Biomedical subjects

David Finkelstein

Publications and source records attributed to David Finkelstein.

6 recordsLinked to original sources

PubMed2005

Differential expression of the GABA transporters GAT-1 and GAT-3 in brains of rats, cats, monkeys and humans.

The homeostasis of GABA is critical to normal brain function. Extracellular levels of GABA are regulated mainly by plasmalemmal gamma-aminobutyric acid (GABA) transporters. Whereas the expression of GABA transporters has been extensively studied in rodents, validation of this data in other species, including humans, has been limited. As this information is crucial for our understanding of therapeutic options in human diseases such as epilepsy, we have compared, by immunocytochemistry, the distributions of the GABA transporters GAT-1 and GAT-3 in rats, cats, monkeys and humans. We demonstrate subtle differences between the results reported in the literature and our results, such as the predominance of GAT-1 labelling in neurons rather than astrocytes in the rat cortex. We note that the optimal localisation of GAT-1 in cats, monkeys and humans requires the use of an antibody against the human sequence carboxyl terminal region of GAT-1 rather than against the slightly different rat sequence. We demonstrate that GAT-3 is localised mainly to astrocytes in hindbrain and midbrain regions of rat brains. However, in species such as cats, monkeys and humans, additional strong immunolabelling of oligodendrocytes has also been observed. We suggest that differences in GAT distribution, especially the expression of GAT-3 by oligodendrocytes in humans, must be accommodated in extrapolating rodent models of GABA homeostasis to humans.

Animals↗

PubMed2005

Dopaminergic innervation of the human striatum in Parkinson's disease.

In Parkinson's disease (PD), dopaminergic input to the caudate nucleus and a band of putaminal tissue abutting the external globus pallidus seems well preserved on immunohistochemical staining for the dopamine transporter. Counting of dopaminergic terminals showed that terminal density in these regions in PD was the same as that in controls, which indicates that input is truly preserved and not a consequence of a compensatory upregulation of metabolism in a reduced pool of surviving terminals. When the branching pattern of dopaminergic axons coursing through the globus pallidus was examined, we found no evidence for increased axonal sprouting in PD that might have contributed to preservation of dopaminergic input to the putamen or caudate nucleus. Although terminal counting indicated that anatomic input was preserved to parts of the striatum, dopamine uptake site density in these regions was reduced significantly. This suggests that the impact of disease in these areas is more profound than was thought previously.

Aged↗

PubMed2005

The effect of non-insulin dependent diabetes mellitus on uncontrolled hemorrhage in a rodent model.

OBJECTIVE: Diabetes mellitus (DM) is an independent risk factor for higher morbidity and mortality rates from trauma. We tested the null hypothesis that there would be no difference in the hemorrhage volumes and hemodynamic responses to uncontrolled hemorrhage between Zucker Diabetic Fat rats (ZDF) and euglycemic Sprague-Dawley rats (SD). METHODS: Twenty-four adult male rats (12 ZDF and 12 SD) were anesthetized with althesin via the intraperitoneal route. The femoral artery was cannulated by cut-down to monitor the heart rate (HR), mean arterial pressure (MAP) and to obtain blood samples for blood gas analysis. Twelve rats (6 ZDF and 6 SD) underwent uncontrolled hemorrhage by 50% tail amputation. Twelve rats (6 ZDF and 6 SD) served as non-hemorrhage controls. The HR, MAP, lactate (LAC), glucose levels (GL) and cumulative hemorrhage volume (CHV) were measured pre-hemorrhage and then every 15 min post-hemorrhage for 120 min. Data were reported as mean+/-S.E.M. Group comparisons were analyzed by ANOVA with repeated values; post hoc testing by Bonferroni (all tests were two-tailed, alpha = 0.05). RESULTS: Pre-hemorrhage the SD and ZDF were evenly matched for LAC, HR and MAP. CHV (cm3/100 g) was significantly (p = 0.008) greater in the ZDF (1.49+/-0.12) as compared to the SD (0.38+/-0.11). The ZDF had significantly (p < 0.001) higher LAC (7.96+/-0.61 mmol/L) than the SD (2.0+/-0.41 mmol/L). CONCLUSION: DM as compared to non-DM rats suffered a greater blood loss with a more severe lactic acidosis after a comparable uncontrolled vascular injury.

Analysis of Variance↗

PubMed2004

Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays.

BACKGROUND: Primary human tissues are an invaluable widely used tool for discovery of gene expression patterns which characterize disease states. Tissue processing methods remain unstandardized, leading to unanswered concerns of how to best store collected tissues and maintain reproducibility between laboratories. We subdivided uterine myometrial tissue specimens and stored split aliquots using the most common tissue processing methods (fresh, frozen, RNALater) before comparing quantitative RNA expression profiles on the Affymetrix U133 human expression array. Split samples and inclusion of duplicates within each processing group allowed us to undertake a formal genome-wide analysis comparing the magnitude of result variation contributed by sample source (different patients), processing protocol (fresh vs. frozen vs. 24 or 72 hours RNALater), and random background (duplicates). The dataset was randomly permuted to define a baseline pattern of ANOVA test statistic values against which the observed results could be interpreted. RESULTS: 14,639 of 22,283 genes were expressed in at least one sample. Patient subjects provided the greatest sources of variation in the mixed model ANOVA, with replicates and processing method the least. The magnitude of variation conferred by processing method (24 hours RNALater vs 72 hours RNALater vs. fresh vs frozen) was similar to the variability seen within replicates. Subset analysis of the test statistic according to gene functional class showed that the frequency of "outlier" ANOVA results within each functional class is overall no greater than expected by chance. CONCLUSIONS: Ambient storage of tissues for 24 or 72 hours in RNALater did not contribute any systematic shift in quantitative RNA expression results relative to the alternatives of fresh or frozen tissue. This nontoxic preservative enables decentralized tissue collection for expression array analysis without a requirement for specialized equipment.

Analysis of Variance↗

PubMed2004

A knowledge-based clustering algorithm driven by Gene Ontology.

We have developed an algorithm for inferring the degree of similarity between genes by using the graph-based structure of Gene Ontology (GO). We applied this knowledge-based similarity metric to a clique-finding algorithm for detecting sets of related genes with biological classifications. We also combined it with an expression-based distance metric to produce a co-cluster analysis, which accentuates genes with both similar expression profiles and similar biological characteristics and identifies gene clusters that are more stable and biologically meaningful. These algorithms are demonstrated in the analysis of MPRO cell differentiation time series experiments.

Algorithms↗

PubMed2002

Microarray data quality analysis: lessons from the AFGC project. Arabidopsis Functional Genomics Consortium.

Genome-wide expression profiling with DNA microarrays has and will provide a great deal of data to the plant scientific community. However, reliability concerns have required the development data quality tests for common systematic biases. Fortunately, most large-scale systematic biases are detectable and some are correctable by normalization. Technical replication experiments and statistical surveys indicate that these biases vary widely in severity and appearance. As a result, no single normalization or correction method currently available is able to address all the issues. However, careful sequence selection, array design, experimental design and experimental annotation can substantially improve the quality and biological of microarray data. In this review, we discuss these issues with reference to examples from the Arabidopsis Functional Genomics Consortium (AFGC) microarray project.

Arabidopsis↗