Pro-fibrotic polymorphisms predictive of advanced liver fibrosis in the severely obese

Background/Aims: Insulin resistance and systemic hypertension are predictors of advanced fibrosis in obese patients with non-alcoholic fatty liver disease (NAFLD). Genetic factors may also be important. We hypothesize that high angiotensinogen (AT) and transforming growth factor-beta1 (TGF-beta1) producing genotypes increase the risk of liver fibrosis in obese subjects with NAFLD. Methods: One hundred and five of 130 consecutive severely obese patients having a liver biopsy at the time of laparoscopic obesity surgery agreed to have genotype analysis. Influence of specific genotype or combination of genotypes on the stage of hepatic fibrosis was assessed after controlling for known risk factors. Results: There was no fibrosis in 70 (67%), stages 1-2 in 21 (20%) and stages 3-4 fibrosis in 14 (13%) of subjects. There was no relationship between either high AT or TGF-beta1 producing genotypes alone and hepatic fibrosis after controlling for confounding factors. However, advanced hepatic fibrosis occurred in five of 13 subjects (odds ratio 5.7, 95% confidence interval 1.5-21.2, P = 0.005) who inherited both high AT and TGF-beta1 producing polymorphisms. Conclusions: The combination of high AT and TGF-beta1 producing polymorphisms is associated with advanced hepatic fibrosis in obese patients with NAFLD. These findings support the hypothesis that angiotensin II stimulated TGF-beta1 production may promote hepatic fibrosis. (C) 2003 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

A convergent solution-phase synthesis of the macrocycle Ac-Phe-[Orn-Pro-D-Cha-Trp-Arg], a potent new antiinflammatory drug

Relatively few cyclic peptides have reached the pharmaceutical marketplace during the past decade, most produced through fermentation rather than made synthetically. Generally, this class of compounds is synthesized for research purposes on milligram scales by solid-phase methods, but if the potential of macrocyclic peptidomimetics is to be realized, low-cost larger scale solution-phase syntheses need to be devised and optimized to provide sufficient quantities for preclinical, clinical, and commercial uses. Here, we describe a cheap, medium-scale, solution-phase synthesis of the first reported highly potent, selective, and orally active antagonist of the human C5a receptor. This compound, Ac-Phe[Orn-Pro-D-Cha-Trp-Arg], known as 3D53, is a macrocyclic peptidomimetic of the human plasma protein C5a and displays excellent antiinflammatory activity in numerous animal models of human disease. In a convergent approach, two tripeptide fragments Ac-Phe-Orn-(Boc)-Pro-OH and H-D-Cha-Trp(For)-Arg-OEt were first prepared by high-yielding solution-phase couplings using a mixed anhydride method before coupling them to give a linear hexapeptide which, after deprotection, was obtained in 38% overall yield from the commercially available amino acids. Cyclization in solution using BOP reagent gave the antagonist in 33% yield (13% overall) after HPLC purification. Significant features of the synthesis were that the Arg side chain was left unprotected throughout, the component Boe-D-Cha-OH was obtained very efficiently via hydrogenation Of D-Phe with PtO2 in TFA/water, the tripeptides were coupled at the Pro-Cha junction to minimize racemization via the oxazolone pathway, and the entire synthesis was carried out without purification of any intermediates. The target cyclic product was purified (>97%) by reversed-phase HPLC. This convergent synthesis with minimal use of protecting groups allowed batches of 50100 g to be prepared efficiently in high yield using standard laboratory equipment. This type of procedure should be useful for making even larger quantities of this and other macrocyclic peptidomimetic drugs.

Assessment of arginine 97 and lysine 72 as determinants of substrate specificity in cytochrome P450 2C9 (CYP2C9)

CYP2C9 is distinguished by a preference for substrates bearing a negative charge at physiological pH. Previous studies have suggested that CYP2C9 residues R97 and K72 may play roles in determining preference for anionic substrates by interaction at the active site or in the access channel. The aim of the present study was to assess the role of these two residues in determining substrate selectivity. R97 and K72 were substituted with negative, uncharged polar and hydrophobic residues using a degenerate polymerase chain reaction-directed strategy. Wild-type and mutant enzymes were expressed in bicistronic format with human cytochrome P450 reductase in Escherichia coli. Mutation of R97 led to a loss of holoenzyme expression for R97A, R97V, R97L, R97T, and R97E mutants. Low levels of hemoprotein were detected for R97Q, R97K, R97I, and R97P mutants. Significant apoenzyme was observed, suggesting that heme insertion or protein stability was compromised in R97 mutants. These observations are consistent with a structural role for R97 in addition to any role in substrate binding. By contrast, all K72 mutants examined (K72E, K72Q, K72V, and K72L) could be expressed as hemoprotein at levels comparable to wild-type. Type I binding spectra were obtained with wildtype and K72 mutants using diclofenac and ibuprofen. Mutation of K72 had little or no effect on the interaction with these substrates, arguing against a critical role in determining substrate specificity. Thus, neither residue appears to play a role in determining substrate specificity, but a structural role for R97 can be proposed consistent with recently published crystallographic data for CYP2C9 and CYP2C5.

Screening for physical activity in family practice - Evaluation of two brief assessment tools

Background: Physical activity (PA) is relevant to the prevention and management of many health conditions in family practice. There is a need for an efficient, reliable, and valid assessment tool to identify patients in need of PA interventions. Methods: Twenty-eight family physicians in three Australian cities assessed the PA of their adult patients during 2004 using either a two- (2Q) or three-question (3Q) assessment. This was administered again approximately 3 days later to evaluate test-retest reliability. Concurrent validity was evaluated by measuring agreement with the Active Australia Questionnaire, and criterion validity by comparison with 7-day Computer Science Applications, Inc. (CSA) accelerometer counts. Results: A total of 509 patients participated, with 428 (84%) completing a repeat assessment, and 415 (82%) accelerometer monitoring. The brief assessments had moderate test-retest reliability (2Q k = 58.0%, 95% confidence interval [CI] = 47.2-68.8%; 3Q k = 55.6%, 95% CI = 43.8-67.4%); fair to moderate concurrent validity (2Q k = 46.7%, 95% CI = 35.657.9%; 3Q k = 38.7%, 95% CI = 26.4-51.1%); and poor to fair criterion validity (2Q k = 18.2%, 95% CI = 3.9-32.6%; 3Q k = 24.3%, 95% CI = 11.6-36.9%) for identifying patients as sufficiently active. A four-level scale of PA derived from the PA assessments was significantly correlated with accelerometer minutes (2Q rho = 0.39, 95% CI = 0.28-0.49; 3Q rho = 0.31, 95% CI = 0.18-0.43). Physicians reported that the assessments took I to 2 minutes to complete. Conclusions: Both PA assessments were feasible to use in family practice, and were suitable for identifying the least active patients. The 2Q assessment was preferred by clinicians and may be most appropriate for dissemination.

Reliability and validity of a brief physical activity assessment for use by family doctors

Objective: To evaluate the reliability and validity of a brief physical activity assessment tool suitable for doctors to use to identify inactive patients in the primary care setting. Methods: Volunteer family doctors (n = 8) screened consenting patients (n = 75) for physical activity participation using a brief physical activity assessment tool. Inter-rater reliability was assessed within one week (n = 71). Validity was assessed against an objective physical activity monitor (computer science and applications accelerometer; n = 42). Results: The brief physical activity assessment tool produced repeatable estimates of sufficient total physical activity, correctly classifying over 76% of cases (kappa 0.53, 95% confidence interval (CI) 0.33 to 0.72). The validity coefficient was reasonable (kappa 0.40, 95% CI 0.12 to 0.69), with good percentage agreement (71%). Conclusions: The brief physical activity assessment tool is a reliable instrument, with validity similar to that of more detailed self report measures of physical activity. It is a tool that can be used efficiently in routine primary healthcare services to identify insufficiently active patients who may need physical activity advice.

The appetitive motivation scale and other BAS measures in the prediction of Approach and Active Avoidance

The Appetitive Motivation Scale (Jackson & Smillie, 2004) is a new trait conceptualisation of Gray's (I 970 199 1) Behavioural Activation System. In this experiment we explore relationships that the Appetitive Motivation Scale and other measures of Gray's model have with Approach and Active Avoidance responses. Using a sample of 144 undergraduate students, both Appetitive Motivation and Sensitivity to Reward (from the Sensitivity to Punishment and Sensitivity to Reward Questionnaire, SPSRQ; Torrubia, Avila, Molto, & Ceseras, 2001), were found to be significant predictors of Approach and Active Avoidance response latency. This confirms previous experimental validations of the SPSRQ (e.g., Avila, 2001) and provides the first experimental evidence for the validity of the Appetitive Motivation scale. Consistent with interactive views of Gray's model (e.g., Corr, 2001), high SPSRQ Sensitivity to Punishment diminished the relationship between Sensitivity to Reward and our BAS criteria. Measures of BIS did not however interact in this way with the appetitive motivation scale. A surprising result was the failure for any of Carver and White's (1994) BAS scales to correlate with RST criteria. Implications of these findings and potential future directions are discussed. (C) 2004 Elsevier Ltd. All rights reserved.

Objective and behavioural assessment of the emergence from post-traumatic amnesia (PTA)

Primary objective: To determine the profile of resolution of typical PTA behaviours and describe new learning and improvements in self-care during PTA. Research design: Prospective longitudinal study monitoring PTA status, functional learning and behaviours on a daily basis. Methods and procedures: Participants were 69 inpatients with traumatic brain injury who were in PTA. PTA was assessed using the Westmead or Oxford PTA assessments. Functional learning capability was assessed using a routine set of daily tasks and behaviour was assessed using an observational checklist. Data was analysed using descriptive statistics. Main outcomes and results: Challenging behaviours that are typically associated with PTA, such as agitation, aggression and wandering resolved in the early stages of PTA and incidence rates of these behaviours were less than 20%. Independence in self-care and bowel and bladder continence emerged later during resolution of PTA. New learning in functional situations was demonstrated by patients in PTA. Conclusions: It is feasible to begin active rehabilitation focused on functional skills-based learning with patients in the later stages of PTA. Formal assessment of typically observed behaviours during PTA may complement memory-based PTA assessments in determining emergence from PTA.

Dynamic assessment of articulation during lingual fatigue in myasthenia gravis

The present study aimed to investigate how induced lingual fatigue affected lingual strength, articulatory kinematics, and perceptual speech features in CS, a 51-year-old female with active myasthenia gravis (MG), and three age and gender matched control participants, Lingual fatigue was elicited via a series of endurance tasks using a tongue pressure bulb. Following each endurance task, the participants performed a speech task containing the phonemes /k/, /t/, and /j/ that was recorded with an electromagnetic articulograph, followed by a lingual strength assessment using a tongue pressure bulb. Participants repeated this schedule over five phases and kinematic and strength changes during each phase were compared to baseline measurements. All of CSs significant kinematic changes occurred during the final fatigue phase compared to 27.3% of the control group's kinematic changes occurring during this phase, suggesting the kinematic changes associated with fatigue were not accelerated in CS. The endurance tasks also elicited different kinematic effects for CSs anterior and posterior tongue segments. While CS exhibited mostly similar kinematic and perceptual changes to the control group, some of CS's perceptual transcriptions for /k/ and kinematic changes were not replicated, indicating that some different perceptual and physiological consequences to CS's speech were elicited by the endurance tasks.

Population-based detection of systolic and diastolic dysfunction with amino-terminal pro-B-type natriuretic peptide

Background There is limited information regarding the clinical utility of amino-terminal pro-B-type natriuretic pepticle (NT-proBNP) for the detection of left ventricular (LV) dysfunction in the community. We evaluated predictors of circulating NT-proBNP levels and determined the utility of NT-proBNP to detect systolic and diastolic LV dysfunction in older adults. Methods. A population-based sample of 1229 older adults (mean age 69.4 years, 50.1% women) underwent echocardiographic assessment of cardiac structure and function and measurement of circulating NT-proBNP levels. Results Predictors of NT-proBNP included age, female sex, body mass index, and cardiorenal parameters (diastolic dysfunction [DID] severity; LV mass and left atrial volume; right ventricular overload; decreasing ejection fraction [EF] and creatinine clearance). The performance of NT-proBNP to detect any degree of LV dysfunction, including mild DID, was poor (area under the curve 0.56-0.66). In contrast, the performance of NT-proBNP for the detection of EF 0.90 regardless of age and sex; history of hypertension, diabetes, coronary artery disease; or body mass category. The ability of NT-proBNP to detect EF

Cardiovascular risk estimation by professionally active cardiovascular nurses: Results from the Basel 2005 Nurses Cohort

Background Nurses play a key role in the prevention of cardiovascular disease (CVD) and one would, therefore, expect them to have a heightened awareness of the need for systematic screening and their own CVD risk profile. The aim of this study was to examine personal awareness of CVD risk among a cohort of cardiovascular nurses attending a European conference. Methods Of the 340 delegates attending the 5th annual Spring Meeting on Cardiovascular Nursing (Basel, Switzerland, 2005), 287 (83%) completed a self-report questionnaire to assess their own risk factors for CVD. Delegates were also asked to give an estimation of their absolute total risk of experiencing a fatal CVD event in the next 10 years. Level of agreement between self-reported CVD risk estimation and their actual risk according to the SCORE risk assessment system was compared by calculating weighted Kappa (κw). Results Overall, 109 responders (38%) self-reported having either pre-existing CVD (only 2%), one or more markedly raised CVD risk factors, a high total risk of fatal CVD (≥ 5% in 10 years) or a strong family history of CVD. About half of this cohort (53%) did not know their own total cholesterol level. Less than half (45%) reported having a 10-year risk of fatal CVD of < 1%, while 13% reported having a risk ≥ 5%. Based on the SCORE risk function, the estimated 10-year risk of a fatal CVD event was < 1% for 96% of responders: only 2% had a ≥ 5% risk of such an event. Overall, less than half (46%) of this cohort's self-reported CVD risk corresponded with that calculated using the SCORE risk function (κw = 0.27). Conclusion Most cardiovascular nurses attending a European conference in 2005 poorly understood their own CVD risk profile, and the agreement between their self-reported 10-year risk of a fatal CVD and their CVD risk using SCORE was only fair. Given the specialist nature of this conference, our findings clearly demonstrate a need to improve overall nursing awareness of the role and importance of systematic CVD risk assessment.

Assessing the Conservation Status and Threats to Priority Plants: a threat assessment approach and case study in South-East Queensland, Australia

bstract: During the Regional Forest Agreement (RFA) process in south-east Queensland, the conservation status of, and threats to, priority vascular plant taxa in the region was assessed. Characteristics of biology, demography and distribution were used to assess the species' intrinsic risk of extinction. In contrast, the threats to the taxa (their extrinsic risk of extinction) were assessed using a decision-support protocol for setting conservation targets for taxa lacking population viability analyses and habitat modelling data. Disturbance processes known or suspected to be adversely affecting the taxa were evaluated for their intensity, extent and time-scale. Expert opinion was used to provide much of the data and to assess the recommended protection areas. Five categories of intrinsic risk of extinction were recognised for the 105 priority taxa: critically endangered (43 taxa); endangered (29); vulnerable (21); rare (10); and presumed extinct (2). Only 6 of the 103 extant taxa were found to be adequately reserved and the majority were considered inadequately protected to survive the current regimes of threatening processes affecting them. Data were insufficient to calculate a protection target for one extant taxon. Over half of the taxa require all populations to be conserved as well as active management to alleviate threatening processes. The most common threats to particular taxa were competition from weeds or native species, inappropriate fire regimes, agricultural clearing, forestry, grazing by native or feral species, drought, urban development, illegal collection of plants, and altered hydrology. Apart from drought and competition from native species, these disturbances are largely influenced or initiated by human actions. Therefore, as well as increased protection of most of the taxa, active management interventions are necessary to reduce the effects of threatening processes and to enable the persistence of the taxa.