In an earlier report, we described a DNA helicase that is specifically induced upon infection of Vero cells with herpes simplex virus 1. We have purified this enzyme to near homogeneity and found it consist three polypeptides molecular weights 120,000, 97,000, 70,000. Immunochemical analysis has shown these be the products genes UL52, UL5, UL8 are required for replication plasmid containing 1 origin (oriS). addition activity, contains tightly associated primase. Thus, three-subunit...

Herpes simplex virus 1 contains seven genes that are necessary and sufficient for origin-dependent DNA synthesis in cultured cells. We have expressed the product of one these genes, UL9, insect cells by using a baculovirus expression vector. The apparent size UL9 protein, both herpes virus-infected Vero cells, is 82,000 Da. By an immunoassay protein-DNA interaction, we shown protein binds specifically to origins replication, oriS oriL. DNase I "footprint" analysis has interacts with two...

Seven herpes simplex virus (HSV) genes have been shown recently to be necessary and sufficient support the replication of origin-containing plasmids. Two these (pol dbp) encode well-known DNA proteins (the polymerase major single-stranded binding protein), a third gene (UL42) encodes previously identified infected-cell protein which binds tightly double-stranded DNA. The products four remaining not identified. Using predicted amino acid sequence data (D.J. McGeoch, M.A. Dalrymple, A. Dolan,...

The prevalence and clinical relevance of thyroid stimulating hormone (TSH) receptor (TSHR) blocking antibodies (TBAb) in patients with autoimmune disease (AITD) was investigated. Serum TBAb were measured a reporter gene bioassay using Chinese hamster ovary cells. Blocking activity defined as percentage inhibition luciferase expression relative to induction bovine TSH alone (cut-off 40% inhibition). All samples for TSHR stimulatory antibody (TSAb) binding inhibiting immunoglobulins (TBII). A...

Abstract Context Scarce data exist regarding the relevance of stimulatory (TSAb) and blocking (TBAb) thyrotropin receptor antibodies in management Graves disease (GD). Objective To evaluate clinical utility predictive value TSAb/TBAb. Design Prospective 2-year trial. Setting Academic tertiary referral center. Patients One hundred consecutive, untreated, hyperthyroid GD patients. Methods TSAb was reported as percentage specimen-to-reference ratio (SRR) (cutoff SRR < 140%). Blocking...

The measurement of TSH receptor (TSHR) antibodies is warranted for diagnosis Graves' disease (GD).The performance, detection sensitivity, and specificity 6 TSHR immunoassays were compared.Two bioassays 4 binding assays (Kronus, Immulite, Kryptor, Dynex) compared in a dilution study performed patients with autoimmune thyroid disease. Both to 2 using stimulatory (M22) blocking (K1-70) monoclonal antibody (MAb) mixtures.Thirty samples from (TSAb)-positive/blocking (TBAb)-negative GD diluted...

Abstract Context Serum TSH receptor autoantibody (TSH-R-Ab) is a biomarker of Graves disease (GD). Studies have shown that the levels this TSH-R-Ab clinical significance. Objective To differentiate between thyroidal GD only and orbitopathy (GD + GO). Design Controlled, follow-up study. Setting Academic tertiary referral center for GO. Subjects Sixty patients with GD, GO, controls. Intervention Serial serum dilution analyses six automated, ELISA, cell-based assays TSH-R-Ab. Main Outcome...

Summary Thyroid-stimulating immunoglobulins (TSI) are a functional biomarker of Graves' disease (GD). To develop novel TSI bioassay, cell line (MC4-CHO-Luc) was bio-engineered to constitutively express chimeric TSH receptor (TSHR) and constructed with cyclic adenosine monophosphate (cAMP)-dependent luciferase reporter gene that enables quantification. Data presented as percentage specimen-to-reference ratio (SRR%) were obtained from 271 patients various autoimmune thyroid diseases 180...

Background: Cell-based bioassays for functional thyroid stimulating autoantibodies (TSAb) are sensitive diagnostic tools. However, there is no bioassay available that standardized with international reference material. We aimed to promote the standardization of test results among laboratories perform TSAb and calibrate levels against second standard (IS) 08/204 from National Institute Biological Standards Control (NIBSC). Methods: Serum activity was measured a FDA-cleared utilizes CHO cells...

Summary Autoantibodies to the thyrotrophin (TSH) receptor (anti-TSHR) are unique, in that they involved directly pathophysiology of certain autoimmune thyroid diseases (AITD). Thyroid-stimulating antibodies (TSAb) act as agonists activate gland and cause Graves' disease. Other anti-TSHR block TSH can hypothyroidism. Thyroid-blocking (TBAb) have not been studied extensively TSAb. We developed a TBAb bioassay based on cell line expresses chimeric TSHR. The 50% inhibitory concentration Chinese...

The analytical performance and the clinical utility of a thyrotropin receptor (TSHR)-stimulating immunoglobulin (TSI) bioassay were compared with those TSHR-binding inhibitory (TBII) assay. Limits detection (LoD) quantitation (LoQ), assay cutoff, half-maximal effective concentration (EC(50)) measured. Dilution analysis was performed in sera hyperthyroid patients Graves disease (GD) during antithyroid treatment (ATD). Titer defined as first dilution step at which measurement TSI or TBII fell...

We hypothesized that TSH-receptor (TSHR) stimulating antibodies (TSAbs) are involved in oxidative stress mechanisms patients with Graves disease (GD). Nicotinamide adenine dinucleotide phosphate oxidase, isoform 2 (NOX2); parameters; and burst were measured serum, urine, whole blood from GD control subjects. Superoxide production was investigated human embryonic kidney (HEK)-293 cells stably overexpressing the TSHR. Lipid peroxidation determined by immunodot-blot analysis for protein-bound...

Background and Objective: A cell-based bioassay for the measurement of thyroid blocking autoantibodies (TBAb) has been recently reported. The analytical performance validation this is assessed described. Methods: Chinese hamster ovary cells expressing a chimeric thyrotropin receptor were treated with bovine (b) TSH different concentrations an immunoglobulin G (IgG) monoclonal human TBAb (K1-70). was measured as function luciferase activity relative to bTSH alone expressed percent inhibition....

Biochemical characterization of the herpes simplex virus (HSV) DNA polymerase, a model polymerase and an important target for antiviral drugs, has been limited by lack pure enzyme in sufficient quantity. To overcome this limitation, HSV gene was introduced into baculovirus, Autographa californica nuclear polyhedrosis virus, under control polyhedrin promoter to give rise recombinant BP58. BP58-infected Spodoptera frugiperda insect cells expressed polypeptide that indistinguishable from...

Journal Article Mitochondrial DNA Polymorphism in Candida albicans Get access Paul D. Olivo, Olivo Research Service, Middleton Veterans Hospital, and the Department of Medicine, University Wisconsin Medical School, Madison, Wisconsin; Microbiology Graduate Program Cellular, Molecular, Developmental Biology, Tennessee, Knoxville, Tennessee Search for other works by this author on: Oxford Academic PubMed Google Scholar Edward J. McManus, McManus W. Stuart Riggsby, Riggsby Jeffrey M. Jones...

This report describes the clinical evaluation of a novel fluorescent immunoassay (FIA), Sofia Influenza A+B FIA (Quidel, San Diego, CA), for rapid detection and differentiation influenza A B viruses. total 2,047 subjects provided nasal swabs nasopharyngeal or aspirates. The overall sensitivity specificity virus vs culture were 94% 95%, respectively, 89% 96%, respectively. Fourteen hundred sixty-one specimens available testing with reverse transcriptase-polymerase chain reaction (RT-PCR)....

A novel and rapid cell-based bioassay, Turbo TSI, for measurement of thyroid-stimulating immunoglobulins (TSI) was recently reported. An assessment the analytical performance this TSI bioassay is described herein. Thawed cells from kits were treated with different concentrations a World Health Organization (WHO) international standard (IS) TSI-positive serum. measured as function luciferase activity relative light units (RLU) converted into per liter (IU/L). Analytical studies performed on...

Vaccines are effective tools to improve public health. The effectiveness of vaccines is, however, dependent on the overall level protection in a population. Antibodies vaccine-related antigens good biomarkers and serosurveillance can help target vaccination programs. An integrated approach perform multiple vaccine-preventable diseases (VPDs) has been advocated would be facilitated by standardized multiplex immunoassay. In this report, we describe evaluation performance multi-lyte bead-based...