General Anesthetics Regulate Autophagy via Modulating the Inositol 1,4,5-Trisphosphate Receptor: Implications for Dual Effects of Cytoprotection and Cytotoxicity.

General anesthetics are both neuroprotective and neurotoxic with unclear mechanisms. General anesthetics may control cell survival via their effects on autophagy by activation of type 1 inositol triphosphate receptor (InsPR-1). DT40 or SH-SY5Y cells with only or over 99% expression of InsPR-1 were treated with isoflurane or propofol.

Elevated Autophagy and Mitochondrial Dysfunction in the Smith-Lemli-Opitz Syndrome.

Smith-Lemli-Opitz syndrome (SLOS) is a congenital, autosomal recessive metabolic and developmental disorder caused by mutations in the enzyme which catalyzes the reduction of 7-dehydrocholesterol (7DHC) to cholesterol. Herein we show that dermal fibroblasts obtained from SLOS children display increased basal levels of LC3B-II, the hallmark protein signifying increased autophagy. The elevated LC3B-II is accompanied by increased beclin-1 and cellular autophagosome content.

Alterations in membrane caveolae and BKCa channel activity in skin fibroblasts in Smith-Lemli-Opitz syndrome.

The Smith-Lemli-Opitz syndrome (SLOS) is an inherited disorder of cholesterol synthesis caused by mutations in DHCR7 which encodes the final enzyme in the cholesterol synthesis pathway. The immediate precursor to cholesterol synthesis, 7-dehydrocholesterol (7-DHC) accumulates in the plasma and cells of SLOS patients which has led to the idea that the accumulation of abnormal sterols and/or reduction in cholesterol underlies the phenotypic abnormalities of SLOS. We tested the hypothesis that 7-DHC accumulates in membrane caveolae where it disturbs caveolar bilayer structure-function.

Clostridium botulinum toxin A inhibits contractility in pregnant human myometrium in vitro.

Studies were undertaken to evaluate the effect of Botulinum neurotoxin type-A (BoNTA) preparation on oxytocin-induced contractions of pregnant human myometrium in vitro. Human myometrial tissue was exposed to increasing concentrations (1-50 000 U/mL) of BoNT/A. Isometric contractions were measured using a force displacement transducer.

Modification of YACs by Homologous Recombination and Transfer of a 330 kb YAC into Mammalian Cells.

YAC clones were modified via homologous recombination by transfecting yeast cells containing YAC with plasmid PRAN4 DNAs. The integration of neo gene into YAC DNA was identified using Southern blotting and PCR methods. A modified YAC that carried 330 kb human DNA was introduced into L929 cells through PEG mediated spheroplast fusion, and G418 resistant colonies were obtained.