Validation of the Applied Biosystems Prism 377 automated sequencer for the forensic short tandem repeat analysis

The Applied Biosystems (ABI) Prism 377 DNA sequencer has been evaluated in an attempt to increase the throughput of samples for short tandem repeat (STR) analysis, in both forensic casework and the UK National Criminal Intelligence DNA Database. The gel system assessed consisted of 0.2 mm, 4% acrylamide 6 M urea gels, with a well-to-read distance of 36 cm. Gels were run at a constant voltage of 3 kV and constant temperature of 51 degrees C. The run time of our second generation multiplex (SGM) STR system was achieved in less than 2 h. Rigorous validation has been performed on the instrument hardware and software. Complete resolution of 1 base differences was obtained, up to and beyond 350 bases; sizing precision across gels was more than 2-fold higher than the 373A and the sensitivity was increased by one third.     

Antibiotic resistance patterns of metal-tolerant bacteria isolated from an estuary

Estuarine bacteria isolated on metal-containing media were also found to be antibiotic resistant; ampicillin and chloramphenicol were the antibiotics to which resistance was most common. Patterns of antibiotic resistance were found associated with a variety of taxa.     

A 76-amino acid disulfide loop in the Yersinia pseudotuberculosis invasin protein is required for integrin receptor recognition

The Yersinia pseudotuberculosis invasin protein is a 986-amino acid protein that promotes bacterial penetration into mammalian cells by avidly binding multiple beta 1-chain integrins. A 192-amino acid carboxyl-terminal domain of invasin was previously shown to be sufficient for binding. Evidence is presented here that a 76-amino acid disulfide loop in the integrin binding domain of invasin is required for invasin-mediated cell binding and entry. Bacterial mutants that were altered at either of 2 cysteine residues in the binding domain of invasin were completely defective for entry. Purified invasin protein derivatives altered at either of these cysteines, in contrast to the wild-type invasin, did not promote either cell binding or penetration. Analysis of proteolytic products of invasin in the presence or absence of reducing agent provided evidence of an intra-chain disulfide bond near the carboxyl terminus of the protein. Alkylation of invasin derivatives with [3H]iodoacetate indicated that these 2 cysteines were normally disulfide-bonded. A treatment that resulted in the maximal reduction of the disulfide bond also resulted in maximal loss of cell attachment activity. These results indicate that the 76-amino acid disulfide loop at the carboxyl terminus of invasin is required for recognition by integrins.     

Follow-up of argon laser trabeculoplasty: is a day-one postoperative IOP check necessary?

BACKGROUND AND OBJECTIVES: To evaluate the benefit of measuring the intraocular pressure (IOP) on the first postoperative day after argon laser trabeculoplasty (ALT). PATIENTS AND METHODS: We retrospectively reviewed 407 ALT procedures with perioperative apraclonidine performed on 226 patients between January 1991 and December 1993. Data analyzed included type of glaucoma, extent of treatment, whether the procedure was initial or repeat, laser parameters, and IOP preoperatively and at 1 hour, 1 day, and 1 month postoperatively. RESULTS: The percentage of patients with an IOP rise of greater than 3 mm Hg at 1 hour, 1 day, and 1 month following ALT was 11.3%, 4.2%, and 5.2% respectively. The incidence of IOP elevations greater than 10 mm Hg was 2.2%, 1.0%, and 1.5% at 1 hour, 1 day and 1 month, respectively. Of 17 cases with an IOP elevation greater than 3 mm Hg at 1 day, four eventually required a trabeculectomy. However, there were no consistent factors that distinguished which cases with elevated IOP at 1 day ultimately needed further therapy, nor did the 1-day postoperative examination predict which patients would have IOP elevation at 1 month. CONCLUSION: IOP 1 day after ALT is rarely elevated and does not correlate with IOP elevation at 1 month. Therefore, an IOP check at 1 day is not felt to be necessary for most patients.     

Identification of 130 kDa cell surface LDL-binding protein from smooth muscle cells as a partially processed T-cadherin precursor

Atypical cell surface lipoprotein-binding proteins of 105 kDa and 130 kDa are present in membranes of vascular smooth muscle cells. We recently identified the 105 kDa protein from human aortic media as T-cadherin, an unusual glycosylphosphatidylinositol (GPI)-anchored member of the cadherin family of cell adhesion proteins. The goal of the present study was to determine the identity of 130 kDa lipoprotein-binding protein of smooth muscle cells. We applied different approaches that included protein sequencing of purified protein from human aortic media, the use of human T-cadherin peptide-specific antisera, and enzymatic treatment of cultured cells with trypsin and GPI-specific phospholipase C. Our results indicate that the 130 kDa protein is a partially processed form of T-cadherin which is attached to the membrane surface of smooth muscle cells via a GPI anchor and contains uncleaved N-terminal propeptide sequence. Our data disclose that, in contrast to classical cadherins, T-cadherin is expressed on the cell surface in both its precursor (130 kDa) and mature (105 kDa) forms.