ABSTRACT
During operations, neurosurgeons usually perform multiple temporary occlusions of parental artery, possibly resulting in the neuronal damage. It is generally thought that neuronal damage by cerebral ischemia is associated with extracellular concentrations of the excitatory amino acids. In this study, we measured the dynamics of extracellular glutamate release in 11 vessel occlusion (VO) model to compare between single occlusion and repeated transient occlusions within short interval. Changes in cerebral blood flow were monitored by laser-Doppler flowmetry simultaneously with cortical glutamate level measured by amperometric biosensor. From real time monitoring of glutamate release in 11 VO model, the change of extracellular glutamate level in repeated transient occlusion group was smaller than that of single occlusion group, and the onset time of glutamate release in the second ischemic episode of repeated occlusion group was delayed compared to the first ischemic episode which was similar to that of single 10 min ischemic episode. These results suggested that repeated transient occlusion induces less glutamate release from neuronal cell than single occlusion, and the delayed onset time of glutamate release is attributed to endogeneous protective mechanism of ischemic tolerance.
Subject(s)
Humans , Arteries , Biosensing Techniques , Brain Ischemia , Excitatory Amino Acids , Glutamic Acid , Glycosaminoglycans , Ischemia , Laser-Doppler Flowmetry , Neurons , ParentsABSTRACT
We investigated the effect by the chemical fixative on human fibroblast cells (HFCs) in order to make nano-scale images using by the atomic force microscopy (AFM). The cell fixation needed to be optimized as prerequisite step for the preparation before analysis. AFM imaging after optimal wet fixation can provide practical, simple and fast technique for scanning living cells. In this study, AFM images - topography and amplitude - and the optic images of HFCs which were fixed with phosphate buffered saline (PBS), 2:1 ethanol:acetic acid, 4% glutaraldehyde and 37% formaldehyde were compared respectively. The final effect by washing with PBS or distilled water (D.W.) was examined after 4% glutaraldehyde fixation. To determine the optimal fixation method for HFCs, we performed quantitative and qualitative analysis by the height profile, the presence of artifacts and the morphology of well-conserved fibroblastic topography image by AFM. From AFM image which showed fibroblastic cellular morphology and differential height value of cytoplasm (670+/-47 nm, n=10) and nucleus (847+/-32 nm, n=10) in HFCs, we proposed that wet fixation by 4% glutaraldehyde, followed by final washing with PBS, could be the most suitable preparation for AFM imaging of HFCs, which enable us to approach easily on living cells with the least shrinkage.
Subject(s)
Humans , Artifacts , Cytoplasm , Fibroblasts , Formaldehyde , Glutaral , Microscopy, Atomic Force , WaterABSTRACT
Numerous studies have reported that genes with similar expression patterns are co-regulated. From gene expression data, we have assumed that genes having similar expression pattern would share similar transcription factor binding sites (TFBSs). These function as the binding regions for transcription factors (TFs) and thereby regulate gene expression. In this context, various analysis tools have been developed. However, they have shortcomings in the combined analysis of expression patterns and significant TFBSs and in the functional analysis of target genes of significantly overrepresented putative regulators. In this study, we present a web-based A Functional Clustering Analysis Tool for Predicted Transcription Regulatory Elements and Gene Ontology Terms (FCAnalyzer). This system integrates microarray clustering data with similar expression patterns, and TFBS data in each cluster. FCAnalyzer is designed to perform two independent clustering procedures. The first process clusters gene expression profiles using the K-means clustering method, and the second process clusters predicted TFBSs in the upstream region of previously clustered genes using the hierarchical biclustering method for simultaneous grouping of genes and samples. This system offers retrieved information for predicted TFBSs in each cluster using Match(TM) in the TRANSFAC database. We used gene ontology term analysis for functional annotation of genes in the same cluster. We also provide the user with a combinatorial TFBS analysis of TFBS pairs. The enrichment of TFBS analysis and GO term analysis is statistically by the calculation of P values based on Fisher's exact test, hypergeometric distribution and Bonferroni correction. FCAnalyzer is a web-based, user-friendly functional clustering analysis system that facilitates the transcriptional regulatory analysis of co-expressed genes. This system presents the analyses of clustered genes, significant TFBSs, significantly enriched TFBS combinations, their target genes and TFBS-TF pairs.
Subject(s)
Binding Sites , Cluster Analysis , Gene Expression , Gene Ontology , Transcription Factors , TranscriptomeABSTRACT
We carried out a validation-calibration study of the food frequency questionnaire (FFQ) that we had previously developed for a community-based cohort of the Korean Genome and Health Study of the Korea National Genome Research Institute. We have collected a total of 254 3-day diet records (DRs) from 400 subjects, 200 each randomly selected from the two study cohorts of Ansung and Ansan. FFQ was administered at the time of cohort recruitment in 2001, and DRs were collected during a two month period from January through February of 2002. The mean age was 52.2 years. Farming for men and housewife for women were the most common occupations. The majority of the subjects had undergone 6~12 years of education. The general characteristics including demographic and other data were not different from the total cohort subjects. Absolute levels of consumed nutrients including total energy (energy), protein, fat, carbohydrate, calcium, phosphorus, sodium, potassium, iron, retinol, carotene, vitamin A, thiamin, riboflavin, niacin and vitamin C were compared. The average of energy intake was not significantly different between the data collected by the 2 methods. However, consumptions of protein and fat were higher in data of DRs, whereas that of carbohydrate was higher in FFQ data. Significant correlation of each nutrient consumption between the data sets was observed (p <0.05) except in the case of iron, while the average correlation coefficient between them was 0.22 ranging from 0.33 for energy to 0.11 for iron. The results of cross classification by quantile for exact classification ranged from 25.2% (carotene) to 35.0% (phosphorus), and from 64.6% (vitamin A) to 76.4% (retinol) for adjacent classification. The proportion of completely opposite classification was 8.1% in average. Calibration slope was estimated by regression and calibration parameters ranged from 0.025 for carotene to 0.423 for niacin. We conclude that the FFQ we have developed is an appropriate tool for assessing the nutrient intakes as ranking exposures in epidemiology studies in view that amounts of consumed nutrients obtained by FFQ were similar to those collected by DRs, that correlations between consumed nutrients collected by these methods were significant, and that classification results were relatively fair. The correlation coefficients, however, were lower than expected, which may be mainly due to the survey season. In fact, any short-term dietary survey cannot accurately reflect the overall dietary intakes that change heavily depending on seasons. Further studies including the analysis of chemical indices would be helpful for the studies of causal relationship between the diet and disease.