Age adjusted D-dimer in the Belfast Health and Social Care Trust: A retrospective study.

D-dimers combined with clinical pre-test probability (PTP) scores are used to determine the likelihood of a venous thromboembolic event (VTE). It is recognised that with advancing age, d-dimer values increase, leading to a cohort of patients with a d-dimer above the standard cut-off of 500µg/L. A recent systemic review, examined the accuracy of an age-adjusted D-dimer in those aged > 50 years with a low clinical risk of a VTE. This showed an increase in specificity without loss of sensitivity. Our study, aimed to examine a population of patients, who between 2011 and 2014 underwent ultrasound Doppler studies of lower limbs. By applying a corresponding age-adjusted D-dimer, we determined the sensitivity and specificity and compared this to use of conventional D-dimer.

The role of specialist neuroradiology second opinion reporting: is there added value?

AIM: To assess the impact on patient management of formal neuroradiology "second reading" of computed tomography (CT) and magnetic resonance imaging (MRI) images initially interpreted by general radiologists. MATERIALS AND METHODS: Second opinion reports during the calendar year 2004 were compared with the original report and assessed for major or minor discrepancies. A major discrepancy was separated from a minor discrepancy whereby a change in opinion significantly affected patient management. RESULTS: There were 506 second opinions during 2004 given by three consultant neuroradiologists. Incomplete data were found in 141. Forty-one percent were CT images and the remainder MRI. The majority of second opinions were requested by neurologists. Most of the remaining referrals were from neurosurgeons or the primary radiologist. There was a 13% major and a 21% minor discrepancy rate. The remaining 66% were in complete agreement. There was a mixture of overcalls, misinterpretation, and undercalls. There were similar rates of minor and major discrepancies in both CT and MRI. CONCLUSION: There is a significant major discrepancy rate between specialist neuroradiology second opinion and general radiologists. The benefit of a formal specialist second opinion service is clearly demonstrated; however, it is time-consuming.

Failure of aneurysm sac shrinkage after endovascular repair; the effect of mural calcification.

AIM: To evaluate the effect of abdominal aortic aneurysm wall calcification on subsequent sac shrinkage after endovascular repair. MATERIALS AND METHODS: Seventy-three patients underwent endovascular aneurysm repair. The degree of sac wall calcification on pre-procedural computed tomography (CT) examination was graded from 1 to 4 according to the degree of circumferential involvement. On follow-up CT imaging, the maximum transverse diameter (MTD) of the sac was recorded, as well as the presence or absence of endoleak. In those patients with a non-shrinking aneurysm, but no CT evidence of endoleak, contrast-enhanced ultrasound (USS) was performed. Any patient with an endoleak, however diagnosed, was excluded from the study. Kruskal-Wallis and Spearman's rank correlation coefficient testing was applied to compare the degree of calcification and change in MTD. RESULTS: Sixty-three pre-procedural CT images were available for calcification grading. Six of this group had endoleaks resulting in 57 sets of data being available for the study. A reduction in MTD occurred in 68.25% of these patients by 1 year post-procedure. Our figures show aortic calcification is inversely associated with MTD reduction at 6 months (p = 0.01), 1 year (p = 0.05) and 2 years (p = 0.05). CONCLUSION: This study indicates that the degree of aortic wall calcification is significant in predicting MTD reduction post-endovascular repair. The possible mechanisms and implications of this are discussed.

Hodgkin' s disease with fulminant non-alcoholic steatohepatitis.

We report a woman with daily febrile episodes who developed fulminant hepatic failure. A percutaneous liver biopsy demonstrated non-alcoholic steatohepatitis, with no evidence of neoplastic infiltration. Post-mortem examination revealed stage IV Hodgkin's disease with trivial liver involvement. Rapidly progressive steatohepatitis causing acute liver failure may be a paraneoplastic presentation of Hodgkin's disease, possibly mediated by cytokines.

Cloning of random oligonucleotides to create single-insert plasmid libraries.

Random double-stranded oligonucleotides are useful reagents to identify the optimal binding sites for DNA-binding proteins, such as transcriptional activators. Some applications require ligation of random oligonucleotides to form plasmid-based libraries such as the yeast one-hybrid system, where the activation of a cloned DNA sequence from a library of random DNA-binding sequences activates a reporter gene. Current theories do not account for the low efficiencies of oligonucleotide-based plasmid library construction methods. We developed a technique to clone single oligonucleotides into plasmid vectors with high efficiency that predictably results in only one oligonucleotide insert per colony and used this method to clone a yeast one-hybrid library. This method, either as presented or with modifications, should be suitable for any situation where high-efficiency cloning of single oligonucleotide inserts is desired.

Characterization of a human plasma membrane heme transporter in intestinal and hepatocyte cell lines.

Heme is the most bioavailable form of dietary iron and a component of many cellular proteins. Controversy exists as to whether heme uptake occurs via specific transport mechanisms or passive diffusion. The aims of this study were to quantify cellular heme uptake with a fluorescent heme analog and to determine whether heme uptake is mediated by a heme transporter in intestinal and hepatic cell lines. A zinc-substituted porphyrin, zinc mesoporphyrin (ZnMP), was validated as a heme homolog in uptake studies of intestinal (Caco-2, I-407) and hepatic (HepG2) cell lines. Uptake experiments to determine time dependence, heme inhibition, concentration dependence, temperature dependence, and response to the heme synthesis inhibitor succinylacetone were performed. Fluorescence microscope images were used to quantify uptake and determine the cellular localization of ZnMP; ZnMP uptake was seen in intestinal and hepatic cell lines, with cytoplasmic uptake and nuclear sparing. Uptake was dose- and temperature dependent, inhibited by heme competition, and saturated over time. Preincubation with succinylacetone augmented uptake, with an increased initial uptake rate. These findings establish a new method for quantifying heme uptake in individual cells and provide strong evidence that this uptake is a regulated, carrier-mediated process.

Transfection assays for transformation of model colonic cell lines.

The recognition of cancer as a genetic disease has changed the approach investigators take to understanding the mechanisms of carcinogenesis. The discovery of oncogenes, and the recognition of the inactivation of tumor suppressor genes, DNA repair enzymes, and of apoptotic pathways have provided a clearer picture of the dysregulation which is required for a cell to become a cancer.

Functional properties of transfected human DMT1 iron transporter.

Recently, mutation of the DMT1 gene has been discovered to cause ineffective intestinal iron uptake and abnormal body iron metabolism in the anemic Belgrade rat and mk mouse. DMT1 transports first-series transition metals, but only iron turns on an inward proton current. The process of iron transport was studied by transfection of human DMT1 into the COS-7 cell line. Native and epitope-tagged human DMT1 led to increased iron uptake. The human gene with the Belgrade rat mutation was found to have one-fifth of the activity of the wild-type protein. The pH optimum of human DMT1 iron uptake was 6.75, which is equivalent to the pH of the duodenal brush border. The transporter demonstrates uptake without saturation from 0 to 50 microM iron, recapitulating earlier studies of isolated intestinal enterocytes. Diethylpyrocarbonate inhibition of iron uptake in DMT1-transfected cells suggests a functional role for histidine residues. Finally, a model is presented that incorporates the selectivity of the DMT1 transporter for transition metals and a potential role for the inward proton current.

A case of Balamuthia mandrillaris meningoencephalitis.

Balamuthia mandrillaris is a newly described pathogen that causes granulomatous amebic encephalitis, an extremely rare clinical entity that usually occurs in immunosuppressed individuals. We report a case of pathologically proven Balamuthia encephalitis with unusual laboratory and radiologic findings. A 52-year-old woman with idiopathic seizures and a 2-year history of chronic neutropenia of unknown cause had a subacute illness with progressive lethargy, headaches, and coma and died 3 months after the onset of symptoms. Cerebrospinal fluid (CSF) glucose concentrations were extremely low or unmeasurable, a feature not previously described (to our knowledge). Cranial magnetic resonance imaging scans showed a single large temporal lobe nodule, followed 6 weeks later by the appearance of 18 ring-enhancing lesions in the cerebral hemispheres that disappeared after treatment with antibiotics and high-dose corticosteroids. The initial brain biopsy specimen and analysis of CSF samples did not demonstate amebae, but a second biopsy specimen and the postmortem pathologic examination showed Balamuthia trophozoites surrounded by widespread granulomatous inflammation and vasculitis. The patient's neutropenia and antibiotic use may have caused susceptibility to this organism. Amebic meningoencephalitis should be considered in cases of subacute meningoencephalitis with greatly depressed CSF glucose concentrations and multiple nodular lesions on cerebral imaging. Arch Neurol. 2000;57:1210-1212

Multicenter evaluation of the Abbott LCx Mycobacterium tuberculosis ligase chain reaction assay.

Four Australian hospital laboratories evaluated the performance of the Abbott LCx Mycobacterium tuberculosis assay with 2,347 specimens (2,083 respiratory and 264 nonrespiratory specimens) obtained from 1, 411 patients. A total of 152 specimens (6.5%) were culture positive for Mycobacterium tuberculosis complex (MTBC); of these, 79 (52%) were smear positive. After resolution of discrepant data, the overall sensitivity, specificity, and positive and negative predictive values for the LCx assay were 69.7, 99.9, 99.1, and 97.7% respectively. For smear-positive respiratory specimens that were culture positive for MTBC, the values were 98.5, 100, 100, and 98.4%, respectively, while the values for smear-negative respiratory specimens were 41.5, 99.9, 96.4, and 98%, respectively. Relative operating characteristic curves were constructed to demonstrate the relationship between sensitivity and specificity for a range of possible cutoff values in the LCx assay. These graphs suggested that the assay sensitivity for respiratory samples could be increased from 70.2 to 78.6%, while the specificity would be reduced from 99.9 to 99.4% by inclusion of a grey zone (i.e., LCx assay values of between 0.2 and 0.99). An algorithm is presented for the handling of specimens with LCx assay values within this grey zone.

Fosphenytoin y-site stability studies with Lorazepam and midazolam hydrochloride.

The treatment of status epilepticus may require in certain situations the concurrent administration of lorazepam or midazolam with fosphenytoin. Simulated Y-site fosphenytoin/lorazepam and fosphenytoin/midazolam hydrochloride admixtures, respectively, in 0.9% sodium chloride injection were analyzed using a stability-indicating high-performance liquid chromatography (HPLC) method. Each drug was analyzed for stability by HPLC from three simulated Y-site samples over an eight-hour period. The HPLC assay results indicate that both fosphenytoin and lorazepam are stable together at a Y site over an eight-hour period. In addition, there was neither a change in sample clarity nor a change in pH over the same period. The results indicate that, when medically necessary, fosphenytoin and lorazepam in 0.9% sodium chloride injection can be administered via the same intravenous (IV) line. Midazolam free base was precipitated upon admixture of midazolam hydrochloride and fosphenytoin solutions. Therefore, midazolam hydrochloride and fosphenytoin should not be given via the same IV line.

Associations of smoking, alcohol and physical activity with risk factors for coronary heart disease and diabetes in the first follow-up cohort of the Heart Disease and Diabetes Risk Indicators in a Screened Cohort study (HDDRISC-1).

OBJECTIVES: To investigate the associations between risk factors for cardiovascular disease and cigarette smoking, alcohol intake, and physical activity in a group of predominantly healthy men. DESIGN: Cohort study with baseline characterisation, clinical follow-up, and identification of predictors of coronary artery disease and diabetes. SETTING: University hospital metabolic day ward. SUBJECTS: Participants in a company health programme (n=742). MAIN OUTCOME MEASURES: Routine haematology and biochemistry, cholesterol, triglycerides, high density lipoprotein (HDL) cholesterol (on a subset of 522 subjects), and glucose and insulin levels during a 3 h oral glucose tolerance test (OGTT). RESULTS: Independent associations with previous cigarette smoking included high uric acid and low HDL cholesterol, and with current cigarette smoking, high haemoglobin and white cell count and low OGTT insulin. Increasing alcohol intake was associated with increasing blood pressure, uric acid, HDL cholesterol and fasting glucose. The moderate range of exercise intensity in this cohort was associated with decreasing systolic blood pressure, fasting insulin and OGTT glucose and insulin. Factor analysis distinguished principal factors comprising features of the metabolic syndrome with low physical activity, and high white cell count, high haemoglobin concentration and low HDL cholesterol with increasing previous and current cigarette smoking and alcohol intake. CONCLUSIONS: Some characteristics of the metabolic syndrome were seen with previous but not current smoking habit. Regular alcohol consumption was associated with mainly unfavourable metabolic characteristics, although there was an independent beneficial association with HDL cholesterol. The improved metabolic syndrome profile seen with increasing exercise is consistent with even moderate degrees of physical activity having beneficial effects on metabolism.

New solution for prolonged myocardial preservation for transplantation.

BACKGROUND: A solution for prolonged cold storage of the heart has been developed. The Jerusalem-Cape Town Solution (JCT) is an "intracellular" type cardioplegic solution and is formulated to (1) minimize hypothermic-induced cell swelling, (2) diminish intracellular acidosis, (3) prevent the expansion of the interstitial space during the reperfusion, (4) protect against oxygen free radical injury during early reperfusion, and (5) provide substrates for regenerating high-energy phosphates. METHODS: With a Langendorff model, rat hearts were subjected to 15 minutes of perfusion with Krebs-Henseleit, 10 minutes of cardioplegic infusion and 20 hours of cold storage (5 degrees to 6 degrees C). Hearts were reperfused for 60 minutes and hemodynamic recovery was assessed. The hearts were assigned to three groups (eight hearts in each), according to the cardioplegic solution used: group 1, JCT; group 2, Bretschneider's HTK cardioplegic solution; and group 3 University of Wisconsin cold storage solution. RESULTS: After 60 minutes of reperfusion, the recovery of the coronary artery flow in group 1 (JCT) was significantly better than in group 2, and slightly better than in group 3 (64% +/- 8.9%, 47.2% +/- 11.6%, 52.5% +/- 19.9%, mean +/- SD, respectively; group 1 versus group 2, p < 0.01). The recovery of the left ventricular developed pressure (LVDP) was significantly better in group 1 compared with group 2 and group 3 (60.2% +/- 14.5%, 41.1% +/- 12.6% and 36.5% +/- 10.1%, respectively; p < 0.01). The recovery of the heart contractility expressed by the product of LVDP and the heart rate (LVDP x heart rate) was significantly higher in group 1 than in group 2 and group 3 (47.5% +/- 3.4%, 23.6% +/- 9.6%, and 28.7% +/- 8.3%, respectively, p < 0.001). In hearts stored for 12 hours in JCT or HTK, the recovery of the heart contractility did not differ significantly (73.4% +/- 12.7% or 70.8% +/- 30.8%, respectively). Modified reperfusion aimed to improve postischemic heart recovery did not bring significant changes in cardiac mechanical function but resulted in an increase in postischemic coronary artery flow recovery in hearts reperfused with amino acid-enriched buffer. CONCLUSIONS: The JCT solution is effective (as well as HTK) in preserving the ischemic hearts for up to 12 hours. It is superior to HTK or University of Wisconsin solution at 20 hours of isolated ischemic storage.

Factors of the metabolic syndrome: baseline interrelationships in the first follow-up cohort of the HDDRISC Study (HDDRISC-1). Heart Disease and Diabetes Risk Indicators in a Screened Cohort.

Syndromes of risk factor disturbance may contribute to the development of coronary heart disease and non-insulin-dependent diabetes mellitus, but their definition and quantification remain problematic. Using factor analysis, constellations of risk factor variables that could indicate distinct syndromes of metabolic disturbance were explored in the baseline data of the first follow-up cohort of 742 men from the Heart Disease and Diabetes Risk Indicators in a Screened Cohort (HDDRISC) study. The primary analysis considered 16 intercorrelated variables measured in more than 90% of cohort participants. A missing-values estimation routine was used to ensure inclusion of all participants in the analysis. Subanalyses were undertaken, including a repeat of the primary analysis on the 522 individuals who had received measurement of HDL cholesterol, an oblique rather than orthogonal factor rotation procedure performed on primary and HDL subset analyses, a repeat of these two primary and HDL subset analyses using only those participants with complete measurements, and a repeat of these six analyses including only the seven variables conventionally associated with the metabolic syndrome. The principal factor that emerged in all analyses undertaken comprised oral glucose tolerance test insulin and glucose response, serum uric acid, and body mass index. Fasting serum triglyceride concentration was included in this factor in 11 of the 12 analyses undertaken, fasting plasma insulin in 8, fasting plasma glucose in 5, and mean arterial pressure in 3. HDL cholesterol factored in isolation from insulin in all analyses undertaken. These findings provide strong support for a core metabolic cluster, which is unlikely to include blood pressure and does not include HDL. The factor scores relating to this cluster will provide a means of assessing its quantitative importance in prospective analysis of the development of CHD and diabetes in this cohort.

Metal binding properties and secondary structure of the zinc-binding domain of Nup475.

Nup475 is a nuclear zinc-binding protein of unknown function that is induced in mammalian cells by growth factor mitogens. Nup475 contains two tandemly repeated sequences YKTELCX8CX5CX3H (Cys3His repeats) that are thought to be zinc-bindin domains. Similar sequences have been found in a number of proteins from various species of eukaryotes. To determine the metal binding properties and secondary structure of the putative zinc-binding domains of Nup475, we have used synthetic or recombinant peptides that contain one or two domain sequences. The peptide with a single domain bound 1.0 +/- 0.1 equivalents of Co2+, and the peptide with two domains bound 1.7 +/- 0.4 equivalents of Co2+. Both peptides bound Co2+ and Zn2+ with affinities similar to those of classical zinc finger peptides. In each case, the Co2+ complex exhibited strong d-d transitions characteristic of tetrahedral coordination. For structural studies by nuclear magnetic resonance spectroscopy, we used a more soluble two-domain peptide that had a single amino acid substitution in a nonconserved amino acid residue in the second Cys3His repeat. The mutant peptide unexpectedly showed loss of one of its metal binding sites and displayed ordered structure for only the first Cys3His sequence. On the basis of the nuclear magnetic resonance data, we propose a structure for the Nup475 metal-binding domain in which the zinc ion is coordinated by the conserved cysteines and histidine, and the conserved YKTEL motif forms a parallel sheet-like structure with the C terminus of this domain. This structure is unlike that of any previously described class of metal binding domain.

Cytomegalovirus colitis mimicking colon carcinoma in an HIV-negative patient with chronic renal failure.

Cytomegalovirus (CMV) colitis is thought to occur almost exclusively in immunosuppressed persons. Colonoscopy in patients with CMV colitis usually shows diffuse or localized ulceration, although mucosal friability, erosions, hemorrhage, and plaque-like pseudomembranes may be observed. We report on a patient with chronic renal failure undergoing hemodialysis therapy who had abdominal symptoms, including bloody diarrhea, along with colonoscopic findings suggestive of carcinoma of the colon. The patient was not infected with the human immunodeficiency virus and had normal lymphocyte subset numbers. He was subsequently found to have invasive CMV disease of the colon. CMV colitis can occur in persons who are not severely immunosuppressed, and its colonoscopic appearance may mimic that of colon cancer.