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Daniel Lamoureux,1 David T Wong,2,3 Tina Felfeli2,4 1Northern Ontario School of Medicine University, Thunder Bay, ON, Canada; 2Department of Ophthalmology and Vision Sciences, University of Toronto, Toronto, ON, Canada; 3Department of Ophthalmology, St. Michael’s Hospital, Unity Health Toronto, Toronto, ON, Canada; 4The Institute of Health Policy, Management and Evaluation, University of Toronto, Toronto, ON, CanadaCorrespondence: Tina Felfeli, University Health Network, Toronto General Hospital, Eaton Building, 10th Floor, Toronto, ON M5G 2C4, Canada, Tel +1 647 678 1634, Fax +1 416 340 3459, Email tina.felfeli@mail.utoronto.caPurpose: Certain factors such as instrumental and sample processing errors may contribute to variability of ocular biofluid samples when they are run as replicates with multiplex assays. There is a paucity of literature on the variability of replicates in multiplex assays. This study aims to evaluate whether there is significant variability in replicate analyses of multiplex assays.Methods: A total of 152 human ocular biofluid samples (51 aqueous humor and 101 vitreous) were collected and assayed for 27 cytokine biomarker concentrations (pg/mL). Samples were evaluated as replicates (duplicate analysis) at four different time points. Statistical methods including paired samples t-test, 3-way ANOVA, intraclass correlation coefficient (ICC; < 0.5– 0.75=poor-moderate, 0.75-> 0.90 =good-excellent reliability), and coefficients of variation (CV) were employed to evaluate for statistical significance, with Bonferroni corrected P=0.002.Results: Among the 4104 biomarker replicate assays for aqueous humor and vitreous, two analytes (PDGF-BB and IL-7) had a statistically significant difference between the sampled concentrations of the replicates in vitreous samples (mean (diff)=2.05, P< 0.001, mean (diff)=1.56, P< 0.001, respectively). Majority of the ICC values fell within the good-excellent range (86% of samples) with a minority falling in the poor-moderate range (14% of samples). More variability was noted in the vitreous humour, with five analytes (IL-2, IL-10, IL-12(p70), IL-13, IL-17) demonstrating an average ICC of less than 0.5. The CV calculated for each set of replicates suggested that 93% of replicates had an acceptable level of quantitative assay variability (CV< 20%).Conclusion: This study demonstrates that the analysis of most biomarkers in ocular fluids may not require the use of replicates. However, certain analytes such as PDGF-BB and IL-7 may require the use of replicates to ensure reliable results. Caution should be taken when applying these findings to other laboratory settings as our study was conducted by an experienced technician using a standardized protocol. In less standardized settings, replicates may be required in order to ensure accuracy of results. These findings may guide researchers with the design of their studies on ophthalmic biomarker analysis.Keywords: cytokines, aqueous, vitreous, biomarkers, assay

Sinny Delacroix,1,2 Ramesh G Chokka,1,3 Adam J Nelson,1,2 Dennis T Wong,4 Samuel Sidharta,1,2 Stephen M Pederson,5 Adil Rajwani,1,2 Joanne Nimmo,1,2 Karen S Teo,1,2 Stephen G Worthley1,2 1Cardiovascular Research Centre, Royal Adelaide Hospital, 2Department of Medicine, University of Adelaide, Adelaide, SA, 3South Australian Health and Medical Research Institute, University of Adelaide, Adelaide, SA, 4Monash Medical Centre, Clayton, VIC, 5Bioinformatics Hub, School of Biological Sciences, University of Adelaide, Adelaide, SA, Australia Aim: Preclinical studies have demonstrated improvements in renal blood flow after renal sympathetic denervation (RSDN); however, such effects are yet to be confirmed in patients with resistant hypertension. Herein, we assessed the effects of RSDN on renal artery blood flow and diameter at multiple time points post-RSDN. Methods and results: Patients (n=11) with systolic blood pressures ≥160 mmHg despite taking three or more antihypertensive medications at maximum tolerated dose were recruited into this single-center, prospective, non-blinded study. Magnetic resonance imaging indices included renal blood flow and renal artery diameters at baseline, 1 month and 6 months. In addition to significant decreases in blood pressures (p

In this 2-part review series, the current uses of saliva as a diagnostic fluid are reviewed, first with a focus on known measurements of systemic conditions. In Part 2, the role of saliva to measure bone turnover with a special emphasis on osteoporosis will be discussed. [ABSTRACT FROM AUTHOR]

In vivo assessment of ventricular function in rodents has largely been restricted to transthoracic echocardiography (TTE). However 1.5 T cardiac magnetic resonance (CMR) and transoesophageal echocardiography (TOE) have emerged as possible alternatives. Yet, to date, no study has systematically assessed these three imaging modalities in determining ejection fraction (EF) in rats. Twenty rats underwent imaging four weeks after surgically-induced myocardial infarction. CMR was performed on a 1.5 T scanner, TTE was conducted using a 9.2 MHz transducer and TOE was performed with a 10 MHz intracardiac echo catheter. Correlation between the three techniques for EF determination and analysis reproducibility was assessed. Moderate-strong correlation was observed between the three modalities; the greatest between CMR and TOE (intraclass correlation coefficient (ICC) = 0.89), followed by TOE and TTE (ICC = 0.70) and CMR and TTE (ICC = 0.63). Intra- and inter-observer variations were excellent with CMR (ICC = 0.99 and 0.98 respectively), very good with TTE (0.90 and 0.89) and TOE (0.87 and 0.84). Each modality is a viable option for evaluating ventricular function in rats, however the high image quality and excellent reproducibility of CMR offers distinct advantages even at 1.5 T with conventional coils and software. [ABSTRACT FROM AUTHOR]

Background: A higher oropharyngeal leak pressure (OLP) is a marker of efficacy and safety when using laryngeal mask airway devices. The new disposable laryngeal mask airway (LMA™) Supreme™ has lower OLP compared with the LMA ProSeal™. Increased intracuff pressure of laryngeal mask airway devices may improve OLP but may result in more postoperative pharyngolaryngeal adverse events. This study was designed to compare the OLP of the LMA Supreme at varying intracuff pressures.Methods: One hundred and twenty-three patients were divided randomly into three groups. General anesthesia was standardized using a propofol-fentanyl induction and desflurane in air-oxygen for maintenance. Intracuff pressures of the LMA Supreme were adjusted to 80 cm H(2)O, 60 cm H(2)O, and 40 cm H(2)O according to group allocation. The primary outcome was OLP. Secondary outcomes included postoperative pharyngolaryngeal adverse events and the satisfaction scores of patients and anesthesiologists. The OLP was compared amongst groups using analysis of variance with Bonferroni correction. All reported P values are two-sided.Results: The OLP with an intracuff pressure of 80 cm H(2)O was significantly higher compared with 60 cm H(2)O and 40 cm H(2)O (26 [6] vs 20 [6] vs 18 [5] cm H(2)O, respectively; P < 0.001). The incidence of postoperative pharyngolaryngeal adverse events (P = 0.6), patient satisfaction scores (P = 0.2), and anesthesiologist satisfaction scores (P = 0.8) were comparable amongst the three groups.Conclusion: An intracuff pressure of 80 cm H(2)O with the LMA Supreme is associated with a higher OLP compared with 60 cm H(2)O or 40 cm H(2)O without a greater incidence of postoperative pharyngolaryngeal adverse events. For a superior glottic seal when using the LMA Supreme, we recommend intracuff pressures up to 80 cm H(2)O. [ABSTRACT FROM AUTHOR]

The protein compositions, or the proteomes, found in human salivary and plasma fluids are compared. From recent experimental work by many laboratories, a catalogue of 2290 proteins found in whole saliva has been compiled. This list of salivary proteins is compared with the 2698 proteins found in plasma. Approximately 27% of the whole-saliva proteins are found in plasma. However, despite this apparent low degree of overlap, the distribution found across Gene Ontological categories, such as molecular function, biological processes, and cellular components, shows significant similarities. Moreover, nearly 40% of the proteins that have been suggested to be candidate markers for diseases such as cancer, cardiovascular disease, and stroke can be found in whole saliva. These comparisons and correlations should encourage researchers to consider the use of saliva to discover new protein markers of disease and as a diagnostic non-proximal fluid to detect early signs of disease throughout the body. [ABSTRACT FROM AUTHOR]

Background and Objective: The Supreme laryngeal mask airway (LMA) is a new single-use polyvinyl chloride supraglottic device that combines the functionality of the ProSeal and Fastrach airways. High oropharyngeal leak pressures are important as they indicate airway protection, feasibility of positive pressure ventilation and likelihood of successful LMA placement. The oropharyngeal leak pressure of the LMA Supreme is not well established versus the LMA ProSeal. This study was designed to compare the safety and efficacy of the LMA Supreme versus the LMA ProSeal in elective ambulatory procedures.Method: Hospital ethics board approval was obtained. One hundred and five patients were consented and randomly allocated to LMA Supreme or ProSeal groups. Anaesthesia was induced with intravenous propofol 2-3 mg kg(-1) and fentanyl 1-2 microg kg(-1) and maintained with desflurane in an air-oxygen mixture. Anaesthesiologists with more than 5 years of experience performed all of the LMA insertions. Manometry was used to standardize intracuff pressure at 60 cmH2O. The primary outcome was the oropharyngeal leak pressure. Secondary outcomes were the time and number of attempts for insertion, ease of insertion and the anaesthesiologist's satisfaction score of the airway device. The success on first attempt insertion was measured. Patients were interviewed postoperatively for any pharyngolaryngeal adverse events.Results: A total of 99 patients were analysed for the primary outcome. The baseline demographic data for both groups were comparable. The mean oropharyngeal leak pressure with the LMA Supreme was 21 +/- 5 cmH2O (95% confidence interval 20-22). This was significantly lower than that of the LMA ProSeal, 25 +/- 6 cmH2O (95% confidence interval 23-27; P<0.001). The success rate of the first attempt insertion was higher for the LMA Supreme than for the LMA ProSeal (98 and 88%, respectively; P=0.04). There was no difference in the median time taken for insertion with the LMA Supreme versus the LMA ProSeal: 26 s (interquartile range 23-45) versus 30 s (interquartile range 20-38), respectively (P=0.16). The ease of insertion, postoperative pharyngolaryngeal adverse events, patient satisfaction scores and anaesthesiologist's satisfaction scores were comparable in both groups. There were no complications of aspiration or nerve injuries.Conclusion: The LMA Supreme has lower oropharyngeal leak pressures than the LMA ProSeal. The success of the first attempt insertion was higher for the LMA Supreme. The LMA Supreme is a safe, efficacious and easy-to-use disposable supraglottic airway device in elective ambulatory procedures. The higher rate of success on first attempt insertion may make it more suitable as an airway rescue device. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To identify key target genes and activated signaling pathways associated with the pathogenesis of Sjögren's syndrome (SS) by conducting a systems analysis of parotid glands manifesting primary SS or primary SS/mucosa-associated lymphoid tissue (MALT) lymphoma phenotypes. METHODS: A systems biology approach was used to analyze parotid gland tissue samples obtained from patients with primary SS, patients with primary SS/MALT lymphoma, and subjects without primary SS (non-primary SS controls). The tissue samples were assessed concurrently by gene-expression microarray profiling and proteomics analysis, followed by weighted gene-coexpression network analysis. RESULTS: Gene-coexpression modules related to primary SS and primary SS/MALT lymphoma were significantly enriched with genes known to be involved in the immune/defense response, apoptosis, cell signaling, gene regulation, and oxidative stress. Detailed functional pathway analyses indicated that primary SS-associated modules were enriched with genes involved in proteasome degradation, apoptosis, signal peptides of the class I major histocompatibility complex (MHC), complement activation, cell growth and death, and integrin-mediated cell adhesion, while primary SS/MALT lymphoma-associated modules were enriched with genes involved in translation, ribosome biogenesis and assembly, proteasome degradation, class I MHC signal peptides, the G13 signaling pathway, complement activation, and integrin-mediated cell adhesion. Combined analyses of gene expression and proteomics data implicated 6 highly connected 'hub' genes for distinguishing primary SS from non-primary SS, and 8 hub genes for distinguishing primary SS/MALT lymphoma from primary SS. CONCLUSION: Systems biology analyses of the parotid glands from patients with primary SS and those with primary SS/MALT lymphoma revealed pathways and molecular targets associated with disease pathogenesis. The identified gene modules/pathways provide further insights into the molecular mechanisms of primary SS and primary SS/MALT lymphoma. The identified disease-hub genes represent promising targets for therapeutic intervention, diagnosis, and prognosis. [ABSTRACT FROM AUTHOR]

Objective: For clinical applications of biomarkers, there is a need for multiplex assays using high throughput platforms. The objective of this study was to determine the efficacy of Luminex Multianalyte Profiling (xMAP) technology for measurement of salivary proteins and to evaluate whether multiplex assays are as effective as single-plex assays and enzyme-linked immunosorbent assay (ELISA). Results: The average levels of interleukin-8 (IL-8) from the single-plex assay were 3313.2 ± 3759.8 pg ml−1 [oral squamous cell carcinoma (OSCC), n = 20] and 1061.7 ± 1978.8 pg ml−1 (control, n = 20). The IL-1β average levels from the single-plex assay were 945.2 ± 1134.8 pg ml−1 (OSCC, n = 20) and 314.2 ± 444.8 pg ml−1 (control, n = 20). The average levels of IL-8 from the multiplex assay were 2834.9 ± 3385.6 pg ml−1 (OSCC, n = 20) and 947.3 ± 2036.8 pg ml−1 (control, n = 20). The IL-1β average levels from the multiplex assay were 1013.5 ± 1221.1 pg ml−1 (OSCC, n = 20) and 376.3 ± 576.3 pg ml−1 (control, n = 20). The correlation coefficient between Luminex and ELISA assay for IL-8 ( n = 19) and IL-1β ( n = 19) was 0.91 and 0.84, respectively. Conclusion: Luminex xMAP single-plex and multiplex assays are as effective as ELISA assays for quantification of proteins in saliva. Both IL-8 and IL-1β were expressed at significantly higher levels in OSCC subjects than in the matched healthy control subjects. [ABSTRACT FROM AUTHOR]

Purpose: To summarize the published literature related to two, new supraglottic airways (SGAs): the Cobra Perilaryngeal Airway (CobraPLA) and the Streamlined Liner of Pharyngeal Airway (SLIPA).Source: We identified original articles, through searches on Medline and PubMed, using the keywords 'Cobra', 'CobraPLA', 'Cobra-PLA', 'PLA', and 'SLIPA'.Principal Findings: Searches identified 28 publications evaluating the CobraPLA, six of which were randomized, controlled trials. Five publications evaluated the SLIPA, two of which were randomized, controlled trials. Studies compared the CobraPLA with a number of SGAs, in regards to both efficacy and safety. While the CobraPLA is comparable to the laryngeal mask airway (LMA) Classic, with respect to insertion times and the incidence and severity of sore throat, the CobraPLA is superior, with respect to airway sealing pressure, and has been used successfully in patients with limited mouth opening and limited head extension. Insertion times, first insertion success rates, recovery times, and hemodynamic responses, associated with insertion of the SLIPA, are similar to those of the LMA Proseal. Although the SLIPA has a unique reservoir chamber to contain regurgitated fluid, the extent of its protection against pulmonary aspiration has not been established in the clinical setting.Conclusion: Both the CobraPLA and the SLIPA are intended for use during short procedures under general anesthesia, and have comparable efficacy and complication rates in comparison to the LMA Classic. Both airways may be considered as primary SGA devices, and the CobraPLA may be also be used as a rescue airway device. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To identify a panel of protein and messenger RNA (mRNA) biomarkers in human whole saliva (WS) that may be used in the detection of primary Sjögren's syndrome (SS). METHODS: Mass spectrometry and expression microarray profiling were used to identify candidate protein and mRNA biomarkers of primary SS in WS samples. Validation of the discovered mRNA and protein biomarkers was also demonstrated using real-time quantitative polymerase chain reaction and immunoblotting techniques. RESULTS: Sixteen WS proteins were found to be down-regulated and 25 WS proteins were found to be up-regulated in primary SS patients compared with matched healthy control subjects. These proteins reflected the damage of glandular cells and inflammation of the oral cavity system in patients with primary SS. In addition, 16 WS peptides (10 up-regulated and 6 down-regulated in primary SS) were found at significantly different levels (P < 0.05) in primary SS patients and controls. Using stringent criteria (3-fold change; P < 0.0005), 27 mRNA in saliva samples were found to be significantly up-regulated in the primary SS patients. Strikingly, 19 of 27 genes that were found to be overexpressed were interferon-inducible or were related to lymphocyte filtration and antigen presentation known to be involved in the pathogenesis of primary SS. CONCLUSION: Our preliminary study has indicated that WS from patients with primary SS contains molecular signatures that reflect damaged glandular cells and an activated immune response in this autoimmune disease. These candidate proteomic and genomic biomarkers may improve the clinical detection of primary SS once they have been further validated. We also found that WS contains more informative proteins, peptides, and mRNA, as compared with gland-specific saliva, that can be used in generating candidate biomarkers for the detection of primary SS. [ABSTRACT FROM AUTHOR]

BACKGROUND: Adenosine stress cardiovascular magnetic resonance (CMR) has been proven an effective tool in detection of reversible ischemia. Limited evidence is available regarding its accuracy in the setting of acute coronary syndromes, particularly in evaluating the significance of non-culprit vessel ischaemia. Adenosine stress CMR and recent advances in semi-quantitative image analysis may prove effective in this area. We sought to determine the diagnostic accuracy of semi-quantitative versus visual assessment of adenosine stress CMR in detecting ischemia in non-culprit territory vessels early after primary percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI). METHODS: Patients were prospectively enrolled in a CMR imaging protocol with rest and adenosine stress perfusion, viability and cardiac functional assessment 3 days after successful primary-PCI for STEMI. Three short axis slices each divided into 6 segments on first pass adenosine perfusion were visually and semi-quantitatively analysed. Diagnostic accuracy of both methods was compared with non-culprit territory vessels utilising quantitative coronary angiography (QCA) with significant stenosis defined as ?70%. RESULTS: Fifty patients (age 59 ± 12 years) admitted with STEMI were evaluated. All subjects tolerated the adenosine stress CMR imaging protocol with no significant complications. The cohort consisted of 41% anterior and 59% non anterior infarctions. There were a total of 100 non-culprit territory vessels, identified on QCA. The diagnostic accuracy of semi-quantitative analysis was 96% with sensitivity of 99%, specificity of 67%, positive predictive value (PPV) of 97% and negative predictive value (NPV) of 86%. Visual analysis had a diagnostic accuracy of 93% with sensitivity of 96%, specificity of 50%, PPV of 97% and NPV of 43%. CONCLUSION: Adenosine stress CMR allows accurate detection of non-culprit territory stenosis in patients successfully treated with primary-PCI post STEMI. Semi-quantitative analysis may be required for improved accuracy. Larger studies are however required to demonstrate that early detection of non-culprit vessel ischemia in the post STEMI setting provides a meaningful test to guide clinical decision making and ultimately improved patient outcomes. [ABSTRACT FROM AUTHOR]