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Biomedical subjects

M J Jones

Publications and source records attributed to M J Jones.

At least 55 records · Page 3Linked to original sources

'Fruit-flavoured' mask for isoflurane induction in children.

The effect of disguising the odour of isoflurane with fruit flavouring, upon the quality of inhalational induction, was studied in 41 children aged 3-10 years in a double-blind, randomised trial. Facemasks were either lightly coated with fruit extract or moistened with water so that their appearance was identical to children, anaesthetist and observer. Children allocated to receive fruit flavouring were significantly quieter than the placebo group, but their degree of movement during induction was unchanged. The incidence of respiratory complications including breath-holding, laryngospasm and coughing was similar in both groups. Overall impression of the quality of induction as assessed by the anaesthetist showed no difference between the groups. This simple, cheap modification of inhalational induction warrants further appraisal while the case for disguising the odour of isoflurane remains unproven.

Anesthesia, Inhalation↗

Intercostal block with cryotherapy.

A retrospective survey of 70 patients treated from 1982 to 1984 for chronic intercostal pain with cryotherapy to the intercostal nerves is presented. The results with patients suffering from postherpetic neuralgia are so poor that the authors cannot recommend this form of treatment for this condition. The results with patients suffering from thoracic surgical scar pain are significantly better (P less than 0.005) and support the use of the cryoprobe for this condition.

Cicatrix↗

Effect of membrane perturbation on protein kinase C activation: treatment with exogenous phospholipase C decreases translocation of enzyme to cellular membranes.

Incubation of mouse epidermal HEL-37 cells with C. perfringens phospholipase C for 30 min caused a partial loss of protein kinase C activity after 30 min incubation. Essentially all the kinase activity was present in the cytosolic fraction of both control and treated cells, despite the continued hydrolysis of phospholipid by the exogenous phospholipase. At shorter incubation times with phospholipase C (5 and 10 min) an association of protein kinase with particulate material was observed, presumably reflecting the accumulation of diacylglycerol. It is proposed that further incubation with phospholipase C renders the plasma membrane unable to bind protein kinase C and that already bound enzyme is destroyed by proteolysis.

Animals↗

Phenotypic characteristics of Aeromonas species isolated from adult humans.

The phenotypic characteristics of 89 Aeromonas strains, most of which had been isolated from feces, were examined. Eighty-two percent of the isolates could be placed into one of four groups on the basis of five tests. The relationship of these groups to the three motile species of Aeromonas (Aeromonas caviae, A. hydrophila, and A. sobria) that have been isolated from humans is unclear. Because the means for identification of Aeromonas strains to the species level appear to be imprecise and because the role of each of these species in human diarrhea is unclear at this time, we recommend that identification of enteric Aeromonas isolates to the species level not be done routinely.

Acetoin↗

Microbiological and biochemical characterization of spirochetes isolated from the feces of homosexual males.

Spirochetes were isolated from the feces of 11 homosexual males who had diarrhea. The anaerobic organisms were isolated from a selective medium that consisted of Trypticase soy agar supplemented with either 5% horse or human blood, 400 micrograms of spectinomycin per ml, and 5 micrograms of polymyxin B per ml. Nonselective media that permitted good growth of these fastidious organisms were developed, and selected biochemical tests were performed. The tests included carbohydrate utilization, detection of certain enzymes, and determination of volatile fatty-acid end products of metabolism. Two growth patterns were noted on solid media, a haze of growth and production of small colonies. Based on the results of biochemical tests, patterns of preformed enzymes, and volatile fatty-acid production, we believe that the 11 isolates represent a heterogeneous group of spirochetes. The data suggest that the human colon may harbor unique strains of cultivable spirochetes; additional study of the taxonomy of the organisms and assessment of their virulence for humans are needed.

Culture Media↗

Water and nonelectrolyte permeability of isolated rat hepatocytes.

We have measured the diffusive permeability coefficients of isolated rat hepatocytes to 3H2O, [14C]urea, [14C]erythritol, [14C]mannitol, [3H]sucrose, and [3H]inulin, employing a technique previously developed for erythrocytes (Redwood et al., J. Gen. Physiol 64:706-729, 1974). Diffusion coefficients for the tracer molecules were measured in packed hepatocytes, supernatant fluid, and intracellular medium (lysed hepatocytes) and were calculated assuming one-dimensional semi-infinite diffusion through a homogeneous medium. By applying the series-parallel pathway model, the following permeability coefficients (10(-5) cm/sec) for the hepatocyte plasma membrane were obtained. 3H2O, 98.6 +/- 18.4; [14C]urea, 18.2 +/- 5.3; [14C]erythritol, 4.8 +/- 1.6; [14C]mannitol, 3.1 +/- 1.4; [3H]sucrose, 0; [3H]inulin, 0. These results indicate that isolated rat hepatocytes are highly permeable to water and polar nonelectrolytes, when compared with other transporting epithelia. This relatively high cellular permeability is consistent with a model in which nonelectrolyte permeation is via an aqueous pathway of equivalent pore diameter of 8-12 A. The finding that [14C]erythritol and [14C]mannitol cross the hepatocyte plasma membrane indicates that these molecules enter the bile canaliculus through the transcellular route. Conversely, the failure of [3H]sucrose and [3H]inulin to permeate the hepatocyte in the isolated condition supports the concept that biliary entry of these large carbohydrates, at least that fraction which cannot be accounted for by a vesicular mechanism, must occur via the transjunctional shunt pathway.

Animals↗

Role of inorganic phosphate in total biliary phosphorus determination.

Total phosphorus, inorganic phosphate, and phospholipids were measured in bile of rats and guinea pigs during choleresis and cholestasis produced by taurocholate and taurolithocholate, respectively. Under either experimental condition, total biliary phosphorus concentrations increased significantly in both species, due primarily to an increase in inorganic phosphate. These studies indicate that, if total phosphorus is taken as an estimate of biliary phospholipid concentration, correction for inorganic phosphate is essential under conditions associated with changes in bile secretory function.

Animals↗

Postnatal development of bile secretory physiology in the dog.

To determine whether bile formation in the dog is an immature process at birth, several determinants of bile secretion were studied in anesthetized, bile duct-cannulated puppies of 0-42 days of age and adult dogs. Basal canalicular bile flow rate, estimated by 14C-erythritol biliary clearance, averaged 0.182 microliter/min/g liver in 0-3 day-old puppies and increased to 0.324 and 0.461 microliter/min/g in puppies 7-21 and 28-42 days of age, respectively. The latter two values were not significantly different from that observed in adult dogs (0.348 microliter/min/g). This age-related increase in canalicular bile flow could be accounted for primarily by an increase in bile acid independent bile flow (0.103, 0.277, 0.375, and 0.252 microliter/min/g in 0-3, 7-21, and 28-42 day-old puppies and adult dogs, respectively). Calculated ductular bile water reabsorption (14C-erythritol biliary clearance-bile flow) was virtually absent in 0-3 day-old puppies (0.005 microliter/min/g), and averaged 0.017 and 0.092 microliter/min/g in puppies of 7-21 and 28-42 days of age, respectively. In adult dogs, ductular bile water reabsorption was 0.132 microliter/min/g. These functional deficiencies of the newborn dog were associated with an increased biliary permeability to 3H-inulin which could not be accounted for solely by an increased solute diffusion due to the lower rate of canalicular bile flow. Administration of taurocholate up to 2000 nmol/min/kg produced in all animals a similar increase in canalicular bile flow and bile acid excretion, and was not associated with changes in ductular bile water reabsorption rate. These findings are interpreted to indicate that, in the dog, bile secretory function is immature at birth and develops during postnatal life.

Animals↗

Choleretic effects of differently structured bile acids in the guinea pig.

The effects of 10 differently structured bile acids on bile flow and composition were studied in anesthetized, bile duct-cannulated guinea pigs. At the infusion rates of 2 and 4 mumole/min/kg, all bile acids produced choleresis. The most potent was chenodeoxycholate, which increased bile flow by an average of 31.25 microliters/mumole of bile acids excreted in bile. The weakest choleretic was tauroursodeoxycholate (11.02 mu/mumole). When the choleretic activity was plotted against bile acid hydrophobicity (high-performance liquid chromatography retention factor, obtained from the literature), linearity was observed with similarly conjugated bile acids. The order of potency was deoxycholate greater than chenodeoxycholate greater than cholate greater than ursodeoxycholate, both for the glycine and taurine conjugates, and for the unconjugated bile acids as well. Conjugation was also important, and the rank ordering for the choleretic activity (unconjugated bile acids greater than glycine-conjugates greater than taurine-conjugates) was the same as that for the hydrophobicity. When the choleretic activity was plotted against bile acid micellar aggregation number (in 0.15 M NaCl at 36 degrees C, obtained from the literature), a linear, direct relationship was observed. All bile acids produced similar effects on bile electrolyte concentrations: both bicarbonate and chloride slightly declined during choleresis, whereas bile acid concentrations increased. These studies suggest that, in the guinea pig the differing choleretic activities of differently structured bile acids are not due to their forming micelles in bile of different sizes; either the more hydrophobic bile acids form vesicles, whereas the more hydrophilic form micelles; or bile acids produce choleresis, in part or exclusively, by stimulating an additional secretory mechanism, possibly an inorganic ion pump; or both.

Animals↗

Effect of albumin on the in vitro conjugation of bilirubin by rat liver microsomes.

These studies were carried out to determine whether bovine serum albumin (BSA), which is usually included in the incubation mixture for the in vitro determination of bilirubin-UDP-glucuronyl transferase (GT) activity, affects GT activity. Using bilirubin as substrate, addition of BSA to the enzyme reaction mixture at concentrations varying from 2 to 30 mg/ml resulted in a dose-related inhibition of "native" GT activity of rat liver microsomes. When detergent-activated enzyme was employed, increasing concentrations of BSA also required higher concentrations of deoxycholate, digitonin, or Triton X-100 to produce maximal bilirubin conjugation. Low BSA concentrations (2 mg/ml) prevented enzyme activation by both detergents and UDP-N-acetyl glucosamine. When BSA was omitted and bilirubin dissolved in dimethyl sulfoxide, UDP-N-acetyl glucosamine failed to enhance GT activity, and activation by detergents was only 15-25% of that observed in the presence of optimal concentrations of BSA. When rat albumin was substituted for BSA, a similar dose-related inhibition of in vitro bilirubin conjugation by untreated microsomes was observed, although at any given albumin concentration, GT activity was lower with rat than with bovine albumin. Additionally, both detergents and UDP-N-acetyl glucosamine produced similar GT activation regardless of the rat albumin concentration. Finally, these effects of BSA and rat albumin could not be reproduced when beta-lactoglobulin was employed and/or when p-nitrophenol was the acceptor substrate of GT. These findings indicate that albumin, in particular BSA, profoundly and selectively influences the in vitro activity of microsomal GT toward bilirubin as the acceptor substrate.

Animals↗

Bilirubin diglucuronide formation by rat liver microsomes: demonstration by affinity and thin layer chromatography of bile pigment tetrapyrroles.

The conjugates formed in vitro by bilirubin UDP-glucuronyl transferase were studied by examining reaction products as intact tetrapyrroles, rather than as dipyrrolic azoderivatives. Bile pigments were extracted from conventional microsomal enzyme reaction mixtures by affinity chromatography over albumin-agarose, eluted with 50% ethanol, and separated by a silica gel thin layer chromatographic system. In the presence of UDPGA, native and activated microsomal preparations all formed both bilirubin mono- and diglucuronides from unconjugated bilirubin, and bilirubin diglucuronide from bilirubin monoglucuronide. No significant non-enzymatic conversion of mono- to diglucuronide occurred without UDPGA, or in the presence of denatured enzyme. Hence, bilirubin diglucuronide is a major product of bilirubin-UDP-glucuronyl transferase.

Animals↗

Effect of low-dose phenobarbital on hepatic microsomal UDP-glucuronyl transferase activity.

To determine whether hepatic microsomal enzyme induction occurs in rats following administration of phenobarbital at doses similar to those used in humans (0.5 to 7.5 mg/kg), UDP-glucuronyl transferase (UDPGT) and cytochrome P-450 activities were measured in liver homogenate and microsomal preparations from control rats and rats treated for 6 days with phenobarbital at 1 and 3 mg per kg per day. While no significant increases in liver weight and protein content of homogenate and microsomal preparations were observed with either dose of the drug, both UDPGT and P-450 activities were enhanced significantly following administration of phenobarbital at 3 mg per kg per day. The activity of P-450 was increased by approximately 30% and that of UDPGT by 15-24 and 45-66%, respectively, employing bilirubin and p-nitrophenol as the acceptor substrate. The extent of induction of bilirubin or p-nitrophenol UDPGT was similar when measured with "native" enzyme or with enzyme activated by UDP-N-acetyl glucosamine, digitonin or deoxycholate. These data suggest that the discordant effects of phenobarbital on UDPGT and cytochrome P-450 previously reported in humans and rats may not be attributable solely to differences in the drug doses employed.

Animals↗

Comparison of different activators and diazotization procedures in the assay of bilirubin-UDP-glucuronyl transferase activity in rat liver.

The activity of bilirubin-UDP-glucuronyl transferase (UDPGT) in rat liver microsomes was assayed in standardized incubation mixtures employing two different procedures to quantitate formation of conjugated bilirubin: solvent extraction and diazotization with sulfanilic acid, or selective coupling with diazotized ethyl anthranilate at pH 2.7. Both of these procedures yielded equivalent UDPGT activities with several different microsomal preparations, averaging, respectively, 385 and 372 microU protein with untreated microsomes, 646 and 608 microU protein following activation with UDP-N-acetylglucosamine (3.07 mmol/l), and 1370 and 1478 microU protein after treatment with digitonin (0.65%). In rats pretreated with phenobarbital (75 mg/kg per 6 days), a 70-90% increase in UDPGT activity was observed with either diazotization procedure, irrespective of whether 'native', UDP-N-acetylglucosamine-or digitonin-activated enzyme was employed. These studies indicate that, when other conditions are standardized, equivalent information about UDPGT can be obtained with either of two widely employed diazotization reactions and a variety of enzyme activation procedures.

Animals↗

Decreased [20-3H]phorbol-12,13-dibutyrate binding to phospholipase C pretreated epidermal cells.

Incubation of mouse epidermal cells (HEL-37) with phospholipase C (Clostridium perfringens) caused about a 50% decrease in the specific binding of [20-3H]phorbol-12,13-dibutyrate. Phospholipase C caused a decrease in the apparent number of binding sites from 2.86 X 10(5) to 1.21 X 10(5) per cell with little effect on ligand affinity. The decrease was observed with purified phospholipase containing no detectable protease activity, and when cells were fixed with glutaraldehyde. The phorbol-12,13-dibutyrate binding capacity of treated cells was recovered within 4 h incubation in complete medium. The results suggest that not all phorbol ester binding sites are equivalent, with differences arising either from varying phospholipid requirements or from membrane localisation.

Animals↗

Antimicrobial susceptibilities of bacteria associated with periodontal disease.

A total of 193 bacterial strains were tested for their susceptibilities to 14 antimicrobial agents. Penicillin G was active at 2 U/ml against 98% of the oral isolates. Other antibiotics with good activity were cefoperazone, moxalactam, Sch 29,482, and clindamycin. Metronidazole was active against more than 90% of the anaerobic bacteria and Capnocytophaga but was inactive against most other microaerophilic and facultative strains.

Anti-Bacterial Agents↗