Two different strategies for scanning and screening of mutations in polymerase chain reaction (PCR) products by hybridization analysis are described, employing real-time biospecific interaction analysis (BIA) for detection. Real-time BIA was used to detect differences in hybridization responses between PCR products and different 17-mer oligonucleotide probes. For the analysis using a biosensor instrument, two different experimental formats were investigated based on immobilization of either biotinylated PCR products or oligonucleotide probes onto a sensor chip. Applied on the human tumour suppressor p53 gene, differences in hybridization levels for full-match and mismatch situations employing both formats allowed the detection of point mutations in exon 6 PCR products, derived from a breast tumour biopsy sample. In addition, a mutant sample sequence could be detected in a 50/50 background of wild type exon 6 sequence. The suitability of the different formats for obtaining a regenerable system and a high throughput of samples is discussed.

Bacterial Proteins↗

PubMedDate not supplied

Detection of antigen-antibody interactions by surface plasmon resonance. Application to epitope mapping.

Surface plasmon resonance (SPR) detection requires no labeling of antigen or antibodies and allows quantification of two or more interacting molecular species. The automated SPR instrument used here consists of an optical detection unit, an integrated liquid handling unit, and an autosampler. A first molecule is immobilized to the dextran modified surface of the sensor chip. By sequential introduction, the stepwise formation of multimolecular complexes can then be monitored. A two-site binding assay which allows characterization of MoAb epitope specificities is described. A polyclonal rabbit anti-mouse IgG1 (RAMG1) immobilized to the dextran surface is used to capture the first MoAb from unprocessed hybridoma culture supernatants. After introducing the antigen, the ability of a second MoAb to bind to the antigen is tested. The analysis cycle which is fully automated can be performed more than 100 times using the same RAMG1 surface. Since the detection principle allows monitoring of each reactant in the consecutive formation of a multimolecular complex, multi-site binding experiments can be performed. Five MoAbs recognizing different epitopes on an antigen were shown to bind sequentially, forming a hexamolecular complex. MoAbs were further characterized by inhibition analysis using synthetic peptides derived from the primary structure of their antigen. As a model system MoAbs against recombinant HIV-1 core protein p24 were used in all experiments.

Amino Acid Sequence↗

PubMedDate not supplied

Progress and regression within primary Sjögren's syndrome.

The purpose of this review is to give a modern view and an update of important areas in primary Sjögren's syndrome (SS), which may be the most common of the autoimmune systemic rheumatic diseases. Interest in aspects of primary SS including clinical manifestations, pathogenesis, aetiology, treatment, prognosis, etc has increased during the past three decades, the volume of scientific papers and the number of theses being the indicators. However, only a fraction of the money that is used for research into rheumatoid arthritis (RA) is used for SS, and the statement that SS is under-diagnosed, under-treated and under-researched will still be valid for several years to come. The topics that are focused on in this review are: (a) clinical areas with subsections on signs and symptoms, terminology, predictors for development of non-Hodgkin malignant lymphoma (NHML) and prognosis, (b) treatment, (c) the Danger model (aetiopathogenesis) and (d) pathology, including immunoglobulin G4 (IgG4)-positive cells.

Antibodies, Monoclonal↗

PubMedDate not supplied

Erroneous results due to blood sampling from a permanent intravenous port.

Explore the source record for details and available documents.

Blood Specimen Collection↗

PubMedDate not supplied

The value of neonatal serum thyroglobulin determinations in the follow-up of patients with congenital hypothyroidism.

Serum thyroglobulin was determined in 68 newborn infants with positive screening tests for congenital hypothyroidism. In 38 infants the diagnosis was confirmed (patients), but the remaining 30 were euthyroid at follow-up (controls). The mean thyroglobulin concentration at the age of 2 weeks did not differ significantly between the patients and the controls (179 vs. 125 micrograms/l). Thyroid scintigraphy was performed in 15 patients. All seven with thyroid aplasia, based on 99mTc pertechnetate scintigraphy, had measurable thyroglobulin (greater than 2 micrograms/l) and thyroid hormones in their serum. This indicates that total absence of thyroid tissue is very rare in Swedish patients with congenital hypothyroidism. Scintigraphy based on 99mTc does not permit detection of small amounts of thyroid tissue. The neonatal concentrations of thyroglobulin did not correlate with the results of Griffiths test at 3 years and are therefore not useful for prognosis of psychomotor development. We conclude that neonatal measurement of thyroglobulin is of limited value in the follow-up of patients with congenital hypothyroidism.

Bone Development↗

PubMedDate not supplied

Circulating thyroid antibodies in congenital hypothyroidism.

The role of maternal thyroid antibodies in congenital hypothyroidism is controversial. We have analysed serum thyroid antibodies in patients and their mothers. In a bioassay, antibodies interacting with thyroid cells were analysed by measuring of TSH-stimulated cAMP production in a rat thyroid cell line, FRTL5. Serum antibodies against the TSH receptor, thyroid peroxidase and thyroglobulin were determined by radioreceptor assay and enzyme-linked immunosorbent assays. The bioassay was performed with IgG preparations from 89 mothers of children with congenital hypothyroidism. Analyses for TSH receptor antibodies and thyroid peroxidase/thyroglobulin antibodies were performed on 144 and 118 sera of newborn patients respectively. No evidence of an increased prevalence of thyroid antibodies was found on comparison with controls. One infant had transient neonatal hyperthyrotropinaemia because of TSH receptor blocking antibodies transferred from the mother. Our data indicate that, apart from transplacental transfer of TSH receptor antibodies, maternal immunoglobulins have a limited role in the aetiology of congenital thyroid dysfunction.

Autoantibodies↗

PubMedDate not supplied

Characterization of synergy between ofloxacin, ceftazidime, and tobramycin against Pseudomonas aeruginosa.

The purposes of this study were to investigate the potential for synergy between ceftazidime, tobramycin, and ofloxacin against two clinical isolates (PSA 9258, and PSA 9263) of Pseudomonas aeruginosa utilizing time-concentration-kill curves. A pharmacodynamic model was used to simulate one-compartment pharmacokinetics for single-, double-, or triple-drug combinations utilizing two different elimination half-lives (T1/2). Each duplicate experiment was conducted for 24 h in cation-supplemented Mueller-Hinton broth. Synergy, indifference, and antagonism were defined as reductions of > or = 2.0, > or = 0 to < or = 2, or < or = 0 in mean log10 CFU/ml (CFU = colony-forming units) in bacterial counts at any time point during the 24-hour experiment, respectively. Time-concentration-kill curve studies simulating a peak concentration (CP) = 6 micrograms/ml and T1/2 = 5.5 h for ofloxacin combined with ceftazidime (CP = 80 micrograms/ml; T1/2 = 2 h) resulted in 2.18 and 1.81 log CFU/ml mean decreases in bacterial counts for PSA 9258 and 9263, respectively. Simulated ofloxacin pharmacokinetic parameters (CP = 6 micrograms/ml); T1/2 = 5.5 h) combined with tobramycin (CP = 8 micrograms/ml; T1/2 = 2 h) produced 2.26 and 0.6 log CFU/ml mean reductions in bacterial counts for PSA 9258 and 9263, respectively. Time-concentration-kill curve results were inconsistent with checkerboard synergy experiments which indicated antagonism for ofloxacin/tobramycin combinations (fractional inhibitory concentrations = 2.0/2.5) and indifference for ofloxacin/ceftazidime combinations (fractional inhibitory concentrations = 1.0/1.0). In secondary experiments, tobramycin (T1/2 = 2 h) and ceftazidime (T1/2 = 2 h) at concentrations of one fourth and equal to the minimum inhibitory concentration were combined with ofloxacin (CP = 6 micrograms/ml; T1/2 = 5.5 h). When ofloxacin was combined with tobramycin equivalent to the minimum inhibitory concentration, mean reductions in bacterial counts were 3.74 and 5.59 CFU/ml. These results suggest that an enhanced antipseudomonal activity may result by the combination of clinically achievable concentrations of ofloxacin with minimum inhibitory concentration equivalent concentrations of tobramycin.

Anti-Bacterial Agents↗

PubMedDate not supplied

InAs quantum dots capped by GaAs, In0.4Ga0.6As dots, and In0.2Ga0.8As well.

We have fabricated and characterized three types of InAs quantum dots (QDs) with different InxGa1-xAs capping layers. Post-growth atomic force microscopy measurements show that the In0.2Ga0.8As/InAs structure has a smooth surface (dot-in-well structure), whereas the In0.4Ga0.6As/InAs structure revealed large QDs with a density similar to that underneath InAs QDs on GaAs (dot-in-dot). With increasing In mole fraction of the capping layer and increasing In0.4Ga0.6As thickness, the energy position of the room-temperature photoluminescence (PL) peak is red-shifted. The quantum dot-in-dot structure emits stronger room-temperature PL than does the quantum dot-in-well structure. With a spatially distributed strain in the InAs quantum dot, we have solved the three-dimensional Schrödinger equation by the Green's function theory for the eigenvalues and eigen wave functions. It is concluded that the ground state increases its wave function penetration into the low-barrier InxGa1-xAs capping layer so that its energy position is red-shifted. The reduced PL peak intensity of the dot-in-well (compared with GaAs covered dots) is due to the reduced overlapping between the ground state and the extended states above the GaAs barrier. The overlapping reduction in the dot-in-dot is over compensated for by the reduced relaxation energy (full width at half-maximum), indicating the importance of the sample quality in determining the PL intensity.

Arsenicals↗

PubMedDate not supplied

Serum and cerebrospinal fluid examinations in the diagnosis of Borrelia infection in Bell's palsy.

Seventy-two consecutive patients with Bell's palsy were investigated for evidence of tick-borne Borrelia infection and to compare the merits of serum and cerebrospinal fluid examinations in the diagnosis of Borrelia infection. Serum was taken in the acute and convalescent stages from 72 patients, and cerebrospinal fluid was obtained from 35 of these. An enzyme-linked immunosorbent assay (ELISA) was used to analyse serum and cerebrospinal fluid samples for IgG and IgM antibodies against the Borrelia spirochete. The serum of 8 (11%) of the 72 patients was positive for IgG or both for IgG and IgM. Only 1 of the 35 patients presented an elevated titre of IgG and IgM in the cerebrospinal fluid as well as elevated IgG and IgM titres in the serum. The results, even if limited, indicate that screening of serum is sufficient to exclude Borrelia infection in cases of uncomplicated isolated acute peripheral facial palsy.

Acute Disease↗