pH-sensing nano-crystals of carbonate apatite: effects on intracellular delivery and release of DNA for efficient expression into mammalian cells.

Two unique and fascinating properties of carbonate apatite which are well-known in hard tissue engineering, have been unveiled, for the first time, for the development of the simplest, but most efficient non-viral gene delivery device - ability of preventing the growth of crystals needed for high frequency DNA transfer across a plasma membrane and a fast dissolution rate for effective release of DNA during endosomal acidification, leading to a remarkably high transgene expression (5 to 100-fold) in mammalian cells compared to the widely used transfecting agents. Moreover, by modulating the crystal dissolution rate of carbonate apatite through incorporation of fluoride or strontium into it, transfection activity could be dramatically controlled, thus shedding light on a new barrier in the non-viral route, which was overlooked so far. Thus we have developed an innovative technology with significant insights, that would come as a promising tool for both basic research laboratories and clinical settings.

Ibotenic acid-induced lesions of the medial septum increase hippocampal membrane associated protein kinase c activity and reduce acetylcholine synthesis: prevention by a phosphatidylcholine/vitamin B12 diet.

Ibotenic acid infusion into the medial septum (MS) results in biochemical alterations in the hippocampus. The biochemical events involved in this neuronal lesion are poorly understood. We investigated the effect of a purified diet supplemented with egg phosphatidylcholine (PC) and vitamin B(12) on ibotenic acid-medicated biochemical changes in the rat hippocampus and crude synaptosomal membranes. Male Wistar rats with this MS lesion were fed a purified diet (control diet) or a purified diet supplemented with 5.7 g PC and 125 microg vitamin B(12) per 100 g (experimental diet) for 18 days. Sham-operated rats were fed the control diet. Compared with the sham-operated rats, MS-lesioned rats fed the control diet showed increased activity of membrane-bound protein kinase C (PKC), decreased activity of choline acetyltransferase, and decreased concentrations of acetylcholine in the hippocampus. The ratio of cholesterol to phospholipid in the crude synaptic membrane was lower in the lesioned rats than in the sham-operated rats, but this was not accompanied by any alteration in membrane lipid fluidity. MS-lesioned rats fed the experimental diet showed lowered PKC activity and elevated acetylcholine concentrations than did rats fed the control diet, but there were no significant effects on choline acetyltransferase activity and the lipid ratio. The ibotenic acid-mediated elevation of PKC activity was observed as early as 2 days postinjury in the control diet-fed rats but not in the experimental diet-fed rats. We propose that ibotenic acid mediates pathophysiologic actions through the activation of PKC and that PC combined with vitamin B(12) ameliorates the second messenger-mediated injury.

Synthesis of sulfated colominic acids and their interaction with fibroblast growth factors.

Colominic acid, an alpha (2-->8)-linked poly(sialic acid), was sulfated and characterized by NMR spectroscopy. During sulfation, the secondary hydroxyl group at C-4 had almost the same reactivity as the primary hydroxyl group at C-9, while the secondary hydroxyl group at C-7 was hardly substituted. Analysis by molecular modeling suggested that the lack of substitution at C-7 was due to a steric hindrance. A mobility shift assay indicated that FGF-2 bound to the sulfated colominic acid. Synthetic sulfated colominic acid potentiated the mitogenic activity of FGFs for fibroblasts in the same manner as heparin. Sulfated colominic acid with a low degree of sulfation was able to potentiate FGF activity. Regardless of the degree of sulfation, sulfated colominic acid-induced cytotoxicity was not observed. It was suggested that the carboxyl groups in sulfated colominic acid cooperate with the sulfate groups to reinforce the interaction with FGFs and to reduce the cytotoxicity of sulfated colominic acids.

[An anatomical study of the carpal tunnel for endoscopic carpal tunnel release].

An anatomical study of the carpal tunnel was performed to characterize the transverse carpal ligament, enabling improved endoscopic carpal tunnel release. One hundred wrist joints obtained from 50 cadavers were studied. The width of the transverse carpal ligament (TCL) was found to be narrower in females than in males, and there was no significant difference in the width of the ligament between the right and the left wrist joints. The length of the TCL was not significantly different between males and females or between the right and left wrist joints. On the sagittal plane, the TCL was spindle-shaped and its thickest part was in the distal third. The thickness of the TCL had a significant negative correlation with age. The width of the TCL had a significant positive correlation with both its length (p < 0.01) and the thickness of its distal part (p < 0.01). Neither the distance between the distal edge of the TCL and the superficial palmar arch nor the distance between the radial edge of the pisiform bone and the ulnar artery at the distal edge of the TCL were significantly correlated with sex, laterality, or age. These results indicated that the width and thickness of the TCL and the ulnar artery can be predicted by calculating the length of the TCL percutaneously, thus enabling a safe and reliable endoscopic carpal tunnel release to be performed clinically without any complication.

Effect of sulfated polysaccharides on hepatocyte adhesion.

The effects of sulfated polysaccharides on hepatocyte adhesion on the polystyrene dish and fibronectin-coated dish were investigated. Dextran sulfate and synthetic mannopyranan sulfate slightly inhibited hepatocyte adhesion on fibronectin-coated dish, while heparin showed no effect. On the other hand, hepatocyte adhesion on polystyrene dishes was stimulated in the presence of sulfated polysaccharides in the medium.