Distinct antibody specificities to a 64-kD islet cell antigen in type 1 diabetes as revealed by trypsin treatment.

Type 1 diabetes is associated with antibodies that immunoprecipitate a 64-kD islet cell membrane protein from detergent extracts of pancreatic islets. In this study we have determined whether mild trypsin treatment of islet membranes can release fragments of the antigen that bind antibodies in the serum of Type 1 diabetic patients. Partial tryptic proteolysis of [35S]methionine-labeled 64-kD antigen immunoprecipitated from detergent extracts of rat islets resulted in the formation of 50-, 40-, and 37-kD fragments. Similar sized fragments were recovered when sera from diabetic patients were employed to immunoprecipitate polypeptides solubilized by mild trypsin treatment of a particulate fraction of radiolabeled rat islets. Of 27 diabetic patients, 22 possessed antibodies to the 50-kD polypeptide and 21 to the 40- and 37-kD polypeptides. A positive association was found between 64k antibodies and antibodies to the 50-kD fragment but not between 64k antibodies and antibodies to the 40- or 37-kD fragments. Some 64k antibody negative patients possessed antibodies that efficiently immunoprecipitated the latter fragments. Serum from 25 of 27 (93%) diabetic patients immunoprecipitated at least one of the three tryptic polypeptides. One of 20 nondiabetic controls immunoprecipitated a 50-kD polypeptide and all controls were negative for antibodies to 40- and 37-kD fragments. Thus, Type 1 diabetes is associated with the presence of at least two antibody reactivities to distinct determinants of the 64-kD antigen, and some patients may possess antibodies to a cryptic epitope on the detergent-solubilized molecule. These data suggest that the detection of antibodies (present in 93% of patients) to epitopes on tryptic polypeptides of the 64-kD antigen may be of even greater diagnostic value for the onset of Type 1 diabetes than analyses of antibodies reactive with the intact 64-kD antigen.

High frequency shear horizontal plate acoustic wave devices.

It has been shown recently that shear horizontal acoustic waves propagating in piezoelectric plates whose thickness h is much less than the acoustic wavelength lambda possess a number of attractive properties for use in sensor and signal processing applications. In order to exploit the potential benefits of these waves, however, one needs to fabricate devices on very thin plates. We have developed a suitable fabrication method which can be used to realize devices on such thin plates. In this method, the device is first fabricated on a plate of normal thickness (approximately 500 microm) and the substrate is then lapped from the back side to reduce the thickness. The technique has been utilized to realize devices on plates of thickness less than 70 microm. A shear horizontal plate acoustic wave (SH-PAW) delay line of fundamental resonant frequency greater than 25 MHz and insertion loss less than 7 dB has been realized on a 60 microm thick Y--cut, X--propagation lithium niobate substrate. The device also shows strong response near the third harmonic frequency of 75 MHz.

Shake culture studies for the production of amylases by Thermomyces lanuginosus.

The paper describes shake flask culture optimization studies for the production of amylases by Thermomyces lanuginosus. The culture was found to produce maximally alpha-amylase and glucoamylase (18.4 & 11.2 units/ml), respectively when the medium contained rice flour (2% w/v) as carbon and corn steep liquor as nitrogen source with medium pH adjusted to 5.5 and incubated for 72 h under shaking conditions (150 rpm) at 50 degrees C.

A phylogenetic analysis based on the gene encoding phosphoglycerate kinase.

We have determined the nucleotide sequence of both genomic and complementary DNA (cDNA) for the gene encoding the glycolytic enzyme phosphoglycerate kinase from the ciliated protozoan Tetrahymena thermophila. The amino acid sequence for the enzyme has also been derived from the cDNA sequence. The gene contains an open reading frame of 1260 nucleotides encoding 420 amino acids. Coding sequence in genomic DNA is interrupted by two introns at positions corresponding to introns 3 and 4 in mammalian phosphoglycerate kinase genes. The derived amino acid sequence was used to prepare a phylogeny by aligning the Tetrahymena sequence with 25 other phosphoglycerate kinase amino acid sequences. The Tetrahymena sequence is a typical eukaryotic sequence. There is recognizable and clear homology across species that cover nearly the complete range of life forms. The phylogenetic reconstruction of these sequences generally supports the conclusions that have been reached using rRNA sequences.