signal transduction. However, the consequences of deleting the AK1 gene on cardiac energetic dynamics and performance in the setting of ischemia-reperfusion have not been determined. Here, at the onset of ischemia, AK1 knockout mice hearts displayed accelerated loss of contractile force compared with wild-type controls, indicating reduced tolerance to ischemic stress. On reperfusion, AK1 knockout hearts demonstrated reduced nucleotide salvage, resulting in lower ATP, GTP, ADP, and GDP levels and an altered metabolic steady state associated with diminished ATP-to-P(i) and creatine phosphate-to-P(i) ratios. Postischemic AK1 knockout hearts maintained approximately 40% of beta-phosphoryl turnover, suggesting increased phosphotransfer flux through remaining adenylate kinase isoforms. This was associated with sustained creatine kinase flux and elevated cellular glucose-6-phosphate levels as the cellular energetic system adapted to deletion of AK1. Such metabolic rearrangements, along with sustained ATP-to-ADP ratio and total ATP turnover rate, maintained postischemic contractile recovery of AK1 knockout hearts at wild-type levels. Thus deletion of the AK1 gene reveals that adenylate kinase phosphotransfer supports myocardial function on initiation of ischemic stress and safeguards intracellular nucleotide pools in postischemic recovery.

Adenosine Triphosphate↗

PubMedDate not supplied

Mapping hypoxia-induced bioenergetic rearrangements and metabolic signaling by 18O-assisted 31P NMR and 1H NMR spectroscopy.

Brief hypoxia or ischemia perturbs energy metabolism inducing paradoxically a stress-tolerant state, yet metabolic signals that trigger cytoprotection remain poorly understood. To evaluate bioenergetic rearrangements, control and hypoxic hearts were analyzed with 18O-assisted 31P NMR and 1H NMR spectroscopy. The 18O-induced isotope shift in the 31P NMR spectrum of CrP, betaADP and betaATP was used to quantify phosphotransfer fluxes through creatine kinase and adenylate kinase. This analysis was supplemented with determination of energetically relevant metabolites in the phosphomonoester (PME) region of 31P NMR spectra, and in both aromatic and aliphatic regions of 1H NMR spectra. In control conditions, creatine kinase was the major phosphotransfer pathway processing high-energy phosphoryls between sites of ATP consumption and ATP production. In hypoxia, creatine kinase flux was dramatically reduced with a compensatory increase in adenylate kinase flux, which supported heart energetics by regenerating and transferring beta- and gamma-phosphoryls of ATP. Activation of adenylate kinase led to a build-up of AMP, IMP and adenosine, molecules involved in cardioprotective signaling. 31P and 1H NMR spectral analysis further revealed NADH and H+ scavenging by alpha-glycerophosphate dehydrogenase (alphaGPDH) and lactate dehydrogenase contributing to maintained glycolysis under hypoxia. Hypoxia-induced accumulation of alpha-glycerophosphate and nucleoside 5'-monophosphates, through alphaGPDH and adenylate kinase reactions, respectively, was mapped within the increased PME signal in the 31P NMR spectrum. Thus, 18O-assisted 31P NMR combined with 1H NMR provide a powerful approach in capturing rearrangements in cardiac bioenergetics, and associated metabolic signaling that underlie the cardiac adaptive response to stress.

Animals↗