Presence of single-stranded DNA in PCR products of slow electrophoretic mobility.

PCR products were characterized by electrophoresis, blotting and hybridization. In addition to the bands of expected size, bands of slower electrophoretic mobility were often detected. The slower bands completely disappeared when the PCR products were subjected to slow cooling, treated with S1 nuclease or run on an alkaline gel, whereas the bands of expected size were unaffected. The slower bands are therefore likely to contain single-stranded DNA.     

Specific localization of 2′,3′-cyclic nucleotide 3′-phosphohydrolase, (Ca2+/Mg2+)-ATPase,and acetylcholinesterase in human erythyrocyte membrane

A procedure was developed for the detection of 2′,3′-cyclic nucleotide 3′-phosphohydrolase in myelin. This assay was sufficiently sensitive to detect the low levels of 2′,3′-cyclic nucleotide 3′-phosphohydrolase in human erythrocytes. The 2′,3′-cyclic nucleotide 3′-phosphohydrolase of human erythrocytes was determined to be exclusively associated with the inner (cytosolic) side of the membrane. Leaky ghostsand resealed ghosts were assayed for 2′,3′-cyclic nucleotide 3′-phosphohydrolase, (Ca²⁺/Mg²⁺-ATPase, and acetylcholinesterase activity, and the 2′,3′-cyclic nucleotide 3′-phosphohydrolase profile is the same as that of the (Ca²⁺/Mg²⁺)-ATPase, an established inner membrane maker.     

Effects of azide on gastric mucose

Sodium azide, a classical inhibitor of cytochrome oxidase, is an effective inhibitor of gastric acid secretion in bullfrog and skate gastric mucosae at low concentrations. While a portion of the oxygen uptake in these tissues is sensitive to azide (KI less than 2 mM), there remains a large fraction (25-60%) with a KI more than 10 times this value, suggesting the presence of a second oxidase. The spectra of cytochromes c and b change with oxygen-nitrogen alternation in the presence of high azide concentrations which essentially eliminate the reactivity of cytochrome oxidase. In both species two additional components are observed in the spectra. The first has a peak at 590 nm, is not the cytochrome oxidase-CO complex, is fully reactive in the presence of azide and accounts for the asymmetry of the oxidase peak. The second is a component at 557 nm which can only be separated from cytochromes c and b by spectral deconvolution, and seems to react in a manner similar to cytochrome c. It is suggested that the 590 compound may be the alternate cytochrome oxidase.     

Studies on the structure of the haemagglutinin

The biosynthesis of the haemagglutinin glycoproteins of infectious influenza virus particles involves proteolytic cleavage of the primary translation products and the amino acid sequences at the two sites of processing are presented. In addition, details of the primary structure of the haemagglutinin of A/Japan/305/57 (H2N1) are reported and compared with information available for haemagglutinins of other subtypes.     

Cell culture and in vitro studies of fresh and cryopreserved human insulinoma

Summary Dispersed cells from both fresh and cryopreserved human insulinoma have been maintained in cell culture. Initial yield of viable cells was 50% for fresh and 25% for cryopreserved tissue. Viability of cells in culture was documented by increasing numbers of cells (doubling time approximately 5 d initially and 2 d at the sixth subculture for both fresh and cryopreserved tissue) and continued release of insulin over time (approximately 100 ng/ml per 10⁵ cells at 10 d and 175 ng/ml per 10⁵ cells at 30 d of culture for both fresh and cryopreserved tissue). Evidence that cells growing in culture were beta cells was provided by: (a) recovery of intracellular and extracellular immunoreactive insulin (IRI), (b) electron microscopic morphology, and (c) immunohistochemical staining. Cells from fresh insulinoma incubated with increasing concentrations of extracellular glucose released increasing amounts of IRI up to approximately 15 mM glucose, which paralleled changes in plasma insulin obtained during a preoperative glucose tolerance test. Under an Intergovernmental Personnel Act Exchange from the Department of Surgery, University of California, Davis, Sacramento Medical Center.     

Use of the cylinder osteotome for cancellous bone grafting

An instrument is described for removing iliac crest cancellous bone grafts under local anesthesia. The device has widespread applications for plastic and orthopedic surgery. The convenience of obtaining bone grafts with this device broadens the use of bone-grafting techniques in patients in whom the need for bone graft is unexpected.