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Biomedical subjects

R G Clark

Publications and source records attributed to R G Clark.

At least 73 records · Page 4Linked to original sources
PubMed1993

Counting dead birds: examination of methods.

We studied three methods (line transect, circular quadrat, complete count) for estimating density of dead birds, using models of sparrows and meadowlarks placed at a density of 50 birds of each type/ha. Line transects with a 500-m search line were used in cultivated pasture and native prairie habitats. The number of birds found by individual searchers in line transects varied markedly, particularly, in pasture habitat. More birds were found, and birds were detected at a greater distance, in prairie than in pasture. More meadowlarks than sparrows were found in both habitats but the mean estimated density of meadowlarks was greater than that of sparrows only in prairie. The number of birds found during most searches was less than that suggested for estimating density accurately and longer search lines were required. Density estimates obtained using circular plots to sample 10% of the prairie area ranged from 20 to 80 birds/ha for meadowlarks and from 10 to 60 birds/ha for sparrows in prairie habitat. A complete search by 25 volunteers spaced at 4 m intervals detected 90% of meadowlarks but only 62% of sparrows in pasture habitat. Mean (SD) time for a single search, including searchers and recorders, was 3.0 (0.8), 1.5 (0.3), and 7.5 person-hours, for line transect, circular plot and complete search, respectively.

Animals↗
PubMed1993

Recombinant human insulin-like growth factor binding protein-3 (rhIGFBP-3): effects on the glycemic and growth promoting activities of rhIGF-1 in the rat.

The effects of IGFBPs on IGF-1 bioactivity in vivo remain largely unknown. We have tested the ability of rhIGFBP-3, made in 293 cells or CHO cells, to affect the glycemic and anabolic activities of co-administered rhIGF-1. For glycemic studies female dw/dw rats were anesthetized, a jugular catheter inserted, basal blood samples taken and i.v. injections given of rhIGF-1 (0.25 mg/kg), rhIGFBP-3 (0.50 mg/kg), or rhIGF-1 plus rhIGFBP-3 (0.25 plus 0.50 mg/kg, respectively). The blood glucose (20 min later, as a percent of initial, mean +/- SD) reached its nadir for rhIGF-1 alone (58 +/- 5%) but was not changed by rhIGFBP-3 alone (99 +/- 4%) or rhIGF-1 plus rhIGFBP-3 (93 +/- 5%). In growth studies, young female hypophysectomized rats (90-105 g) were injected s.c. twice daily for 3-4 days, or infused s.c. for 7 days, with excipient, rhIGF-1 or rhIGF-1 plus rhIGFBP-3 at doses similar to those used in the hypoglycemia studies. Weight gain induced by rhIGF-1 was either unchanged, or enhanced, when the rhIGF-1 was delivered with rhIGFBP-3. The hypoglycemic activity of IGF-1 was greatly reduced if IGF-1 was administered bound to 293-cell rhIGFBP-3 but anabolic activity was unchanged or enhanced.

Animals↗
PubMed1992

Cloning and expression of an amylase gene from Streptococcus bovis in Escherichia coli.

An amylase gene was identified in a Streptococcus bovis 033 lambda gtWES lambda B genomic library. Using a starch overlay and a Congo red-iodine staining procedure, amylase positive clones could be identified by zones of clearing. Ten amylase positive clones were identified using this procedure. The clone chosen for further study, lambda SBA105, contained an insert of approximately 7.5 kb. The insert was mapped, and subcloning localized the amylase gene to a region of approximately 3.1 kb. Cloning of the 3.1 kb amylase fragment into pUC18 in both orientations revealed that the amylase gene was transcribed from its own promoter. Amylase activity was expressed by the Escherichia coli subclones and was found to be largely associated with the cytoplasmic fraction. Southern hybridization of genomic DNA from the amylolytic strains, S. bovis 033, S. bovis 077, Butyrivibrio fibrisolvens 194 and 195 revealed a single hybridizing band in S. bovis 033 DNA only. This indicates that the amylase gene from S. bovis may differ from the amylases of these other amylolytic bacteria.

Amylases↗
PubMed1992

Growth hormone (GH)-binding protein in normal and GH-deficient dwarf rats.

There are GH-binding proteins (GHBPs) present in the blood of many species, and these correspond to the extracellular GH-binding domain of the GH receptor. In the rat, GHBP arises by alternative splicing of the GH receptor mRNA, but little is known of the physiological role of circulating GHBP, or its relationship with episodic GH secretion. We have developed a sensitive radioimmunoassay based on recombinant GHBP, and have measured rat GHBP levels in small samples of plasma from normal and GH-deficient dwarf rats. In normal adult rats, GHBP levels were two- to threefold higher in females than in males (16.6 +/- 0.8 vs 6.4 +/- 0.4 micrograms/l, P < 0.001), but this sex difference was not seen in dwarf rats. A continuous infusion of human GH in dwarf males raised plasma GHBP to 23.5 +/- 3.5 micrograms/l compared with 6.7 +/- 0.5 micrograms/l in sham-infused animals, whereas suppression of GH by continuous infusion of a long-acting somatostatin analogue in female dwarf rats had no effect on GHBP. In anaesthetized rats, large changes in plasma GH caused by i.v. administration of rat GH, somatostatin or GH-releasing factor did not affect GHBP acutely. Both GH and GHBP were also measured in serial blood samples from conscious normal and dwarf rats. A sexually dimorphic GH secretory pattern was observed in both strains. Males showed peaks and troughs of GH every 3 h varying over a 100-fold range, whereas females exhibited more continuous GH secretion. Despite the large fluctuations in endogenous GH, GHBP levels remained relatively constant in individual normal or dwarf males, as well as in females of both strains, and there was no significant correlation between GH and GHBP either in individual rats or as a group. Our results suggest that GHBP is GH-dependent in the longer term, and that the higher GHBP levels in female rats require their continuous GH secretory pattern. However, plasma GHBP levels remain stable and are not affected by acute changes in endogenous or exogenous GH.

Animals↗
PubMed1991

Influence of epidural analgesia upon postoperative fatigue.

This study examined the effect of epidural analgesia on the development of postoperative fatigue and the ability to ambulate after surgery. Seventeen patients admitted for elective abdominal surgery were randomized to receive postoperative analgesia with a continuous infusion of epidural fentanyl (epidural group) or intermittent intramuscular morphine (non-epidural group). Fatigue was assessed on an analogue scale of 1 (fit) to 10 (fatigued). Steady state measurements of respiratory gas exchange, respiratory rate, tidal volume and heart rate were made before operation and repeated on the third postoperative day. Energy expenditure was calculated from Weir's formula. Ambulatory measurements were made during treadmill walking at a work rate of 20 kpm min-1 (3.3 W). At rest, patients in both groups had a similar cardiorespiratory response to surgery irrespective of the method of analgesia. Subjective feelings of fatigue were significantly greater in those patients who had received epidural analgesia (P less than 0.01) and patients in this group expended significantly more energy in performing the postoperative exercise test than did those in the non-epidural group (P less than 0.05). The use of epidural opiate analgesia does not limit postoperative fatigue in patients undergoing upper abdominal surgery.

Abdomen↗
PubMed1991

Ligand-mediated immunofunctional assay for quantitation of growth hormone-binding protein in human blood.

Human serum contains a high affinity GH-binding protein (GHBP) whose amino-terminal sequence is identical to the extracellular domain of the GH receptor. Current methods that measure GHBP are laborious, require size or charcoal separation of the GH/GHBP complex, and may be influenced by ambient GH concentrations. We have developed a novel assay method that allows quantitation of the total amount of functional GHBP in serum or plasma. The assay can also be used to measure the concentration of the circulating GH/GHBP complex. An anti-GHBP monoclonal antibody, which recognizes both free GHBP and GH-bound GHBP, is used to capture the GHBP on a microtiter plate. Recombinant human GH is added to saturate all binding sites, and an anti-GH antibody conjugated with horseradish peroxidase is used to detect the amount of GH (endogenous and exogenous) bound to the GHBP. The same procedure, but without incubation with GH, allows measurement of the endogenous GH/GHBP complex. The assay is sensitive (detection range, 31-2000 pmol/L), with average inter- and intraassay precisions of 11.3% and 7.3%, respectively. Measurements in random blood samples from 16 healthy adults showed that all subjects had clearly detectable GHBP concentrations (range, 65.8-305.6 pmol/L). In contrast, GHBP levels were undetectable in samples from 2 patients with Laron-type dwarfism. We believe that this ligand-mediated immunofunctional assay, which combines the simplicity and specificity of an enzyme-linked immunosorbent assay with the ability to detect only biochemically active binding protein, will be useful for studies of the role of the GHBP in health and disease.

Antibodies, Heterophile↗
PubMed1990

The episodic secretory pattern of growth hormone regulates liver carbonic anhydrase III. Studies in normal and mutant growth-hormone-deficient dwarf rats.

Carbonic anhydrase III (CAIII) occurs in male rat liver at concentrations twenty times those in the female, and is sensitive to the pattern of growth hormone (GH) release. Males release GH episodically and have high concentrations of CAIII; females produce GH in a more continuous fashion and have lower CAIII levels. In normal female rats, the endogenous GH secretory pattern was masculinized, either by regular injections of GH-releasing factor (GRF) or by intermittent infusions of somatostatin (90 min on/90 min off). Both treatments induced regular GH pulses and stimulated growth, but only intermittent somatostatin infusions raised CAIII levels (controls, 1.5 +/- 0.5; somatostatin-treated, 9.0 +/- 2.9 micrograms/mg; means +/- S.D.). GRF pulses (4 micrograms every 4 h) did not however raise CAIII levels (controls 1.8 +/- 0.5; GRF-treated 1.4 +/- 0.4 micrograms/mg). Surprisingly, hepatic CAIII is also sexually dimorphic (males, 18.8 +/- 3; females, 2.22 +/- 0.4 micrograms/mg) in a GH-deficient dwarf rat strain which has low plasma GH levels without 3-hourly GH peaks. Intermittent somatostatin infusions in female dwarf rats partially masculinized hepatic CAIII, an effect reduced by co-infusion with GRF. This CAIII response was not secondary to growth induction, since neither somatostatin nor GRF stimulated growth in dwarf rats, and pulses of exogenous GH stimulated growth in female dwarfs without masculinizing CAIII levels. Furthermore, continuous GH infusion in male dwarf rats partially feminized hepatic CAIII levels (to 9.1 +/- 2.4 micrograms/mg), whereas infusions of insulin-like growth factor-1, which induced the same body weight gain, did not affect hepatic CAIII (20.8 +/- 6 micrograms/mg). These results show that hepatic CAIII expression is highly sensitive to the endogenous GH secretory pattern, independent of growth. They also implicate the low basal GH levels between pulses, rather than the peak GH levels, as the primary determinant of the sexually dimorphic hepatic CAIII expression in the rat.

Animals↗
PubMed1990

Changes in cardiorespiratory and muscle function associated with the development of postoperative fatigue.

An understanding of the factors which contribute to postoperative fatigue may benefit the rehabilitation of patients after surgery. Subjective feelings of fatigue and fatigue measured objectively in the adductor pollicis muscle after ulnar nerve stimulation have been studied in relation to changes in cardiorespiratory function and muscular efficiency both at rest and when walking on a treadmill at a work rate of 20 and 56 kpm min-1. Twelve patients admitted for elective abdominal surgery were studied before operation and again on the third postoperative day. The postoperative period was characterized by an increased feeling of fatigue. Surgery had no effect upon fatigue in the adductor pollicis muscle suggesting that the genesis of postoperative fatigue is partly central in origin. Muscular efficiency (s.d.) fell from 34(6) per cent before operation to 22(3) per cent (P less than 0.05) on the third postoperative day and was accompanied by a 19 per cent rise in the net energy (s.d.) expenditure (7.3(0.8) to 8.6(0.3) kJ min-1; P less than 0.03) required to perform a given workload. The increased cardiorespiratory effort and reduced muscular efficiency associated with the performance of low-intensity exercise may limit mobilization after surgery and contribute to a greater feeling of fatigue.

Adult↗
PubMed1990

Use of free fasciocutaneous and muscle flaps for reconstruction of the foot.

Seventeen free flaps were used to reconstruct severe injuries to the foot over the last 36 months at the University of Cincinnati College of Medicine. The type of free flaps used included six fasciocutaneous free flaps and eleven free muscle flaps with split-thickness skin grafts. The fasciocutaneous flaps were either radial forearm or scapular flaps. The muscle flaps used were gracilis, rectus, or latissimus dorsi muscle flaps. Each type, with their specific advantages, disadvantages, and indications for use as they apply to the anatomical areas of the foot, are described. Regardless of the type of free flap used, careful preoperative planning, attention to the size and location of the anatomical defect, and correct contouring and insetting should allow for maximal functional result and minimize postoperative morbidity.

Accidental Falls↗
PubMed1990

Somatostatin and its physiological significance in regulating the episodic secretion of growth hormone in the rat.

Somatostatin (SS) is a powerful inhibitor of growth hormone (GH) secretion both in vitro and in vivo, and is generally regarded as having a negative influence on GH secretion and growth. In the conscious rat, the GH secretory pattern, the response to repeated injections of GH-releasing hormone (GHRH), and the feedback mechanism by which GH regulates its own release are all sexually dimorphic and probably reflect a sexually dimorphic pattern of SS release. Prolonged infusions of SS initially block GHRH-induced GH release but responses begin to break through with time. Withdrawal of SS induces a rebound release of GH in vivo, largely dependent on the release of endogenous GHRH. Prolonged exposure to intermittent infusions of SS in normal female rats produces a paradoxical growth response by inducing regular peaks of GH secretion whereas continuous infusions of SS do not. This response is GH-dependent, and is not due to other effects of SS since intermittent infusions of SS do not increase growth in dwarf rats which are deficient in pituitary GH. The sex differences in GH secretion are also reflected in the expression of several GH-dependent liver enzymes, one of which, carbonic anhydrase III, responds to manipulations of the endogenous secretory pattern by SS in both normal and dwarf rats, and appears to be sensitive to differences in basal, rather than peak, GH levels.

Animals↗
PubMed1990

Sex difference in growth hormone feedback in the rat.

Growth hormone inhibits its own secretion in animals and man but the mechanism for this inhibition is unclear: both stimulation of somatostatin release and inhibition of GH-releasing factor (GRF) release have been implicated. We have now studied the GRF responsiveness of conscious male and female rats under conditions of GH feedback induced by constant infusion of exogenous human GH (hGH). Intravenous infusions of hGH (60 micrograms/h) were maintained for 3 to 6 h whilst serial injections of GRF(1-29)NH2 (0.2-1 microgram) were given at 45-min intervals. The GH responses were studied by assaying blood samples withdrawn at frequent intervals using an automatic blood sampling system. We have confirmed that male and female rats differ in their ability to respond to a series of GRF injections; female rats produced consistent GH responses for up to 13 consecutive GRF injections, whereas male rats showed a 3-hourly pattern of intermittent responsiveness. In female rats, multiple injections of GRF continued to elicit uniform GH responses during hGH infusions, whereas hGH infusions in male rats disturbed their intermittent pattern of responsiveness to GRF, and their regular 3-hourly cycle of refractoriness was prolonged. We suggest that this sex difference in GH feedback may be due to GH altering the pattern of endogenous somatostatin release differentially in male and female rats. Such a mechanism of GH autofeedback could be involved in the physiological control of the sexually differentiated pattern of GH secretion in the rat.

Animals↗
PubMed1989

Reference curves to diagnose cobalt deficiency in sheep using liver and serum vitamin B12 levels.

Reference curves demonstrating the relationship between serum or liver vitamin B12 and weight gain were derived from the examination of 16 published and 48 unpublished N.Z. trials. From these curves probability of obtaining an economic reponse (>10g/day body weight increase) for any serum or liver vitamin B12 can be determined. No significant (P<0.05) weight gain responses occurred to vitamin B12 or cobalt treatment in trials with mean serum vitamin B12 levels above 500 pmol/l or liver vitamin B12 levels greater than 500 nmol/kg. The reference curves were therefore derived from trials with vitamin B12 levels below these levels; 36 trials with serum vitamin B12 and 19 trials with liver vitamin B12 data. The mean vitamin B12 level at the mid point of the weight gain response period was selected from each trial. Examination of serum vitamin B12 reference curves for spring, summer, autumn and winter indicated that curves derived from data closest to the middle of January (summer) adequately reflected response to treatment at any time during the first year of life. Reference curves for liver vitamin B12 also used data closest to middle of January. This was partly because insufficient liver data was available to compare seasonal variations. The fitted response curve approached 0 gram/day at 500 pmol/l for serum vitamin B12 and 375 nmol/kg for liver vitamin B12. The minimum vitamin B12 level at which an economic response to treatment (>10 g/day) is not likely was 336 pmol/l for serum and 282 nmol/kg for liver.

Journal Article↗
PubMed1989

Growth responses in a mutant dwarf rat to human growth hormone and recombinant human insulin-like growth factor I.

A new mutant GH-deficient dwarf rat has been used to study the effects of iv infusions of human GH (hGH) and recombinant human insulin-like growth factor I (hIGF-I). This animal has only about 5% of normal pituitary GH content, low circulating GH levels, and no regular GH surges. The defect seems to be specific for GH. Infusions of hIGF-I at 180 micrograms/day for 9 days elevated serum IGF-I concentrations significantly over those in the saline-infused controls (713 +/- 20 ng/ml vs. 395 +/- 31 ng/ml); hGH infusions did not raise IGF-I levels significantly (435 +/- 20 ng/ml). Gel filtration of serum samples showed that the high-dose hIGF-I infusions increased free IGF concentrations, without apparently altering the pattern of IGF-I binding whereas hGH infusions increased the amount of high mol wt IGF-I binding protein. Neither IGF-I nor hGH infusions affected the small amounts of rat GH present in the dwarf rat pituitary glands. Continuous iv infusions of hGH (200 mU/day for 9 days) stimulated body wt gain (2.1 +/- 0.2 g/day) and bone growth (96 +/- 9 microns/day) significantly compared to saline-infused dwarf rats (1.2 +/- 0.3 g/day and 43 +/- 3 microns/day). Infusions of hIGF-I at 180 micrograms/day produced a body wt gain (2.1 +/- 0.5 g/day) similar to that seen in the hGH-infused group but a significantly smaller stimulation of bone growth (63 +/- 3 microns/day). Infusion of a 5-fold lower dose of hIGF-I (36 micrograms/day for 9 days) had no effect on body wt or bone growth. Food intake was unaffected by either hGH or hIGF-I infusions. The pattern of tissue growth was affected differentially by hGH and IGF-I infusions that produced the same overall body wt gain. hGH induced a relatively proportional growth in most of the organs studied, whereas hIGF-I infusion at 180 micrograms/day stimulated a disproportionately greater growth of the kidney, adrenals, and spleen. In some of the animals, tissues were extracted for RIA of IGF-I; the amounts of IGF-I in the liver were similar in control, hGH, or IGF-I-infused animals, whereas kidney and adrenals from IGF-I infused animals contained larger amounts of immunoreactive IGF-I than did those tissues from hGH-treated rats. Thus, both hGH and hIGF-I can promote growth in the mutant dwarf rat, but they differ both quantitatively and qualitatively in their pattern of actions.

Animals↗
PubMed1989

Growth hormone and growth in diabetic rats: effects of insulin and insulin-like growth factor-I infusions.

The effects of streptozotocin-induced diabetes on weight gain, bone growth and GH secretion have been studied in conscious chronically cannulated male rats. In addition to the classic diabetic symptoms (hyperphagia, polydipsia, polyuria, glycosuria and hyperglycaemia), the slow body weight gain (0.95 +/- 0.5 compared with 2.63 +/- 0.5 g/day in non-diabetic controls) was associated with a reduction in bone growth (from 162 +/- 9 to 48 +/- 4 microns/day) and a reduced pituitary GH content (from 1.5 +/- 0.2 to 0.6 +/- 0.06 mg/gland). Serial blood sampling during the day or overnight showed that the normal male episodic GH secretory pattern was obliterated in the diabetic animals. The constant osmotic stimulation of hyperglycaemia and high fluid turnover was reflected in a significant reduction in pituitary oxytocin and arginine vasopressin (AVP) stores. Intravenous insulin infusions (67-1340 pmol/h for 4 or 7 days) caused a large initial weight gain (greater than 20 g in 2 days) followed by a slower increase, and stimulated tibial bone growth (to 100 +/- 16 and 126 +/- 8 microns/day after 4 or 7 days respectively). Insulin infusion for 7 days also increased pituitary GH content (to 1 +/- 0.15 mg/gland), and the normal episodic GH secretory pattern returned. Intravenous infusions of insulin which reduced, but did not completely normalize, blood glucose levels, allowed the resumption of growth and pulsatile GH secretion. Continuous infusion of recombinant human insulin-like growth factor-I (hIGF-I) at 1110 pmol/h for 54 h also caused a large initial rise in body weight in diabetic rats (17.1 +/- 1.6 compared with 7.5 +/- 2.8 g in saline-infused controls) due primarily to increased fluid retention. This effect of hIGF-I occurred without any significant changes in pituitary GH, AVP, oxytocin, blood glucose or bone growth over this short-term infusion, nor was there any obvious effect on spontaneous GH secretion, monitored over the entire infusion period. We conclude that the diabetic rat is not a good model to study growth stimulation by short-term insulin or IGF-I treatments because the insulin-like effects of these peptides obscure their specific growth-promoting activities in this model.

Animals↗
PubMed1989

Concentration of creatine kinase and aspartate aminotransferase in the blood of wild mallards following capture by three methods for banding.

The concentration of the enzymes creatine kinase (CK) and aspartate aminotransferase (AST), that are released from damaged muscle, was measured in the blood of wild adult male mallards (Anas platyrhynchos) captured for banding in decoy and bait traps and by rocket net. The concentration of CK and to a lesser extent AST was elevated markedly in ducks captured by all methods, indicating muscle injury. The level of CK was significantly greater in ducks captured in decoy traps and rocket net than in those captured in bait traps. This elevation appeared to be related to the length of time the birds were restrained and to the method of restraint.

Animals↗
PubMed1988

The effect of sporidesmin toxicity on ovine serum vitamin B12 levels.

A field outbreak of facial eczema occurred during a vitamin B12 response trial in young growing sheep. Pasture cobalt levels were in the low range for sheep (<0.08 mg/kg, 1.358 micromol/kg) and mean (of 3) liver vitamin B12 levels in the sheep were low (<400 nmol/kg) during the period in which facial eczema occurred. Mean serum vitamin B12 levels of the untreated group were low (<485 pmol/l) for the two months (January and February) preceding the period of facial eczema. However, levels showed an approximate 3.5 fold increase in both cobalt supplemented and unsupplemented groups with the onset of facial eczema in March. From February to March the mean serum vitamin B12 and glutamyl transferase (GGT) activity showed parallel increases with a positive correlation (r = 0.73) between log serum vitamin B12 and log serum GGT activity during the period January to July for both groups. This finding suggested that the increase in serum vitamin B12 was due to sporidesmin induced liver damage. The diagnostic implication is that, in areas where facial eczema is a problem, liver is the sample of choice for determining vitamin B12 status. because sporidesmin toxicity can elevate low serum vitamin B12 levels to diagnostically normal levels.

Journal Article↗
PubMed1988

Paradoxical growth-promoting effects induced by patterned infusions of somatostatin in female rats.

In previous studies with iv infusions of GH or its releasing factor (GRF), we showed that a pulsatile pattern of GH was more effective than continuous GH exposure in stimulating growth in the rat. Since GH release is profoundly affected by its inhibitory factor, somatostatin (SS), we were interested to know whether the effects of SS on GH secretion and growth were also dependent on its pattern of administration. SS infusions were given iv to conscious chronically cannulated female rats through programmable multichannel infusion pumps. Multiple blood samples were obtained with the use of an automated system of pumps, solenoid fluid valves, and a fraction collector, all controlled by a microcomputer. SS infusions (5, 25, or 50 micrograms/h) suppressed GH secretion and elicited a rapid, short-lived rebound release of GH after stopping the infusion. Sinusoidal SS infusions in female rats produced cyclic episodes of GH secretion, but a male type of regular 3-hourly secretory pulses of GH was best achieved by prolonged infusions in which the delivery of SS was interrupted for a short period every 3 h. This intermittent SS infusion pattern elicited a repetitive series of rebound bursts of GH secretion, which increased body weight gain and pituitary GH content. In contrast, continuous infusions of equivalent amounts of SS had no effect on body weight gain and reduced bone growth significantly. Thus the effects of SS on growth do depend on the pattern in which it is administered, and this peptide, which itself powerfully inhibits GH secretion, paradoxically stimulates weight gain in a normal animal when given in a manner that promotes a more pulsatile GH secretory pattern.

Animals↗