Effects of Cold Stimulation on Mitochondrial Activity and VEGF Expression in vitro.

We aimed to clarify the effects of cold stimulation at various temperatures on mitochondrial activity and vascular endothelial growth factor (VEGF) expression in vitro. Human fibroblast, human mesenchymal stem cell, and rat skeletal muscle myoblast cell lines were used. For each cell type, cells were divided into 4 groups and stimulated in various cold temperatures (0, 4, 17 and 25°C) 3 times for 15 min each by placement on crushed ice or floating on cold water set at each temperature. Control cells were subjected to warm water at 37°C. Factors related to mitochondrial activity, mitochondrial DNA copy numbers, and VEGF expression were analyzed 24 h after the last cold stimulation. In all cell types, significant increases of factors related to mitochondrial activity and mitochondrial DNA copy numbers were seen in the 4°C and 17°C-stimulated cells compared with control cells. In rat skeletal muscle cells stimulated at 4°C, VEGF expression significantly increased compared to the control cells. Our data suggest that cold stimulation at certain temperatures promotes mitochondrial activity, biogenesis and VEGF expression.

A linkage study with DNA markers (D4S95, D4S115, and D4S111) in Japanese Huntington disease families.

Attempts to isolate the Huntington disease (HD) gene based on its position have been frustrated by apparently contradictory recombination events in HD pedigrees that have predicted two non-overlapping candidate regions: 100 kb at the telomere of the short arm of chromosome 4, and a 2.2 Mb region located internally at 4p16.3. The proximal location is also supported by the detection of a linkage disequilibrium between HD and some restriction fragment length polymorphisms (RFLPs) at the D4S95, D4S98, and D4S127 loci. In the present study, a proximal marker D4S95 showed tight linkage to the disease locus in Japanese pedigrees (Zmax = 3.31, theta max = 0.00), while distal markers D4S115 and D4S111 did not. Particularly, a two point linkage analysis between D4S111 and HD yielded a lod score -2.01 for theta = 0.015. This result leads to the exclusion, as a possible region of localization of the HD gene, of more than 3 cM of the genome around D4S111 locus. At the same time our results favor aforementioned proximal location as a candidate location for the HD gene.

Nuclear distribution of glutamate, gamma-aminobutyrate and aspartate within the normal human thalamus.

Detailed mapping of amino acid neurotransmitter candidates, glutamate, gamma-aminobutyrate and aspartate within the human thalamus, was performed with a grid microdissection method using two horizontal slices of the right thalamus from four neurologically normal cases. Although the fine distribution of these amino acids varied considerably from case to case, characteristic patterns of distribution for each amino acid could be demonstrated by superimposing maps of each case. Glutamate was high around a caudal part of nucleus medialis and a rostral part of nucleus pulvinaris. Interestingly enough, gamma-aminobutyrate distributed in a similar manner to glutamate, suggesting the possible interaction between glutamate and gamma-aminobutyrate in these areas of the thalamus. The distribution of aspartate was different from the former two amino acids and highly concentrated areas were patchy and scattered in the middle part of the thalamus. The results were discussed in relation to the recently accumulated evidence obtained from animal brains for neurotransmitter roles of these amino acids in the thalamus.