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Biomedical subjects

R J Williams

Publications and source records attributed to R J Williams.

At least 145 records · Page 8Linked to original sources
PubMed1995

Inhibition of ADP-ribosyltransferase increases synthesis of Gs alpha in neuroblastoma x glioma hybrid cells and reverses iloprost-dependent heterologous loss of fluoride-sensitive adenylate cyclase.

Exposure of NG108-15 cells to 50 mM nicotinamide [an inhibitor of mono(ADP-ribosyl)transferase] for 18 hr led to an increase in membrane associated Gs alpha measured either as cholera toxin substrate or by immunoblotting with a specific antiserum. Prolonged exposure of NG108-15 cells to iloprost is followed by homologous loss of iloprost sensitivity, and heterologous loss of fluoride-dependent activation of adenylate cyclase. Nicotinamide reversed the loss of fluoride sensitivity, but failed to restore iloprost-dependent activation of adenylate cyclase. These results with nicotinamide in NG108-15 cells contrasted with those from platelets, which also exhibit heterologous desensitization of fluoride sensitivity following prolonged exposure to iloprost. Treatment of platelets with 50 mM nicotinamide for 18 hr led to an increase of 75.0 +/- 19.4% in the amount of membrane associated cholera toxin substrate. However, there was no associated increase in the abundance of Gs alpha as determined by immunoblotting. Furthermore, in platelets there was no restoration by nicotinamide of the iloprost-dependent loss of fluoride-sensitive adenylate cyclase activity. It follows that heterologous desensitization in platelets is accompanied by inactivation of Gs alpha, which is retained within the plasma membrane in its inactive state. The nicotinamide-dependent increase in the abundance of membrane associated cholera toxin substrate and immunoreactive Gs alpha in NG108-15 cells is associated with an increase of 72.0 +/- 20.3% in the levels of mRNA encoding Gs alpha. The capacity of nicotinamide to increase the abundance of membrane associated Gs alpha was reversed when the cells were cultured in the presence of 20 micrograms/mL cycloheximide. These results suggest that the ability of nicotinamide to increase the abundance of Gs alpha in NG108-15 cells is mediated by de novo protein synthesis.

ADP Ribose Transferases↗
PubMed1995

Nitric oxide activates metalloprotease enzymes in articular cartilage.

Nitric oxide (NO.) is a multifunctional messenger molecule generated by a family of enzymes, collectively termed the nitric oxide synthases. We investigated the role of NO. in the modulation of two metal-dependent proteolytic enzymes (collagenase and stromelysin) which are activated during inflammatory and infective arthritis. The inflammatory mediators interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha) and the bacterial cell wall fragment endotoxin, induced both nitric oxide synthase activity and stromelysin and collagenase activity in whole cell preparations and in conditioned media from explants of bovine and human cartilage. Both NO2- (the stable end-product of NO.) and metalloprotease activity were inhibited by competitive inhibitors of nitric oxide synthase. The NO. donor, S-nitroso-N-acetyl-D,L-penicillamine (SNAP) also induced metalloprotease activity in a dose-dependent fashion. These data provide evidence that NO. plays a regulatory role in the activation of metal-dependent proteases in articular chondrocytes and cartilage.

Amino Acid Oxidoreductases↗
PubMed1995

A locking endurance test for hinged sections of removable partial dentures.

In this study an apparatus is described that will open and close a hinged section of a removable partial denture. A metal framework based on a "swinglock" type of hinge and latch design was constructed. An electric motor provided the drive for the opening and closing mechanism, which when operated set the device in motion. The device and results of a test run of 12,000 opening and closing operations are discussed.

Dental Clasps↗
PubMed1995

A new type of flexor tendon repair. Biomechanical evaluation by cyclic loading, ultimate strength and assessment of pulley friction in vitro.

Experiments were performed to evaluate biomechanical aspects of the performance of a "deep-biting peripheral suture" for flexor tendon repair, either when used alone or with a square or modified Kessler core stitch, and the technique was compared to the Kleinert repair. Tests included progressively increasing cyclic loads, force to pull the repair into the A2 pulley, and ultimate failure strength. 50% of the Kleinert repairs failed under 30 N cyclic loading, while 100% of the DBPS plus Kessler core stitch repairs survived. There was no discernable difference in gliding function or repair bulk between these sutures, but ultimate strength increased significantly with the DBPS repairs. We concluded that the DBPS plus Kessler-type core stitch will survive active mobilization better than the Kleinert method.

Biomechanical Phenomena↗
PubMed1995

Glucose regulates glutamate-evoked arachidonic acid release from cultured striatal neurons.

L-Glutamate stimulates the liberation of arachidonic acid from mouse striatal neurons via the activation of N-methyl-D-aspartic acid (NMDA) receptors and by the joint stimulation of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and metabotropic receptors. In this study, we investigated whether starving cultured mouse striatal neurons of glucose would modify glutamatergic receptor-mediated arachidonic acid release. Glucose deprivation for 30 min led to enhancement of the NMDA-evoked release of arachidonic acid, compared with that observed in the presence of glucose. This enhanced response depended on both the concentration of glucose and the length of time of glucose deprivation. The enhanced NMDA response appeared to result from both a release of glutamate and the subsequent additional release of arachidonic acid due to the activation of AMPA and metabotropic receptors. Indeed, the increased NMDA response was completely reversed when extracellular glutamate was enzymatically removed. Moreover, glucose deprivation potentiated the combined AMPA/metabotropic receptor-evoked release of arachidonic acid, even in the absence of extracellular glutamate. However, removing glucose did not improve the calcium rise induced by AMPA or NMDA. The ATP-evoked release of arachidonic acid from striatal astrocytes was not altered by glucose starvation. In summary, glucose deprivation affected two properties of striatal neurons: (a) it induced an NMDA-evoked release of glutamate from striatal neurons and (b) it selectively potentiated the AMPA/(1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid-evoked release of [3H]arachidonic acid without altering the authentic NMDA-mediated response.

Adenosine Triphosphate↗
PubMed1995

Effects of acute and chronic ethanol on cyclic AMP accumulation in NG108-15 cells: differential dependence of changes on extracellular adenosine.

1. This study investigated the effects of acute and chronic ethanol on basal, agonist- and forskolin-stimulated cyclic AMP formation in NG108-15 mouse neuroblastoma x rat glioma hybrid cells, and examined the role of changes in extracellular adenosine concentrations on the effects observed. 2. NG108-15 cells incubated acutely with ethanol (1-200 mM) displayed concentration-dependent increases in basal and iloprost-stimulated (300 nM; a prostanoid IP receptor agonist) cyclic AMP accumulation but a concentration-dependent decrease in forskolin-stimulated (10 microM) accumulation. 3. Cells treated chronically with ethanol (200 mM) for 48 h displayed increases over control in basal, iloprost- (0.001-10 microM) and forskolin (0.01-100 microM)-stimulated cyclic AMP formation. However, chronic ethanol did not affect [3H]-iloprost binding to cell membranes. 4. Inclusion of adenosine deaminase (ADA; 1 unit ml-1) during the incubation period to measure cyclic AMP accumulation completely abolished the increase in basal accumulation following chronic ethanol, but did not affect the increase in iloprost stimulation. On the other hand ADA partially reversed the increase in forskolin stimulation following chronic ethanol, but even in the presence of high concentrations of ADA (5 units ml-1) the forskolin stimulation remained elevated above control. 5. Cells treated chronically with the adenosine receptor agonist 5'-(N-ethylcarboxamido)-adenosine (NECA; 10 microM for 48 h) displayed a reduction in subsequent NECA- and forskolin-stimulated cyclic AMP accumulation, but iloprost stimulation was not affected. ADA included acutely during the incubation period to measure cyclic AMP accumulation abolished the reduction in forskolin but not NECA stimulation produced by the chronic NECA pretreatment. 6. We have previously noted that ethanol inhibits NG108-15 cell proliferation and alters cell morphology.To mimic this, cells were incubated in the absence of foetal calf serum for 48 h. Following this time, basal, iloprost- and forskolin-stimulated cyclic AMP formation was enhanced over that in cells grown in the presence of serum.7. These results indicate that chronic ethanol enhances cyclic AMP formation in intact NG108-15 cells by more than one mechanism: one involves increased extracellular adenosine concentrations and the other a change in the transduction system beyond the receptor, possibly involving the adenylyl cyclase enzyme. Furthermore the ethanol-induced changes in cyclic AMP accumulation may relate to alterations in NG108-15 cell growth and development.

Adenosine↗
PubMed1995

Significant pneumothorax complicating a fractured clavicle.

Pneumothorax has been described as a complication of a fractured clavicle only three times. It is an important and potentially serious complication. This case report describes a fractured clavicle complicated by a significant pneumothorax which required chest drain insertion.

Adult↗
PubMed1995

Medullary carcinoma of the thyroid metastatic to breast.

Medullary carcinoma of the thyroid commonly spreads to the lymphatics and later on in the disease process, to distant sites such as lung, liver and bone. Spread to the breast is rare (only two previous reported cases) and presents a major diagnostic dilemma. We report a case of metastatic medullary carcinoma of the thyroid which presented as a primary breast nodule with ipsilateral axillary lymphadenopathy. Clearly it was important to distinguish this tumour from a breast primary as each are managed differently. Both cytological and radiological investigations were inconclusive but excision biopsy was diagnostic.

Breast Neoplasms↗
PubMed1995

Surgical shock presenting as lower limb ischaemic rest pain.

Ischaemic rest pain affecting the lower limb is characteristically constant, severe and distressing. Attention is thereby concentrated on the affected leg and its vascular supply which may distract the attending clinician from a precipitating cause. We present two patients with shock that led to acute onset of ischaemic leg pain.

Acute Disease↗
PubMed1994

Synthetic proteins. Designer proteins.

Attempts to synthesize de novo proteins with particular properties should improve our understanding of the long-range cooperative interactions that are so crucial to the structure and function of proteins.

Protein Structure, Secondary↗
PubMed1994

Instrument to detect near-infrared fluorescence in solid-phase immunoassay.

The construction of a near-infrared (near-IR) fluorescence detector for measuring picomolar levels of near-IR laser dyes is described. The detector is designed for use in an immunoassay technique that employs antibodies labeled with near-IR polymethine cyanine dyes. These dyes possess spectral properties that are exclusive to the near-IR region (650-1100 nm). The instrumentation is characterized, including its hardware and data acquisition software components. The detector is capable of measuring fluorescence in both solution and solid-phase environments. Data on the detector's performance is presented.

Carbocyanines↗
PubMed1994

Persistent organochlorine residues in foodstuffs from Australia, Papua New Guinea and the Solomon Islands: contamination levels and human dietary exposure.

Concentrations of polychlorinated biphenyls (PCBs), DDT and its metabolites (DDTs), HCH isomers (HCHs), chlordane compounds (CHLs), aldrin, dieldrin, heptachlor, heptachlor epoxide and HCB were determined in a wide variety of foodstuffs collected from different locations in Australia, Papua New Guinea and the Solomon Islands. Elevated levels of PCBs, CHLs, DDTs and dieldrin were detected in most of the foodstuffs of animal origin. Fish samples collected near the coast of Sydney recorded noticeable concentrations of PCBs, CHLs and DDTs. PCB contamination was generally prominent in samples collected in urban areas, whereas organochlorine pesticides were distributed uniformly throughout Australia. The widespread usage of CHLs was evident from the composition of its compounds in various foods while HCHs and DDTs were found to have been used sporadically in space and time. For the most part, residue levels of organochlorines in foodstuffs were below the tolerance limits established by national and international health organizations. The exception was CHLs in some fish samples, which exceeded the maximum residue limits recommended by the National Health and Medical Research Council of the Australian government. Although the number of samples analysed in each class of foods is small for the purpose of estimating human dietary exposure, we tentatively conclude that the dietary intake of PCBs in Australia was higher than those observed in most developed and developing nations, whereas the intake of organochlorine pesticides was higher than in developed nations but lower than in developing countries. Meat products contributed to the higher dietary intakes of most of the organochlorines. The contamination pattern and the residual concentrations of organochlorines in foodstuffs of Papua New Guinea and the Solomon Islands resembled those observed in Australia.

Animals↗
PubMed1994

Gs alpha-dependent and -independent desensitisation of prostanoid IP receptor-activated adenylyl cyclase in NG108-15 cells.

NG108-15 mouse neuroblastoma x rat glioma cells were treated with the prostanoid IP receptor agonist iloprost (1 microM) and the time course of changes in the levels of prostanoid IP receptors, adenylyl cyclase activity, and the alpha-subunit of the stimulatory guanine nucleotide binding regulatory protein, Gs, were measured. Incubation of cells with iloprost produced a biphasic time course of desensitisation of prostanoid IP receptor-activated adenylyl cyclase. Parallel analysis of iloprost-induced loss of membrane Gs alpha, NaF-stimulated adenylyl cyclase and [3H]iloprost binding suggested only monophasic curves, with t0.5 values similar to the initial phase of desensitisation of iloprost-stimulated adenylyl cyclase activity. This suggests that the loss of receptor and Gs alpha occur at the same time and account for the initial period of desensitisation due to iloprost pretreatment. Pretreatment of NG108-15 cells with cholera toxin produced a near complete loss of membrane-associated Gs alpha, but the loss of [3H]iloprost binding due to iloprost treatment was not affected by pretreatment with cholera toxin, suggesting that prostanoid IP receptors can be down-regulated in the absence of any coupling to Gs. The second phase of desensitisation of iloprost-stimulated adenylyl cyclase activity, during which there was no further change in NaF-stimulated adenylyl cyclase or in the membrane levels of Gs alpha, was not due to protein kinase A activation, since elevating intracellular cyclic AMP levels with forskolin did not subsequently decrease iloprost-stimulated adenylyl cyclase activity or [3H]iloprost binding. These results demonstrate that iloprost pretreatment of NG108-15 cells induces two distinct phases of desensitisation; an initial desensitisation due to concurrent loss of prostanoid IP receptors and Gs alpha, and then a further desensitisation by an as yet uncharacterized mechanism during which there is no further loss of Gs alpha.

Adenylyl Cyclases↗
PubMed1994

Dose-dependent metabolism of 2,2-dichloro-1,1,1-trifluoroethane: a physiologically based pharmacokinetic model in the male Fischer 344 rat.

2,2-Dichloro-1,1,1-trifluorethane (HCFC-123) is used industrially as a refrigerant, as a foam blowing agent, and as a solvent. It is also being considered as a replacement for halons and chlorinated fluorocarbons which have been banned by the Montreal Protocol because they deplete atmospheric ozone. Male Fischer 344 rats were exposed to 1.0, 0.1, and 0.01% HCFC-123 by inhalation. Parent compound was measured in blood, fat, and exhaled breath and trifluoroacetic acid (TFA) was measured in blood and urine. A physiologically based pharmacokinetic (PBPK) model was developed which included a gut compartment and a variable size fat compartment in addition to the standard flow-limited compartments. Compartment volumes and flows were chosen from the literature, partition coefficients were measured in the laboratory, and metabolic parameters were optimized from experimental data using model simulations. Laboratory experiments showed that the TFA blood concentration during the 1.0% exposure was more than 50% less than the TFA blood concentration during the 0.1% exposure. After cessation of the 4-hr exposure, TFA blood concentrations from the 1.0% exposure rebounded and peaked between 12 and 26 hr after the exposure at about the same concentration as the 0.1% peak. This rebound phenomenon suggested that it was not killing of the metabolic enzymes but substrate inhibition that made the TFA blood concentrations lower than expected. Substrate inhibition by halothane, a structural analog of HCFC-123, has been described in the literature. Only by including a term for substrate inhibition in the PBPK model could pharmacokinetic data for TFA in blood be simulated adequately. This combination of laboratory experimentation and PBPK modeling can be applied to relate the levels of parent and metabolite to toxic effects with some hope of elucidating the toxic species. This work is the first step toward developing models that can be used to predict the toxicokinetics of HCFC-123 in humans throughout various potential use scenarios.

Adipose Tissue↗
PubMed1994

Multicentric granular cell tumor of the heart presenting with aortic dissection.

A 43-year-old woman with a history of cutaneous granular cell tumor presented with severe aortic regurgitation due to a type A aortic dissection. During repair, multiple masses were found on the surface of the heart, which later were identified to be granular cell tumor. We present an extremely rare case of multicentric granular cell tumor of the skin and heart, manifested over a 23-year period.

Adult↗