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Biomedical subjects

J Briggs

Publications and source records attributed to J Briggs.

At least 19 recordsLinked to original sources

Stereotactic brachytherapy for malignant glioma using a relocatable frame.

Interstitial brachytherapy for recurrent gliomas normally necessitates the invasive application of a stereotactic frame by screw-fixation which must be kept on for several hours. The use of a relocatable stereotactic frame offers many advantages over conventional systems. We present our experience in 18 patients and verify that the frame used is accurate, comfortable, well tolerated and associated with no major disadvantages.

Adult

Quantitation of DNA and protein impurities in biopharmaceuticals.

The development of drugs and biologicals for human injection generated from recombinant DNA and hybridoma technologies has resulted in new standards for product purity. We discuss the regulatory position relative to impurities in these biopharmaceuticals, focusing on the analytical goals for quantitation. Current methods for making these measurements are reviewed, and a new system designed for improved analysis is described. Assay results for both contaminating DNA and proteins are presented.

Chemistry, Pharmaceutical

Temporal change in diagnostic criteria as a cause of the increase of malignant melanoma over time is unlikely.

To assess whether the increase in malignant melanoma incidence could be due, at least in part, to changes in histological criteria of malignancy, pathologists in Australia, France, Italy, New Zealand, Norway, Sweden, the United Kingdom, the United States and the USSR reviewed diagnoses of 50 consecutive pigmented naevi (40 junctional and compound; 10 intradermal) and 20 consecutive malignant melanomas made in each participating centre around 1930, around 1955 and around 1980. Collaborating pathologists re-read the material, 2,665 cases in all, either from the original slide (82%) or from a recut block (17%), gave their diagnosis and indicated whether the lesion was benign (B), dubious benign (DB), dubious malignant (DM) or malignant (M). As the distribution of review diagnoses was much the same whether the original slide or one made from a recut block was read, the material was pooled. Overall, 2.8% of cases originally reported as B/DB were reviewed as DM/M, while 4.4% of the DM/M diagnoses were held to be B/DB. The shifts between categories were greatest around 1955 and least around 1980, suggesting increasing uniformity of interpretation. All available blocks were recut and sections sent to IARC for review: 1.7% (22) of 1293 B/DB diagnoses were considered to be DM/M and 3.3% (18) of 551 DM/M diagnoses were considered to have been B/DB. The consistently low frequency of shift in diagnostic category, whether the material was reviewed in the collaborating laboratories or by one pathologist at IARC, in a study designed to give maximum attention to those lesions--the junctional and compound naevi--in which a change in opinion as to malignancy would be most likely to arise, suggests that pathologists, irrespective of geographical location, are using common criteria. These findings argue against changes in histological interpretation as being responsible for more than a small portion of the continuous increase of some 3% to 8% per annum observed in malignant melanoma incidence. Other explanations, such as an increase in the frequency or potential for malignant transformation of precursor lesions, must be sought. The anatomical distribution of the malignant melanomas examined followed the usual site pattern by sex, and their thickness was observed to decrease over the period of the study in most centres.

Female

Picogram quantitation of total DNA using DNA-binding proteins in a silicon sensor-based system.

We report a rapid and reproducible method to quantify total DNA at picogram levels. Two high-affinity DNA-binding proteins are used to construct a sandwich assay and a semiconductor sensor is used for quantitation. Single-stranded DNA-binding protein (SSB) from Escherichia coli is conjugated with a linker molecule, biotin, for specific capture of the DNA complex onto a membrane. Monoclonal anti-DNA antibody is conjugated with an enzyme, urease, for signal generation. To detect DNA, a sample is denatured to form single-stranded DNA and then incubated with a reagent containing both DNA-binding protein conjugates and streptavidin. After incubation of the reagent with the DNA sample for 1 h at 37 degrees C to form a complex of streptavidin--biotin--SSB--DNA--anti-DNA--urease, the mixture is filtered through a biotin-coated nitrocellulose membrane which binds the streptavidin component of the complex. The unbound reagent is washed off the membrane, and then the captured DNA complex is detected with a light-addressable potentiometric sensor which measures the pH change catalyzed by the urease in the complex. This assay can detect 2 pg of DNA with a quantitation coefficient of variation of less than 10% in the range 10 to 200 pg.

Animals

Sub-femtomole quantitation of proteins with Threshold, for the biopharmaceutical industry.

The Threshold system provides for rapid quantitation of a variety of analytes at sub-femtomole levels, which is fewer than 6 x 10(8) molecules. The operating principles of the measurement system will be described, including the means for chemical modulation of the signal and the subsequent signal detection utilizing a proprietary silicon sensor. The Immuno-Ligand Assay is a universal ligand-binding assay system which provides quantitative measurement of proteins (including antibodies) in a variety of biological media. Assay performance will be described, along with data demonstrating sensitivity, precision and accuracy, for a variety of analytes of interest to the biopharmaceutical industry.

Animals

Fast biosensor-based system for measuring total DNA in protein solutions.

We report the quantitation of picogram levels of DNA, with a fast, reproducible, and non-isotopic assay system. Two DNA binding proteins are used, one to link DNA in the sample to a membrane and the other to label DNA with an enzyme. Enzyme levels are quantitated with very high sensitivity, by monitoring surface potential changes with a proprietary biosensor, as the enzyme changes its local pH. With this configuration, the assay is generic, detecting the presence of any DNA in the sample; that is, there is no specific hybridization probe involved. The prototype system involves instrumentation, software, plastic disposables, and reagents. With a simple protocol, it allows the user to determine the DNA levels, in the range of 2 to 100 pg, concurrently in multiple samples, with standards, with a total processing time of less than one hour, after sample pretreatment. Representative data will be shown. Essentially the same material presented here appeared in a poster session at the 2nd Annual Symposium on Analytical Biotechnology, Baltimore, Maryland, May 1988.

Automation

Are malignant melanoma time trends explained by changes in histopathological criteria for classifying pigmented skin lesions?

The incidence of malignant melanoma in developed countries has been rising rapidly in recent years. Its causes and the reasons for this trend are uncertain. Changes in histopathological criteria for classifying pigmented skin lesions could explain the rising incidence rates. With support from the International Agency for Research on Cancer, this hypothesis is explored but not substantiated for time trends in the South Western Region, United Kingdom.

England

Glucagon stimulation of brown adipose tissue growth and thermogenesis.

Despite long-standing observations of a whole-body thermogenic effect of glucagon, the role of glucagon in activating thermogenesis in brown adipose tissue has not often been studied. We investigated the ability of administered glucagon to produce alterations in brown adipose tissue similar to changes produced by accepted stimuli of brown fat activity: cold, norepinephrine, and overfeeding. Eighteen days of glucagon injections (1 mg/kg) to male Sprague-Dawley rats produced, relative to saline-injected controls, decreases in feed efficiency and increases in brown adipose tissue weight, protein content, DNA content, and mitochondrial mass as reflected in cytochrome oxidase activity. The observed changes were similar, though of lesser magnitude, to changes produced in these same parameters induced by administration of norepinephrine (250 micrograms/kg) for a positive control group. Four days of glucagon administration (1 mg/kg) produced increases in specific activity of cytochrome oxidase and lipoprotein lipase. After 8 days of glucagon administration, changes in whole-pad activity similar to those seen with 18 days of administration were present. Glucagon also increased whole-pad lipoprotein lipase activity after 4 and 8 days. Surgically denervated interscapular brown adipose tissue retained its ability to respond to exogenous glucagon, though the magnitude of the response was diminished. Guanosine 5'-diphosphate (GDP) binding to brown adipose tissue mitochondria was measured as an assessment of functional state after 5 days of glucagon (1 mg/kg). There was an increase in GDP binding relative to controls whether expressed as picomoles per milligram mitochondrial protein or nanomoles per pad.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue, Brown

Tubuloglomerular feedback responses with native and artificial tubular fluid.

Experiments were performed to compare the tubuloglomerular feedback response to native and artificial tubular fluid. The change in early proximal flow rate produced by changes in loop of Henle flow rate was measured in anesthetized rats using micropuncture techniques. Loop perfusion fluid was either an artificial solution with an electrolyte composition similar to that of proximal fluid (ATF) or native tubular fluid (NTF) collected from the late proximal tubule. In control rats, in rats on a low NaCl diet, in rats on restricted food intake, and in acutely saline-expanded rats no differences were detected between ATF- and NTF-perfused nephrons. In rats receiving 10 g NaCl/100 g, diet, responses with ATF and NTF to a flow change from 0 to 15 nl/min did not differ significantly; maximum feedback responses (flow change from 0 to 40 nl/min) were, however, significantly greater with ATF (-25.4%) than NTF (-12.9%). Chloride absorption was not different with the two perfusates. With both ATF and NTF a significant negative correlation was found between maximum responses and NaCl intake, with the slope being steeper with NTF. We conclude that some unidentified constituent of tubular fluid affects maximum feedback responses during very high NaCl intake, but feedback responses to physiological flow rate changes appear to be independent of luminal factors.

Animals

Role of the renin-angiotensin system in tubuloglomerular feedback.

The link between the renal tubule and glomerular vasculature comprised of the juxtaglomerular apparatus appears to serve two functions: the regulation of filtration rate and of renin secretion. Elevation of macula densa NaCl concentration stimulates a vasoconstrictor response, which results in a fall in filtration rate, a response that has been termed tubuloglomerular feedback (TGF). Simultaneously, renin secretion is suppressed. The two responses appear to be initiated by a furosemide-sensitive transport step probably located in the macula densa. Both show a pattern of anion specificity identical to Na/K/Cl cotransport mechanisms. An increase in intracellular calcium in the effector cells, the vascular smooth muscle, and the renin-containing granular cells is a likely effector mechanism for both reactions. Angiotensin probably does not mediate the vasoconstrictive feedback response, because changes in local (intracellular) angiotensin concentration would have to be opposite from systemic changes. However, acute changes in angiotensin levels appear to be an important modulator of the magnitude of the TGF response.

Angiotensin II

Fiber optic probe cytometer.

An optical fiber probe is used to both excite and collect fluorescence from a suspension of cells. The configuration of the probe is such that one or a few cells are sensed at a time, with a convenient cell concentration. With fluorescently labeled antibodies to cellular antigens, the fiber optic cytometer is able to identify the presence of a specific set of cells with high sensitivity.

ABO Blood-Group System

Alterations in the pyruvate dehydrogenase complex during adaptation to glucose by Neurospora.

A 20-fold induction of the pyruvate dehydrogenase complex, pyruvate dehydrogenase (EC 1.2.4.1) plus dihydrolipoate S-acetyltransferase, (lipoyltransacetylase) (EC 2.3.1.12) plus dihydrolipoyl dehydrogenase, NADH : lipoamide oxidoreductase, (EC 1.6.4.3), from a specific activity of 3.5-65.0 was observed in mitochondrial extracts during adaptation of Neurospora to glucose from acetate media. The extent of ATP-dependent, time-dependent inactivation of the pyruvate dehydrogenase complex was approximately the same in both acetate- and glucose-grown cells, thereby indicating that the low pyruvate dehydrogenase complex activities in acetate-grown cells did not represent phosphorylated pyruvate dehydrogenase complex molecules. High levels of dihydrolipoyl transacetylase (EC 2.3.1.12) were observed in mitochondrial extracts from acetate-grown cells; this lipoyltransacetylase was analyzed on sucrose density gradients and found to be associated with the pyruvate dehydrogenase complex. Digitonin fractionation of mitochondria revealed that both the pyruvate dehydrogenase complex and lipoyltransacetylase were primarily associated with the mitochondrial outer membrane.

Acetates