Levels of C-reactive protein associated with high and very high cardiovascular risk are prevalent in patients with rheumatoid arthritis.

ObjectiveC-reactive protein (CRP) levels>3 mg/L and>10 mg/L are associated with high and very high cardiovascular risk, respectively, in the general population. Because rheumatoid arthritis (RA) confers excess cardiovascular mortality, we determined the prevalence of these CRP levels among RA patients stratified on the basis of their RA disease activity.MethodsWe evaluated physician and patient global assessments of disease activity, tender and swollen 28 joint counts, erythrocyte sedimentation rate (ESR), and CRP measured in a single clinic visit for 151 RA patients. Disease activity was calculated using the Clinical Disease Activity Index (CDAI) and the Disease Activity Score 28 Joints (DAS28-ESR and DAS28-CRP).ResultsMedian CRP level was 5.3 mg/L. 68% of patients had CRP>3 mg/L, and 25% had CRP>10 mg/L. Of those with 0-1 swollen joints (n = 56), or 0-1 tender joints (n = 81), 64% and 67%, respectively, had CRP>3 mg/L, and 23% and 20%, respectively, had CRP>10 mg/L. Of those with remission or mildly active disease by CDAI (n = 58), DAS28-ESR (n = 39), or DAS28-CRP (n = 70), 49-66% had CRP>3 mg/L, and 10-14% had CRP>10 mg/L. Of patients with moderate disease activity by CDAI (n = 51), DAS28-ESR (n = 78), or DAS28-CRP (n = 66), 67-73% had CRP>3 mg/L, and 25-33% had CRP>10 mg/L.ConclusionEven among RA patients whose disease is judged to be controlled by joint counts or standardized disease scores, a substantial proportion have CRP levels that are associated high or very high risk for future cardiovascular events in the general population

A systematic review of the association between circulating concentrations of C reactive protein and cancer.

The objective of this study was to review and summarise the published evidence for an association between circulating concentrations of C reactive protein (CRP) and cancer through a systematic review. 90 discrete studies were identified. 81 (90%) were prevalent case-control or cross-sectional studies, and only 9 studies had a prospective design. In most prevalent studies, CRP concentrations were found to be higher in patients with cancer than in healthy controls or controls with benign conditions. Of the nine large prospective studies identified in this review, four reported no relationship between circulating CRP levels and breast, prostate or colorectal cancers, and five studies found that CRP was associated with colorectal or lung cancers. Most of the studies evaluating CRP as a diagnostic marker of cancer did not present relevant statistical analyses. Furthermore, any association reported in the prevalent studies might reflect reverse causation, survival bias or confounding. The prospective studies provided no strong evidence for a causal role of CRP in cancer. Instead of further prevalent studies, more large prospective studies and CRP gene-cancer association studies would be valuable in investigating the role of CRP in cancer

Identification of the CRP regulon using in vitro and in vivo transcriptional profiling

The Escherichia coli cyclic AMP receptor protein (CRP) is a global regulator that controls transcription initiation from more than 100 promoters by binding to a specific DNA sequence within cognate promoters. Many genes in the CRP regulon have been predicted simply based on the presence of DNA-binding sites within gene promoters. In this study, we have exploited a newly developed technique, run-off transcription/microarray analysis (ROMA) to define CRP-regulated promoters. Using ROMA, we identified 176 operons that were activated by CRP in vitro and 16 operons that were repressed. Using positive control mutants in different regions of CRP, we were able to classify the different promoters into class I or class II/III. A total of 104 operons were predicted to contain Class II CRP-binding sites. Sequence analysis of the operons that were repressed by CRP revealed different mechanisms for CRP inhibition. In contrast, the in vivo transcriptional profiles failed to identify most CRP-dependent regulation because of the complexity of the regulatory network. Analysis of these operons supports the hypothesis that CRP is not only a regulator of genes required for catabolism of sugars other than glucose, but also regulates the expression of a large number of other genes in E.coli. ROMA has revealed 152 hitherto unknown CRP regulons

Cost-Effectiveness of Point-of-Care C-Reactive Protein Tests for Respiratory Tract Infection in Primary Care in England

Introduction Despite recommendations that general practitioners (GPs) delay antibiotic prescribing for respiratory tract infections (RTIs), antibiotic prescriptions in primary care in England increased by 4.1% from 2010 to 2013. C-reactive protein (CRP) point-of-care tests (POCT), for example, the Afinion™ Analyzer (Alere Ltd, Stockport, UK) device, are widely used in several countries in the European Union. Studies suggest that CRP POCT use, either alone or in combination with communication training, reduces antibiotic prescribing and improves quality of life for patients presenting with RTI symptoms. The aim of this study is to evaluate the cost-effectiveness of CRP POCT for RTIs in primary care in England over 3 years for three different strategies of care compared to standard practice. Methods An economic evaluation was carried out to compare the costs and benefits of three different strategies of CRP testing (GP plus CRP; practice nurse plus CRP; and GP plus CRP and communication training) for patients with RTI symptoms as defined by National Institute for Health and Care Excellence guideline CG69, compared with current standard GP practice without CRP testing. Analysis consisted of a decision tree and Markov model to describe the quality-adjusted life years (QALYs) and cost per 100 patients, together with the number of antibiotic prescriptions and RTIs for each group. Results Compared with current standard practice, the GP plus CRP and practice nurse plus CRP test strategies result in increased QALYs and reduced costs, while the GP plus CRP testing and communication training strategy is associated with increased costs and reduced QALYs. Additionally, all three CRP arms led to fewer antibiotic prescriptions and infections over 3 years. Conclusion The additional cost per patient of the CRP test is outweighed by the associated cost savings and QALY increment associated with a reduction in infections in the long term

Critical appraisal of CRP measurement for the prediction of coronary heart disease events: new data and systematic review of 31 prospective cohorts.

BACKGROUND: Non-uniform reporting of relevant relationships and metrics hampers critical appraisal of the clinical utility of C-reactive protein (CRP) measurement for prediction of later coronary events. METHODS: We evaluated the predictive performance of CRP in the Northwick Park Heart Study (NPHS-II) and the Edinburgh Artery Study (EAS) comparing discrimination by area under the ROC curve (AUC), calibration and reclassification. We set the findings in the context of a systematic review of published studies comparing different available and imputed measures of prediction. Risk estimates per-quantile of CRP were pooled using a random effects model to infer the shape of the CRP-coronary event relationship. RESULTS: NPHS-II and EAS (3441 individuals, 309 coronary events): CRP alone provided modest discrimination for coronary heart disease (AUC 0.61 and 0.62 in NPHS-II and EAS, respectively) and only modest improvement in the discrimination of a Framingham-based risk score (FRS) (increment in AUC 0.04 and -0.01, respectively). Risk models based on FRS alone and FRS + CRP were both well calibrated and the net reclassification improvement (NRI) was 8.5% in NPHS-II and 8.8% in EAS with four risk categories, falling to 4.9% and 3.0% for 10-year coronary disease risk threshold of 15%. Systematic review (31 prospective studies 84 063 individuals, 11 252 coronary events): pooled inferred values for the AUC for CRP alone were 0.59 (0.57, 0.61), 0.59 (0.57, 0.61) and 0.57 (0.54, 0.61) for studies of 10 years follow up, respectively. Evidence from 13 studies (7201 cases) indicated that CRP did not consistently improve performance of the Framingham risk score when assessed by discrimination, with AUC increments in the range 0-0.15. Evidence from six studies (2430 cases) showed that CRP provided statistically significant but quantitatively small improvement in calibration of models based on established risk factors in some but not all studies. The wide overlap of CRP values among people who later suffered events and those who did not appeared to be explained by the consistently log-normal distribution of CRP and a graded continuous increment in coronary risk across the whole range of values without a threshold, such that a large proportion of events occurred among the many individuals with near average levels of CRP. CONCLUSIONS: CRP does not perform better than the Framingham risk equation for discrimination. The improvement in risk stratification or reclassification from addition of CRP to models based on established risk factors is small and inconsistent. Guidance on the clinical use of CRP measurement in the prediction of coronary events may require updating in light of this large comparative analysis

A new high-sensitive nephelometric method for assaying serum C-reactive protein based on phosphocholine interaction

Background: The measurement of C-reactive protein (CRP) concentrations has been of interest as a classical marker of acute phase response; in addition, it has been of particular interest in cardiovascular risk stratification where high-sensitive measurements are necessary. Since CRP is able to bind phospholipids (mainly phosphocholine) in the presence of calcium ions, we explored the possibilities of developing a high-sensitive affordable nephelometric CRP assay based on diluted soy oil emulsions. Methods: Serum (or heparinized plasma) was mixed with Intralipid 20% in Tris-calcium buffer (pH 7.5). After 12 min of incubation at 37 degrees C, the CRP-phospholipid complexes were measured by nephelometry (840 nm) using a BN II nephelometer (Siemens). Results (n=97) were compared with those obtained using a typical immunoturbidimetric method (Roche). Results: Imprecision of the functional nephelometric assay was evaluated using three human serum pools. Within-run coefficients of variation (CVs) for level 1, 2 and 3 were 6.1%, 4.7% and 4.5%, respectively, and between-run CVs were 17.6%, 18.8% and 11.3%, respectively. Good agreement was obtained between the functional nephelometric and the immunoturbidimetric CRP assay in a concentration range from 0.1 mg/L to 50 mg/L (r=0.884). A logit-log calibration curve was made between 0.056 mg/L and 1.785 mg/L. The limit of detection was 0.5 mg/L. Conclusions: The functional nephelometric CRP assay allowed high-sensitive CRP determinations in serum and plasma. Since the assay is species independent, the described functional CRP assay could be used for veterinary purposes as well

A novel interdigitated capacitor based biosensor for detection of cardiovascular risk marker

C-reactive protein (CRP) is a potential biomarker whose elevated levels in humans determine cardiovascular disease risk and inflammation. In this study, we have developed a novel capacitive biosensor for detection of CRP-antigen using capacitor with interdigitated gold (GID) electrodes on nanocrystalline diamond (NCD) surface. The NCD surface served as a dielectric layer between the gold electrodes. GID-surface was functionalized by antibodies and the immobilization was confirmed by Fourier transform spectroscopy (FT-IR) and contact angle measurements. The CRP-antigen detection was performed by capacitive/dielectric-constant measurements. The relaxation time and polarizability constants were estimated using Cole-Cole model. Our results showed that the relaxation time constant (tau) of only CRP-antibody was within 10(-16)-10(-13) s, which was increased to 10(-11) s after the incubation with CRP-antigen, suggesting that the CRP-antigen was captured by the antibodies on GID-surface. In addition, polarizability constant(m) of CRP was also increased upon incubation with increasing concentration of CRP-antigen. Our results showed that the response of GID-NCD-based capacitive biosensor for CRP-antigen was dependent on both concentration (25-800 ng/ml) as well as frequency (50-350 MHz). Furthermore, using optimized conditions, the GID-NCD based capacitive biosensor developed in this study can potentially be used for detection of elevated levels of protein risk markers in suspected subjects for early diagnosis of disease

Chemoenzymatic Synthesis of Cryptophycin Anticancer Agents by an Ester Bond-Forming Non-ribosomal Peptide Synthetase Module

Cryptophycins (Crp) are a group of cyanobacterial depsipeptides with activity against drug-resistant tumors. Although they have been shown to be promising, further efforts are required to return these highly potent compounds to the clinic through a new generation of analogues with improved medicinal properties. Herein, we report a chemosynthetic route relying on themultifunctional enzyme CrpD-M2 that incorporates a 2-hydroxy acid moiety (unit D) into Crp analogues. CrpD-M2 is a unique nonribosomal peptide synthetase (NRPS) module comprised of condensation-adenylation-ketoreduction-thiolation (C-A-KR-T) domains. We interrogated A-domain 2-keto and 2-hydroxy acid activation and loading, and KR domain activity in the presence of NADPH and NADH. The resulting 2-hydroxy acid was elongated with three synthetic Crp chain elongation intermediate analogues through ester bond formation catalyzed by CrpD-M2 C domain. Finally, the enzyme-bound seco-Crp products were macrolactonized by the Crp thioesterase. Analysis of these sequential steps was enabled through LC-FTICR-MS of enzyme-bound intermediates and products. This novel chemoenzymatic synthesis of Crp involves four sequential catalytic steps leading to the incorporation of a 2-hydroxy acid moiety in the final chain elongation intermediate. The presented work constitutes the first example where a NRPS-embedded KR domain is employed for assembly of a fully elaborated natural product, and serves as a proof-of-principle for chemoenzymatic synthesis of new Crp analogues

DETERMINANTS OF PARTICIPATION BEHAVIOR OF LIMITED RESOURCE FARMERS IN CONSERVATION RESERVE PROGRAM IN ALABAMA

This study investigated factors that affect participation behavior of limited resource farmers (LRF) in Conservation Reserve Program (CRP) in Alabama. A binary logit model was employed to analyze data obtained from 611 respondents. Results indicated that males, part-time occupation, total acres, participation in non-conservation related programs, direct method of receiving CRP information, and respondents' interest in receiving CRP information, were strong predictors of participation in the CRP. Key words: Limited resource, CRP, minorities, participation behaviorLimited resource, CRP, minorities, participation behavior, Environmental Economics and Policy,