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Biomedical subjects

A White

Publications and source records attributed to A White.

At least 379 records · Page 21Linked to original sources

Enzyme-linked immunosorbent assay for detection of Staphylococcus epidermidis antibody in experimental S. epidermidis endocarditis.

The immune response against Staphylococcus epidermidis, as determined by an enzyme-linked immunosorbent assay, was evaluated in experimental S. epidermidis infections in rabbits. Antigens from 8 of 10 clinical S. epidermidis strains detected significant antibody production in five rabbits immunized with different strains of S. epidermidis and in five of six rabbits with experimental endocarditis caused by four different strains. The antigens from two strains detected antibody production in all rabbits, and strain ATCC 14990 discriminated best between positive and negative samples. Consecutive blood samples from rabbits with endocarditis and control rabbits with bacteremia, which were successfully prevented from developing endocarditis by using prophylactic antibiotics, were examined by using an enzyme-linked immunosorbent assay and an ultrasonic extract of strain ATCC 14990 as the antigen. This assay discriminated between rabbits with endocarditis and rabbits with uncomplicated bacteremia. Antibody production was detected as early as 3 days after the onset of infection in rabbits with endocarditis.

Animals↗

Demonstration of specific high affinity receptors for aldosterone in cytosol of rat colon.

Since both aldosterone and glucocorticoids increase cation transport in rat distal colon, and a specific glucocorticoid high affinity cytosolic receptor has been identified in this tissue, it was possible that the action of aldosterone was dependent on interaction with the glucocorticoid receptor. Studies were, therefore, performed to determine whether a specific high affinity receptor for aldosterone was present in rat distal colon. At 4 C, aldosterone binding was saturable and exhibited a high affinity site with an apparent Kd of 6.2 +/- 0.9 X 10(-10) M and a calculated number of binding sites of 57.2 +/- 10.8 fmol/mg cytosol protein. Scatchard plot analysis also revealed a low affinity site with a Kd of 5.9 +/- 1.1 X 10(-8) M and 961 +/- 191 fmol/mg cytosol protein-binding sites. Competitive binding studies demonstrated that the high affinity binding protein was specific for aldosterone, compared to either dexamethasone or RU-28362. Since a specific high affinity receptor protein for aldosterone is present in rat distal colon, these data are consistent with a direct action of aldosterone that is independent of the glucocorticoid receptor system.

Aldosterone↗

Effect of corticosteroids on electrolyte transport in the descending colon of the rat.

The effect of corticosteroids on electrolyte movement was studied in an everted sac model from distal rat colon. Corticosterone had a significant stimulatory effect on sodium and fluid transfer in a dose-related manner. At the low dose of 10(-9) M corticosterone increased sodium absorption by 29.4 microEq X g-1 X h-1, a value that was statistically greater than the effect of 10(-9) M dexamethasone. At the higher concentration of 10(-7) M the effect of corticosterone on sodium movement was twice that of aldosterone or dexamethasone in equimolar concentrations. By contrast, the effect of corticosterone on potassium movement was biphasic: at lower concentration (10(-9) M) a significant net secretion occurred, whereas potassium absorption was observed at 10(-7) M corticosterone. Dexamethasone significantly increased net potassium secretion, meanwhile the effect of aldosterone on potassium movement was not significantly different from controls. These data suggest that the native glucocorticoid, corticosterone, exerts regulatory control of colonic fluid and electrolyte function.

Aldosterone↗

Characterisation of monoclonal antibodies to adrenocorticotrophin.

We have produced 8 monoclonal antibodies to adrenocorticotrophin (ACTH) derived from immunisation with ACTH (1-24) conjugated to bovine serum albumin or ACTH (1-39) linked to chicken immunoglobulin by a novel method. Antibody specificity was assessed by studying the binding of purified human ACTH, synthetic ACTH (1-24), fragments of ACTH and peptides from the ACTH precursor molecule which have sequence homology. A wide range of specificities was demonstrated. Thus antibody 3H9 recognises the extreme N-terminal sequence (ACTH 4-10), antibody 1A12 is specific for residues 10-18 including alpha MSH, antibody 1D1 is specific for the mid N-terminal sequence but not alpha MSH, and antibody 2A3 is specific for the C-terminal portion (ACTH 18-39). These monoclonal antibodies can be easily purified and labelled and their defined specificities make it possible to select antibody combinations which provide the basis for 2-site immunometric assays for ACTH.

Adrenocorticotropic Hormone↗

Extracts of feverfew inhibit granule secretion in blood platelets and polymorphonuclear leucocytes.

Extracts of feverfew (Tanacetum parthenium) inhibited secretory activity in blood platelets and polymorphonuclear leucocytes (PMNs). Release of serotonin from platelets induced by various aggregating agents (adenosine diphosphate, adrenaline, sodium arachidonate, collagen, and U46619) was inhibited. Platelet aggregation was consistently inhibited but thromboxane synthesis was not. Feverfew also inhibited release of vitamin B12-binding protein from PMNs induced by the secretagogues formyl-methionyl-leucyl-phenylalanine, sodium arachidonate, and zymosan-activated serum. Feverfew did not inhibit the secretion induced in platelets or PMNs by the calcium ionophore A23187. The pattern of the effects of the feverfew extracts on platelets is different from that obtained with other inhibitors of platelet aggregation and the effect on PMNs is more pronounced than has been obtained with very high concentrations of non-steroidal anti-inflammatory agents.

Blood Platelets↗

A study of 19-O-carboxymethyl ether and 19-hemisuccinate derivatives of testosterone: their immunogenicity and use as iodinated radioligands for radioimmunoassay of testosterone.

Testosterone 19-O-carboxymethyl ether (T19C) and 19-hemisuccinate (T19H) derivatives were synthesised, and coupled to bovine serum albumin (BSA) or porcine thyroglobulin (PT) for immunogens or to iodohistamine for radioligands. The immunogenicity of these conjugates in mice was compared with those of testosterone 3-O-carboxymethyloxime and 15 beta-thioethyl conjugates. Of 10 immunogens studied, those linked to PT gave the highest antiserum titres and more sensitive standard curves. The site of conjugation (19 or 3 position) had little effect on immunogenicity. Cross-reactivity with 5 alpha-dihydrotestosterone (DHT) was in the range 22-100%, for the 19-linked immunogens. Antisera to T19C and T19H conjugated to PT were then raised in rabbits and characterised with 4 radioligands. Homologous assay systems in which the chemical bridge was identical in immunogen and 125I-radioligand gave the highest antiserum titres but the poorest assay sensitivity while those heterologous with respect to bridge or site gave the most sensitive standard curves. Rabbit antisera to both T19C and T19H immunogens showed good specificities with respect to DHT (range of cross reactions 0.78-21.1%), androstenedione, AN (range 0.45-2.3%) and progesterone, Po (range 0.05-1.4%) with all radioligands. The best assay system employed an antiserum to the T19H-PT immunogen with heterologous radioligand [125I]T19C. It had a detection limit of 15pg/tube and low cross-reactivity with DHT (0.78%), Po (0.3%) and AN (0.43%). We conclude that 19-linked derivatives of testosterone conjugated to PT are good immunogens and the antisera when combined with [125I]testosterone radioligands which are heterologous with respect to the chemical bridge, provide highly specific assays for testosterone with potential for clinical application.

Animals↗

Monoclonal antibodies to testosterone: the effect of immunogen structure on specificity.

A 15 beta-thioalkyl derivative of testosterone conjugated to bovine serum albumin (BSA) was synthesised and used to produce monoclonal antibodies. These antibodies were evaluated using 15 beta-(2-carboxyphenylthio)-testosterone [125I]histamine as radioligand. Out of 1368 hybrids, 5 secreted anti-testosterone antibodies. These were compared with monoclonal antibodies derived from immunisation with testosterone-3-carboxymethyloxime-BSA. The first group of monoclonal antibodies all showed very low cross-reactivity with 5 alpha-dihydrotestosterone (less than 2.8%) indicating that this site of linkage is a good choice for discriminating between differences at the 4-5 position in the A-ring on the testosterone molecule. However they generally showed much higher cross-reactivity with progesterone and androstenedione than monoclonal antibodies raised to the 3-linked immunogen. Nevertheless within each fusion there were monoclonal antibodies with markedly different specificities. None of these antibodies could be considered suitable for use in a testosterone immunoassay, but it does suggest that an antibody with an improved specificity profile could be found using the monoclonal antibody approach.

Animals↗

Multiple hormone secretion by a human pancreatic glucagonoma in culture.

A patient presenting clinically with the glucagonoma syndrome had high plasma glucagon levels (1920 ng/l) and at laparotomy, a pancreatic islet cell tumour was removed. The tumour was dispersed and placed in culture where it remained viable for 63 days. The tumour cells secreted immunoreactive (IR) glucagon at levels up to 2400 ng/l as detected by a C-terminal glucagon specific antibody and 85 400 ngequiv./l as measured by an N-terminal glucagon specific antibody. The difference between these two levels was attributed to the presence of different molecular forms of glucagon measured with the N-terminal specific antibody. IR insulin (up to 302 mU/l) and IR somatostatin (up to 2500 ng/l) were also detected. There was no direct or inverse correlation between different hormone levels. Small but significant levels of N-terminal and C-terminal vasoactive intestinal peptide (VIP) were detected in some cultures but there was no evidence of gastrin or ACTH. Glucagon and somatostatin secretion persisted for the duration of the culture (63 days) but insulin concentrations declined. Incubation of cultures with somatostatin (1 ng/ml) caused a 75% decrease in glucagon levels, while insulin (1000 mU/l) produced a 70% inhibition of somatostatin.

Adenoma, Islet Cell↗

The effects of homogeneous human prealbumin on in vitro and in vivo immune responses in the mouse.

A highly purified preparation of human prealbumin was shown to potentiate the sensitivity of rosette spleen forming cells of adult thymectomized mice to azathioprine in vitro and in vivo and to induce the appearance of the Thy 1, 2 antigen in vitro on spleen cells of adult thymectomized mice. Prealbumin also enhanced IgM antibody synthesis to sheep red blood cells (SRBC) in vitro in 12 week old mice and in vivo in aged (45 - 58 week old) and nude (nu/nu) mice. In vivo administration, to mice that had been pre-treated with hydrocortisone, resulted in a decrease in the specific activity of thymocyte terminal deoxynucleotidyl transferase. The data indicate that the prealbumin molecule possesses immunopotentiating properties in a number of in vitro and in vivo immunocompromised murine models and that the immuno-enhancing properties of the partially purified preparation previously described were in fact due to the prealbumin component and not to other contaminating proteins.

Adjuvants, Immunologic↗

The influence of hormones and inflammatory agents on C-reactive protein, cortisol and alanine aminotransferase in the plaice (Pleuronectes platessa L.).

Endotoxin stimulates production of both C-reactive protein (CRP) and cortisol in the plaice within 24 hr. Cortisol alone (optimum dose i.p. 500 micrograms/300 g wt fish) also stimulates CRP production and the possibility that endotoxin acts through cortisol was examined. Dexamethasone suppresses cortisol production but elevates CRP. Cortisol levels are restored to normal within 24 hr of endotoxin injection. Turpentine and ACTH which stimulate cortisol do not affect CRP. Endotoxin and cortisol have no significant effect on alanine aminotransferase activity in the serum and liver although it is elevated in the serum within 24 hr of the administration of adrenalin or turpentine.

Adrenocorticotropic Hormone↗

Analgesia following femoral neck surgery. Lateral cutaneous nerve block as an alternative to narcotics in the elderly.

In a prospective controlled randomised trial on patients undergoing operative repair of fractured neck of femur via a lateral incision, the postoperative analgesic requirements of one group of patients who received a lateral cutaneous nerve block were compared with a second group who received no block. The former group were found to need significantly less intramuscular pethidine in the first 24 hours, and 44% required no supplementary analgesia whatsoever during this period. The time to first dose of opioid in the remainder was greatly increased. No untoward sequelae associated with the nerve block were seen.

Aged↗

Effect of theophylline on nuclear retention of oestrogenreceptor complexes: correlation with oestrogen responses.

In the ovariectomized adult rat uterine oedema induced by 0.01 and 0.1 micrograms oestradiol-17 beta/100g body weight increased further in the presence of theophylline. Nuclear retention of oestrogen-receptor complexes also increased in response to theophylline both in vivo and in vitro. Theophylline decreased the number of eosinophils in the blood and concurrently decreased oestrogen-induced uterine eosinophilia at doses of 0.001, 0.01, 0.1, 1, 10 or 30 micrograms oestradiol/100 g body weight, through a mechanism independent of glucocorticoids. There was, therefore, no correlation between changes in the number of uterine eosinophils and changes in uterine wet weight induced by theophylline and oestrogen. It is suggested that the presence of oestrogen-receptor complexes in the nucleus for at least 4 h is a prerequisite for the induction of uterine oedema and growth in the presence of theophylline and oestradiol-17 beta.

Animals↗

A model of ovine endotoxemia characterized by an increased cardiac output.

High cardiac output sepsis is a major clinical problem. We have designed a sheep endotoxin model to simulate this condition and have evaluated how closely it approximates the clinical situation. The animals were prepared for chronic study by the implantation of cardiopulmonary catheters. One week later, endotoxin (0.75 microgram/kg/30 min) was administered following baseline measurements, and the animals were studied for an additional 15 hr. From 6-15 hr after the administration of endotoxin, there was a statistically significant twofold increase in cardiac output. Simultaneously, the total peripheral vascular resistance and mean arterial pressure was reduced. Eicosanoid measurements made at this time indicated that the vasodilator prostanoid, prostacyclin, was not elevated. A high cardiac output (hyperdynamic) model of sepsis has thus been established by the 30 min infusion of a small quantity of endotoxin. Prostacyclin is not a mediator of this response.

Animals↗

Prevention of infections of skin and skin structures.

Treatment of nasal carriers of coagulase-positive staphylococci with a wide variety of topical systemic antibiotics suppressed these organisms only during treatment. Treatment of methicillin-sensitive or methicillin-resistant staphylococci with oral rifampin plus cloxacillin, vancomycin, fusidic acid, or trimethoprim-sulfamethoxazole eradicated the colonizing Staphylococcus aureus in approximately 80 percent of studies. An alternative approach is replacement of virulent S.aureus with the 502 A S. aureus strain. The latter approach has aborted epidemics of staphylococcal diseases in newborn nurseries and has prevented recurrent furunculosis.

Adult↗

Wound sepsis after low risk elective cholecystectomy: the effect of cefuroxime.

A prospective randomized double-blind study was performed to assess the value of a single intravenous injection of 1.5 g cefuroxime in the prevention of wound sepsis after low risk elective cholecystectomy. Wound infection developed in 10 of 81 control patients compared with 2 of 79 treated patients (P less than 0.05). Wound sepsis occurred more commonly in patients with infected bile or with a positive culture from a closing wound swab. Cefuroxime significantly reduced the isolation of staphylococci but not other organisms from the closing wound swab and also produced a significant reduction in wound infection in patients with sterile bile.

Cefuroxime↗

The bacteriology of the urine and renal calculi.

The bacteriological status of the pre-operative urine, removed calcium and the pelvic urine was studied in 63 consecutive patients undergoing removal of intrarenal calculi. The overall infection rates in the pre-operative urine, removed calculi and pelvic urine were 29%, 38% and 30% respectively. In patients with staghorn calculi, 58% of patients had an infected pre-operative M.S.S.U., and 82.5% of removed calculi demonstrated significant bacterial growth, with Proteus mirabilis being the predominant organism isolated. In patients with single or multiple oxalate calculi, 17% had an infected pre-operative M.S.S.U., and 22% of removed calculi demonstrated a significant bacterial growth. The findings would indicate that the presence of a positive M.S.S.U. in a patient with a renal calculus is an indication for surgical removal.

Adult↗

Radioimmunoassay of serum cortisol in the plaice (Pleuronectes platessa L.).

The validity of a radioimmunoassay kit for direct measurement of cortisol in the plaice is described. This technique was used to determine the cortisol concentration in adult male and female plaice in 100 microliter of either plasma or serum. There was no significant difference in the serum cortisol values determined by radioimmunoassay either with or without chromatography. Storage of whole blood at 4 degrees for varying periods up to 24 hr, before removal of the serum, did not affect the cortisol concentration. Decreases also did not occur in serum samples kept at room temperature for up to 5 days, although losses of 1-12% occurred after 7 days and 11-39% after 8 days. The daily variations in serum cortisol levels were examined in fish exposed at 11 degrees to 12 hr light: 12 hr dark over a 32-hr period and the existence of a 24-hr cortisol rhythm was observed. Blood samples were taken from the same six fish every 4 hr, each fish being bled nine times during the experiment. Blood samples were also taken every 4 hr from groups of seven fish, each fish during this experiment being bled only once. Measurement of monthly serum samples throughout 1 year showed cortisol concentrations at a maximum in April, during the peak spawning period. There was no significant difference between cortisol levels in male and female plaice, except in January, and no difference between serum and plasma values in either sex.

Animals↗