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Biomedical subjects

A Lambert

Publications and source records attributed to A Lambert.

At least 145 records · Page 8Linked to original sources
PubMed1984

Computerized continuous monitoring of cytochemical enzyme reaction product formation by the Vickers M85A microdensitometer.

An experimental set-up for the continuous monitoring of cytochemical enzyme reaction rates is described. The design consists of a 32K Commodore PET microcomputer, upgraded to Basic 4 and containing a graphics facility. The microcomputer is interfaced with a Vickers M85A microdensitometer, which has been modified and equipped with a Zeiss X63 water immersion objective. The software allows for the automatic transfer of density readings from the M85A to the microcomputer, and for the continuous monitoring of the enzyme reaction as its proceeds at short time intervals (i.e. down to 3.3 s). The relationship between the absorbance change of the enzyme final reaction product and time is printed both numerically and graphically. Initial velocity rates are calculated by regression analysis, and statistical evaluation of grouped data is possible.

20-Hydroxysteroid Dehydrogenases↗
PubMed1984

Cortisol production by dispersed guinea-pig adrenal cells; a specific, sensitive and reproducible response to ACTH....and its fragments.

Naturally occurring steroids and peptide hormones, tested at supraphysiological concentrations, were without effect on basal and human (h) 1-39 ACTH (NIBSC code 74/555, 25 ng/l (5.5 X 10(-12) mol/l] stimulated cortisol production. Further, low concentrations of angiotensin II, N-pro-opiocortin (N terminal fragment 1-76) and gamma-MSH all of which have been reported to synergise with ACTH with regard to cortisol production, were without significant effect alone or in combination with ACTH over the range 2.2 X 10(-13) to 5.5 X 10(-12) mol/l. The activity of h 1-39 was compared with that of the ACTH related peptides 1-24, 1-18, 1-17, 1-16, 1-13-NH2 (alpha MSH), 1-10 and 4-10. The dose responses were parallel and the same maximal cortisol output was observed with all the peptides except the 1-10 fragment. Half maximal stimulation occurred at 3.1 X 10(-12) (1-24), 4.4 X 10(-12) (h 1-39), 1.5 X 10(-11) (1-39), 3.3 X 10(-10) (1-18), 5 X 10(-9) (1-13-NH2), 8 X 10(-9) (1-17), 2 X 10(-7) (1-16) and 1 X 10(-5) (4-10) mol/l respectively. Interference by the above ACTH-derived peptides in cortisol secretion by the cells in response to 5.5 X 10(-12) mol/l h 1-39 ACTH was minimal over the range 5.2 X 10(-12)-2.2 X 10(-6) mol/l. The sensitivity of the adrenal cells to h 1-39 ACTH was such that 2 ng/l (4.4 X 10(-13) mol/l) provoked cortisol secretion over the control (P less than 0.05, n = 17). The coefficient of variation within assay for each dose on the full standard curve (2.2 X 10(-13)-1.1 X 10(-10) mol/l) was less than 10% (n = 6). Half maximal stimulation was given by 14.5 ng/l (3.2 X 10(-12) mol/l). Between control and 1.1 X 10(-10) mol/l ACTH there was a 32 +/- 8 (mean +/- SD, n = 9) fold change in cortisol production.

Adrenal Glands↗
PubMed1984

Preliminary experiences with a bioassay for adrenocorticotrophin (ACTH) in unextracted human plasma using dispersed guinea-pig adrenal cells.

A simple, accurate and precise dispersed cell bioassay suitable for the assay of ACTH levels greater than 50 ng/1 in unextracted human plasma is described. Isolated adrenal cells were prepared by tryptic digestion of the guinea-pig adrenal gland and cortisol production by these cells was controlled specifically by ACTH. The cortisol response to 2 ng/1 of human pituitary ACTH (1-39) was always significantly (P less than 0.05, n = 17) different from the control. ACTH concentrations between 1 and 20 ng/1 gave a 14 +/- 3 fold (n = 17) increment in cortisol response. Dilutions of test plasmas were parallel with standard ACTH (1-39) when the plasma concentrations were maintained throughout at 4% (1 in 25 dilution) by the addition of dexamethasone-suppressed plasma ('ACTH free'). Using a 25 fold plasma dilution, the limit of detection of ACTH in unextracted plasma was 50 ng/1. Recovery of ACTH (1-39) spiked into dexamethasone suppressed plasma at concentrations of 250 and 100 ng/1 was 97 +/- 10% (n = 17) and 114 +/- 15% (n = 17) respectively. The within assay coefficient of variation (CV) of both quality control and patients' samples never exceeded 9% while between assay variation was less than or equal to 13%. The sample throughput was 30 plasma samples/week/technician.

Addison Disease↗
PubMed1984

On the site of action of the anti-adrenal steroidogenic effect of etomidate and megestrol acetate.

The sites of action of the anti-steroidogenic action of etomidate and megestrol acetate have been established with a novel in vitro approach based upon the inhibition of cortisol (Co) secretion by dispersed guinea-pig adrenal cells. The cells were challenged with the Co precursor steroids (all at 10(-5) mol/l) pregnenolone (Pe), 17-hydroxy-pregnenolone (17-Pe), progesterone (Po), 17-hydroxyprogesterone (17-Po), 21-deoxycortisol (21-DOC) and 11-deoxycortisol (11-DOC), or 1-24 adrenocorticotrophin (ACTH, 100 ng/l) in the presence or absence of either etomidate, megestrol acetate, metyrapone or trilostane (all at 5 X 10(-5) mol/l). In the absence of drug, the steroid precursors or ACTH provoked a cortisol secretion of greater than 14 times that secreted by cells incubated in their absence. ACTH-stimulated Co secretion was inhibited by greater than 85% by all the drugs employed. In the presence of trilostane and megestrol acetate, Co secretion provoked by the delta 4 3-keto steroids (Po, 17-Po, 21-DOC and 11-DOC) was similar to the controls. However, with the delta 5, 3 beta-hydroxy steroids, 17-Pe and Pe, Co secretion was inhibited by greater than 57% in the presence of these drugs. In contrast, etomidate and metyrapone inhibited Co secretion by greater than 60% when 11-deoxycortisol was employed, indicative of a block at 11 beta-hydroxylase, the final step in the cortisol biosynthetic pathway. Similar results were seen with Pe, 17-Pe, Po and 17-Po, all of which are converted to cortisol via a biosynthetic route which includes catalysis by 11 beta-hydroxylase.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxysteroid Dehydrogenases↗
PubMed1983

A sensitive silver stain for proteins in agarose gels.

A silver stain for proteins in agarose gels which is at least 10 times as sensitive as Coomassie blue is described. The method is simple to use and is particularly useful for the study of protein bands in the gamma region on electrophoresis of fluids such as cerebrospinal fluid in which the protein concentration is low. It readily detects bands of IgG containing 20 to 40 ng/band (approx 3 to 6 ng of IgG/mm2 of gel).

Electrophoresis, Agar Gel↗
PubMed1982

Characteristics of the response of dispersed guinea-pig adrenal cells to low doses (1-50 pg/ml) of alpha 1-24 adrenocorticotrophin.

Isolated adrenal cells prepared by tryptic digestion of the guinea-pig adrenal gland are sensitive to low concentrations (less than 25 pg/ml) of adrenocorticotrophin (ACTH). Cell which have been pre-incubated for 2 h. centrifuged and resuspended in fresh culture medium prior to the introduction of 10 pg/ml ACTH for 60 min show a marked increase (328 +/- 109 nmol/l; mean +/- SD) in cortisol secretion over the control compared to freshly dispersed cells (75 +/- 45 nmol/l). Further potentiation of the ACTH effect was seen with the pre-incubated cells by suplementing the medium with calcium (8 mM) and ascorbate (2 mM) but not with theophylline (1 mM). Basal cortisol secretion was not affected by any of the additives. In the presence of 8 mM calcium and after 60 min incubation 10 pg/ml ACTH stimulated cortisol secretion from 328 nmol/l over the control to 839 +/- 382 nmol/l. The effect of ascorbate (2 mM) was to further increase the effect of ACTH at all dose levels tested (1-25 pg/ml). The concentration of ACTH required to provoke half maximal cortisol secretion decreased from 95 pg/ml with normal medium to 12 pg/ml with calcium -ascorbate supplemented medium. Using this supplemented medium the cells were sensitive to 1 pg/ml and cortisol secretion was stimulated 10-fold over the control with 50 pg/ml, a dose which saturated the system.

Adrenal Glands↗
PubMed1982

In vitro activity of temocillin (BRL 17421), a novel beta-lactam antibiotic.

The minimal inhibitory concentration of temocillin (BRL 17421) against 476 clinical isolates was determined by an agar dilution method. Temocillin was active against most of the Enterobacteriaceae, Haemophilus, and Neisseria strains tested. The compound showed low activity or was inactive against Bacteroides, Campylobacter, Acinetobacter, Pseudomonas, and Staphylococcus aureus strains. Within each species, individual strains showed a high degree of uniformity in their susceptibility to temocillin; the drug concentrations that inhibited the growth of 90% of organisms were the same or close to those which inhibited the growth of 50% of organisms. In contrast, the same strains showed a very wide range of susceptibility to the other antibiotics tested, including third-generation cephalosporins. Against strains of Enterobacteriaceae highly susceptible to third-generation cephalosporins, temocillin was considerably less active than cefotaxime, ceftazidime, and moxalactam, although it was more active than cefazolin and piperacillin. Against certain strains of Enterobacter and Citrobacter resistant to third-generation cephalosporins, temocillin was more active than cefotaxime and ceftazidime. An increase in the inoculum size did not alter the activity of temocillin, indicating that the compound has high stability to beta-lactamases. The minimal lethal concentration was also very similar to the minimal inhibitory concentration when an inoculum of 10(5) colony-forming units was used.

Bacteria↗
PubMed1982

[The oncomiricidium of Diplozoon nipponicum Goto, 1891. A new type of larval haptor for the genus Diplozoon (Monogenea, Discocotylidae)].

Diplozoon nipponicum Goto 1891, a branchial parasite of Cyprinus carpio is mentioned for the first time in France and Western Europe. Carps, along with their Monogenea, were introduced from Hungary. After being obtained experimentally, the oncomiracidium is thoroughly studied (excretory system, ciliated cells, chaetotaxy, haptor). A new haptor for the genus Diplozoon is described: two pairs of larval lateral hooklets are present. The taxonomic and phylogenic implications of these observations are discussed for Diplozoon and Discocotylidae. Mainly as a result of the study of the haptor and the egg structure, it appears that two groups occur, the taxonomic and evolutive significance of which are to be discussed in Diplozoon.

Animals↗
PubMed1981

Inhibition of canine duodenal interdigestive myoelectric complex by nutrient perfusion of jejunal and ileal Thirty-Vella loops.

The mechanisms by which the intestinal interdigestive myoelectric complex (IDMEC), recurring at about 90 minute intervals in the fasted dog, is disrupted by feeding remain unknown. We investigated whether the IDMEC could be disrupted in the duodenum by perfusing a Thiry-Vella loop with glucose in the dog. An intestinal Thiry-Vella loop, measuring one half (80 to 160 cm) of the total length of the small bowel was constructed in four dogs from the jejunum, and in four other dogs from the ileum. Extracellular nichrome electrodes were sewn on the duodenum for recording the electrical activity of the intestine. After three weeks' recovery, electrical recordings were performed in the fasted dogs in order to observe whether the IDMEC persisted in the duodenum when the Thiry-Vella loops were perfused, at different days, for four hours with solutions made of either (1) NaCl 154 mM, (2) NaCl 308 mM, (3) glucose 300 mM, or (4) glucose 600 mM, at a rate of 8 ml/min. NaCl 308 mM and glucose 600 mM were also delivered at a rate of 4 ml/min. Glucose output from the Thiry-Vella loops was measured throughout the experiments over consecutive five minute intervals. Each experiment was performed three times in each dog. The results showed that perfusing the Thiry-Vella loops with NaCl 154 mM or NaCl 308 mM did not suppress the IDMEC in the duodenum whether the flow rate was 4 or 8 ml/min. On the contrary, perfusing the jejunal loops with glucose 300 mM disrupted the IDMEC in 54% of the experiments; perfusing glucose 600 mM disrupted the IDMEC in 83% of the experiments. In the ileal Thiry-Vella loop experiments, the IDMEC was disrupted in 33% of the cases with glucose 300 mM and in 66% of the cases with 600 mM. No significant difference was observed with glucose 300 mM delivered at a rate of 8 ml/min and glucose 600 mM delivered at a rate of 4 ml/min. Finally, the inhibitory effect of perfusing the Thiry-Vella loops with glucose increased as the amount of absorbed glucose increased. These results indicate that interruption of the IDMEC by feeding probably involves extraintestinal factors. These factors do not seem to be specific for any one part of the small intestine, but they seem to be activated by intestinal absorption.

Action Potentials↗