PubMed Health⌕ Search

Biomedical subjects

M D Page

Publications and source records attributed to M D Page.

At least 37 records · Page 2Linked to original sources
PubMed1993

Mutants of Methylobacterium extorquens and Paracoccus denitrificans deficient in c-type cytochrome biogenesis synthesise the methylamine-dehydrogenase polypeptides but cannot assemble the tryptophan-tryptophylquinone group.

Five mutants of Methylobacterium extorquens and four mutants of Paracoccus denitrificans that have a general defect in c-type cytochrome synthesis also failed to assemble an active methylamine dehydrogenase. In all cases methanol dehydrogenase, another periplasmic enzyme, was fully active. All nine mutant strains accumulated both the heavy and light subunits of methylamine dehydrogenase to essentially wild-type levels. In all nine mutants, the heavy-subunit and light-subunit polypeptides were proteolytically processed, suggesting that translocation to the periplasm had occurred; in the case of the P. denitrificans mutants, a periplasmic location for the heavy and light subunits was confirmed experimentally. While specific quinone staining of the methylamine dehydrogenase light subunit in wild-type M. extorquens and P. denitrificans strains could readily be demonstrated, the light subunit polypeptides accumulated by the mutants did not quinone stain, indicating that the methylamine dehydrogenase prosthetic group, tryptophan tryptophylquinone, is not assembled in the absence of functional c-type cytochromes.

Benzoquinones↗
PubMed1993

Insertion of transposon Tn5 into a structural gene of the membrane-bound nitrate reductase of Thiosphaera pantotropha results in anaerobic overexpression of periplasmic nitrate reductase activity.

Chlorate-resistant mutants of the denitrifying bacterium Thiosphaera pantotropha were generated by transposon Tn5 mutagenesis. One class was deficient in membrane-bound nitrate reductase activity but retained a periplasmic nitrate reductase activity. Using transposon marker rescue it was shown that in one such mutant, M-6, the transposon was inserted in the membrane-bound nitrate reductase beta subunit structural gene (termed narH in order to be consistent with the nomenclature of the Escherichia coli major nitrate reductase operon). The translated sequence (total of 106 amino acids) from around the point of transposon insertion showed approximately 90% amino acid identity with the beta subunits of the E. coli nitrate reductases. Under anaerobic growth conditions M-6 overproduced the periplasmic nitrate reductase activity allowing anaerobic growth with nitrate as electron acceptor. A regulatory link was inferred between the presence of the membrane-bound nitrate reductase and expression of the periplasmic nitrate reductase. This is the first demonstration of full denitrification in an organism possessing only a periplasmic nitrate reductase.

Amino Acid Sequence↗
PubMed1993

An audit of dynamic function tests: errors in the timing of blood specimens.

OBJECTIVE: The interpretation of dynamic endocrine investigations is dependent on the timing of blood sampled after a stimulus. We have studied the accuracy with which doctors adhered to a prescribed protocol. DESIGN: A new request form for dynamic tests was designed to include both the planned and the actual time of blood sampling in order to allow assessment of the variance of sample timing. MEASUREMENTS: We followed 152 investigation procedures comprising 791 planned specimens. A total number of 627 timing intervals could be analysed of which 366 timing intervals did not differ from the planned interval. Protocols were divided into long and short (greater or less than 6 hours) and into those governed by clock times or 'stopwatch', i.e. minutes elapsed, intervals. RESULTS: A total of 22% specimens were taken more than 10 minutes earlier or later than planned. Six per cent of specimens were taken more than 30 minutes earlier or later than planned. CONCLUSIONS: The data presented in this study suggest that in order to improve the interpretative accuracy of dynamic tests, it would appear prudent to pay more attention to the accuracy of sample timing. New dynamic test publications should include time tolerance for the normal response.

Endocrine System Diseases↗
PubMed1993

Thyrotoxicosis increases right to left shunt in congenital cyanotic heart disease.

We wish to report two cases of congenital cyanotic heart disease in whom coexisting thyrotoxicosis increased the right to left shunt. We review the literature on the cardiac and respiratory effects of thyroid hormone to try to explain this phenomenon. Two patients, one with Fallot's tetralogy (diagnosed at 68 years of age), the other with a ventricular septal defect, developed thyrotoxicosis which resulted in a dramatic but reversible deterioration in their cardiac condition. We conclude that thyrotoxicosis increases right to left shunt in congenital cyanotic heart disease. This effect is profound but reversible. We suggest that when an unexplained rapid deterioration occurs in patients with congenital cyanotic heart disease, thyrotoxicosis should be excluded.

Adult↗
PubMed1993

A novel device for the rapid in-clinic measurement of haemoglobin A1c.

In order to evaluate a novel technique for measuring haemoglobin A1c using a cassette-based immunoassay method we have undertaken a laboratory evaluation using 48 patient samples and a separate within-clinic prospective study of 59 diabetic patients. Individuals were recruited from general (48 patients), paediatric (19 patients), obstetric (24 patients), and general practice (15 patients) diabetic clinics. Agreement was evaluated between HbA1c results obtained using the new method (AMES DCA 2000) and an established laboratory assay (DIAMAT HPLC system). The mean differences between the two results (AMES DCA 2000-DIAMAT) (95% confidence intervals) were: laboratory -0.69% (-1.42 to 0.04%); paediatric clinic -0.93% (-1.93 to 0.07%); obstetric clinic -0.29% (-1.09 to 0.51%), and general practice clinic -0.77% (-1.3 to -0.24%). For the AMES DCA 2000, the coefficient of variation for HbA1c of 5.2% was 1.6% and for HbA1c of 13%, 2.4%. This instrument was used without difficulty by four different operators. Intra-assay coefficient of variation for each operator was < 3.4%. In 9/18 patients where a specific assessment of clinical utility was made, knowledge of the HbA1c result at the time of consultation lead to a change in treatment. Methodology of this type may be used successfully within a diabetic clinic setting in both primary care and hospital environments.

Adult↗
PubMed1993

Corticosteroid therapy and intercurrent illness: the need for continuing patient education.

In patients receiving long-term therapeutic or replacement corticosteroids, delayed or inappropriate adjustment of steroid dosage during intercurrent illness may be fatal. We used a questionnaire to assess current levels of patient knowledge, awareness of the need for action during intercurrent illness and the frequency with which steroid warning cards and Medic Alert pendants were carried, in 61 patients on long-term replacement corticosteroids and in 40 patients receiving long-term therapeutic corticosteroids. Only 67 of the 101 patients taking corticosteroids were carrying a steroid warning card. Eleven of the 21 Medic Alert owners wore their pendants. Only 18 of the 41 patients in the therapeutic group and 41 of the 60 patients in the replacement group would take appropriate action during an intercurrent illness (P < 0.001). Lack of patient knowledge in this important area emphasizes the need for continuing and effective education of these groups of patients during follow-up. An information sheet detailing the exact changes to be made during intercurrent illness may help reinforce verbal advice.

Acute Disease↗
PubMed1993

Practice and attitudes of primary care physicians in a rural southern state concerning HIV/AIDS patients.

In preparing for the development of a plan of care for persons with AIDS and other HIV related conditions, the HIV Services Planning Program, a joint effort of the Department of Family Medicine of the University of Mississippi Medical Center and the Bureau of Preventive Health of the Mississippi State Department of Health, conducted a survey of all primary care physicians throughout the state of Mississippi. The purpose of the survey was to determine health care services provided to persons with HIV/AIDS. Specific areas of interest included: (a) type of medical specialty; (b) location of primary practice; (c) services provided for persons with HIV/AIDS; and (d) utilization of universal blood and body fluid precautions, i.e., barrier techniques. Descriptive statistics were utilized in analyzing responses.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Immunochemiluminometric assays (ICMA) specific for growth hormone releasing hormone 1-44 NH2 and 1-40 OH.

We describe specific two-site immunochemiluminometric assays able to directly measure human growth hormone-releasing hormone 1-44 NH2 and 1-40 OH concentrations in unextracted plasma. A common N-terminal antibody was purified from polyclonal rabbit antisera to growth hormone-releasing hormone 1-44 NH2 on a growth hormone-releasing hormone 1-29 NH2 linked affinity column and labelled with chemiluminescent acridinium ester. C-terminal specific monoclonal antibodies to growth hormone-releasing hormone 1-44 NH2 and 1-40 OH were raised in Balb/C mice and used as solid phase antibodies. Assay of fasting specimens from normal individuals gave medians (and ranges) of 23 pg/ml (2-200) and 30 pg/ml (3-134) for growth hormone-releasing hormone 1-44 NH2 and 1-40 OH, respectively. Samples from a series of acromegalics showed that most have values in the normal range though median values were higher, 56 pg/ml for growth hormone-releasing hormone 1-44 NH2 (P < 0.001) and 52 pg/ml for 1-40 OH (P < 0.001). Using these assays it will be possible for the first time to directly study the physiology and pathophysiology of these two peptides.

Acromegaly↗
PubMed1992

Differential effects of arginine on growth hormone releasing hormone and insulin induced growth hormone secretion.

OBJECTIVE: We wished to investigate the interaction of arginine, GHRH and insulin stress on GH secretion. DESIGN: Six healthy, non-obese volunteers underwent seven separate studes in random order. They received (1) insulin alone at 0 minutes; (2) GHRH alone at 15 minutes; (3) arginine alone at 0-30 minutes; (4) arginine at 0-30 minutes and GHRH at 15 minutes; (5) insulin at 0 minutes and arginine at 0-30 minutes; (6) insulin at 0 minutes, GHRH at 15 minutes and arginine at 0-30 minutes; (7) insulin at 0 minutes and GHRH at 15 minutes. MEASUREMENTS: GH and PRL were measured from -30 to 150 minutes at intervals of 15 minutes. RESULTS: Arginine increased GH responses to GHRH and decreased GH responses to hypoglycaemia, but this inhibitory effect of arginine was reversed by GHRH. CONCLUSIONS: The findings suggest that arginine-induced GH release is mainly mediated by a decrease in somatostatinergic tone, while GH responses to insulin stress are probably mediated by both an increase in hypothalamic GHRH release and inhibition of somatostatin.

Adult↗
PubMed1992

Low recurrence rate after partial hypophysectomy for prolactinoma: the predictive value of dynamic prolactin function tests.

OBJECTIVE: To determine the factors influencing the outcome of transethmoidal partial hypophysectomy for suspected prolactinoma and the predictive value of pre and post-operative dynamic PRL function tests. DESIGN: A retrospective study of patients undergoing surgery for a suspected prolactinoma in Cardiff between 1979 and 1989. PATIENTS: Eighty-two hyperprolactinaemic patients (75 women, seven men) diagnosed as having a prolactinoma on the basis of dynamic PRL function tests, radiological investigation and exclusion of other causes. MEASUREMENTS: TSH and PRL responses to domperidone (10 mg i.v.) and TRH (200 micrograms i.v.) measured preoperatively, 2 months post-operatively, and annually thereafter. CT scan performed preoperatively in 58 patients. Operative findings, including adenoma size, documented in each case. RESULTS: Forty-two patients (51%) had microadenomas (less than 10 mm), 37 (46%) had macroadenomas and in three no tumour was found at operation. Preoperatively, normal responses of both TSH (incremental rise less than 2.0 mU/l) and PRL (greater than 100% rise) to domperidone were observed in two patients only: both had an abnormal vascular supply to the pituitary rather than an adenoma. Serum PRL was normalized in the early post-operative period (less than 72 h; 'early cure') in 65 patients (79%). The highest early cure rate (96%, n = 26) was in patients with adenomas of 5-9 mm, lower rates being achieved for lesions of 10-19 mm (80%, n = 30), less than 5 mm (63%, n = 19) or greater than or equal to 20 mm (57%, n = 7). The early cure rate was strongly correlated with preoperative PRL, ranging from 100% in patients with PRL less than 1000 mU/l (n = 13) to zero in those with PRL greater than 10,000 mU/l. Dopamine agonist therapy of between 5 weeks and 4 years duration prior to surgery was associated with a significantly reduced early cure rate (60 vs 94%, P less than 0.02) in macroadenoma but not microadenoma patients. Recurrent hyperprolactinaemia during mean follow-up of 51.7 months occurred in eight patients (12%), in five cases within 2 months of surgery and in the others at 26, 48 and 50 months. Recurrence could not be predicted from any preoperative parameter, but a serum PRL greater than 150 mU/l 1-3 days following microadenomectomy was associated with early recurrence and probably indicates failed surgery. An abnormal response of TSH to domperidone was documented 2 months post-operatively in 11/60 patients with normal basal PRL, and preceded all three late recurrences. Of four patients with abnormal responses of both PRL and TSH at this time, two have relapsed to date. CONCLUSIONS: In carefully selected patients, partial hypophysectomy is an acceptable alternative to medical treatment for prolactinoma. Preoperatively, dynamic tests accurately identified those patients whose hyperprolactinaemia was non-adenomatous in origin and, post-operatively, identified a subgroup of patients at increased risk of late recurrence.

Adult↗
PubMed1992

Prescribing and dispensing of insulin: margins for error?

To investigate recent anecdotal reports of incorrect insulin dispensing, we compared retail pharmacists' and doctors' understanding of commonly used insulin brands. Eighty pharmacists, 29 general practitioners (GPs), and 31 junior hospital doctors were interviewed and completed a questionnaire concerning the duration of action and formulation of 17 insulins. Views on dispensing of alternative insulins were sought from the pharmacists. Fifteen insulin pairs in two columns, 'insulin prescribed' and 'dispense instead', were presented and pharmacists and doctors asked if substitution was appropriate. Mixtard, Penmix and Actrapid were the best understood insulins, Insulatard, Humulin Zn and Protaphane the worst. Both GPs (p = 0.014) and hospital doctors (p < 0.001) scored better than pharmacists. No pharmacist stated that they would routinely dispense alternative insulin, 27 out of 80 not even in an emergency, though 37 were prepared to do so in such a situation after checking, usually with a doctor or in the relevant literature. Sixteen pharmacists, 24 GPs, and all 31 hospital doctors completed the section on insulin pairs but accuracy at identifying appropriate substitutions was poor. These findings coupled with confusing insulin nomenclature may lead to dangerous dispensing errors.

Dosage Forms↗
PubMed1991

Cholinergic blockade with pirenzepine induces dose-related reduction in glucose and insulin responses to a mixed meal in normal subjects and non-insulin dependent diabetics.

OBJECTIVE: The aim of the study was to assess the effects of cholinergic blockade with pirenzepine on glucose and insulin responses to a mixed meal in normal subjects and patients with non-insulin dependent diabetes (NIDDM). Further, to assess in normal subjects the relative importance of nocturnal GH suppression by pirenzepine. DESIGN: Placebo, 100 or 200 mg pirenzepine were given to the normal subjects 1 hour before a standard mixed meal. The effects of placebo or 200 mg pirenzepine at night on nocturnal GH secretion and subsequent breakfast carbohydrate tolerance were also studied. NIDDMs were given placebo or 200 mg pirenzepine before the meal. SUBJECTS: We studied six healthy male volunteers (ages 20-22, body mass indices 20.3-23.3) and ten NIDDMs (eight men, ages 42-74); five obese (BMI 25.5-31.8) and five non-obese (BMI 21.2-24.8). MEASUREMENTS: Serial plasma glucose and insulin concentrations were measured for 3 hours after a standard mixed meal. RESULTS: Acute pretreatment of normal male volunteers with pirenzepine produced a dose-related improvement in carbohydrate tolerance. Peak post-prandial plasma glucose levels were delayed and significantly reduced following 200 mg orally (6.5 +/- 0.1 mmol/l), but not following 100 mg (7.3 +/- 0.3), compared with placebo (7.6 +/- 0.3). Peak insulin levels were similarly delayed and reduced by the 200 mg dose only (36.5 +/- 6.1 mU/l, compared with 49.8 +/- 8.7). Suppression of nocturnal GH by 200 mg pirenzepine at night produced a small reduction in fasting plasma glucose (5.0 +/- 0.1 mmol/l, compared with 5.3 +/- 0.1, P less than 0.02) but did not improve subsequent breakfast carbohydrate tolerance. Peak plasma glucose in NIDDMs was reduced following pirenzepine (12.4 +/- 0.9 mmol/l) compared with placebo (14.3 +/- 1.0, P less than 0.01). This reduction was equally significant in obese and non-obese groups. Peak plasma insulin was also reduced by pirenzepine (22.4 +/- 3.9 mU/l) compared with placebo (42.4 +/- 5.3, P less than 0.01). Insulin suppression was quantitatively greater in obese than in non-obese patients. CONCLUSIONS: Improvement in carbohydrate tolerance after pirenzepine in normal subjects is dose related and largely independent of GH suppression. Cholinergic blockade can also improve meal carbohydrate tolerance with simultaneous reduction in plasma insulin concentrations in non-insulin dependent diabetics, particularly those with obesity.

Adult↗
PubMed1991

Effect of pyridostigmine and pirenzepine on GH responses to GHRH in hyperthyroid patients.

OBJECTIVE: We wished to investigate whether thyrotoxicosis can influence the cholinergic modulation of GH secretion. DESIGN: Pyridostigmine was given orally, then GHRH injected i.v., and levels were measured. In a separate study, pirenzepine was injected i.v., then GHRH, and growth hormone levels were measured. PATIENTS: Thyrotoxic patients were compared with normal subjects. MEASUREMENTS: GH was measured from -30 to +120 minutes at intervals of 15 minutes. RESULTS: Pyridostigmine markedly increased GH responses to GHRH in normal subjects, but not in thyrotoxic patients. Pirenzepine abolished the GH response to GHRH in thyrotoxic patients. CONCLUSIONS: GH responses to GHRH in hyperthyroid patients were suppressed by cholinergic muscarinic receptor blockade with pirenzepine. Activation of cholinergic pathways with pyridostigmine did not increase GH responses to GHRH in these patients. This may be a consequence of increased hypothalamic cholinergic function or reduced hypothalamic GHRH activity in hyperthyroidism. Our findings demonstrate a further mechanism by means of which thyroid status may affect the secretory activity of the somatotroph.

Adolescent↗
PubMed1990

The identification of cytochromes involved in the transfer of electrons to the periplasmic NO3- reductase of Rhodobacter capsulatus and resolution of a soluble NO3(-)-reductase--cytochrome-c552 redox complex.

The involvement of cytochromes in the electron-transport pathway to the periplasmic NO3- reductase of Rhodobacter capsulatus was studied in cells grown photoheterotrophically in the presence of nitrate with butyrate as carbon source. The specific rate of NO3- reduction by such cells was five times higher than when malate was carbon source. Reduced minus NO3(-)-oxidized spectra of cells had peaks in the alpha-band region for cytochromes at 552 nm and 559 nm, indicating the involvement of c- and b-type cytochromes in the electron-transport pathway to NO3-. The total ferricyanide-oxidizable cytochrome that was also oxidized in the steady state by NO3- was greater in cells grown with butyrate rather than malate. Low concentrations of cyanide inhibited NO3- reduction. Neither CN-, nor a previously characterized inhibitor of NO3- reduction, 2-n-heptyl-4-hydroxyquinoline N-oxide, prevented the oxidation of the cytochromes by NO3-. This suggested a site of action for these inhibitors on the reducing side of the b- and c-type cytochromes involved in electron transport to the NO3- reductase. The predominant cytochrome in a periplasmic fraction prepared from cells of R. capsulatus grown on butyrate medium was cytochrome c2 but a c-type cytochrome with an alpha-band reduced absorbance maximum at 552 nm could also be identified. The reduced form of this latter cytochrome, but not that of cytochrome c2, was oxidized upon addition of NO3- to a periplasmic fraction. The NO3(-)-oxidizable cytochrome co-purified with the periplasmic NO3- reductase through fractionation procedures that included ammonium sulphate precipitation, gel filtration at low and high salt concentrations, and ion-exchange chromatography. A NO3(-)-reductase-cytochrome-c552 redox complex that comprised two types of polypeptide, a nitrate reductase subunit and a c-type cytochrome subunit, was purified. The polypeptides were separated when the complex was chromatographed on a phenyl-Sepharose hydrophobic chromatography column.

Cell Fractionation↗
PubMed1990

A clinical update on hypothalamic-pituitary control.

There is at present an accumulation of data which indicate the importance of stimulatory and inhibitory growth factors in the paracrine, and perhaps autocrine, regulation of anterior pituitary cell growth and function. There may well be several more specific pituitary growth factors which will be identified in the near future. Preliminary data are also emerging concerning the pattern of oncogene expression in human pituitary tumours. It is now necessary to relate growth factor production, oncogene expression and hypothalamic regulation to specific anterior pituitary cell populations and human pituitary adenomas. In the midst of these studies it should not be forgotten that coordinated vascular and supporting tissue growth is also crucial to the normal development and maintenance of anterior pituitary structure and function.

Adrenocorticotropic Hormone↗
PubMed1990

Apo forms of cytochrome c550 and cytochrome cd1 are translocated to the periplasm of Paracoccus denitrificans in the absence of haem incorporation caused either mutation or inhibition of haem synthesis.

An apo form of cytochrome c550 can be detected by immunoblotting cell-free extracts of a mutant of Paracoccus denitrificans that is deficient in c-type cytochromes. This apoprotein is found predominantly in the periplasm, the location of the holocytochrome in the wild-type organism, indicating that translocation of the polypeptide occurs in the absence of haem attachment. The polypeptide molecular weight, as judged by sodium dodecyl sulphate/polyacrylamide gel electrophoresis, is indistinguishable from that of the holoprotein and the chemically prepared apoprotein; this suggests that the N-terminal signal sequence is removed in the mutant as in the wild-type organism. In the presence of levulinic acid, an inhibitor of haem biosynthesis, apocytochrome c550 and aponitrite reductase (cytochrome cd1) accumulated in the periplasm of wild-type cells. Synthesis of these apoproteins was blocked by chloramphenicol. Thus in P. denitrificans the synthesis of these polypeptides is neither autoregulated nor regulated by the availability of haem. That the apoproteins appear in the periplasm argues against the possibility of polypeptide/haem co-transport from cytoplasm to periplasm. These observations are related to, and contrasted with, the biosynthesis of c-type cytochromes in eukaryotic cells.

Apoenzymes↗
PubMed1990

Long-term treatment of acromegaly with octreotide (Sandostatin).

Sixteen patients with acromegaly have been treated with octreotide [100 micrograms twice daily to 200 micrograms 3 times daily according to growth hormone (GH) response] for between 3 and 44 months. The mean serum GH concentrations fell from 39.3 mU/l before treatment to 10.5 mU/l on day 1 of treatment and, with continued treatment (and higher doses in 8 of 16 patients), to 7.9 mU/l. In 2 patients there was no GH suppression. GH suppression after the first administration of octreotide appeared to predict long-term response. Pre-treatment serum insulin-like growth factor 1 levels were elevated in 11 of 12 patients investigated, but were normalized on continued octreotide therapy in 10 of 12. Octreotide suppressed prolactin secretion in all 6 hyperprolactinaemic patients. Steatorrhoea occurred in 15 of 16 patients at initiation of treatment, but resolved in all within 7 days. Two patients developed gallstones. In summary, octreotide is a safe and effective treatment for acromegaly, producing clinical and biochemical improvement in up to 90% of patients. Octreotide is a valuable adjunctive treatment where surgery has failed and also in those with contra-indications to surgery.

Acromegaly↗