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Biomedical subjects

R C Harris

Publications and source records attributed to R C Harris.

At least 163 records · Page 9Linked to original sources

Evidence for glomerular actions of epidermal growth factor in the rat.

Epidermal growth factor (EGF), an endogenous mitogenic peptide, has recently been shown to be a potent vasoconstrictor of vascular smooth muscle. In view of its potential role in proliferative and inflammatory renal glomerular diseases, we examined the effects of EGF both on cultured rat mesangial cells and on in vivo glomerular hemodynamics. Mesangial cells possess specific, saturable EGF receptors of differing affinities, with Kd's of 0.1 and 1.7 nM, respectively. EGF produced a rapid increase in intracellular pH of 0.12 +/- 0.01 pH U, which was sodium dependent and amiloride inhibitable. The addition of EGF to mesangial cells cultured on either glass or dimethylpolysiloxane substratum induced reproducible cell contraction. Intrarenal EGF infusion did not affect systemic blood pressure or hematocrit but reversibly decreased GFR and renal blood flow from 4.19 +/- 0.33 to 3.33 +/- 0.26 and from 1.17 +/- 0.09 to 0.69 +/- 0.07 ml/min, respectively. Glomerular micropuncture confirmed decreases in single nephron plasma flow and in single nephron GFR (from 142 +/- 9 to 98 +/- 8 and from 51.6 +/- 11.7 to 28.5 +/- 3.5 nl/min, respectively) which were due to significant increases in both pre- and postglomerular arteriolar resistances (from 1.97 +/- 0.31 to 2.65 +/- 0.36 and from 1.19 +/- 0.11 to 2.00 +/- 0.15 10(10) dyn.s.cm-5 respectively) and to a significant decrease in the ultrafiltration coefficient, Kf, which fell from 0.100 +/- 0.019 to 0.031 +/- 0.007 nl/(s.mmHg). These studies demonstrate that mesangial cells possess specific receptors for EGF, and exposure of these cells to physiologic concentrations of EGF results in an in vitro functional response characterized by activation of Na+/H+ exchange and by resultant intracellular alkalinization, as well as by cell contraction. EGF administration in vivo significantly reduces the glomerular capillary ultrafiltration coefficient, Kf, which, in combination with EGF-induced constriction of both preglomerular and postglomerular arterioles, results in acute major reductions in the rates of glomerular filtration and perfusion.

Animals↗

Formation of acetylcarnitine in muscle of horse during high intensity exercise.

To study the changes in carnitine in muscle with spring exercise, two Thoroughbred horses performed two treadmill exercise tests. Biopsies of the middle gluteal were taken before, after exercise and after 12 min recovery. Resting mean muscle total carnitine content was 29.5 mmol.kg-1 dry muscle (d.m.). Approximately 88% was free carnitine, 7% acetylcarnitine and acylcarnitine was estimated at 5%. Exercise did not affect total carnitine, but resulted in a marked fall in free carnitine and almost equivalent rise in acetylcarnitine. The results are consistent with a role for carnitine in the regulation of the acetyl-CoA/CoA ratio during sprint exercise in the Thoroughbred horse by buffering excess production of acetyl units.

Acetylcarnitine↗

The effect of antibiotics on the cell morphology of Legionella pneumophila.

Legionella pneumophila, in Buffered Yeast Extract broth, was treated for 5 h at 37 degrees C with rosaramicin, erythromycin, cefotaxime, dibekacin, penicillin, methicillin, cefoxitin, cephalothin, ticarcillin, carbenicillin or polymyxin B at near-MIC levels and above. Electronmicroscopy demonstrated morphological changes to the bacteria in some, but not all, of the antibiotic-treated suspensions. Penicillin, at 1000 micrograms/ml (40 X MIC) but not less, produced smooth bubble-like structures on cell surfaces; methicillin produced rough bubble-like structures at 100 micrograms/ml (MIC) but not at 1000 micrograms/ml. In each case, these structures resembled spheroplasts. Polymyxin B induced small-bleb formation on the bacterial cell surfaces at all concentrations tested (MIC-10 X MIC). The other eight antibiotics did not induce any morphological changes at any concentration tested.

Anti-Bacterial Agents↗

Metabolic response to maximal exercise of 800 and 2,000 m in the thoroughbred horse.

To define the metabolic response to maximal exercise in the thoroughbred horse under field conditions, muscle biopsies and venous blood samples were taken from five horses after a single 800-m gallop and from four horses after a single 2,000-m gallop. Muscle and blood samples were also collected during 60 min of recovery. After exercise muscle ATP contents were decreased by 30 +/- 7 (SD) and 47 +/- 3% after the 800- and 2,000-m gallops, respectively. As indicators of purine catabolism, ammonia and uric acid increased in plasma, the accumulation being greater after the 2,000-m gallop. Blood ammonia peaked immediately after exercise and uric acid after 40-60 min of recovery. Muscle glycogen utilization over the 800- and 2,000-m gallops averaged 2.68 +/- 0.90 and 1.06 +/- 0.12 mmol glucosyl units.kg dry muscle-1.s-1, respectively, and the total used amounted to 27.3 +/- 6.6 and 32.5 +/- 8.8% of the initial store. Muscle lactate accumulation averaged 123.5 +/- 49.7 and 167.3 +/- 20.7 mmol/kg dry muscle, respectively, and declined during recovery with half times of 22.9 +/- 4.2 and 18.9 +/- 6.6 min. Blood lactate peaked 5-10 min after exercise. Exercise resulted in only a small increase in muscle glycerol content, but this continued to rise during recovery reaching 9-12 mmol/kg dry muscle after 20 min. During this time the increase in muscle glycerol content exactly matched the decline in glycerol 3-phosphate.

Adenine Nucleotides↗

Acetylcarnitine formation during intense muscular contraction in humans.

To study the changes in carnitine during intense muscular effort subjects underwent 4 min intermittent electrical stimulation of the quadriceps femoris muscle and on a separate occasion performed 4 min exercise on a bicycle ergometer. Biopsies of the vastus lateralis muscle were taken at rest and after 2 and 4 min of stimulation or exercise. Resting mean muscle total carnitine content was 20.0 mmol/kg dry muscle. Approximately 77% was free carnitine and 19% acetylcarnitine. Four minutes of stimulation or intense exercise did not effect total carnitine but did result in a marked fall in free carnitine and almost equivalent rise in acetylcarnitine. The results indicate that acetylcarnitine is a major metabolite formed during intense muscular effort and that carnitine may function in the regulation of the acetyl-CoA/CoA ratio by buffering excess production of acetyl units.

Acetylcarnitine↗

Pulmonary complications of acute spinal cord injuries.

The records of 123 consecutive patients admitted with spinal cord injury were examined for the presence of pulmonary complications. Forty-nine had tetraplegia and 23 had paraplegia; the remainder suffered a variety of neurological deficits. Multiple injuries were encountered in 36 patients. Fifty-three pulmonary complications were noted in 44 (35.7%) patients. The most common problems were atelectasis and pneumonia. There were 22 (18%) deaths. Fourteen deaths were related to pulmonary complications. The mean age of patients who died was 52 +/- 13 (SE) compared to 28 +/- 12 for survivors. A mean forced vital capacity (FVC) of 1127 +/- 410 cc in patients suffering respiratory difficulties compared to a FVC of 1865 +/- 85 cc in patients without complications (P less than 0.001). Oxygenation (PaO2 90 +/- 19 torr) was normal in patients without respiratory problems and was abnormal in patients developing problems (PaO2 76 +/- 30 torr; P less than 0.05). Twenty patients were treated with a rotating bed. The complication rate of patients on the bed was only 10%. In conclusion, respiratory problems remain a significant cause of morbidity and mortality in spinal cord injury. The forced vital capacity, blood oxygen tension, and age are predictors of pulmonary complications. The use of a multidisciplinary approach and a rotating bed may minimize these problems.

Adolescent↗

Coupling of Na-H exchange and Na-K pump activity in cultured rat proximal tubule cells.

Na-H exchange was studied using electron probe analysis and microfluorescent pH analysis of individual cells, in 3-day primary cultures of rat proximal tubule cells (RPTC) obtained from 40- to 50-day-old Sprague-Dawley rats. After Na-K pump inhibition, the initial rate of net Na influx was inhibited 87% by 1 mM amiloride. K influx, an estimate of Na-K pump activity, was increased approximately three times in cells containing high Na (0.114 mM K X mM P-1 X min-1) compared with control cells containing low Na (0.038 mM K X mM P-1 X min-1). Single cell measurements of RPTC loaded with the cytoplasmic pH indicator 5- (and -6) carboxy-4',5'-dimethylfluorescein indicated that there was reversible intracellular acidification in the absence of external Na or in the presence of amiloride. When intracellular acidification was induced by the addition and subsequent removal of NH4Cl, recovery of intracellular pH was inhibited in the absence of external Na or in the presence of amiloride. Using a similar protocol, it was found that after intracellular acidification, the rate of Na influx increased at least 5.9 times, and intracellular Na content was increased 3.15 +/- 0.64 times at 60 s. There was an initial 50% inhibition of Na-K pump activity within the first 60 s compared with control (nonacidified) RPTC, secondarily followed by an increase in Na-K pump activity. Amiloride (0.5 mM) inhibited the acidification-induced increase in Na influx, and persistent acidification led to a persistent inhibition of Na-K pump activity compared with control. In summary, these results demonstrate that Na-H exchange mediates the majority of net Na influx into RPTC under our basal conditions and is necessary for maintenance of intracellular pH homeostasis. In RPTC, Na-H exchange is further activated by intracellular acidification, leading to a net increase in intracellular Na content, which secondarily stimulates Na-K pump activity. The initial inhibition of Na-K pump activity may be due to a direct effect of intracellular acidification.

Amiloride↗

Sodium-hydrogen exchange and glucose transport in renal microvillus membrane vesicles from rats with diabetes mellitus.

Diabetes mellitus is associated with important changes in renal hemodynamics and transport function. Disturbances in solute transport have also been characterized in nonrenal tissues during hyperglycemia and insulinopenia. The purpose of this study was to determine if diabetes is associated with adaptive changes in function of the brush-border membrane of the proximal tubule. We studied Na+ and glucose transport in rat microvillus membrane vesicles isolated from the renal cortex of streptozotocin-induced and BB/W autoimmune diabetic rats. Untreated diabetes was associated with an increase in pH-stimulated total and amiloride-sensitive 22Na+ uptake into vesicles. Insulin treatment returned vesicle 22Na+ uptake to control levels. The increased Na+/H+ exchange was shown to be a result of increased net renal acid production rather than a specific response to insulinopenia because treatment with NaHCO3 also returned 22Na+ uptake to control levels. On the other hand, Na+-glucose cotransport, which was depressed in vesicles from untreated diabetics, returned to control levels with insulin but not NaHCO3 administration. This decreased Na+-glucose cotransport was not secondary to reduction in transport sites in untreated diabetics. These results show that in diabetes mellitus, increased Na+/H+ exchange activity is not the direct result of insulinopenia. However, the diabetic state appears to alter the functioning of the luminal Na+-glucose cotransporter.

Acid-Base Equilibrium↗

Two similar but atypical strains of coryneform group A-4 isolated from patients with endophthalmitis.

Corynebacterium species and other coryneform organisms isolated from clinical specimens are frequently considered contaminants. We isolated two strains of a gram-positive organism from the vitreous fluid of two patients with endophthalmitis who had previously received intraocular lens transplants. The biochemical characteristics and gas chromatographic patterns of both isolates were similar to those of coryneform group A-4 strains. Major differences included esculin hydrolysis, nitrate reduction, growth pigment, and lactic acid production. These two strains along with a limited number of strains collected at the Special Bacterial Pathogens Laboratory (Division of Bacterial Diseases, Centers for Disease Control, Atlanta, Ga.) may represent a subgroup of coryneform group A-4. Results of in vitro susceptibility testing performed with antimicrobial agents commonly used to treat patients with bacterial endophthalmitis underscore the importance of determining MBCs for slow-growing organisms. This report cautions microbiologists not to discard organisms frequently considered contaminants when isolated from body fluids that are normally sterile and from patients receiving local steroids.

Actinomycetales↗

Metabolic response of equine muscle to intermittent maximal exercise.

Four thoroughbred horses performed 4 gallops (G1-G4) with intervals of 5 min. With one exception, gallops were sustained at maximal speed over 620 m. Muscle biopsy samples of the middle gluteal and brachiocephalicus were taken before, during, and after exercise and assayed for ATP and intermediary metabolites. The results showed a major involvement of the brachiocephalicus, in addition to the middle gluteal, during galloping. In three horses, who were clearly fatigued, muscle ATP decreased by up to 50% by the end of G4. This was matched by an equal rise in inosine 5'-monophosphate. Pronounced accumulations of glycerol 3-phosphate, glycerol, and lactate (up to 204 mmol X kg dry muscle-1) occurred with exercise. In the fourth horse, which was less fatigued, a decrease in ATP and increases in intermediary metabolites were much less. Postexercise there was little or no recovery in muscle ATP or lactate during 30 min. The decreases in ATP are consistent with a high activity of adenosine 5'-monophosphate deaminase in horse muscle and indicative also of the high level of anaerobic stress of the exercise program. There was evidence to suggest that the increase in muscle glycerol resulted from hydrolysis of glycerol 3-phosphate and not from the utilization of triglyceride.

Adenosine Triphosphate↗

Another alternative in leg revascularization.

Prosthetic grafts of various types have been the only alternative to revascularization of the leg when the saphenous vein is not used. Our complication rate with prosthetic grafts has led us to search for a different alternative. Long-segment endarterectomy of the iliac, femoral, and popliteal arteries has become a realistic procedure with the use of the Hall oscillating endarterectomy instrument. Although it may be difficult technically and more time-consuming than a usual bypass graft procedure, the results have been good. Fifty-eight procedures have been reported in 53 patients. Eighty-eight percent of the procedures were performed for limb salvage with a 5 percent operative mortality rate. There was an overall cumulative limb salvage rate of 88 percent. The cumulative patency rate with a follow-up of 5 to 24 months was 55 percent. Special emphasis has been placed on technical considerations and pitfalls of the procedure. The procedure is versatile and can be used in combination with a variety of both inflow and outflow procedures.

Adult↗

Adaptation of Na+-H+ exchange in renal microvillus membrane vesicles. Role of dietary protein and uninephrectomy.

The ablation of renal mass and institution of a high protein diet both lead to renal cortical hypertrophy and increased glomerular filtration rate (GFR). We studied Na+ transport in rat microvillus membrane vesicles isolated from uninephrectomized or sham operated rats fed 6% (low), 24% (standard), or 40% (high) protein diets. The feeding of high protein, as compared with low protein, was associated with a 50% increase in rates of pH-stimulated 22Na+ transport in isolated vesicles from sham and uninephrectomized animals. Values for the standard protein diet were intermediate to values for high and low protein. At each level of dietary protein intake, vesicular Na+ transport was greater in the uninephrectomized than in sham rats. The high protein diet was also associated with increased vesicular 22Na+ flux inhibitable by 1 mM amiloride. Increases in total and amiloride sensitive flux were also noted in the absence of a pH gradient. Conductive Na+ and H+ transport were not altered, nor were sodium-glucose and sodium-alanine cotransport. Kinetic studies revealed evidence for an increased Vmax of Na+-H+ exchange in uninephrectomized animals fed a 40 vs. a 6% protein diet whereas Km was unchanged. Supplements of NaHCO3 in the 40% protein diet, to adjust for an increased rate of net acid excretion, did not prevent the increased rates of Na+-H+ exchange. However, treatment with actinomycin D (0.12 mg/kg) prevented the increased Na+-H+ activity as well as the increased renal mass and GFR noted 24 h after unilateral nephrectomy. Na+-H+ exchange rate was closely correlated with GFR (r = 0.961; P less than 0.005) and renal mass (r = .986; P less than 0.001). These observations provide evidence for modification of the luminal membrane Na+-H+ exchanger in response to changes in dietary protein content and nephron number.

Animals↗

A single step polarographic assay of glycogen in muscle tissue and a note on the action of amyloglucosidase.

A rapid, single step polarographic assay of glycogen in KOH digests of muscle is presented. Amyloglucosidase is used to hydrolyse the glycogen to glucose, which is then immediately reacted with glucose oxidase. The method has a good reproducibility, is rapid, and requires as little as 0.7 mg freeze-dried muscle powder. From comparison of the rate of reaction of glycogen in the complete assay mixture with those of alpha- and beta-D-glucose, it was concluded that in the hydrolysis of glycogen--a polymer of alpha-D-glucose--by amyloglucosidase, the immediate and only product is beta-D-glucose. This suggests that during hydrolysis, the pyranose ring is opened at carbon 1, and that carbon 1 is rotated to the beta-configuration before closure.

Glucan 1,4-alpha-Glucosidase↗

Epidermoid cysts of the liver.

To the four recorded cases of epidermoid cysts of the liver are added two cases recently seen in children. One was in a 4-yr-old girl and associated with biliary cirrhosis. The other was in a 5-mo-old boy. The former was treated by roux-en-y cystjejunostomy, the latter by enucleation. The origin of these cysts is unknown. One theory suggests derivation from accessory foregut buds. Because of their malignant potential, treatment should be by excision. Where this is impossible, roux-en-y cystjejunostomy offers satisfactory palliation.

Child, Preschool↗

The cyclic-AMP concentration in plasma and in muscle in response to exercise and beta-blockade in man.

At rest the cAMP concentration in (muscle samples of) the quadriceps femoris ranged from 1.55 to 3.00 mumol per kg dry muscle and in plasma from 15.3 to 32.3 nmol per 1. Blockade of the beta adrenoreceptors with propranolol resulted in a significant decrease in the concentration in muscle at rest, the magnitude of the fall being related to the initial level. Similarly in plasma there was a trend towards lower levels of cAMP in those with the highest pretreatment levels, but the overall change was not statistically significant. There was no relation between the concentrations in muscle and plasma, before or after beta-blockade. Maximum dynamic exercise for 4-8 min resulted in an approximate doubling in the cAMP concentration in both muscle and blood. The increase in plasma was closely related to that in muscle. Beta-blockade inhibited totally the rise in cAMP in muscle during exercise but was marginally less effective in preventing the increase in blood. No increase in plasma or muscle cAMP levels during 40-70 s isometric contraction were observed.

Adolescent↗

Effects of beta-blockade on glycogen metabolism in human subjects during exercise.

The effect of beta-blockade on glycogen metabolism during isometric and dynamic exercise in humans has been investigated. Isometric exercise increased the glycogenolytic rate in muscle but had no effect on the cAMP content. Neither the metabolic pattern nor the time of contraction was affected by beta-blockade. Dynamic exercise increased the cAMP content in muscle by about 100%. The cAMP content at rest was significantly reduced after propranolol infusion and did not increase during exercise. Total hexosemonophosphates increased sixfold during exercise but little or no increase occurred after administration of propranolol. The accumulation of lactate in muscle was slightly reduced during exercise following beta-blockade. The fraction of phosphorylase in the alpha form was 22.5% of the total at rest but decreased to 16% at exhaustion. Synthetase I was similarly decreased. During exercise with propranolol phosphorylase alpha decreased further to 3%, whereas synthetase I was unchanged. It is concluded that beta-blockade has no effect on muscle glycogenolysis during isometric contraction but decreases the rate of glycogen degradation during dynamic exercise at high work loads due to changes in the phosphorylase-synthetase system.

Adolescent↗