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Biomedical subjects

Stuart Smith

Publications and source records attributed to Stuart Smith.

At least 19 recordsLinked to original sources
PubMed2007

Proteomic comparison of Mycobacterium avium subspecies paratuberculosis grown in vitro and isolated from clinical cases of ovine paratuberculosis.

Paratuberculosis (Johne's disease) poses a significant economic problem to beef, dairy and sheep industries worldwide, and is caused by Mycobacterium avium subspecies paratuberculosis. In this study, 2D PAGE was used as a tool to investigate the virulent state of M. avium subsp. paratuberculosis, incorporating the technique of beating the organism with zirconium/silica beads to provide a comprehensive representation of its proteome. A direct comparison of the proteomes of M. avium subsp. paratuberculosis scraped from the terminal ileum of ovine paratuberculosis cases, and the identical strain grown in vitro, is presented. These analyses identified a set of 10 proteins whose expression is upregulated during natural infection: 1-pyrroline-5-carboxylate dehydrogenase (RocA), a putative acyl-CoA dehydrogenase (FadE14), 2-methylcitrate dehydratase (2-mcd), arginosuccinate synthase (ArgG), universal stress protein (usp), 30S ribosomal protein S2 (RpsB), peptidyl-prolyl cis-trans isomerase (PpiA), luciferase-like monooxygenase (lmo), thiosulfate sulfurtransferase (SseA) and ATP-dependent Clp protease (ClpB). Most of the proteins identified do not have obviously related functions; however, ArgG and RocA function in the same pathway, and may have a concerted action for energy production in vivo.

1-Pyrroline-5-Carboxylate Dehydrogenase↗
PubMed2006

Worldsid assessment of far side impact countermeasures.

Far side impact trauma has been demonstrated as a significant portion of the total trauma in side impacts. The objective of the study was to assess the potential usefulness of countermeasures and assess the trade-offs associated with generic countermeasure design. Because the WorldSID dummy has demonstrated promise as a potential far side impact dummy, it was chosen to assess countermeasures in this mode. A unique far side impact buck was designed for a sled test system that included, as a standard configuration, a center console and outboard three-point belt system. This configuration assumed a left side driver with a right side impact. The buck allowed for additional options of generic restraints including shoulder or thorax plates or an inboard shoulder belt. The entire buck could be mounted on the sled in either a 90-degree (3-o'clock PDOF) or a 60-degree (2-o'clock PDOF) orientation. A total of 19 WorldSID tests were completed. The inboard shoulder belt configuration produced high shear forces in the lower neck (2430 N) when the belt position was placed over the mid portion of the neck. Shear forces were reduced and of opposite sign when the inboard belt position was horizontal and over the shoulder; forces were similar to the standard outboard belt configuration (830 - 1100 N). A shoulder or thorax restraint was effective in limiting the head excursion, but each caused significant displacement at the corresponding region on the dummy. A shoulder restraint resulted in shoulder displacements of 30 - 43 mm. A thorax restraint caused thorax deflections of 39 - 64 mm. Inboard restraints for far side impacts can be effective in reducing head excursion but the specific design and placement of these restraints determine their overall injury mitigating characteristics.

Accidents, Traffic↗
PubMed2006

An attempt to estimate the global burden of disease due to fluoride in drinking water.

A study was conducted to examine the feasibility of estimating the global burden of disease due to fluoride in drinking water. Skeletal fluorosis is a serious and debilitating disease which, with the exception of one area in China, is overwhelmingly due to the presence of elevated fluoride levels in drinking water. The global burden of disease due to fluoride in drinking water was estimated by combining exposure-response curves for dental and skeletal fluorosis (derived from published data) with model-derived predicted drinking water fluoride concentrations and an estimate of the percentage population exposed. There are few data with which to validate the output but given the current uncertainties in the data used, both to form the exposure-response curves and those resulting from the prediction of fluoride concentrations, it is felt that the estimate is unlikely to be precise. However, the exercise has identified a number of data gaps and useful research avenues, especially in relation to determining exposure, which could contribute to future estimates of this problem.

Adolescent↗
PubMed2005

Effect of modification of the length and flexibility of the acyl carrier protein-thioesterase interdomain linker on functionality of the animal fatty acid synthase.

A natural linker of approximately 20 residues connects the acyl carrier protein with the carboxy-terminal thioesterase domain of the animal fatty acid synthase. This study examines the effects of changes in the length and amino acid composition of this linker on catalytic activity, product composition, and segmental motion of the thioesterase domain. Deletion of 10 residues, almost half of the interdomain linker, had no effect on either mobility of the thioesterase domain, estimated from fluorescence polarization of a pyrenebutyl methylphosphono moiety bound covalently to the active site serine residue, or functionality of the fatty acid synthase; further shortening of the linker limited mobility of the thioesterase domain and resulted in reduced fatty acid synthase activity and an increase in product chain length from 16 to 18 and 20 carbon atoms. Surprisingly, however, even when the entire linker region was deleted, the fatty acid synthase retained 28% activity. Lengthening of the linker, by insertion of an unusually long acyl carrier protein-thioesterase linker from a modular polyketide synthase, increased mobility of the thioesterase domain without having any significant effect on catalytic properties of the complex. Interdomain linkers could also be used to tether, to the acyl carrier protein domain of the fatty acid synthase, a thioesterase active toward shorter chain length acyl thioesters generating novel short-chain fatty acid synthases. These studies reveal that although truncation of the interdomain linker partially impacts the ability of the thioesterase domain to terminate growth of the acyl chain, the overall integrity of the fatty acid synthase is quite tolerant to moderate changes in linker length and flexibility. The retention of fatty acid synthesizing activity on deletion of the entire linker region implies that the inherent flexibility of the phosphopantetheine "swinging arm" also contributes significantly to the successful docking of the long-chain acyl moiety in the thioesterase active site.

Acyl Carrier Protein↗
PubMed2005

Structure and molecular organization of mammalian fatty acid synthase.

De novo synthesis of fatty acids in the cytosol of animal cells is carried out by the multifunctional, homodimeric fatty acid synthase (FAS). Cryo-EM analysis of single FAS particles imaged under conditions that limit conformational variability, combined with gold labeling of the N termini and structural analysis of the FAS monomers, reveals two coiled monomers in an overlapping arrangement. Comparison of dimeric FAS structures related to different steps in the fatty acid synthesis process indicates that only limited local rearrangements are required for catalytic interaction among different functional domains. Monomer coiling probably contributes to FAS efficiency and provides a structural explanation for the reported activity of a FAS monomer dimerized to a catalytically inactive partner. The new FAS structure provides a new paradigm for understanding the architecture of FAS and the related modular polyketide synthases.

Animals↗
PubMed2005

Cloning, expression, and characterization of the human mitochondrial beta-ketoacyl synthase. Complementation of the yeast CEM1 knock-out strain.

A human beta-ketoacyl synthase implicated in a mitochondrial pathway for fatty acid synthesis has been identified, cloned, expressed, and characterized. Sequence analysis indicates that the protein is more closely related to freestanding counterparts found in prokaryotes and chloroplasts than it is to the beta-ketoacyl synthase domain of the human cytosolic fatty acid synthase. The full-length nuclear-encoded 459-residue protein includes an N-terminal sequence element of approximately 38 residues that functions as a mitochondrial targeting sequence. The enzyme can elongate acyl-chains containing 2-14 carbon atoms with malonyl moieties attached in thioester linkage to the human mitochondrial acyl carrier protein and is able to restore growth to the respiratory-deficient yeast mutant cem1 that lacks the endogenous mitochondrial beta-ketoacyl synthase and exhibits lowered lipoic acid levels. To date, four components of a putative type II mitochondrial fatty acid synthase pathway have been identified in humans: acyl carrier protein, malonyl transferase, beta-ketoacyl synthase, and enoyl reductase. The substrate specificity and complementation data for the beta-ketoacyl synthase suggest that, as in plants and fungi, in humans this pathway may play an important role in the generation of octanoyl-acyl carrier protein, the lipoic acid precursor, as well as longer chain fatty acids that are required for optimal mitochondrial function.

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase↗
PubMed2005

Learning from the past to inform the future--a survey of consultant nurses in emergency care.

This paper reports the findings of a survey of UK consultant nurses in emergency care. The purpose of the survey was to elicit information regarding level of preparation for the consultant nurse role, the use of formal competency frameworks, current clinical scope of practice and perspectives on future preparation for the role. A semi-structured questionnaire was e-mailed to consultant nurses in emergency care. Respondents had an average of only 2 years in post and for 24% of respondents this was their second post as a consultant nurse. The survey identified that three quarters of the respondents had no specific preparation for the consultant nurse role. The remainder had varying levels of preparation ranging from brief induction to 6-month clinical training. It could be argued that this diversity of preparation is a reflection of the lack of clarity regarding the consultant nurse role and the ill-defined organisational frameworks within which some consultant nurse posts were established. With the exception of the expert practice domain and clinical leadership, the majority of respondents felt under prepared for one or more elements of the consultant nurse role. Clinically their scope of practice ranged from managing patients with minor illness or injury, to leading resuscitation teams. There was great inequity in the level of preparation for the role, particularly in the transformational leadership, education and training, and practice and service development domains. Strategies for addressing these deficiencies are identified.

Consultants↗
PubMed2005

Insulin acutely decreases hepatic fatty acid synthase activity.

Insulin is viewed as a positive regulator of fatty acid synthesis by increasing fatty acid synthase (FAS) mRNA transcription. We uncover a new mechanism by which insulin acutely reduces hepatic FAS activity by inducing phosphorylation of the carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) and its interaction with FAS. Ceacam1 null mice (Cc1(-/-)) show loss of insulin's ability to acutely decrease hepatic FAS activity. Moreover, adenoviral delivery of wild-type, but not the phosphorylation-defective Ceacam1 mutant, restores the acute effect of insulin on FAS activity in Cc1(-/-) primary hepatocytes. Failure of insulin to acutely reduce hepatic FAS activity in hyperinsulinemic mice, including L-SACC1 transgenics with liver inactivation of CEACAM1, and Ob/Ob obese mice, suggests that the acute effect of insulin on FAS activity depends on the prior insulinemic state. We propose that this mechanism acts to reduce hepatic lipogenesis incurred by insulin pulses during refeeding.

Animals↗
PubMed2005

Replacing the academic medical center's teaching hospital.

Addressing the need for updated teaching hospital facilities is one of the most significant issues that an academic medical center faces. The authors describe the process they underwent in deciding to build a new facility at the Medical University of South Carolina (MUSC). Initial issues included whether or not the teaching hospital would continue to play a role in clinical education and whether to replace or renovate the existing facility. Once the decision to build was reached, MUSC had to choose between an on-campus or distant site for the new hospital and determine what the function of the old hospital would be. The authors examine these questions and discuss the factors involved in different stages of decision making, in order to provide the academic medicine community guidance in negotiating similar situations. Open communication within MUSC and with the greater community was a key component of the success of the enterprise to date. The authors argue that decisions concerning site, size, and focus of the hospital must be made by developing university-wide and community consensus among many different constituencies. The most important elements in the success at MUSC were having unified leadership, incorporating constituent input, engaging an external consultant, remaining unfazed by unanticipated challenges, and adhering to a realistic, aggressive timetable. The authors share their strategies for identifying and successfully managing these complex and potentially divisive aspects of building a new teaching hospital.

Academic Medical Centers↗
PubMed2005

Brushing with a potassium nitrate dentifrice to reduce bleaching sensitivity.

OBJECTIVE: This research systematically evaluated the use of a clinically proven desensitizing dentifrice prior to a bleaching regimen in a randomized, multi-center, parallel group, open label clinical study following Good Clinical Practice guidelines. METHODOLOGY: Fourteen dental offices in West Palm Beach, Florida participated in the study during April/May 2004. Fourteen days prior to bleaching, impressions and oral soft tissue assessments were performed, and patients were randomized to either a KNO3 plus fluoride dentifrice (Sensodyne Fresh Mint), or a standard fluoride dentifrice (Crest Regular), brushing 2x per day. On Day 14, patients returned to the dental office for their custom tray and the dispensation of a bleaching kit (Day White Excel 3; 9.5% hydrogen peroxide and KNO3). This was used daily according to the manufacturer's instructions for 30 minutes, and normal oral hygiene continued to be performed using the assigned toothbrush and dentifrice, brushing 2x per day. At the end of each bleaching day, patients answered diary questions about the occurrence and intensity of sensitivity. At the conclusion of the 14-day bleaching period (Day 28), patients returned to their dental office for re-examination, returning all products and diaries. Within seven days of completing the study, patients answered a telephone patient satisfaction survey. RESULTS: A total of 202 patients in fourteen (14) dental offices completed all aspects of the study and were used for the analysis. The professionally dispensed bleaching product provided an improvement of approximately 4.4 Vita shades, regardless of whether it was used with the KNO3 plus fluoride (Sensodyne) or a standard fluoride (Crest) dentifrice. The patient perception of increased sensitivity caused by the bleaching treatment was low but measurable. In the first week of the bleaching, significantly more patients using the KNO3 plus fluoride dentifrice were free from sensitivity (58%) than the standard fluoride dentifrice group (42%). During the 14-day bleaching treatment period, the KNO3 dentifrice patients experienced significantly more "sensitivity free days" (average = 10.1) compared to the standard fluoride dentifrice group (average = 8.6). CONCLUSION: The use of the KNO3 plus fluoride dentifrice (Sensodyne), two weeks prior to and throughout bleaching, may be a useful adjunct for the management of sensitivity caused by professionally dispensed bleaching products. With the bleaching-induced tooth sensitivity, those patients in the KNO3 plus fluoride toothpaste group were significantly more satisfied with their whitening experience and willing to repeat the bleaching treatment.

Adult↗
PubMed2004

Characterization of the beta-carbon processing reactions of the mammalian cytosolic fatty acid synthase: role of the central core.

The properties of the beta-ketoacyl reductase, dehydrase, and enoyl reductase components of the animal fatty acid synthase responsible for the reduction of the beta-ketoacyl moiety formed at each round of chain elongation have been studied by engineering and characterizing mutants defective in each of these three catalytic domains. These "beta-carbon processing" mutants leak the stalled four-carbon intermediates by direct transfer to CoA. However, enoyl reductase mutants leak beta-ketobutyryl, beta-hydroxybutyryl, and crotonyl moieties, a finding explained, at least in part, by the observation that the equilibrium and rate constant for the dehydrase reaction favor the formation of beta-hydroxy rather than enoyl moieties. In this regard, the type I animal fatty acid synthase resembles its type II counterpart in Escherichia coli in that both systems rely on the enoyl reductase to pull the beta-carbon processing reactions to completion. Kinetic and nucleotide binding measurements on fatty acid synthases mutated in either of the two nucleotide binding domains revealed that the NADPH binding sites are nonidentical, the enoyl reductase exhibiting higher affinity. Surprisingly, NADPH binding is also completely compromised by certain deletions and mutations in the central core region distant from the nucleotide binding sites. Comparable central core sequences are present in the structurally related modular polyketide synthases, except in those modules that lack all three beta-carbon processing enzymes. These findings suggest that the central core region of fatty acid and polyketide synthases plays an important role in facilitating the beta-carbon processing reactions.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed2004

Transcriptional regulation of the cardiac-specific MLC2 gene during Xenopus embryonic development.

The mechanisms by which transcription factors, which are not themselves tissue restricted, establish cardiomyocyte-specific patterns of transcription in vivo are unknown. Nor do we understand how positional cues are integrated to provide regionally distinct domains of gene expression within the developing heart. We describe regulation of the Xenopus XMLC2 gene, which encodes a regulatory myosin light chain of the contractile apparatus in cardiac muscle. This gene is expressed from the onset of cardiac differentiation in the frog embryo and is expressed throughout all the myocardium, both before and after heart chamber formation. Using transgenesis in frog embryos, we have identified an 82 bp enhancer within the proximal promoter region of the gene that is necessary and sufficient for heart-specific expression of an XMLC2 transgene. This enhancer is composed of two GATA sites and a composite YY1/CArG-like site. We show that the low-affinity SRF site is essential for transgene expression and that cardiac-specific expression also requires the presence of at least one adjacent GATA site. The overlapping YY1 site within the enhancer appears to act primarily as a repressor of ectopic expression, although it may also have a positive role. Finally, we show that the frog MLC2 promoter drives pan myocardial expression of a transgene in mice, despite the more restricted patterns of expression of murine MLC2 genes. We speculate that a common regulatory mechanism may be responsible for pan-myocardial expression of XMLC2 in both the frog and mouse, modulation of which could have given rise to more restricted patterns of expression within the heart of higher vertebrates.

Animals↗
PubMed2004

Head-to-head coiled arrangement of the subunits of the animal fatty acid synthase.

The role of the beta-ketoacyl synthase domains in dimerization of the 2505 residue subunits of the multifunctional animal FAS has been evaluated by a combination of crosslinking and characterization of several truncated forms of the protein. Polypeptides containing only the N-terminal 971 residues can form dimers, but polypeptides lacking only the N-terminal 422 residue beta-ketoacyl synthase domain cannot. FAS subunits can be crosslinked with spacer lengths as short as 6 A, via cysteine residues engineered near the N terminus of the full-length polypeptides. The proximity of the N-terminal beta-ketoacyl synthase domains and their essential role in dimerization is consistent with a revised model for the FAS in which a head-to-head arrangement of two coiled subunits facilitates functional interactions between the dimeric beta-ketoacyl synthase and the acyl carrier protein domains of either subunit.

Animals↗
PubMed2004

A multi-centre evaluation of the intra-assay and inter-assay variation of commercial and in-house anti-cardiolipin antibody assays.

AIMS: To assess the intra-assay (intra-run) and inter-assay (inter-run) variation of commercial and in-house IgG and IgM anti-cardiolipin antibody (aCL) assays/kits, and to determine an appropriate maximum value for inclusion in consensus guidelines. METHODS: Frozen aliquots of two patient specimens and one commercial control were sent to nine laboratories for the evaluation of eight commercial kits and one in-house assay. Intra-assay and inter-assay evaluations were performed with all three samples for IgG aCL, and one patient specimen for IgM aCL. RESULTS: The IgG and IgM aCL values varied considerably between the nine assays/kits. The majority of assays/kits demonstrated less than 20% intra-assay and inter-assay variation, with lower intra-assay and inter-assay variation observed with the commercial control. Single calibrator assays were not consistently associated with higher inter-assay variation than multi-point calibrator assays. CONCLUSIONS: An inter-assay coefficient of variation of 20% was determined to be an appropriate maximum value for inclusion in the Australasian aCL Working Party consensus guidelines. Improved standardisation between different assay/kits is still required.

Antibodies, Anticardiolipin↗
PubMed2004

Heart transplantation at the Ottawa heart institute: comparison with Canadian and international results.

Heart transplantation has been carried out in 340 patients in Ottawa, including seventy-one who required mechanical circulatory support as a bridge to transplant. Survival in Ottawa was compared with other Canadian centers based on data from the Canadian Organ Replacement Register up to the year 2000 and with the International Society of Heart and Lung Transplantation (ISHLT) registry 2001. For survival analysis, the number of adult patients at risk at year 0 was 303 (87 transplanted from 1985 to 1990, 105 from 1990 to 1994, and 111 from 1995 to 2000). The Statistical Analysis System (SAS) life test procedure was used. Survival was not adjusted for comorbidities or heart failure class. For the year of transplant 1985-1989, one-, five-, and ten-year patient survival in Ottawa was 83%, 70%, and 60%, respectively, compared to 82%, 71%, and 54%, respectively, for Canada (Wilcoxon test, P = 0.71), and compared to one- and five-year survival for ISHLT from 1980 to 1987 at 76% and 60%, respectively. For 1990-1994, one-, five-, and ten-year patient survival in Ottawa was 88%, 81%, and 74%, respectively, compared to 80%, 71%, and 61%, respectively, for Canada (P = 0.05), and compared to one- and five-year survival for ISHLT from 1998 to 1992 at 80% and 68%, respectively. For 1995-2000, one- and five-year patient survival in Ottawa was 90% and 82%, respectively, compared to 85% and 76%, respectively, for Canada (P = 0.09), and compared to one- and five-year survival for ISHLT from 1993 to 1996 at 82% and 68%, respectively. Survival after heart transplantation in Ottawa compares favorably with Canadian and international data.

Academies and Institutes↗
PubMed2004

A preliminary evaluation of child restraints and anchorage systems for an Australian car.

This study examined the performance of three rear-facing and two forward-facing child restraints (CRS) with three anchorage systems: standard seatbelt, LATCH (flexible) and ISOFIX (rigid). Frontal (64 km/h) and side impact (15 km/h) HyGe sled tests were conducted using a sedan buck. Overall, the preliminary findings suggested superior performance of rigid over seatbelt and flexible anchorages, particularly in side impacts. The results also suggest a need for design improvement for CRS with flexible anchorages to increase stability in side impacts. The findings have important implications for the proposed introduction of changes to Australian Standards for CRS to permit both flexible and rigid systems to coexist with conventional seatbelt anchorage systems.

Accidents, Traffic↗
PubMed2003

The use of oral tacrolimus in a case of short bowel syndrome.

Achieving adequate oral immunosuppression in a renal transplant patient who develops short bowel syndrome provides a significant challenge. We report a case where oral tacrolimus has been used to provide immunosuppression in an established renal transplant patient who developed short bowel syndrome secondary to mesenteric infarction. After reviewing the literature, we conclude that tacrolimus can be used as first-line immunosuppression in patients with short bowel syndrome.

Administration, Oral↗