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Protein-xenobiotic adducts are byproducts of xenobiotic metabolism. While there is a correlation between protein adduction and target organ toxicity, a cause and effect relationship is not often clear. Naphthoquinone (NQ) and monocrotaline pyrrole (MCTP) are two pneumotoxic electrophiles that form covalent adducts with a similar select group of proteins rich in reactive thiols. In this study, we treated human pulmonary artery endothelial cells (HPAEC) with NQ, MCTP, or preformed NQ or MCTP adducts to the protein galectin-1 (gal-1) and examined indicators of reactive oxygen species (ROS) oxidative injury, markers of apoptosis (caspase-3 and annexin V), and gene responses of cellular stress. ROS production was assayed fluorescently using CM-H(2)DCFDA. NQ adducts to gal-1 (NQ-gal) produced 183% more intracellular ROS than gal-1 alone (p < 0.0001). Caspase-3 activity and annexin V staining of phosphatidylserine were used to assess apoptotic activity in treated cells. HPAEC exposed to MCTP-gal had increases in both caspase-3 activation and membrane translocation of annexin V relative to gal-1 alone (p < 0.0001). Direct application of NQ produced significantly more ROS and induced significant caspase-3 activation, whereas MCTP did not. Human bronchial epithelial cells were also exposed to MCTP-gal and found to have significant increases in both caspase-3 activation and annexin V staining in comparison to that of gal-1 (p < 0.05). Western blot analysis showed that both NQ and MCTP significantly induced the Nrf2 mediated stress response pathway despite differences in ROS generation. ER stress was not induced by either adducts or parent compounds as seen by quantitative RT-PCR, but HOX-1 expression was significantly induced by NQ-gal and MCTP alone. Electrophile adduction to gal-1 produces different cytotoxic effects specific to each reactive intermediate.

Objective: Abnormalities in circulating ghrelin have been reported in chronic liver disease. This study assessed the response of anabolic peptides ghrelin, growth hormone (GH) and insulin-like growth factor 1 (IGF-1) in patients with alcoholic cirrhosis and healthy subjects to oral glucose. In a previous study, using oral glucose we identified loss of ghrelin regulation in nonalcoholic steato-hepatitis. PATIENTS/DESIGN/MEASUREMENTS: Fourteen patients with alcoholic cirrhosis were compared with 11 healthy subjects. Patients with cirrhosis were studied when adjudged clinically stable in hospital. After an overnight fast, they ingested 100-g glucose dissolved in 250 ml of water. Blood was sampled before and every 20 minutes after ingestion for 120 minutes. Plasma acylated and des-acyl ghrelin, GH, IGF-1 and insulin were assayed by ELISA., Results: Expressed as median (95% CI): 120-minutes integrated acylated ghrelin was 26 (19-66) in controls compared to 170 (129-252) pg/ml per hour in patients with cirrhosis; P < 0.001. Both groups exhibited a normal postglucose plasma total ghrelin profile. Among patients with cirrhosis (compared to controls), growth hormone was increased 15-fold and IGF-1 decreased 4-fold. Acylated ghrelin correlated with GH (Spearman r = 0.69, P = 0.0015) in control subjects but not in patients with cirrhosis., Conclusions: Acylated ghrelin is markedly increased in alcoholic cirrhosis, with apparent preservation of normal postprandial mechanisms of gastric ghrelin secretion. GH is also increased; however, its correlation with acylated ghrelin (confirmed in healthy subjects) is absent in patients with cirrhosis. Despite increased ghrelin and GH, patients with alcoholic cirrhosis remain anorexic and catabolic suggesting potential tissue resistance to the actions of these anabolic peptides.

Aims: Anaemia is highly prevalent and associated with poor prognosis in patients with chronic heart failure (CHF). Reduced erythroid proliferation capacity of haematopoietic progenitor cells is associated with reduced telomere length, a marker of cellular ageing. We hypothesize that short telomere length contributes to the susceptibility to develop anaemia in patients with CHF., Methods and Results: We studied 875 CHF patients, of whom 254 (29%) fulfilled the WHO criteria of anaemia. Telomere length in DNA from peripheral leucocytes was measured with real-time quantitative polymerase chain reaction. Age, gender, and baseline differences adjusted telomere length was correlated with haemoglobin levels (partial r = 0.130; P = 0.011). One standard deviation shorter telomere length was associated with an increased risk of having anaemia [odds ratio (OR), 1.31; 95% confidence interval (CI), 1.12-1.53; P = 0.001]. This observation was not affected by adjustment for potential confounders (OR, 1.38; 95% CI, 1.05-1.81; P = 0.021 after adjustment for age, gender, erythropoietin levels, renal function, left ventricular ejection fraction, age of CHF onset, blood pressure, history of stroke, diabetes, and B-type natriuretic peptide levels)., Conclusion: Shorter telomere length increases the odds of having anaemia in CHF patients. This finding supports the hypothesis that cellular ageing in CHF contributes to the susceptibility to develop anaemia.

With advancing age and the affluent, low-fiber Western diet, the incidence of diverticular disease is increasing. Fortunately, most cases can be managed conservatively without resorting to surgical intervention. Life-threatening complications such as perforation, especially when it is associated with gross fecal contamination, requires urgent aggressive surgical intervention. A 75-year-old man with absolute constipation and pain in the left iliac fossa underwent urgent laparotomy following fluid and antibiotic resuscitation. A posterior perforated sigmoid diverticulitis associated with myofascial necrosis and generalized pelvic emphysema was identified. In cases where perforation occurs posteriorly and the only external manifestation is surgical emphysema, the outcome is generally favorable.

The immobilisation of proteins on to nanoparticles has a number of applications ranging from biocatalysis through to cellular delivery of biopharmaceuticals. Here we describe a phosphopantetheinyl transferase (Sfp)-catalysed method for immobilising proteins bearing a small 12-mer "ybbR" tag on to nanoparticles functionalised with coenzyme A. The Sfp-catalysed immobilisation of proteins on to nanoparticles is a highly efficient, single step reaction that proceeds under mild conditions and results in a homogeneous population of proteins that are covalently and site-specifically attached to the surface of the nanoparticles. Several enzymes of interest for biocatalysis, including an arylmalonate decarboxylase (AMDase) and a glutamate racemase (GluR), were immobilised on to nanoparticles using this approach. These enzymes retained their activity and showed high operational stability upon immobilisation.

The synthesis of the complete family of phosphatidylinositol phosphate analogues (PIPs) from five key core intermediates A-E is described. These core compounds were obtained from myo-inositol orthoformate 1 via regioselective DIBAL-H and trimethylaluminium-mediated cleavages and a resolution-protection process using camphor acetals 10. Coupling of cores A-E with phosphoramidites 34 and 38, derived from the requisite protected lipid side chains, afforded the fully-protected PIPs. Removal of the remaining protecting groups was achieved via hydrogenolysis using palladium black or palladium hydroxide on carbon in the presence of sodium bicarbonate to afford the complete family of dipalmitoyl- and amino-PIP analogues 42, 45, 50, 51, 58, 59, 67, 68, 76, 77, 82, 83, 92, 93, 99 and 100. Investigations using affinity probes incorporating these compounds have identified novel proteins involved in the PI3K intracellular signalling network and have allowed a comprehensive proteomic analysis of phosphoinositide interacting proteins.

Background: Telomeres are causally involved in senescence. Senescence is a potential factor in the pathogenesis and progression of heart failure. In heart failure telomeres are shorter, but the prognostic value associated with telomere length has not been defined., Methods: Telomere length was prospectively determined by quantitative polymerase chain reaction in 890 patients with New York Heart Association (NYHA) functional class II to IV heart failure. After 18 months, we examined the association between telomere length and the predefined primary end-point: time to death or hospitalization for heart failure., Results: Mean age of the patients was 71 years, 39% were women, 51% were in NYHA class II, and 49% were in class III/IV. A total of 344 patients reached the primary end-point (130 deaths and 214 hospitalizations). Patients with shorter telomeres were at an increased risk of reaching the primary end-point (hazard ratio 1.79; 95% confidence interval (CI) 1.21-2.63). In multivariate analysis shorter telomere length remained associated with a higher risk for death or hospitalization (hazard ratio, 1.74; 95% CI 1.07-2.95) after adjustment for age of heart failure onset, gender, hemoglobin, renal function, and N-terminal pro-B-type natriuretic peptide level, a history of stroke, atrial fibrillation, and diabetes., Conclusions: Shorter length of telomeres predicts the occurrence of death or hospitalization in patients with chronic heart failure.

Introduction: POSSUM and its variants Portsmouth POSSUM (P-POSSUM) and Colorectal POSSUM (CR-POSSUM) equations were derived from a heterogeneous general surgical population, which have been used successfully to provide risk-adjusted operative mortality rates. CR-POSSUM utilises fewer parameters, allowing ease of use. The aim of this study was to predict the mortality outcome in colorectal surgery using these scoring systems compared to the observed mortality and to devise a new scoring system with improved accuracy., Methods: The study was conducted prospectively on all consecutive patients requiring elective and emergency colorectal surgery between April 2002 and May 2005. The outcome parameter was defined as 30-day mortality. The observed mortality was compared with predicted mortality by the scoring systems. Hosmer and Lemeshow test was used to assess statistical accuracy of POSSUM., Results: Eight hundred ninety-nine patients underwent colorectal surgery during the study period. There were 619 elective and 281 emergency patients. Observed 30-day mortality rate was 9%, compared with predicted mortality rate of 13.5% with POSSUM, 5% with P-POSSUM and 9.5% with CR-POSSUM., Conclusion: POSSUM's mortality rate was overestimated, while P-POSSUM's mortality rate was underestimated. CR-POSSUM, the simplest system of all three, most accurately predicted mortality in our unit.

Background: Renal dysfunction is a frequent comorbidity associated with high mortality in patients with chronic heart failure (CHF). The intrinsic biological age might affect the ability of the kidney to cope with the challenging environment caused by CHF. We explored the association between leukocyte telomere length, a marker for biological age, and renal function in patients with CHF., Methods and Results: Telomere length was determined by a real-time quantitative polymerase chain reaction in 866 CHF patients. Renal function was estimated with the simplified Modification of Diet in Renal Disease equation. The median age was 74 (interquartile range 64-79) years, 61% male, left ventricular ejection fraction of 30 (23-44)%, and the estimated glomerular filtration rate was 53 (40-68) ml/min/1.73 m(2). Telomere length was associated with renal function (correlation coefficient 0.123, P < 0.001). This relationship remained significant after adjustment for age, gender, age of CHF onset (standardized-beta 0.091, P = 0.007). Also additionally adjusting for the severity of CHF and baseline differences did not change our findings., Conclusion: The association between shorter leukocyte telomere length and reduced renal function in heart failure suggests that intrinsic biological aging affects the ability of the kidney to cope with the systemic changes evoked by heart failure.

Cardiolipin (1) is a dimeric phospholipid found in the mitochondrial membranes of both plants and animals. In order to understand better its role, we report the preparation of an immobilised analogue (2) using phosphoramidite chemistry; the probe has been used successfully to bind a recombinant protein containing a cardiolipin-binding domain.

Immobilizing chemically synthesized analogues of PI(3,4,5)P3 onto Affi-10 beads and incorporating them into liposomes allowed their use as affinity absorbents in the comprehensive analysis of the phosphoinositide interactome using cytosolic cell extracts of the LIM1215 colon cancer cell line. This led to the identification of 282 proteins that either interact with PI(3,4,5)P3 or are indirectly captured as part of a complex containing a PI(3,4,5)P3 binding partner. Identification of the proteins was achieved using affinity/LC-MS/MS experiments.

Introduction: An involved circumferential resection margin (CRM) following surgery for rectal cancer is the strongest predictor of local recurrence and may represent a failure of the multidisciplinary team (MDT) process., Aim of Study: The study analyses the causes of positive CRM in patients undergoing elective surgery for rectal cancer with respect to the decision-making process of the MDT, preoperative rectal cancer staging and surgical technique., Method: From March 2002 to September 2005, data were collected prospectively on all patients undergoing elective rectal cancer surgery with curative intent. The data on all patients identified with positive CRM were analysed., Results: Of 158 patients (male:female = 2.2:1) who underwent potentially curative surgery, 16 (10%) patients had a positive CRM on postoperative histology. Four were due to failure of the pelvic magnetic resonance imaging (MRI) staging scans to predict an involved margin, two with an equivocal CRM on MRI did not have preoperative radiotherapy, one had an inaccurate assessment of the site of primary tumour and in one intra-operative difficulty was encountered. No failure of staging or surgery was identified in the remaining eight of the 16 patients. Abdominoperineal resection (APR) was associated with a 26% positive CRM, compared with 5% for anterior resection., Conclusion: No single consistent cause was found for a positive CRM. The current MDT process and/or surgical technique may be inadequate for low rectal tumours requiring APR.

Background: To evaluate whether the risk of methotrexate (MTX) exposure through skin contamination using parenteral doses of 25 mg warrants special oncology handling precautions during administration., Methods: We conducted a study with six human volunteers deliberately exposed to an entire dose of 25 mg MTX solution on their skin for 30 min. Serum levels of MTX were measured at baseline, 2, 4, 8, 12 and 24 h as well as serum homocysteine at baseline and 24 h after clinical exposure. Twenty-four-hour urinary excretion of MTX and possible local or systemic signs of toxicity were also recorded., Results: All MTX serum concentrations were less than 0.02 micromol/L within the 24-h period. This is 500 times below the recommended serum concentration for which folinic acid supplementation is recommended. There was also no significant increase in homocysteine level to suggest MTX toxicity. The only adverse effects were mild local dermal reactions in three female volunteers., Conclusion: Deliberate skin contamination and possible inhalation of a 25 mg MTX solution failed to show significant or quantifiable serum and urine concentrations to suggest MTX toxicity. Precautions to prevent contact with MTX designed for oncology protocols are unnecessary for our rheumatology patients or their carers using these much lower immunosuppressant doses for autoimmune diseases.

Background: Renal failure aggravates pathological cardiac remodelling induced by myocardial infarction (MI). Cardiac remodelling is associated with telomere shortening, a marker for biological ageing. We investigated whether mild and severe renal failure shorten cardiac telomeres and excessively shorten telomeres after MI., Methods: Rats were subjected to sham, unilateral (UNX) or 5/6th nephrectomy (5/6NX) to induce none, mild or severe renal failure. MI was induced by left coronary artery ligation. Renal function parameters and blood pressure were measured. DNA was isolated from non-infarcted cardiac tissue. Telomere length was assessed by quantitative polymerase chain reaction (PCR)., Results: Proteinuria was unchanged in UNX and MI compared with control, but strongly increased in 5/6NX, UNX+MI and 5/6NX+MI. Serum creatinine levels were increased fourfold in 5/6NX and tenfold in 5/6NX+MI. 5/6NX and groups with both renal failure and MI showed an approximate 20% reduction of telomere length, similar to the MI group. No excess telomere shortening was observed in hearts from rats with renal ablation after MI., Conclusion: Severe renal failure, but not mild renal failure, leads to shortening of cardiac telomeres to a similar extent as found after MI. Renal failure did not induce excessive telomere shortening after MI. (Neth Heart J 2009;17:190-4.).

Purpose: Cell cycle dysregulation resulting in expression of antiapoptotic genes and uncontrolled proliferation is a feature of undifferentiated nasopharyngeal carcinoma. The pharmacodynamic effects of seliciclib, a cyclin-dependent kinase (CDK) inhibitor, were studied in patients with nasopharyngeal carcinoma., Experimental Design: Patients with treatment-naïve locally advanced nasopharyngeal carcinoma received seliciclib at 800 mg or 400 mg twice daily on days 1 to 3 and 8 to 12. Paired tumor samples obtained at baseline and on day 13 were assessed by light microscopy, immunohistochemistry, and transcriptional profiling using real-time PCR low-density array consisting of a panel of 380 genes related to cell cycle inhibition, apoptosis, signal transduction, and cell proliferation., Results: At 800 mg bd, one patient experienced grade 3 liver toxicity and another had grade 2 vomiting; no significant toxicities were experienced in 13 patients treated at 400 mg bd. Seven of fourteen evaluable patients had clinical evidence of tumor reduction. Some of these responses were associated with increased tumor apoptosis, necrosis, and decreases in plasma EBV DNA posttreatment. Reduced protein expression of Mcl-1, cyclin D1, phosphorylated retinoblastoma protein pRB (T821), and significant transcriptional down-regulation of genes related to cellular proliferation and survival were shown in some patients posttreatment, indicative of cell cycle modulation by seliciclib, more specifically inhibition of cdk2/cyclin E, cdk7/cyclin H, and cdk9/cyclin T., Conclusions: Brief treatment with this regimen of seliciclib in patients with nasopharyngeal carcinoma is tolerable at 400 mg bd and associated with tumor pharmacodynamic changes consistent with cdk inhibition, and warrants further efficacy studies in this tumor.

An approach to nanopatterning is reported in which a scanning near-field optical microscope coupled to a near-UV laser is used to selectively deprotect 2-nitrophenylpropyloxycarbonyl (NPPOC)-protected aminosiloxane monolayers on glass. UV deprotection was studied for unpatterned samples using X-ray photoelectron spectroscopy (XPS) and contact angle measurements. Highly efficient photodeprotection of the NPPOC moiety was observed upon irradiation at both 325 and 364 nm, and complete deprotection was found to occur within minutes. The resulting amine-terminated surfaces were then derivatized using trifluoroacetic anhydride (TFAA) and aldehyde-functionalized polymer nanoparticles. Contact angle and XPS measurements postderivatization indicated that surface functionalization was extensive, with the NPPOC-deprotected surfaces and aminopropylsiloxane control materials exhibiting essentially identical characteristics. Micrometer-scale patterns were fabricated using mask-based exposure, functionalized with polymer nanoparticles, and characterized by atomic force microscopy. Nanometer-scale patterns were fabricated using near-field exposure and characterized by friction force microscopy. The nanopatterns were derivatized with TFAA. The resulting images exhibited a clear contrast inversion that was due to an inversion of surface polarity in the patterned areas and confirmed that high spatial resolution (ca. 100 nm) was readily achievable.

Atherosclerosis and heart failure are major causes of morbidity and mortality in Western countries. Recent studies are suggesting involvement of telomere biology in the development and progression of age-associated conditions, including hypertension, atherosclerosis, and heart failure. Whether any of these reported associations are based on causal relationships remains to be elucidated. The construction of telomerase-deficient (telomerase RNA component, TERC(-/-)) mice might provide a potential instrumental model to study the involvement of telomere biology in cardiovascular disease. Here, we review the current available information from all studies performed in TERC(-/-) mice providing information on the cardiovascular phenotypic characteristics. Although this mouse model has proven its value in the understanding of the role of telomere biology in cancer, stem cell, and basic telomere research, only few studies were specifically designed to answer cardiovascular-related questions. The TERC(-/-) mice provide exciting opportunities to expand our knowledge of telomere biology in cardiovascular disease and the potential identification of novel targets of treatment.

The incidence and prevalence of cardiovascular disease increases progressively with advancing age. Cardiovascular disease is a major cause of morbidity and mortality in Western Countries. In the near future, as the population ages, it is expected that the population prevalence of cardiovascular disease will increase dramatically, imposing a major social and economical burden on society. Not only is age closely related to the development and progression of cardiovascular disease, but genetic and environmental factors also play an important role. Recently, a chromosomal mechanism, telomere shortening, has been considered a driving force by which genetic and environmental factors jointly affect biological aging, and possibly the risk for developing age-associated diseases. Telomeres are the extreme ends of chromosomes and shorten progressively during every cell cycle and therefore can be considered an indicator of biological age. In heart failure, telomere length is severely reduced. In the current review, we will discuss the emerging role of telomere biology in the pathophysiology of heart failure.

Immobilization of proteins onto solid supports is important in the preparation of functional protein microarrays and in the development of bead-based bioassays, biosensors, and industrial biocatalysts. In order to generate the stable, functional, and homogeneous materials required for these applications, attention has focused on methods that enable the efficient and site-specific covalent immobilization of recombinant proteins onto a wide range of platforms. To this end, the phosphopantetheinyl transferase Sfp was employed to catalyze the direct immobilization of recombinant proteins bearing the small, genetically encoded ybbR tag onto surfaces functionalized with CoA. Using mass spectrometry, it was shown that the Sfp catalyzes immobilization of a model acyl carrier protein (ACP) onto CoA-derivatized PEGA resin beads through specific covalent bond formation. Luciferase (Luc) and glutathione-S-transferase (GST) ybbR-fusion proteins were similarly immobilized onto PEGA resin retaining high levels of enzyme activity. This strategy was also successfully applied for the immobilization of the ACP, as well as ybbR-Luc, -GST, and -thioredoxin fusion proteins, on hydrogel microarray slides. Overall, the Sfp-catalyzed surface ligation is mild, quantitative, and rapid, occurring in a single step without prior chemical modification of the target protein. Immobilization of the target proteins directly from a cell lysate mixture was also demonstrated.

Renal function impairment relates to poor outcome in patients with chronic heart failure (HF). Differences in biological aging could affect the susceptibility to develop renal dysfunction in chronic HF. In the present study, we explored the association of leukocyte telomere length with renal function in patients with chronic HF. We studied 610 patients with HF, aged 40 to 80 years, NYHA class II-IV, with left ventricular ejection fraction of 0.40 or less. Glomerular filtration rate was estimated by the Modification of Diet in Renal Diseases (MDRD) formula, and telomere length of leukocytes was determined by a validated quantitative polymerase chain reaction-based method. Age-and gender-adjusted telomere length ratio decreased steadily with decreasing quartile of the MDRD formula (mean 0.80, 95% confidence interval [CI] 0.73 to 0.88; mean 0.74, 95% CI 0.68 to 0.81; 0.70 mean, 95% CI 0.63 to 0.76; mean 0.67, 95% CI 0.61 to 0.73; p <0.01). Telomere length of leukocytes correlated positively with the MDRD formula (correlation coefficient 0.141; p <0.001). These findings remained significant after adjustment for baseline differences and sensitivity analysis based on propensity score one-to-one matching. In conclusion, shorter leukocyte telomere length is associated with decreased renal function as estimated by the MDRD formula in patients with HF. Further studies will be needed to determine whether shorter leukocyte telomere length is the cause or consequence in this population and whether it plays a role in the prognosis of renal dysfunction in HF.

The aim of the study was to compare the standard technique of cardiac output determination by pulmonary artery catheter thermodilution (PAC-TD) with a noninvasive ultrasound Doppler monitor (USCOM Pty., Ltd., Coffs Harbour, Australia) in surgery for liver transplantation. We wished to determine if the degree of accuracy would allow the ultrasound cardiac output monitor (USCOM) to be used as an alternative monitor in a clinical setting in which wide fluctuations in cardiac output could be expected. This was a prospective method comparison study, with 71 paired measurements obtained in 12 patients undergoing liver transplantation in a university teaching hospital. Bland-Altman analysis of the 2 techniques showed a bias of 0.39 L/minute, with the USCOM cardiac output lower compared with that of PAC-TD. The bias was small and did not vary with the magnitude of the cardiac output. The 95% limits of agreement were -1.47 and 2.25 L/minute. There was good repeatability for USCOM measurements, with a repeatability coefficient of 0.43 for USCOM versus 0.77 for PAC-TD. We conclude that USCOM is acceptable for the clinical determination of noninvasive cardiac output, particularly in situations in which tracking changes over time is more important than knowing the precise value. However, the utility of USCOM is limited by its inability to measure pulmonary artery pressure.

Objective: The English arm of the UK Bowel Cancer Screening Pilot has recently concluded its third round. The primary aim of this study was to assess the effects of this programme on the emergency and elective cancer workload at University Hospitals Coventry and Warwickshire NHS Trust; the largest trust within the screened region. The secondary aim was to assess its effect upon Dukes staging, mortality and stoma formation for emergency colorectal cancer (CRC) admissions., Design: A retrospective analysis of validated data for CRC admissions over a period of 6 years from 1999 to 2004 was performed. The first year, 1999, represented the pre-screening year (PSY) which was taken as a baseline. Data for the next 5 years, screening years 1-5 (SY1-SY5), were recorded for the mode of admission, occurrence of emergency surgery, 30-day mortality and Dukes staging., Results: In the PSY (1999), 29.4% of CRCs were admitted as an emergency, decreasing to 15.8% by 2004 (p = 0.001). As a consequence, there was a significant decrease in the number of emergency CRC procedures performed over the same period (p<0.05). There was also a significant reduction in the 30-day mortality from 48% in 1999 to 13% in 2004 (p<0.05). Dukes stage C carcinomas, however, remained the predominant stage presenting as emergencies throughout the studied period (SY3, 53%; SY4, 38%; SY5, 50%)., Conclusion: Following commencement of the UK Bowel Cancer Screening Pilot, there has been a significant decline in emergency CRC workload with a marked improvement in 30-day mortality and decreased stoma formation, in Coventry and North Warwickshire. It is postulated that the witnessed and notable positive impact over such a short time period is the result of increased detection of asymptomatic malignancies within the screening programme, increased public awareness of the symptoms of CRC, together with a change in attitudes and referral patterns of general practitioners within Coventry and North Warwickshire.

Natural products have evolved, at least in part, to bind to biological macromolecules, particularly proteins. As a result, natural products are able to interact with many specific targets within the cell. Indeed for many years this has been central in the drug development process. Today, however, natural products are finding increasing use as probes to interrogate biological systems as part of chemical genomics and related research. In order to demonstrate the utility of natural products in these efforts, the biological activities of many of the major classes of natural products is discussed, according to the cellular organelle and localisation of their specific molecular targets. Emphasis is given to newly discovered compounds and activities that either provide interesting insights into a specific biological function, or that form the basis for potentially new therapeutic approaches.

Background: Atherosclerosis is the leading cause of death in western societies and cigarette smoke is among the factors that strongly contribute to the development of this disease. The early events in atherogenesis are stimulated on the one hand by cytokines that chemoattract leukocytes and on the other hand by decrease in circulating molecules that protect endothelial cells (ECs) from injury. Here we focus our studies on the effects of "second-hand" smoke on atherogenesis., Methods: To perform these studies, a smoking system that closely simulates exposure of humans to second-hand smoke was developed and a mouse model system transgenic for human apoB100 was used. These mice have moderate lipid levels that closely mimic human conditions that lead to atherosclerotic plaque formation., Results: "Second-hand" cigarette smoke decreases plasma high density lipoprotein levels in the blood and also decreases the ratios between high density lipoprotein and low density lipoprotein, high density lipoprotein and triglyceride, and high density lipoprotein and total cholesterol. This change in lipid profiles causes not only more lipid accumulation in the aorta but also lipid deposition in many of the smaller vessels of the heart and in hepatocytes. In addition, mice exposed to smoke have increased levels of Monocyte Chemoattractant Protein-1 in circulation and in the heart/aorta tissue, have increased macrophages in the arterial walls, and have decreased levels of adiponectin, an EC-protective protein. Also, cytokine arrays revealed that mice exposed to smoke do not undergo the switch from the pro-inflammatory cytokine profile (that develops when the mice are initially exposed to second-hand smoke) to the adaptive response. Furthermore, triglyceride levels increase significantly in the liver of smoke-exposed mice., Conclusion: Long-term exposure to "second-hand" smoke creates a state of permanent inflammation and an imbalance in the lipid profile that leads to lipid accumulation in the liver and in the blood vessels of the heart and aorta. The former potentially can lead to non-alcoholic fatty liver disease and the latter to heart attacks.

Among most patients attending a rectal clinic, rectal bleeding is a common presenting feature. In most patients, the cause is attributed to a benign lesion. In a small percentage, the cause is neoplastic, and for this reason, rectal bleeding merits further study. Left-sided tumors account for the majority of these tumors and are within the reach of a flexible sigmoidoscopy. This study aimed at examining the diagnostic performance of the one stop rectal clinic in Coventry. Between November 2001 and May 2002, 250 consecutive patients were seen in the one stop rectal bleeding clinic of a tertiary referral hospital. Patients were asked of the nature of rectal bleed and altered bowel habits and were examined by digital rectal examination, with a proctoscopy and rigid sigmoidoscopy before either a full colonoscopic examination or flexible sigmoidoscopy with a completion Barium enema. During the study period, colorectal cancer was detected in 4 patients (1.6%), adenomatous polyps in 36 patients (14.4%), and ulcerative colitis in 8 patients (3.2%). In 98 patients (39.2%), no abnormality was present, and in the remaining patients, diverticulosis (n = 60; 24%) and hemorrhoids were present (n = 44; 17.6%).

Aim of this explorative study was to determine the prognostic value of protein expression of the DNA damage repair enzymes ERCC1, hRad51, and BRCA1 for tumour response and survival of non-small-cell lung cancer patients treated with chemotherapy. Patients with either a short or long overall survival were selected from a randomized phase III trial comparing cisplatin-gemcitabine and epirubicin-gemcitabine. Tumour biopsies were assessed for differences in immunohistochemical staining using antibodies against ERCC1, hRad51, and BRCA1. A total of 33 patients were included. A positive nuclear staining for ERCC1, hRad51, and BRCA1 was observed in 44, 12, and 90% of biopsies, respectively. In large cell carcinoma nuclear hRad51 staining was absent. In five biopsies stained for hRad51 an unexpected membrane-like staining was observed; these biopsies showed no nuclear staining. DNA damage repair protein expressions were not significantly different in responders versus non-responders, or in patients with a short or long overall survival. In conclusion, immunohistochemical staining of ERCC1, hRad51, and BRCA1, in tumour biopsies from non-small-cell lung cancer patients was not predictive for tumour response and survival after chemotherapy.

The construction and design of a microscope coupled with a miniature UV-vis spectrometer is described. This was applied to the study of dyes linked to solid supports and displayed good correlation in spectral shape and lambda(max) values when compared to the dyes in solution, as well as showing a linear relationship between dye loading and UV-vis absorbance. The spectral profiles of these dyes at various pH's were measured and used to determine the pK(a) of the dyes on the beads, which were compared with the pK(a) values of the dyes in solution, thus enabling the dye-loaded beads to act as pH sensors.

Cell cycle progression is both regulated and accompanied by periodic changes in the expression levels of a large number of genes. To investigate cell cycle-regulated transcriptional programs in the fission yeast Schizosaccharomyces pombe, we developed a whole-genome oligonucleotide-based DNA microarray. Microarray analysis of both wild-type and cdc25 mutant cell cultures was performed to identify transcripts whose levels oscillated during the cell cycle. Using an unsupervised algorithm, we identified 747 genes that met the criteria for cell cycle-regulated expression. Peaks of gene expression were found to be distributed throughout the entire cell cycle. Furthermore, we found that four promoter motifs exhibited strong association with cell cycle phase-specific expression. Examination of the regulation of MCB motif-containing genes through the perturbation of DNA synthesis control/MCB-binding factor (DSC/MBF)-mediated transcription in arrested synchronous cdc10 mutant cell cultures revealed a subset of functional targets of the DSC/MBF transcription factor complex, as well as certain gene promoter requirements. Finally, we compared our data with those for the budding yeast Saccharomyces cerevisiae and found approximately 140 genes that are cell cycle regulated in both yeasts, suggesting that these genes may play an evolutionarily conserved role in regulation of cell cycle-specific processes. Our complete data sets are available at http://giscompute.gis.a-star.edu.sg/~gisljh/CDC.

Background: Nasal strips are used by athletes, people who snore, and asthmatics to ease the burden of breathing. Although there are some published studies that demonstrate higher flow with nasal strips, none had directly measured the effect of the strips on nasal resistance using the airflow perturbation device (APD). The APD is an inexpensive instrument that can measure respiratory resistance based on changes in mouth pressure and rate of airflow., Method: This study tested forty-seven volunteers (14 men and 33 women), ranging in age from 17 to 51. Each volunteer was instructed to breathe normally into the APD using an oronasal mask with and without nasal strips. The APD measured respiratory resistance during inhalation, exhalation, and an average of the two., Results: Results of a paired mean t-test comparing nasal strip against no nasal strip were statistically significant at the p = 0.05 level. The Breathe Right nasal dilator strips lowered nasal resistance by an average of 0.5 cm H20/Lps from an average nasal resistance of 5.5 cm H20/Lps., Conclusions: Nasal strips reduce nasal resistance when measured with the APD. The effect is equal during exhalation and during inhalation.

pH indicating resins were prepared by covalently attaching carboxylic acid derivatives of sulfthalein dyes, synthesized using a Suzuki cross-coupling, onto resin beads. The resin-bound indicators showed the expected colour changes according to pH and their behaviour was analysed using a micro UV/Vis spectrometer.

Although it is known that high levels of cigarette smoke lead to cell death, little is known about the effects of low-to-moderate levels of smoke components that are found in vivo, such as those experienced by cells in tissues. Clinical studies and experimental data show that smokers heal poorly and are more prone to develop fibrotic diseases. Here we show the effects of first-hand cigarette smoke on fibroblasts, cells that are critically involved in these processes. Using doses of smoke found in the tissues of smokers and a variety of cell and molecular approaches, we show that these doses of cigarette smoke do not cause cell death but rather stimulate fibroblasts to produce stress response and survival proteins such as interleukin-8, PKB/Akt, p53, and p21 that in turn contribute to an increase in cell survival. In addition, smoke-treated cells show a decrease in cell migration, which can be explained by the increased cell adhesion and alterations in cytoskeletal elements. We also show that these levels of smoke cause changes in mitochondrial morphology with a minimum loss of function and these changes are the result of exposure to reactive oxygen species. We conclude that the increase in cell survival may lead to a build-up of connective tissue in the area of a wound, potentially leading to delayed healing and/or fibrosis and that the alterations in the cytoskeleton and in cell adhesion result in inhibition of cell migration, a process that could lead to nonclosure of the wound for lack of proper fibroblast migration to form the healing tissue.

Background: Evaluation of acute appendicitis in the elderly presents a diagnostic challenge to the surgeon. The diagnosis is often difficult to establish on account of general poverty of history and clinical signs, comorbidity and the unreliability of diagnostic procedures. Due to this, morbidity and mortality statistics in this group of patients is unacceptably higher than the general population., Methods: Individual case report and literature review., Results: We present an unusual case of perforated appendicitis masquerading as acute coronary syndrome., Conclusions: Patient's education, high index of suspicion, and an aggressive surgical approach is needed if the outcome among the elderly population is to be improved.

Background: It is known that "second-hand" cigarette smoke leads to abnormal tissue repair and remodelling but the cellular mechanisms involved in these adverse effects are not well understood. Fibroblasts play a major role in repair and remodelling. They orchestrate these processes by proliferating, migrating, and secreting proteins such as, cytokines, growth factors and extracellular matrix molecules. Therefore, we focus our studies on the effects of "second-hand" cigarette smoke on the structure and function of these cells., Results: We used sidestream whole (SSW) smoke, a major component of "second-hand" smoke, primary embryonic fibroblasts, cells that behave very much like wound fibroblasts, and a variety of cellular and molecular approaches. We show that doses of smoke similar to those found in tissues cause cytoskeletal changes in the fibroblasts that may lead to a decrease in cell migration. In addition, we also show that these levels of cigarette smoke stimulate an increase in cell survival that is reflected in an increase and/or activation of stress/survival proteins such as cIL-8, grp78, PKB/Akt, p53, and p21. We further show that SSW affects the endomembrane system and that this effect is also accomplished by nicotine alone., Conclusions: Taken together, our results suggest that: (i) SSW may delay wound repair because of the inability of the fibroblasts to migrate into the wounded area, leading to an accumulation of these cells at the edge of the wound, thus preventing the formation of the healing tissue; (ii) the increase in cell survival coupled to the decrease in cell migration can lead to a build-up of connective tissue, thereby causing fibrosis and excess scarring.

[reaction: see text] A formal total synthesis of the selective muscarinic receptor antagonist himbacine is presented. Key C-C bond-forming steps include an intramolecular Diels-Alder reaction, Stille coupling reactions, and a 6-exo-trig acyl radical cyclization to a conjugated enyne. An unexpected secondary alcohol to chloride conversion is witnessed during attempted thionocarbonate formation.

This case is presented to illustrate the imaging and clinical findings of a condition of interest to orthopaedic surgeons. The initial findings are noted on this page. The clinical and imaging diagnoses are presented on the following pages.

[reaction: see text] The syntheses of 4,4a-didehydrohimbacine and 4,4a-didehydrohimandravine are presented. Key steps include an intramolecular Diels-Alder reaction of a bromopentadienyl acrylate and Suzuki-Miyaura and Stille coupling reactions.

Experimental conditions for the intramolecular cycloaddition of four related pentadienyl acrylates 3, 4, 5 and 6 are reported. In contrast with several previous reports, pentadienyl acrylates do undergo synthetically useful intramolecular Diels-Alder reactions: 3, 4, 5 and 6 cyclise at reasonable rates at temperatures of 132-180 degrees C at atmospheric pressure in moderate to good yields. The stereochemical outcome of each of these reactions was accurately measured and the results are in good agreement with transition structure populations predicted using B3LYP/6-31+G(d) theory. The parent system 3 cyclises with moderate endo selectivity; the presence of either a C5-methyl substituent or a C3-bromine atom results in a slight shift towards the trans-fused exo stereoisomer but--overall--a less selective reaction. The presence of both C3-Br and C5-CH3 substituents results in a marked improvement in stereoselectivity with the exo,lk-product predominating. Interpretation of B3LYP/6-31+G(d) transition structures allows insights into the improvement in stereoselectivity obtained by incorporating a removable "steric directing group" into a 5-methyl-1,3,8-nonatriene precursor.

An IBM PC-based real-time data acquisition, monitoring and analysis system for experimental haemodynamic studies was developed. Comprehensive haemodynamic signals, such as aortic and left ventricular pressures, aortic and coronary blood flows, two segmental lengths, two segmental thicknesses, electrocardiogram and airway pressure, were acquired and monitored to assess cardiac function. The system performs computer-aided analysis and derivations on a number of haemodynamic parameters and cardiac function indices. The system has been tested and validated extensively over a number of series of experimental haemodynamic studies to investigate the effects of anaesthetic agents, cardiovascular drugs, and changes in loading on normal and critically ischaemic myocardium of anaesthetised laboratory subjects. Without this specialised and automated system, the analysis of the data acquired from the haemodynamic studies would be too time-consuming and could not be fully performed.

We assessed the Bird T-Bird VS ventilator using a model lung constructed to standard 10651-1 of the International Standards Organization. We used different combinations of lung compliance and airway resistance to simulate normal and diseased adult and paediatric lungs. Different preset tidal volumes at various respiratory rates were delivered to the model lung and compared with the actual measured tidal volumes. The results showed that the delivered tidal volumes were within the manufacturer's specification of +/- 10% of the preset tidal volumes in normal adult and paediatric combinations of lung compliance and airway resistance. The ventilator can be powered by mains electricity supply or battery and requires only optional compressed oxygen. The ventilator is suitable for the provision of advanced ventilatory support during prolonged patient transfer.

We report a rare case of a non-insulin secreting malignant insulinoma of the pancreas and discuss its medical and surgical management. Surgical excision of malignant insulinoma produces good palliation and can increase survival., (Copyright 2000 Harcourt Publishers Ltd.)

Objective: An IBM PC-based real-time data acquisition, monitoring and analysis system was developed for the assessment of cardio-respiratory function, i.e. airway dead space, alveolar volume and pulmonary blood flow, using oscillating inert inspired gas forcing signals., Methods: The forcing gas mixture was generated by an in-house sinusoid gas delivery unit. The system interfaced with a mass spectrometer and an airway flow transducer, and performed real-time tracking of the breath-by-breath end-inspired, end-expired and mixed-expired concentrations. It calculated the cardiorespiratory parameters using two, i.e. continuous and tidal, in-house mathematical models of the lungs. The system's performance was evaluated using a mechanical bench lung, laboratory subjects and awake adults breathing spontaneously. Its predictive accuracy was compared with the measured volumes of the bench lung; single breath CO2 test for airway dead space and N2 washout for alveolar volume in laboratory subjects and awake adults; and thermal dilution technique for pulmonary blood flow in laboratory subjects., Results: Close agreements were found between the true and predicted airway dead space, i.e. mean differences of -12.39%, 14.47% and -17.49%, respectively, and that of alveolar volume, i.e. -8.03%, -3.62% and 7.22%, respectively, in the bench lung, laboratory subject and awake adult studies; and that of pulmonary blood flow (-23.81%) in the laboratory subjects using the continuous lung model. Even closer agreements were observed for airway dead space (-5.8%) and alveolar volume (-4.01%) of the bench lung and for pulmonary blood flow (-8.47%) in the laboratory subjects using the tidal lung model., Conclusions: A system was developed to deliver, monitor and analyse on-line, and in real-time, output data from the sinusoid forcing technique. The technique was administered using the system in various subjects, and produced favourable predictions.

Intrabiliary rupture of a hydatid liver cyst is infrequently reported, but may present with symptoms of choledocholethiasis or cholangitis. We report a case of hydatid liver disease presenting as recurrent pancreatitis, and discuss its clinical, radiological and surgical treatments. Hydatid liver disease has a diverse clinical spectrum, and a diagnosis of acute pancreatitis should be considered in patients with hydatid liver disease presenting with unexplained abdominal pain.

The aim of this study was to compare the ability of artificial neural networks and the Acute Physiology and Chronic Health Evaluation II score to predict mortality in adult intensive care units. The same physiological variables were used in both predictive models to predict hospital mortality from a data set of 8796 patients collected from 26 adult intensive care units in the United Kingdom and Ireland as part of the Intensive Care Society study. The results from the two models were compared with the actual outcome. The overall prediction accuracy and the overall goodness-of-fit of all the models were assessed. Both predictive models showed similar goodness-of-fit and prediction discrimination. The overall predictive and classification performance of the artificial neural network developed matched and in some aspects was better than that of Acute Physiology and Chronic Health Evaluation II.

Metastatic melanoma of the small bowel is a pathological entity that is not frequently reported but may present with features of unexplained anaemia. We report a case of a 51-year-old man with occult intestinal bleeding due to metastatic melanoma of the small bowel. Although the diagnosis was somewhat delayed, the patient was managed successfully by small bowel resection. Careful investigation of melanoma patients with gastrointestinal symptoms is important as surgical intervention often results in improved quality of life and survival.

The military and some civilian rescue agencies have a requirement for portable ventilators in the anaesthetic, resuscitation and pre-hospital environment. This paper describes two new portable ventilators specifically designed to satisfy the military requirement for lightweight, robust and versatile equipment which can also be used in a contaminated environment.

Hypothesis: Patients with melanoma and histologically negative sentinel lymph nodes identified by lymphatic mapping have a very good prognosis., Design: Cohort study with follow-up information obtained from medical records and telephone interviews., Setting and Patients: Of all patients with cutaneous melanoma who underwent intraoperative sentinel lymph node mapping between November 15, 1993, and April 18, 1997, at the Massachusetts General Hospital, Boston, 89 were found to have no evidence of melanoma in their sentinel nodes. Forty-six lesions (51%) were on an extremity and 44 (49%) were of axial location. The median tumor thickness was 1.8 mm (range, 0.36-12.0 mm) and 11 tumors (12%) were ulcerated., Interventions: Patients underwent intraoperative sentinel lymph node mapping with lymphazurin and radiolabeled sulfur colloid. Sentinel lymph nodes were analyzed by standard hematoxylin-eosin staining. Only 2 patients received adjuvant therapy following wide excision of the primary lesion., Main Outcome Measures: Site of initial recurrence and time to initial recurrence., Results: The median follow-up for all patients was 23 months (range, 2-54 months). Eleven patients (12%) developed melanoma recurrences, and 78 (88%) patients remain disease free. Regional lymph nodes were the initial site of recurrence in 7 (8%) of 89 patients, and 7 (7%) of 106 mapped basins. Four patients had recurrence without involvement of regional lymph nodes: 2 with distant metastases and 2 with in transit metastases. The median time to recurrence was 12 months (range, 2-35 months). Sentinel lymph nodes were reanalyzed using serial sections and immunoperoxidase stains in 7 patients with recurrence and metastatic melanoma was identified in 3 (43%)., Conclusions: The risk for melanoma recurrence is relatively low in patients with histologically negative sentinel nodes identified by lymphatic mapping. Longer follow-up will improve our understanding of the prognostic value of this procedure.