Saccharomyces cerevisiae cells secreting an Aspergillus niger beta-galactosidase grow on whey permeate.

We describe the construction of a lactose-utilizing Saccharomyces cerevisiae that expresses the cDNA for a secreted, thermostable beta-galactosidase (lacA) from Aspergillus niger. Yeast cells expressing the lacA gene from the yeast ADH1 promotor on a multicopy plasmid secrete up to 40% of the total beta-galactosidase activity into the growth medium. The secreted product is extensively N-glycosylated, and cells expressing the lacA gene grow on whey permeate (4% w/v lactose) with a doubling time of 1.

Systemic lupus erythematosus in adolescents.

Once considered a rare, fatal illness in children and adolescents, systemic lupus erythematosus (SLE), or lupus, is now understood to be a relatively common chronic disorder in these age groups. This article provides a brief overview of the disease, describing its etiology, pathology, clinical manifestations, diagnosis, and treatment. A case history of an adolescent girl with lupus is presented, and the role of the nurse is discussed in relation to promoting four developmental tasks of adolescents: developing a positive body image, establishing a sexual identity, achieving independence, and acquiring formal thought processes.

D-Xylose (D-glucose) isomerase from Arthrobacter strain N.R.R.L. B3728. Gene cloning, sequence and expression.

Arthrobacter strain N.R.R.

D-Xylose (D-glucose) isomerase from Arthrobacter strain N.R.R.L. B3728. Purification and properties.

D-Xylose (D-glucose) isomerase was purified to homogeneity in yields of approx. 1 g/kg of wet cells from a strain of Arthrobacter that produces it as about 10% of total soluble protein. It is a tetramer of identical 43,114 Da subunits containing a preponderance of acidic residues and no cysteine.

Evaluation of light microscopy to localise the site of haematuria.

Red cell morphology was examined by light microscopy in 122 urine specimens from 99 patients with haematuria in an attempt to define the site of origin. Altogether 84% of glomerular bleeding and 91% of non-glomerular bleeding was correctly assigned according to diagnoses determined by renal biopsy in 51 patients and clinically in 48. The test should be interpreted with caution as both false positive and false negative results were observed, some in the same individual with unchanged pathology at different times.

Kinetics and pathogenicity of autoantibodies induced by mercuric chloride in the brown Norway rat.

Repeated low-dose injections of mercuric chloride (HgCl2) in the brown Norway (BN) rat result in polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) autoantibodies. We examined the kinetics of various autoantibodies produced in vivo, general features of polyclonal activation such as total IgG levels and immune complex formation, and the relationship between organ specific autoimmunity and tissue injury in the kidney and thyroid. The production of immune complexes and autoantibodies to GBM and thyroglobulin was short lived, and the increase in levels of total IgG and antibodies to ssDNA and dsDNA was prolonged; the antibody response to collagen types I and II was intermediate in duration.

Number of interstitial capillary cross-sections assessed by monoclonal antibodies: relation to interstitial damage.

Using thin Plexiglass sections stained with silver, the peritubular capillary area and number of capillary cross-sections can predict both interstitial damage and plasma creatinine concentrations. This technique is both difficult to perform and time-consuming. We have restudied this topic using conventional cryostat sections from 46 biopsies with chronic glomerulonephritis and tubulointerstitial nephritis and two monoclonal antibodies (MoAb) recognising capillary endothelium.

The relevance of induced class II HLA antigens and macrophage infiltration in early renal allograft biopsies.

Using a panel of monoclonal antibodies, immunohistological analysis was performed on frozen sections taken from 14 peritransplant renal biopsies and 42 biopsies taken 6 +/- 2 days posttransplantation. The following parameters were examined: tubular expression of HLA-DR, DP, and DQ and infiltration with T lymphocytes and macrophages. Of the 42 posttransplant biopsies, 26 were diagnosed as rejecting and 16 as nonrejecting according to clinical and histopathological criteria.

Increase in proteinuria and reduction in number of anionic sites on the glomerular basement membrane in rabbits by infusion of human nephrotic plasma in vivo.

1. Intraortic suprarenal infusion in vivo of New Zealand White rabbits with plasma from patients with the nephrotic syndrome reduced the number of anionic sites labelled by the cationic polymer polyethyleneimine in the lamina rara iterna to 14.0 +/- 2.

Linked scans of peptides and protein digests: amino acid sequence determination of components of complex mixtures.

Two-sector linked-scan analysis of an unpurified proteolytic digest of a pyruvate decarboxylase enzyme (60,000 Da) has allowed the discovery and assignment of an amino-terminal post-translational modification and processing event. A difference in amino acid sequence from that predicted by a recently published nucleotide sequence has also been found. These results illustrate both the use and considerable potential of linked-scan methods for the analysis of complex biopolymer mixtures.

Identification, cloning and characterisation of a new gene required for full pyruvate decarboxylase activity in Saccharomyces cerevisiae.

Biochemical evidence that pyruvate decarboxylase in S. cerevisiae might be constituted from two independently encoded subunits led us to question genetic evidence for a single structural gene. The main evidence for this was that three "structural" mutations appeared to be alleles of the same gene, PDC1 (Schmitt and Zimmermann 1982).

Extraction and rapid inactivation of proteins from Saccharomyces cerevisiae by trichloroacetic acid precipitation.

Methods currently used for the extraction of proteins from yeast involve relatively long time periods between sampling cells from a culture and analysis of their proteins by polyacrylamide gel electrophoresis-sodium dodecylsulphate. Often it is desirable to inactivate cellular metabolism rapidly after sampling and here we show that trichloroacetic acid precipitation techniques, often used for rapid extraction and inactivation of proteins from higher eukaryotes, can be adapted for use with organisms which have cell walls.

HLA-DR display by renal tubular epithelium and phenotype of infiltrate in interstitial nephritis.

Renal tubular display of HLA-DR was estimated semi-quantitatively in 28 biopsies from 27 patients with various forms of tubulointerstitial nephritis (10 following use of non-steroidal anti-inflammatory drugs) using a monoclonal anti-MHC class II non-polymorphic antibody (DK-22). Normal donor kidneys and biopsies from patients with minimal-change nephrotic syndrome were examined as controls. The phenotype of infiltrating cells was also studied quantitatively, using monoclonal antibodies on frozen biopsy tissue; the number of cells infiltrating the interstitium was counted per tubular cross-section.

The structure and regulation of phosphoglucose isomerase in Saccharomyces cerevisiae.

We have cloned and sequenced the PGI1 gene, encoding phosphoglucose isomerase (E.C.5.

A host-vector system for an Arthrobacter species.

An efficient host-vector system has been developed for an industrial strain of Arthrobacter sp. (NRRL B3728)used for glucose isomerase production. Protoplasts of Arthrobacter were generated by treating the cells with 0.

Improved prognosis of renal microscopic polyarteritis in recent years.

To determine if the short- and long-term prognosis of renal micropolyarteritis (including Wegener's granulomatosis) had improved in our Unit in the last 5 years compared to our previous series from 1965 to 1980, we examined data from 26 recent patients, 1981-1986. Twenty were admitted during acute phase and six after a mean of 7.1 months of disease.

Respiratory depressant effects of GABA alpha- and beta-receptor agonists in the cat.

The aim of this study was to evaluate the cardiorespiratory effects of intravenously administered gamma-aminobutyric acid (GABA) alpha-(4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol, THIP) and beta-(baclofen) receptor agonists and to locate the site of action of these drugs in the brain. THIP and baclofen were administered to alpha-chloralose-anesthetized cats while minute ventilation (VE), arterial blood pressure (AP), and heart rate were monitored. THIP, in doses of 0.

Intraglomerular T cells and monocytes in nephritis: study with monoclonal antibodies.

Intraglomerular T cells, monocytes, total leucocytes and other mononuclear subsets were sought in renal biopsies from patients with glomerulonephritis, using monoclonal antibodies and immunoperoxidase techniques. Twenty-four biopsies with no significant glomerular proliferation on optical microscopy, thirty-two with only endocapillary hypercellularity, and twenty-one with extra capillary crescentic glomerular disease were studied. Few intra-glomerular leucocytes were seen in the non-proliferative group.

Abnormal podocyte CR-1 expression in glomerular diseases: association with glomerular cell proliferation and monocyte infiltration.

The expression of CR-1 complement receptors on glomerular epithelial cells, was studied in 77 renal biopsies from patients with (74) or without (3) glomerular diseases, employing an anti-CR-1 monoclonal antibody, and an indirect immunoperoxidase technique. Four patterns of CR-1 expression were recognised: normal (18); generally decreased (6); focal/segmental partial loss (44); and complete loss (9). Normal expression was detected in all three biopsies with non-glomerular diseases, and in glomerular diseases with normal glomeruli on light microscopy, but also in several glomerulonephritic biopsies (13), including diffuse proliferative lupus nephritis (1 of 7) and idiopathic membranous nephritis (5 of 14).