Biomarkers In Subjects Research Articles

A11713 Prognostic Value of Cardiovascular Risk Factors in Hypertensive versus Non-hypertensive Middle-aged and Elderly Subjects

Objectives: The prognostic impact of each of the CV risk factors is normally modified and defined by the background risk burden. Hypertension is the most significant and frequent risk factor in the population. Thus, the burden of other risk factors may be different in subjects with versus without hypertension. The objectives were to evaluate the 14-year prognostic impact of conventional cardiovascular (CV) risk factors and biomarkers in subjects with versus without hypertension. Methods: 678 middle aged and elderly subjects with no previous or known CV disease from Copenhagen Holter Study were included. All were studied by interview, clinical examination, 48 hour Holter recording and laboratory testing. Fourteen years rate of CV endpoints (CV death, stroke, or acute myocardial infarction) were evaluated by meticulous evaluation of patient files and registry search. Hypertension was defined as blood pressure > 140/90 or the use of antihypertensive medication. Cox proportional models were used to evaluate the impact of risk factors in each group. Results: There were 491 hypertensive and 187 normotensive subjects, with 111 (22.5%) and 28 (15%) CV-endpoints respectively (p = 0.02). Table 1a and 1b shows the prognostic value of CV risk factors in hypertensives and normotensives. Conclusion: While most CV risk factors have an impact in hypertensives, only inflammation and BP do have a prognostic impact in normotensives.

Present and Future of Ultra-High Field MRI in Neurodegenerative Disorders.

With a high signal-to-noise ratio, unparalleled spatial resolution, and improved contrasts, ultra-high field MR (≥ 7T) has great potential in depicting the normal radiological anatomy of smaller structures in the brain and can also provide more information about morphological, quantitative, and metabolic changes associated with a wide range of brain disorders. By focusing attention on specific brain regions believed to be associated with early pathological change, or by more closely inspecting recognized foci of brain pathology, ultra-high field MR can improve the accuracy and sensitivity of neuroimaging. This article reviews recent studies at ultra-high field about Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). The research on AD has mainly focused on detecting the thinning of hippocampal layers and the susceptibility effect supposed to be related to beta-amyloid deposition. In patients with PD, atypical parkinsonisms and subjects at risk of developing motor symptoms of Parkinson's disease, the main aim was to detect changes in the substantia nigra, probably related to increased iron deposition. In patients with ALS, both brain and spinal cord were investigated, with the aim of finding changes in the primary motor cortex and corticospinal tract which reflect neurodegeneration and neuroinflammation. Ultra-high field MR was shown to be useful for detecting subtle brain changes in patients with AD, and possible new diagnostic biomarkers in patients with PD and ALS were discovered. The ability of 7T MR to provide prognostic biomarkers in subjects at risk for developing synucleinopathies is currently under evaluation.

Enhanced inflammation in subgroups of metabolic syndrome irrespective of glycaemic status

The objectives of this study were toinvestigate the inflammation and endothelial activation biomarkers in subjects with metabolic syndrome (MS)subgroups irrespective of status. Subjects (n=319) were categorized into MS subgroups [diabetes mellitus (MSDM), impaired fastig glucose (MSIFG) and normoglycemia (MSNG)] and normal controls (NC). Inflammatory biomarkers (high-sensitivity C-reactive protein and interleukin-6) and endothelial activation (soluble intercellular adhesion molecule-1, vascular cell adhesion molecule-1 and E-selectin) were measured and compared between the groups. MS subjects had significant increase in all biomarkers compared to NC subjects. Glycaemic status in MS subgroups did not show any association with the quartiles of all biomarkers, except for interleukin-6. This may partially explain their increased coronary risk. Keywords : metabolic syndrome; glycaemic status; inflammation; endothelial activation

A Conservative Weight Loss Intervention Relieves Bowel Symptoms in Morbidly Obese Subjects with Irritable Bowel Syndrome: A Prospective Cohort Study.

Background Irritable bowel syndrome (IBS) is common in subjects with morbid obesity; the effect of weight loss programs on bowel symptoms is largely unknown. Methods This prospective cohort study explored bowel symptoms, health scores, and biomarkers in subjects with morbid obesity during a six-month-long conservative weight loss intervention. Bowel symptoms were assessed with IBS-severity scoring system (IBS-SSS) and Gastrointestinal Symptom Rating Scale-IBS. Changes in all variables and associations between the changes in bowel symptoms and the other variables were analysed. Results Eighty-eight subjects (81% females) were included. Body mass index was reduced from 42.0 (3.6) to 38.7 (3.5) (p < 0.001). IBS-SSS was reduced from 116 (104) to 81 (84) (p=0.001). In all, 19 out of 25 variables improved significantly. In subjects with and without IBS at inclusion, the improvement in IBS-SSS was 88 (95% CI 55 to 121) and 10 (95% CI −9 to 29), respectively. Improved bowel symptoms were associated with improved subjective well-being, sense of humour, and vitamin D and negatively associated with reduced body mass index. Conclusion Body mass index and health scores improved during a conservative weight loss intervention. Subjects with IBS before the intervention had a clinically significant improvement in bowel symptoms.

Comparable Enhanced Prothrombogenesis in Simple Central Obesity and Metabolic Syndrome.

Objective There is limited data comparing prothrombogenic or fibrinolysis biomarkers (tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1)) simultaneously in subjects with Metabolic Syndrome (MS), simple central obesity without MS (COB) and normal controls (NC). We investigated the concentrations of fibrinolysis biomarkers in subjects with MS, COB and NC. Methods A cross-sectional study involving 503 drug naive subjects (163 males, aged 30–65 years old (mean age ± SD = 47.4 ± 8.3 years)) divided into MS, COB and NC groups. COB was defined as central obesity (waist circumference (WC) males ≥90 cm, females ≥80 cm) in the absence of MS according to the International Diabetes Federation 2006. Fasting blood levels of tPA and PAI-1were analyzed. Results MS and COB had significantly higher concentration of all biomarkers compared to NC. The MS group had significantly higher concentration of tPA and PAI-1 compared to COB. WC and HDL-c had significant correlation with all biomarkers (tPA p < 0.001, PAI-1 p < 0.001). Fasting plasma glucose and diastolic blood pressure were independent predictors after correcting for confounding factors. Conclusion Central obesity with or without MS both demonstrated enhanced prothrombogenesis. This suggests that simple obesity possibly increases the risk of coronary artery disease in part, via increased susceptibility to thrombogenesis.

Cartilage oligomeric protein, matrix metalloproteinase-3, and Coll2-1 as serum biomarkers in knee osteoarthritis: a cross-sectional study.

Biochemical markers reflecting joint remodeling in osteoarthritis (OA) are a promising diagnostic tool. The aim of this study was to investigate serum levels of candidate biomarkers in subjects with and without knee OA and assess their correlation with clinical parameters and knee structural damage. 56 patients with primary knee OA and 31 healthy controls participated in this study. Patients were separated into two groups: isolated knee OA and generalized OA. Clinical parameters were obtained by validated self-reported questionnaires and a visual analogue scale. Serum levels of cartilage oligomeric protein (COMP), matrix metalloproteinase-3 (MMP-3), and Coll2-1 were quantified by enzyme-linked immunosorbent assay. Knee structural damage was determined by plain X-ray and 1.5T magnetic resonance imaging (MRI), using Kellgren-Lawrence (KL) grading scale and Whole-Organ Magnetic Resonance Imaging Score (WORMS), respectively. Compared to controls, patients had significantly higher median serum COMP (985 vs. 625ng/ml; p < 0.001) and MMP-3 (36.85 vs. 22.10ng/ml; p = 0.003) levels. Patients with radiographic evidence of KLII/III knee OA had greater median COMP levels than KLI patients (1095 vs. 720ng/ml; p = 0.001). In the generalized OA group, mean MMP-3 levels were higher than in the isolated knee OA group (30.40 vs. 55.13ng/ml; p < 0.001). COMP correlated positively with WORMS (r s = 0.454, p < 0.001) and MMP-3 (r s = 0.337, p = 0.003). Cut-off values for serum COMP and MMP-3 were determined. We observed higher serum COMP and MMP-3 levels in knee OA patients compared to controls. COMP may reflect knee structural damage, while MMP-3-OA "generalization".

Association between hemoglobin glycation index and hepatic steatosis in non-diabetic individuals

Hemoglobin glycation index (HGI), which is the difference between the observed value of HbA1 and the predicted HbA1c based on plasma glucose levels, represents a measure of the degree of non-enzymatic glycation of hemoglobin and it has been found to be positively associated with diabetic complications. Herein we investigated whether HGI is associated with hepatic steatosis and related biomarkers in subjects without diabetes. 1120 White individuals without diabetes were stratified in quartiles according to HGI levels. Hepatic steatosis was diagnosed by ultrasonography. As compared with subjects in the lowest quartile of HGI those in the intermediate and high HGI groups displayed an unfavorable cardio-metabolic risk profile having significantly higher values of body mass index (BMI), waist circumference, % fat mass, total cholesterol, triglycerides, inflammatory markers such as high sensitivity C reactive protein, erythrocytes sedimentation rate, complement C3, platelets and white blood cell count, hepatic insulin resistance assessed by the liver IR index and lower concentrations of high-density lipoprotein. HGI was positively associated with the biomarker of liver damage alanine aminotransferase, and fatty liver index, an indicator of hepatic steatosis. In a logistic regression analysis adjusted for age, gender and BMI individuals in the highest quartile of HGI exhibited a 1.6-fold increased odd of having hepatic steatosis (95% CI: 1.03-2.41; p=0.03) as compared with subjects in the lowest quartile of HGI. Higher levels of HGI may identify subjects without diabetes at increased risk of having hepatic steatosis.

Coronary risk biomarkers in centrally obese subjects with and without metabolic syndrome

Aim: Metabolic Syndrome (MS) is associated with an approximate doubling of coronary risk; and the risk for diabetes is five times higher in individuals with MS compared to those without the syndrome. However, the underlying mechanisms to explain these elevated risks are not well understood. Therefore, the objectives of this study were to examine the inflammation and endothelial activation biomarkers in subjects with MS and central obesity without MS, compared to lean normal controls; and to investigate the influence of glycaemic status in MS on these biomarkers.

Single and multiple cardiovascular biomarkers in subjects without a previous cardiovascular event.

Aims To assess the incremental value of biomarkers, including N-terminal prohormone of brain natriuretic peptide (NT-proBNP), high-sensitivity troponin T (hs-TnT), high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), growth differentiation factor 15 (GDF-15), and procollagen type 1 N-terminal propeptide (P1NP), in predicting incident cardiovascular events and mortality among asymptomatic individuals from the general population, beyond traditional risk factors, including fasting glucose and renal function (cystatin C), medication use, and echocardiographic measures. Methods and results Prospective population-based cohort study of 1324 subjects without a previous cardiovascular event, who underwent baseline echocardiography and biomarker assessment between 2002 and 2006. The clinical endpoint was the composite of myocardial infarction, invasively treated stable/unstable ischemic heart disease, heart failure, stroke, or all-cause mortality. Predictive capabilities were evaluated using Cox proportional-hazards regression, Harrell's concordance index (C-index), and net reclassification improvement. Median age was 66 (interquartile range: 60-70) years, and 413 (31%) were female. During median 8.6 (interquartile range: 8.1-9.2) follow-up years, 368 (28%) composite events occurred. NT-proBNP, hs-TnT, GDF-15, and IL-6 were significantly associated with outcome, independently of traditional risk factors, medications, and echocardiography ( p < 0.05 for all). Separate addition of NT-proBNP and GDF-15 to traditional risk factors, medications, and echocardiographic measurements provided significant improvements in discriminative ability (NT-proBNP: C-index 0.714 vs. 0.703, p = 0.03; GDF-15: C-index 0.721 vs. 0.703, p = 0.02). Both biomarkers remained significant predictors of outcome upon inclusion in the same model ( p < 0.05 for both). Conclusions NT-proBNP and GDF-15 each enhance prognostication beyond traditional risk factors, glucose levels, renal function, and echocardiography in individuals without known cardiovascular disease.

From mild cognitive impairment to subjective cognitive decline: conceptual and methodological evolution.

Identification of subjects at the early stages of Alzheimer’s disease (AD) is fundamental for drug development and possible intervention or prevention of cognitive decline. The concept of mild cognitive impairment (MCI) evolved during the past two decades to define subjects at the transitional stage between normal aging and dementia. Evidence from cross-sectional and longitudinal studies has shown that MCI is associated with an increased risk of positive AD biomarkers and an increased annual conversion rate of 5%–17% to AD. The presence of AD biomarkers in subjects with MCI was associated with an even higher risk of progression to dementia. However, earlier clinical trials for pharmacotherapy in subjects with MCI were disappointing. To extend the spectrum of AD to an earlier stage before MCI, subjective cognitive decline (SCD) was introduced and was defined as self-reported cognitive decline before the deficits could be detected by cognitive tests. Subjects with SCD have an increased risk of underlying AD pathology. However, SCD can also develop secondary to other heterogeneous etiologies, including other neurodegenerative and psychiatric diseases, personality traits, physical conditions, and medication use. Several clinical and biomarker features were proposed to predict risk of conversion to AD in subjects with SCD. Further longitudinal studies are needed to support the validity of these high-risk features.

Associations of Circulating PYY3-36 Concentrations with Metabolic Syndrome in Extremely Obese Subjects.

The gut hormone peptide YY3-36 (PYY3-36) plays major roles in regulation of appetite and energy metabolism, mediates beneficial effects of bariatric surgery, and may be a potential weight-reducing and glucose-modulating therapy. Obesity may influence the metabolic expression of circulating PYY3-36 and metabolic markers. We studied the relationship of PYY3-36 concentrations with metabolic syndrome (MetSyn) components, lipids, insulin resistance, and inflammatory biomarkers in subjects with extreme obesity. We measured MetSyn components and PYY3-36, lipids, hormones, homeostasis model assessment (HOMA) index, and inflammatory biomarkers in consecutively referred patients (180 women and 111 men) aged 18-78 years with body mass index (BMI) ≥40 kg/m2. Associations of PYY3-36 to components, insulin resistance, and biomarkers were examined with partial correlations and linear regression. PYY3-36 concentrations were not related to MetSyn components, HOMA index, or to inflammatory biomarker or leptin concentrations. PYY3-36 concentrations correlated with systolic blood pressure (r = 0.21; P < 0.0001) after adjustment for age and gender. In linear regression analysis, PYY3-36 concentrations were associated with systolic blood pressure after adjustment for age, gender, and central obesity in the entire sample (Beta 0.21; 95% CI 0.09-0.34) as well as in subjects not taking blood pressure-lowering medication (Beta 0.19; 95% CI 0.04-0.36). These associations were not statistically significant in the small subset of participants (22%) with type 2 diabetes. In extremely obese patients, fasting PYY3-36 concentrations were linked to systolic blood pressure, but not to other components of MetSyn, suggesting divergence between pathways of blood pressure and glucose/body weight regulation. However, this finding will need to be further investigated.

Reductions in biomarkers of exposure (BoE) to harmful or potentially harmful constituents (HPHCs) following partial or complete substitution of cigarettes with electronic cigarettes in adult smokers

Changes in fifteen urine, blood and exhaled breath BoEs of HPHCs representing classes of compounds reported by FDA to be significant contributors to smoking-associated disease risks were measured in 105 clinical-confined subjects following randomization and a five-day forced-switch from usual brand conventional combustible cigarettes to: (i) exclusive commercial e-cigarette use; (ii) dual-use of commercial e-cigarettes and the subject’s usual cigarette brand; or (iii) discontinued use of all tobacco or nicotine products. Levels of urinary biomarkers in subjects that completely substituted their usual cigarette with e-cigarettes were significantly lower (29–95%) after 5 days. Percent reductions in eight of nine urinary BoEs were indistinguishable to smokers who had quit smoking, except for nicotine equivalents, which declined by 25–40%. Dual users who halved self-reported daily cigarette consumption with e-cigarettes exhibited reductions (7–38%) in eight of nine urinary biomarkers, but had increase (1–20%) in nicotine equivalents. Reductions were broadly proportional to the reduced numbers of cigarettes smoked. Dual user urinary nicotine equivalents were slightly higher, but not statistically significant. After 5 days, blood nicotine biomarker levels were lower in the cessation (75–96%) and exclusive use groups (11–83%); with dual users experiencing no significant reductions. All subjects experienced significant decreases in exhaled CO. Decreases in the cessation and exclusive groups ranged from 88–89% and 27–32% in dual users. Exhaled NO increased in the cessation and exclusive groups (46–63% respectively), whereas the dual users experienced minimal changes. Overall, smokers who completely or partially substituted conventional cigarettes with e-cigarettes over five days, experienced reductions in HPHCs.

RELATION BETWEEN LIFESTYLE FACTORS AND ALZHEIMER'S DISEASE BIOMARKERS IN SUBJECTS WITH SCI OR MCI

RELATION BETWEEN LIFESTYLE FACTORS AND ALZHEIMER'S DISEASE BIOMARKERS IN SUBJECTS WITH SCI OR MCI

Increased serological cancer-associated biomarker levels at large bowel endoscopy and risk of subsequent primary cancer

Background: Frequently, subjects offered colonoscopy due to symptoms of colorectal neoplasia are diagnosed with diverticula. The symptoms may, however, also be related to extra-colonic neoplasia. The present retrospective study evaluated a possible association between increased levels of predefined biomarkers in subjects diagnosed with diverticula and risk of developing a primary malignant disease.Methods: During 2004/2005, about 4509 subjects were included in a multicenter study with collection of blood samples before bowel endoscopy. The aim was to evaluate a relation between the protein biomarkers CEA, TIMP-1, CA19-9 and YKL-40 and findings at endoscopy. Diverticula were diagnosed in 1021 subjects. By 31 December 2012, subjects who had developed primary malignancy were identified retrospectively and relation between biomarker levels at endoscopy and risk of developing primary malignancy was calculated. The relation with the four biomarkers was divided into three groups: 0 = none increased; 1 = one increased and 2 = two or more increased.Results: In the observation period, 148 subjects developed a primary malignant disease. Univariable analyzes of the biomarker levels showed that CEA, TIMP-1 and CA19-9 were significantly associated with development of primary malignancy. A multivariable analysis showed that increased levels were associated with development of malignancy (p < 0.0001). The 1- and 5-year cumulative risks of being diagnosed with a primary malignancy were: group 0: 1.1%/5.5%; group 1: 4.2%/10.1% and group 2: 11.4%/18.8%, respectively.Conclusion: Increased levels of CEA, TIMP-1 and CA19-9 at endoscopy with findings of diverticula were associated with a significantly increased risk of being diagnosed with a subsequent primary malignant disease.

Serum Biomarkers of COPD

To the Editor: Ambade et al[1][1] conducted an observational study to evaluate the potential serum biomarkers in subjects with COPD. The authors assessed superoxide dismutase-3, glutathione peroxidase, catalase, ceruloplasmin ferroxidase activity, C-reactive protein, and surfactant protein D in

Modulation of Myotilin and Fylamin C in Various Muscle Diseases: A Microarray Analysis

The cytoskeletal protein plays a significant role in the stability of thin filaments during muscle contraction. Mutations in these genes have been associated with various muscles diseases. Myotilin (MYOT) and Fylamin C (FLNC) belong to the cytoskeleton protein family and are associated with different myopathies. We analyzed two microarray datasets obtained from the NCBI Gene Expression Omnibus databank (accession number GDS2855 and GDS1956) in order to verify the modulation of MYOT and FLNC in eight human skeletal muscle diseases. For these studies we also used: the open source tools the Human Protein Atlas to confirm by Immunohistochemistry (IH) the MYOT and FLNC tissue expression; Genome-scale Integrated Analysis of gene Networks in Tissues (GIANT) to identify the genes network; COMPARTMENT to identify the localization in cells. We showed that both MYOT and FLNC were significantly modulated in various muscle diseases. In particular, MYOT and FLNC mRNA were significantly downregulated in Acute quadriplegic myopathy (AQM) and Amyotrophic Lateral Sclerosis (ALS) compared to normal human skeletal muscle. Furthermore, the GIANT analysis showed a relationship confidence of 0.23 to MYOT and FLNC, confirming their strong correlation. These data provide to support our hypothesis that a positive correlation exists between MYOT and FLNC. Larger studies are needed to evaluate if MYOT and FLNC may be a promising clinical biomarker in subjects with diseases of the muscle.

Abstract A06: Elucidating the associations of inflammation with cancer risk and mortality using an inflammatory score: Results from a prospective cohort study

Abstract Introduction: Experimental and clinical studies have demonstrated that inflammation is associated with cancer risk and mortality. However, there are conflicting reports on the associations of biomarkers of inflammation with cancer. Because the biomarkers are systemic in origin, associations between individual biomarkers and cancer may not be always apparent. One inflammatory biomarker may correctly classify some cases and misclassify others and this could contribute to the conflicting results reported in previous studies. To further elucidate the relationship between inflammation and cancer, we investigated the associations of C-reactive protein (CRP) and leukocyte count with cancer risk and mortality using the individual biomarkers and an inflammatory score derived from the two biomarkers. We hypothesize that any observed association would be more apparent in analysis conducted using an inflammatory score compared with analysis using individual biomarkers. Methods: We conducted this analysis among men enrolled in a prospective cohort study from Finland who had baseline examination and blood draw between March 1984 and December 1989. During an average follow-up period ranging from 23-28 years (till December 2012), 653 men developed cancer and 287 cancer deaths occurred. Cancer cases and deaths were ascertained through linkage to nationwide Finnish cancer Registry and Statistics Finland, respectively. Participants were categorized into quartiles based on the distribution of the biomarkers within the study population. In addition to individual biomarker analysis, the biomarkers were analyzed together using a standardized combined z-score. We computed the score for each biomarker based on the subject's biomarker concentration (x), study mean (μ) and study standard deviation (σ) and computed a z-score with z = ( x - μ )/σ . The combined z-score was the sum of the subject's individual CRP and leukocyte z-scores. We used Cox proportional hazard models to evaluate associations of the biomarkers, including the z-scores, with cancer risk and mortality, adjusting for confounders (age at baseline, examination year, smoking history (pack-years), alcohol consumption, energy intake, and cardiorespiratory fitness). Results: In multivariable-adjusted analyses using individual biomarkers, elevated leukocyte count (HR= 1.31, 95%CI 1.04-1.66, p-trend=0.19) and CRP concentration (HR= 1.23, 95%CI 0.97-1.55, p-trend=0.18) were not associated with statistically significantly increased risk of cancer and cancer mortality (HR=1.15, 95%CI 0.81-1.64, p-trend=0.43 for CRP and HR=1.39, 95%CI 0.98-1.97, p-trend=0.42 for leukocyte count). However, in analyses conducted using the derived z-scores, men with elevated z-scores had significantly increased risk of cancer. The HRs comparing men within the highest z-score quartile to those within the lowest z-score quartile were 1.47 (95%CI 1.16-1.88, p-trend&amp;lt;0.01) and 1.48 (95% CI 1.03-2.14, p-trend=0.11) for cancer risk and mortality, respectively. Conclusions: In this prospective cohort study with long follow-up, an inflammatory biomarker score was more predictive of cancer risk than individual biomarkers alone. Our results indicate that combining biomarkers of inflammation may provide better clues to elucidating the associations of inflammation with cancer risk and mortality. Citation Format: Leavitt Morrison, Mark Ziegler, Jari Laukkanen, Sudhir Kurl, Kimmo Ronkainen, Jussi Kauhanen, Adetunji T. Toriola. Elucidating the associations of inflammation with cancer risk and mortality using an inflammatory score: Results from a prospective cohort study [abstract]. In: Proceedings of the Thirteenth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2014 Sep 27-Oct 1; New Orleans, LA. Philadelphia (PA): AACR; Can Prev Res 2015;8(10 Suppl): Abstract nr A06.

A nonparametric fiducial interval for the Youden index in multi-state diagnostic settings.

The Youden index is a commonly employed metric to characterize the performance of a diagnostic test at its optimal point. For tests with three or more outcome classes, the Youden index has been extended; however, there are limited methods to compute a confidence interval (CI) about its value. Often, outcome classes are assumed to be normally distributed, which facilitates computational formulas for the CI bounds; however, many scenarios exist for which these assumptions cannot be made. In addition, many of these existing CI methods do not work well for small sample sizes. We propose a method to compute a nonparametric interval about the Youden index utilizing the fiducial argument. This fiducial interval ensures that CI coverage is met regardless of sample size, underlying distributional assumptions, or use of a complex classifier for diagnosis. Two alternate fiducial intervals are also considered. A simulation was conducted, which demonstrates the coverage and interval length for the proposed methods. Comparisons were made using no distributional assumptions on the outcome classes and for when outcomes were assumed to be normally distributed. In general, coverage probability was consistently met, and interval length was reasonable. The proposed fiducial method was also demonstrated in data examining biomarkers in subjects to predict diagnostic stages ranging from normal kidney function to chronic allograph nephropathy. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Abstract 3433: Identification of potential salivary response biomarkers in subjects practicing yogic breathing

Abstract Yogic Breathing (YB; Pranayama) is a form of Yoga technique practiced since ancient times for healthy living. In the United States, the practice of Yoga has markedly increased in recent years. Recent data suggest that adjunctive Yoga in cancer patients may help patients with symptom management. Whereas there are data on Asana (Postures) that involve physical movements in cancer patients, there are limited data on YB. Considering the ease of practice when compared to Asana, YB might benefit patients who are already physically overwhelmed with symptom burden. Current data measure the outcomes of Yoga using subjective measures, but biomarkers in an easily accessible biospecimen might provide key information as to the effects of YB among practitioners. We hypothesized that YB stimulates alterations in the salivary biomarker expression. We conducted an Institutional Review Board approved pilot randomized controlled trial in 20 subjects (n = 10 in each group) to compare the salivary expression of biomarkers between a 20 min YB group vs. Attention Control (AC) group. The YB group performed two exercises (Pranava Pranayama, and Thirumoolar Pranayama) each for 10 min, and the AC group read a scientific article for the same amount of time. Tandem Mass Spectrometric (LC-LC-MS-MS), Multiplex, and Western blot analysis showed the differential expression of 22 salivary proteins associated with immune response, stress and cancer. As compared with the AC group, YB increased Deleted in Malignant Brain Tumor 1 (DMBT1) as early as 5 min and maintained significantly high levels in all time points tested. In contrast, YB reduced IL-1beta (at 10, 15 and 20 min) and IL-8 levels (at 5, 10, 15 and 20 min) in participants when compared to AC controls. Further analysis of salivary lipidomic profiles demonstrated marginal reductions in sphingosine and sphingosine-1-phosphate levels in YB group. Collectively, our data provide the first evidence that YB could alter salivary biomarkers associated with cancer, inflammation, and stress. These markers may be useful to evaluate symptom management via stress reduction and immune regulation. Citation Format: Sundaravadivel Balasubramanian, Michael G. Janech, Graham W. Warren. Identification of potential salivary response biomarkers in subjects practicing yogic breathing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3433. doi:10.1158/1538-7445.AM2015-3433

Production and characterisation of recombinant human chaperonin 10 for treatment of inflammatory disease

Human chaperonin 10 (Cpn10) and chaperonin 60 (Cpn60) fulfil an essential role in mitochondrial protein folding. Cpn10 is located in the mitochondrial matrix; however, it has also been detected in extra-mitochondrial compartments where it has demonstrated anti-inflammatory and immunoregulatory activity, suggesting a potential therapeutic value for the treatment of inflammatory and autoimmune disorders that corresponds with its recently proposed role as a resolution-associated molecular patterns (RAMPs) molecule. Ala-Cpn10, a recombinant, minimally modified Cpn10 synthesised in Escherichia coli, is formulated for use in clinical trials and pre-clinical studies with intravenous or subcutaneous administration. Herein, we report the development of a bioprocess for the production of ∼115g of recombinant human Ala-Cpn10 from a 100L E. coli fermentation with >99% purity (SDS-PAGE), <0.6EUmg−1 endotoxin, <18pgmg−1 DNA and 3.2% molecular variants. This bioprocess was achieved through a careful optimisation of the gene construct, the promoter system and the fermentation process. Recombinant Ala-Cpn10 produced in this process is active as a molecular chaperone indicating correct tertiary and quaternary structure and the stability profile indicates no significant changes with storage as a liquid for at least 3 years at 2–8°C. The results of validated characterisation assays demonstrate that purified Ala-Cpn10 produced using the optimised process reported here is suitable for its intended purpose as an investigational drug product, and this material is currently being tested in a phase II study for efficacy, safety and impact on biomarkers in subjects with mildly active Systemic Lupus Erythematosus (SLE) under US IND 116156.