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A J Smith

Publications and source records attributed to A J Smith.

At least 163 records · Page 9Linked to original sources
PubMed1998

Characterization of the murine fatty acid transport protein gene and its insulin response sequence.

Fatty acid transport protein (FATP) was identified by expression cloning strategies (Schaffer, J. E., and Lodish, H. F. (1994) Cell 79, 427-436) and shown by transfection analysis to catalyze the transfer of long-chain fatty acids across the plasma membrane of cells. It is expressed highly in tissues exhibiting rapid fatty acid metabolism such as skeletal muscle, heart, and adipose. FATP mRNA levels are down-regulated by insulin in cultured 3T3-L1 adipocytes and up-regulated by nutrient depletion in murine adipose tissue (Man, M. Z., Hui, T. Y., Schaffer, J. E., Lodish, H. F., and Bernlohr, D. A. (1996) Mol. Endocrinol. 10, 1021-1028). To determine the molecular mechanism of insulin regulation of FATP transcription, we have isolated the murine FATP gene and its 5'-flanking sequences. The FATP gene spans approximately 16 kilobases and contains 13 exons, of which exon 2 is alternatively spliced. S1 nuclease and RNase protection assays revealed the presence of multiple transcription start sites; the DNA sequence upstream of the predominant transcription start sites lacks a typical TATA box. By transient transfection assays in 3T3-L1 adipocytes, the inhibitory action of insulin on FATP transcription was localized to a cis-acting element with the sequence 5'-TGTTTTC-3' from -1347 to -1353. This sequence is very similar to the insulin response sequence found in the regulatory region of other genes negatively regulated by insulin such as those encoding phosphoenolpyruvate carboxykinase, tyrosine aminotransferase, and insulin-like growth factor-binding protein 1. Fluorescence in situ hybridization analysis revealed that the murine FATP gene is localized to chromosome 8, band 8B3.3. Interestingly, this region of chromosome 8 contains a cluster of three other genes important for fatty acid homeostasis, lipoprotein lipase, the mitochondrial uncoupling protein 1 (UCP1) and sterol regulatory element-binding protein 1. These results characterize the murine FATP gene and its insulin responsiveness as well as present a framework for future studies of its role in lipid metabolism, obesity, and type II diabetes mellitus.

Adipocytes↗
PubMed1998

Feasibility of combined percutaneous transluminal angioplasty and minimally invasive direct coronary artery bypass in patients with multivessel coronary artery disease.

BACKGROUND: Angioplasty has become an accepted treatment of patients with coronary artery disease and is now commonly used to treat patients with multivessel disease. The major disadvantage of angioplasty has been restenosis requiring repeat interventions with resultant loss of initial cost savings. Compared with the right and the circumflex coronary arteries, the left anterior descending artery (LAD) has been more adversely affected by restenosis. Recently, minimally invasive direct coronary artery bypass (MIDCAB) to the LAD through a small left anterior thoracotomy using the left internal mammary artery has been performed in some centers with excellent early results and with reduced costs compared with standard bypass surgery. METHODS AND RESULTS: We retrospectively reviewed the first 31 consecutive patients treated in our institution with integrated coronary revascularization (ICR): MIDCAB to the LAD combined with PTCA of the other diseased vessels in patients with multivessel disease. Postoperative angiography in 84% of patients revealed a patent anastomosis and normal flow in the graft and bypassed vessel. Thirty-eight (97%) of 39 vessels were successfully treated percutaneously. At a mean follow-up of 7 months, all patients are currently asymptomatic. There have been 2 adverse clinical events, both related to angioplasty and not to MIDCAB. The average length of stay at the hospital after MIDCAB was 2.79+/-1.05 days. CONCLUSIONS: These preliminary results with ICR are encouraging and suggest that a randomized, prospective clinical trial comparing ICR with standard coronary artery bypass surgery for the revascularization of symptomatic patients with multivessel disease involving the LAD is warranted.

Angioplasty, Balloon, Coronary↗
PubMed1998

Availability of PSC833, a substrate and inhibitor of P-glycoproteins, in various concentrations of serum.

BACKGROUND: P-glycoproteins are membrane-associated transporters that can render cells resistant to a variety of chemotherapeutic drugs. Reversal agents are (preferably nontoxic) drugs that can inhibit these P-glycoproteins and thereby overcome multidrug resistance. PSC833, a cyclosporin A analog, is a reversal agent that has shown potential in in vitro experiments and in clinical trials. We tested PSC833 to determine whether it is a transported substrate of human and murine P-glycoproteins associated with multidrug resistance (encoded by the human MDR1 gene and its murine homolog, mdr1a) and whether it can completely inhibit these P-glycoproteins under simulated in vivo conditions. METHODS: Monolayers of polarized LLC-PK1 pig kidney cells transfected with complementary DNA containing either MDR1 or mdr1a sequences were used to measure the directional transport of P-glycoprotein substrates under various serum conditions. RESULTS: In contrast to two previous studies, we found that PSC833 is transported by both the MDR1 and the mdr1a P-glycoproteins, albeit at a low rate. PSC833 has a very high affinity for the MDR1 P-glycoprotein, and its Michaelis constant (Km) for transport is 50 nM, fourfold lower than for cyclosporin A. Inhibition of drug transport by PSC833 is approximately eightfold less effective in 100% fetal bovine serum than in tissue culture medium containing 10% serum. The concentration of PSC833 necessary to fully inhibit transport of digoxin and paclitaxel (Taxol) under complete (i.e., 100%) serum conditions is higher than the plasma concentrations achieved in clinical trials. CONCLUSIONS: Although PSC833 binds efficiently to the MDR1 P-glycoprotein and is released only sluggishly, the high concentrations of PSC833 necessary to inhibit this P-glycoprotein under complete serum conditions in our in vitro system suggest that it may be difficult for PSC833 alone to produce total inhibition of P-glycoprotein activity in patients.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Comparison of cardiorespiratory reflexes in NK1 receptor knockout, heterozygous and wild-type mice in vivo.

Neurokinin-1 receptors (NK1) are present within the nucleus of the solitary tract, a nucleus which plays a vital role in cardiovascular and respiratory homeostasis. We compared the efficacy of the baroreceptor and pulmonary chemoreflexes between NK1 knockout, heterozygous and wild-type urethane-anaesthetised mice. The magnitude of the baroreceptor reflex mediated bradycardia, induced by a phenylephrine induced pressor response, was significantly greater in NK1 knockout mice (P < 0.001) compared to heterozygous and wild-type animals. In comparison, administration of an NK1 antagonist, CP-99,994 (1.5 mg/kg i.v.) to wild-type animals, had no significant effect on baroreceptor reflex performance. In contrast to the baroreceptor reflex, there were no significant differences in the magnitude of the reflex evoked falls in heart rate, arterial pressure, or respiratory depression between the three groups of mice when the pulmonary chemoreflex was evoked with right atrial injections of phenylbiguanide. It is concluded that the baroreceptor reflex pathway over-compensates for the lack of NK1 receptors in knockout mice. Plausible mechanisms accounting for the enhanced baroreceptor reflex responsiveness in NK1 knockout animals are discussed.

Animals↗
PubMed1998

Altered nociception, analgesia and aggression in mice lacking the receptor for substance P.

The peptide neurotransmitter substance P modulates sensitivity to pain by activating the neurokinin-1 (NK-1) receptor, which is expressed by discrete populations of neurons throughout the central nervous system. Substance P is synthesized by small-diameter sensory 'pain' fibres, and release of the peptide into the dorsal horn of the spinal cord following intense peripheral stimulation promotes central hyperexcitability and increased sensitivity to pain. However, despite the availability of specific NK-1 antagonists, the function of substance P in the perception of pain remains unclear. Here we investigate the effect of disrupting the gene encoding the NK-1 receptor in mice. We found that the mutant mice were healthy and fertile, but the characteristic amplification ('wind up') and intensity coding of nociceptive reflexes was absent. Although substance P did not mediate the signalling of acute pain or hyperalgesia, it was essential for the full development of stress-induced analgesia and for an aggressive response to territorial challenge, demonstrating that the peptide plays an unexpected role in the adaptive response to stress.

Aggression↗
PubMed1998

VH gene analysis of clonally related IgM and IgG from human lymphoplasmacytoid B-cell tumors with chronic lymphocytic leukemia features and high serum monoclonal IgG.

An unusual group of human B-cell tumors with cellular features of chronic lymphocytic leukemia or lymphoplasmacytoid leukemia, together with high levels of a monoclonal IgG serum protein, has been investigated. Analysis of tumor-derived VH genes of neoplastic B lymphocytes was used to determine the clonal relationship between the IgM expressed or secreted by the tumor cells and the IgG serum paraprotein. In all five cases, VH gene sequences showed transcripts of IgM and IgG of common clonal origin. Sequences were derived from VH3 (4 of 5) and VH1 (1 of 5) families and were all highly somatically mutated with strong evidence for antigen selection. There was no intraclonal variation detectable in either IgM or IgG sequences. In 3 of 5 cases, in which monoclonal IgM and IgG were found in serum, the VH genes combined to Cmu or Cgamma showed identical mutational patterns. However, in 2 of 5 cases, in which IgM was confined to cell expression with only monoclonal IgG in serum, sequences of the VH transcripts of IgM and IgG showed many shared mutations but also numerous differences. In these cases, the level of mutation was similar in IgM and IgG and both appeared to be antigen selected. In summary, the final neoplastic event in this group of tumors has apparently occurred at the point of isotype switch from IgM to IgG, leading to dual isotype synthesis. In the group that secreted both isotypes, the mutation pattern was identical, indicating either synthesis by a single cell, or silencing of mutational activity before switching. In the group that did not secrete IgM, cells of each isotype were distinct and reflected a divergent mutational history.

Adult↗
PubMed1998

Hepatocyte-specific expression of the human MDR3 P-glycoprotein gene restores the biliary phosphatidylcholine excretion absent in Mdr2 (-/-) mice.

Mice homozygous for a disruption in the Mdr2 gene (Mdr2 (-/-) mice) lack the Mdr2 P-glycoprotein (P-gp) in the canalicular membrane of the hepatocyte and are unable to excrete phosphatidylcholine into the bile. These mice develop a nonsuppurative cholestatic liver disease, presumably caused by the high concentrations of free cytotoxic bile acids in bile. We generated transgenic mice that express the human homolog of Mdr2, MDR3, specifically in the liver by the use of an albumin promoter. In these mice the MDR3 P-gp is exclusively located in the canalicular membrane of hepatocytes and phospholipid excretion into bile is restored. Mice that contain the same amount of MDR3 P-gp as that of Mdr2 P-gp in wild-type mice, also excrete the same amount of phospholipids. No histopathological abnormalities were observed in the livers of these mice. In mice that express MDR3 at a higher or lower level, the phospholipid excretion correlated with the amount of MDR3 P-gp. We conclude that the human MDR3 P-gp is functionally homologous to the murine Mdr2 P-gp and that it can fully replace this P-gp in Mdr2 (-/-) mice, restoring the excretion of phospholipids into the bile. The phospholipid excretion is limited by the amount of MDR3 or Mdr2 P-gp. The excretion of cholesterol is not tightly coupled to the excretion of phospholipids in these mice, because a very low phospholipid excretion level is sufficient to give almost wild-type cholesterol excretion into the bile.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Blood lead-urine lead relationships in adults and children.

To determine the potential for using instead of blood as an indicator of lead exposure, especially in infants, lead concentrations and high-precision lead isotopic measurements have been compared in venous blood and "spot" urine (n > 260 from 182 different subjects) collected within the same 24-h period. Physiological conditions for the children and most of the adults were considered to be in a steady-state between body stores and lead in the environment. In the case of some adults, conditions were initially not steady-state because exposure conditions changed (for example, subjects moved to a country with lead of different isotopic composition.) There was a high correlation (r2 = ) between the blood and urine measurements of the isotope ratios but about 10% of measurements were outliers--the blood and urine measurements were further apart than was consistent with the measurement error that was generally obtained. The discrepancy was usually found to be associated with the urine measurement and was attributed to contamination during sampling. Weekly urine and monthly blood monitoring of an adult male over a 24-month period showed and excellent correlations, although the standard deviations were about an order of magnitude higher than the precision measured for replicate analyses of a single blood or urine sample. "Spot" urine analyses for two male subjects gave excellent agreement with 24-h urine samples. Standard deviations of the spot analyses were of similar order to those in the 24-month monitored subject. In cases where female adults from Eastern Europe migrated to Australia, there was generally a more rapid exchange of skeletal lead with Australian environmental lead in urine compared with blood. These data do not support a differential partitioning of endogenous lead into the plasma. At this stage, isotopic measurements of urine can be used as a proxy for isotopic measurements in blood. However, lead concentrations in blood and in urine are only weakly related. Concentrations of lead in urine cannot serve to predict concentrations of lead in blood, particularly at the lower range of exposures, for example, at blood concentrations less than 10 microgram/d1.

Adult↗
PubMed1998

Immunochemical detection of the proteoglycans decorin and biglycan in human gingival crevicular fluid from sites of advanced periodontitis.

This study characterized proteoglycan metabolites present in gingival crevicular fluid (GCF) collected from sites with clinical evidence of advanced periodontal disease. The metabolites were purified by anion-exchange chromatography from which a chondroitin sulphate rich fraction was identified by cellulose acetate electrophoresis. Sodium dodecylsulphate-polyacrylamide gel electrophoresis of this fraction revealed a broad silver-staining band with mol. wt 55-65 k and Western blotting suggested that this band was immunoreactive with CS-56, a monoclonal antibody for chondroitin sulphate. Digestion of the metabolite with chondroitinase ABC (protease-free) led to the loss of the silver-staining band. Dot-blot analysis identified components in this fraction that were immunoreactive for the monoclonal/polyclonal antibodies against the C-termino of decorin and biglycan. Amino acid analysis revealed the composition of the proteoglycan metabolite to be rich in glycine, serine and glutamic acid. Immunochemical and biochemical analyses were compared with those of proteoglycan purified from human alveolar bone. Changes in the amino acid composition were noted, suggesting the proteoglycan metabolite has undergone extensive modification and fragmentation to the protein core. The results suggest that the proteoglycan metabolite from GCF represented a degradation product originating from the active destruction of the alveolar bone. They provide further support for the proposal that the appearance of proteoglycan metabolites in GCF is a biomarker for active destruction of alveolar bone, the biochemical analysis of which provides important information on mechanisms involved in the pathology of periodontal diseases.

Adult↗
PubMed1998

An in vitro approach for the study of dentinogenesis by organ culture of the dentine-pulp complex from rat incisor teeth.

Culture of the developing dental tissues has contributed to understanding of developmental processes during early odontogenesis. However, to understand fully the mechanisms involved during dentinogenesis and tissue repair there is a need to develop culture models for the dentine-pulp complex from more mature dental tissues. This study describes the development of a system for the organ culture of mature rodent teeth. Slices of incisors from 28-day-old rats were embedded in a semisolid, agar-based medium and cultured on floating Millipore filters at the liquid-gas interface for up to 14 days. Preservation of cell and tissue morphology was observed throughout the entire dentine-pulp complex after each culture period and autoradiographic studies showed that the odontoblasts were actively synthesizing and secreting extracellular matrix during culture. Transmission electron microscopy confirmed that the phenotypic morphology of the odontoblasts had been maintained during culture. These results demonstrate that the dentine-pulp complex from mature rodent tissues can be cultured successfully for substantial periods of time and will provide a useful model for the study of dentinogenesis and tissue repair.

Animals↗
PubMed1998

Preoperative risk models for minimally invasive coronary bypass: a preliminary study.

OBJECTIVE: Available risk assessment models are designed for standard coronary artery bypass grafting. We hypothesized that minimally invasive coronary bypass could improve on predicted outcome in extremely high-risk patients (Parsonnet score > 20%) by the current risk models. METHODS: From September 1996 to September 1997, 27 consecutive extremely high-risk patients underwent minimally invasive coronary bypass. Seventeen patients were male; age was 73 +/- 12 years, and 63% of patients were older than 75 years. Left ventricular ejection fraction was 33.7% +/- 15% and 63% had an ejection fraction of less than 35%. The predicted 30-day mortality according to the System 97 model was 25.6% +/- 11.3%. The Parsonnet risk score was 36.2% +/- 11%; the predicted length of stay in the hospital was 15.3 +/- 3 days. The predicted risk of stroke according to the Multicenter Perioperative Stroke Risk Index was 22.3% +/- 11.7%. RESULTS: Minimally invasive coronary bypass was isolated in 20 patients and integrated with angioplasty and stenting in 7 patients. The observed 30-day mortality was 0% (P < .01 vs predicted): at an average follow-up of 10.8 +/- 4.1 months, 26 patients (96.3%) are alive without angina; one patient with acquired immunodeficiency syndrome died on postoperative day 40 of acute pancreatitis. No patient had a stroke or neurologic deficit (P < .01 vs predicted). Patency of internal thoracic artery anastomosis was confirmed by angiography in all 27 patients. No patient required reoperation. Eighteen patients (67%) were extubated in the operating room. The observed length of hospital stay after minimally invasive coronary bypass was 3.8 +/- 2.6 days (P < .01 vs predicted). CONCLUSION: On the basis of our results on a relatively small series of patients, we suggest that risk models geared for standard coronary bypass grafting may not be appropriate for minimally invasive coronary bypass.

Aged↗
PubMed1998

Brief interventions for patients with alcohol-related trauma.

Patients who have sustained alcohol-related injuries are frequently treated in departments of oral and maxillofacial surgery. Often, an alcohol intervention will not be possible in accident and emergency departments due to intoxication but, when attending out-patient clinics for follow-up, patients are usually sober. This presents a unique opportunity for encouraging patients to review their alcohol consumption at a time when their facial injury may make them more receptive to advice. This article reviews the convincing evidence of the effectiveness of advice and brief interventions designed to be incorporated into standard out-patient consultations and describes practical screening of patients for harmful drinking, the Stages of Change Model of behaviour change and motivational interviewing for facilitating behavioural change.

Alcohol Drinking↗
PubMed1998

Micromechanical devices for intravascular drug delivery.

Microfabrication technology, more commonly applied to the manufacture of integrated circuits, can be used to build devices useful for mechanical delivery of drugs and genes. Microprobes fabricated using silicon micromachining have been used to deliver DNA into cells as an alternative to bombardment and microinjection. This idea can be extended to intravascular stents with integrated microprobes capable of piercing compressed plaque and delivering anti-restenosis therapies into coronary arteries. Preliminary experiments using filleted rabbit arteries have demonstrated transection of the internal elastic lamina. New nonplanar microfabrication technologies are necessary for creating practical devices with cylindrical symmetry; a promising possibility is to use microfabricated structures of anodic metal oxides.

Animals↗
PubMed1998

Calcium-dependent clustering of inositol 1,4,5-trisphosphate receptors.

Rat basophilic leukemia (RBL-2H3) cells predominantly express the type II receptor for inositol 1,4,5-trisphosphate (InsP3), which operates as an InsP3-gated calcium channel. In these cells, cross-linking the high-affinity immunoglobulin E receptor (FcepsilonR1) leads to activation of phospholipase C gamma isoforms via tyrosine kinase- and phosphatidylinositol 3-kinase-dependent pathways, release of InsP3-sensitive intracellular Ca2+ stores, and a sustained phase of Ca2+ influx. These events are accompanied by a redistribution of type II InsP3 receptors within the endoplasmic reticulum and nuclear envelope, from a diffuse pattern with a few small aggregates in resting cells to large isolated clusters after antigen stimulation. Redistribution of type II InsP3 receptors is also seen after treatment of RBL-2H3 cells with ionomycin or thapsigargin. InsP3 receptor clustering occurs within 5-10 min of stimulus and persists for up to 1 h in the presence of antigen. Receptor clustering is independent of endoplasmic reticulum vesiculation, which occurs only at ionomycin concentrations >1 microM, and maximal clustering responses are dependent on the presence of extracellular calcium. InsP3 receptor aggregation may be a characteristic cellular response to Ca2+-mobilizing ligands, because similar results are seen after activation of phospholipase C-linked G-protein-coupled receptors; cholecystokinin causes type II receptor redistribution in rat pancreatoma AR4-2J cells, and carbachol causes type III receptor redistribution in muscarinic receptor-expressing hamster lung fibroblast E36(M3R) cells. Stimulation of these three cell types leads to a reduction in InsP3 receptor levels only in AR4-2J cells, indicating that receptor clustering does not correlate with receptor down-regulation. The calcium-dependent aggregation of InsP3 receptors may contribute to the previously observed changes in affinity for InsP3 in the presence of elevated Ca2+ and/or may establish discrete regions within refilled stores with varying capacity to release Ca2+ when a subsequent stimulus results in production of InsP3.

Animals↗
PubMed1998

Microbial factors and gingival crevicular fluid aspartate aminotransferase levels. A cross-sectional study.

The aim of this cross-sectional study was to investigate the clinical application of chairside tests for gingival crevicular fluid (GCF) aspartate aminotransferase (AST) levels and plaque BANA hydrolysis activity with the presence of the periodontal pathogens Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. The study comprised 100 periodontitis sites (pocket depths > or =4 mm, GI=3) from 10 patients with chronic adult periodontitis and 100 control sites (pocket depths <4 mm, GI<3) from 10 periodontally healthy patients comprising 55 healthy sites (pocket depths <4 mm, GI=0) and 45 gingivitis sites (pocket depths <4 mm, GI=1 or 2). The values for both BANA hydrolysis and AST levels were significantly higher in samples from periodontitis compared with gingivitis and healthy sites (p<0.001). A. actinomycetemcomitans was identified in 45% and P. gingivalis in 17% of periodontitis sites but neither pathogen was recovered from control sites and there was no significant correlation with the clinical parameters measured. There was no significant relationship between the presence of P. gingivalis and/or A. actinomycetemcomitans with BANA hydrolysis or AST levels. A significant correlation (p=0.0017) was observed between BANA hydrolysis and pocket depth and between AST hydrolysis and the GI (p=0.01). This study failed to demonstrate a positive association between chairside analysis of GCF metabolites for AST levels and/or BANA hydrolysis with P. gingivalis and A. actinomycetemcomitans. However, the GCF metabolites had a significant correlation with periodontally diseased sites in patients with chronic adult periodontitis and may help confirm clinical observations.

Adult↗
PubMed1998

Transforming growth factor-beta1 (TGF-beta1) in dentine matrix. Ligand activation and receptor expression.

Transforming Growth Factor-beta (TGF-beta) and other members of this family of growth factors have been implicated in tooth development and dental tissue repair. This study aimed to investigate regulatory factors for TGF-beta1 in rabbit incisor dentine matrix and the expression of receptors for TGF-beta in dental tissues better to understand the control of its biological activity. Approximately half of the TGF-beta1 in dentine matrix was present in active form. TGF-beta1 was found in association with latency associated peptide (LAP), betaglycan and decorin in an isolated dentine matrix preparation. Immunohistochemistry showed strong staining for TGF-beta type I and II receptors in odontoblasts with more variable and weaker staining of other pulpal cells. Association of TGF-beta1 with betaglycan, decorin and LAP may regulate the availability and biological activity of this growth factor and influence its presentation to the TGF-beta type I and II receptors on odontoblasts. During dental tissue repair, such control processes will be important in regulating the biological effects of TGF-beta1 on cells of the dentine-pulp complex.

Animals↗
PubMed1998

Human enamel as a substrate for in vitro acid dissolution studies: influence of tooth surface and morphology.

The variation in susceptibility of enamel to acid dissolution has been quantitatively assessed in vitro by examination of different surfaces of both maxillary and mandibular teeth of different morphology obtained from a pool of sound human teeth. The lingual/palatal surfaces of all teeth showed greater susceptibility to acid dissolution than the corresponding labial/buccal surfaces. The palatal surface of the maxillary teeth appeared more susceptible to acid than the lingual surface of the corresponding teeth in the mandible. The palatal surfaces of the maxillary teeth showed least variation in acid susceptibility, whilst the lingual surfaces of mandibular teeth and buccal/labial surfaces of teeth from both jaws showed much more variation between teeth of different morphology. These differences in susceptibility to acid dissolution probably reflect both developmental differences and post-eruptive changes in enamel and may contribute to the clinical presentation of dental disease. Importantly, the data indicates that careful selection of teeth for in vitro study of enamel dissolution processes is essential to minimise inter-sample variation. For such purposes, the palatal surface of the maxillary teeth appears to be most suitable.

Adult↗