Guidance on Noncorticosteroid Systemic Immunomodulatory Therapy in Noninfectious Uveitis : Fundamentals Of Care for UveitiS (FOCUS) Initiative

Supplemental material available at www.aaojournal.org. Supported by AbbVie, Inc., and the Fundamentals of Care for Uveitis Initiative National Faculty. This manuscript was developed subsequent to an AbbVie-sponsored literature review of noninfectious, nonanterior uveitis. The meeting was conducted to understand the available literature regarding the management of patients with noninfectious, nonanterior uveitis. The program involved a total of 139 experts from 28 countries, who were selected for participation by AbbVie. However, AbbVie was not involved in the development of the manuscript. The authors maintained complete control over the content and this manuscript reflects the opinions of the authors. AbbVie selected the discussion participants and reviewed the final manuscript draft for scientific accuracy, but the authors determined the final content. All authors made substantial contributions to the article or critically revised it for important intellectual content and approved the final manuscript. AbbVie provided funding to invited participants, including honoraria for their attendance at the meetings. Travel to and from the meetings was reimbursed. No payments were made to the authors for the development of this manuscript. Dhinakaran Sambandan, PhD, and Shula Sarner, PhD, of Lucid Partners, Burleighfield House, Buckinghamshire, United Kingdom, provided medical writing and editorial support to the authors in the development of this manuscript; financial support for these services was provided by AbbVie. AbbVie reviewed the manuscript, but was not involved in the methodology, data collection and analysis, or completion of this manuscript.Peer reviewedPublisher PD

Guidance on noncorticosteroid systemic immunomodulatory therapy in noninfectious uveitis: fundamentals of care for uveitis (focus) initiative

Topic: An international, expert-led consensus initiative to develop systematic, evidence-based recommendations for the treatment of noninfectious uveitis in the era of biologics. Clinical Relevance: The availability of biologic agents for the treatment of human eye disease has altered practice patterns for the management of noninfectious uveitis. Current guidelines are insufficient to assure optimal use of noncorticosteroid systemic immunomodulatory agents. Methods: An international expert steering committee comprising 9 uveitis specialists (including both ophthalmologists and rheumatologists) identified clinical questions and, together with 6 bibliographic fellows trained in uveitis, conducted a Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol systematic reviewof the literature (English language studies from January 1996 through June 2016; Medline [OVID], the Central Cochrane library, EMBASE,CINAHL,SCOPUS,BIOSIS, andWeb of Science). Publications included randomized controlled trials, prospective and retrospective studies with sufficient follow-up, case series with 15 cases or more, peer-reviewed articles, and hand-searched conference abstracts from key conferences. The proposed statements were circulated among 130 international uveitis experts for review.Atotal of 44 globally representativegroupmembersmet in late 2016 to refine these guidelines using a modified Delphi technique and assigned Oxford levels of evidence. Results: In total, 10 questions were addressed resulting in 21 evidence-based guidance statements covering the following topics: when to start noncorticosteroid immunomodulatory therapy, including both biologic and nonbiologic agents; what data to collect before treatment; when to modify or withdraw treatment; how to select agents based on individual efficacy and safety profiles; and evidence in specific uveitic conditions. Shared decision-making, communication among providers and safety monitoring also were addressed as part of the recommendations. Pharmacoeconomic considerations were not addressed. Conclusions: Consensus guidelines were developed based on published literature, expert opinion, and practical experience to bridge the gap between clinical needs and medical evidence to support the treatment of patients with noninfectious uveitis with noncorticosteroid immunomodulatory agents

A Fragment of the LG3 Peptide of Endorepellin Is Present in the Urine of Physically Active Mining Workers: A Potential Marker of Physical Activity

Biomarker analysis has been implemented in sports research in an attempt to monitor the effects of exertion and fatigue in athletes. This study proposed that while such biomarkers may be useful for monitoring injury risk in workers, proteomic approaches might also be utilised to identify novel exertion or injury markers. We found that urinary urea and cortisol levels were significantly elevated in mining workers following a 12 hour overnight shift. These levels failed to return to baseline over 24 h in the more active maintenance crew compared to truck drivers (operators) suggesting a lack of recovery between shifts. Use of a SELDI-TOF MS approach to detect novel exertion or injury markers revealed a spectral feature which was associated with workers in both work categories who were engaged in higher levels of physical activity. This feature was identified as the LG3 peptide, a C-terminal fragment of the anti-angiogenic/anti-tumourigenic protein endorepellin. This finding suggests that urinary LG3 peptide may be a biomarker of physical activity. It is also possible that the activity mediated release of LG3/endorepellin into the circulation may represent a biological mechanism for the known inverse association between physical activity and cancer risk/survival

Guidance on Noncorticosteroid Systemic Immunomodulatory Therapy in Noninfectious Uveitis: Fundamentals Of Care for UveitiS (FOCUS) Initiative

Topic: An international, expert-led consensus initiative to develop systematic, evidence-based recommendations for the treatment of noninfectious uveitis in the era of biologics. Clinical Relevance: The availability of biologic agents for the treatment of human eye disease has altered practice patterns for the management of noninfectious uveitis. Current guidelines are insufficient to assure optimal use of noncorticosteroid systemic immunomodulatory agents. Methods: An international expert steering committee comprising 9 uveitis specialists (including both ophthalmologists and rheumatologists) identified clinical questions and, together with 6 bibliographic fellows trained in uveitis, conducted a Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol systematic review of the literature (English language studies from January 1996 through June 2016; Medline [OVID], the Central Cochrane library, EMBASE, CINAHL, SCOPUS, BIOSIS, and Web of Science). Publications included randomized controlled trials, prospective and retrospective studies with sufficient follow-up, case series with 15 cases or more, peer-reviewed articles, and hand-searched conference abstracts from key conferences. The proposed statements were circulated among 130 international uveitis experts for review. A total of 44 globally representative group members met in late 2016 to refine these guidelines using a modified Delphi technique and assigned Oxford levels of evidence. Results: In total, 10 questions were addressed resulting in 21 evidence-based guidance statements covering the following topics: when to start noncorticosteroid immunomodulatory therapy, including both biologic and nonbiologic agents; what data to collect before treatment; when to modify or withdraw treatment; how to select agents based on individual efficacy and safety profiles; and evidence in specific uveitic conditions. Shared decision-making, communication among providers and safety monitoring also were addressed as part of the recommendations. Pharmacoeconomic considerations were not addressed. Conclusions: Consensus guidelines were developed based on published literature, expert opinion, and practical experience to bridge the gap between clinical needs and medical evidence to support the treatment of patients with noninfectious uveitis with noncorticosteroid immunomodulatory agents

The highly abundant urinary metabolite urobilin interferes with the bicinchoninic acid assay

Estimation of total protein concentration is an essential step in any protein- or peptide-centric analysis\ud pipeline. This study demonstrates that urobilin, a breakdown product of heme and a major constituent of\ud urine, interferes considerably with the bicinchoninic acid (BCA) assay. This interference is probably due\ud to the propensity of urobilin to reduce cupric ions (Cu2+) to cuprous ions (Cu1+), thus mimicking the\ud reduction of copper by proteins, which the assay was designed to do. In addition, it is demonstrated that\ud the Bradford assay is more resistant to the influence of urobilin and other small molecules. As such, urobilin\ud has a strong confounding effect on the estimate of total protein concentrations obtained by BCA\ud assay and thus this assay should not be used for urinary protein quantification. It is recommended that\ud the Bradford assay be used instead

Urinary biomarker values for mine site employees.^α

α<p>Values are means<u>±</u>SE.</p>*<p>Indicates values are significantly greater than PRE value (P<0.01).</p>#<p>Indicates value significantly greater than PRE value (P<0.05).</p

LC- MS/MS identifies the LG3 peptide of endorepellin, a C-terminal bioactive fragment of Perlecan.

<p>a) Perlecan (<b><u>underlined bold lower case</u></b>), the C terminal of Perlecan containing Endorepellin (lowercase text) and the LG3 Peptide of endorepellin (<b>BOLD CAPITALS</b>). Individual peptides identified by LC-MS/MS of tryptic in-gel digest in </p><p><b>LIGHT GREY</b></p> and <p><b><u>DARK GREY</u></b></p> highlights. Sequence coverage includes the LG3 peptide, however, the first 25 residues of the LG3 peptide were not detected. <b>b</b>) Western blot analysis confirmed that the ∼20 kDa protein observed by SDS-PAGE and the spectral feature at m/z 16881 are derived from endorepellin. Western Blot of worker urine samples using goat anti-human endorepellin polyclonal antibody (1∶10,000).<p></p

The spectral feature at m/z 16881 is a broad tri-phasic peak, visible by SDS-PAGE.

<p><b>a</b>) The hypothesised pattern of intensity of m/z 16881 in stacked replicate spectra, expected to be observed in an SDS-PAGE gel. <b>b</b>) A band which matched the expected pattern of intensity for the feature at m/z 16881 was detected at ∼20 kDa by SDS-PAGE (<b>arrow)</b> suggesting that the bands at ∼20 kDa in the gel were the proteins which constituted m/z 16881 in the spectra. <b>c</b>) The protein at ∼20 kDa was extracted from excised bands from a non-stained replicate SDS-PAGE gel. Examination of the extracted protein by SELDI-TOF MS confirmed that the ∼20 kDa band was the feature originally detected at m/z 16881.</p

Urinary urea and cortisol levels trend toward recovery in operators but not in maintenance crew.

<p><b>a</b>) Urinary, urea levels were determined by an automated kinetic assay (analytic coefficient of variation being <4%). <b>b</b>) Urinary cortisol levels were determined by competitive immunoassay (analytic coefficient of variation being<4%). Both urea and cortisol measurements were standardised for dieresis against urinary creatinine levels which were determined by the Jaffe method (analytic coefficient of variation being<3%).</p

The ∼20 kDa band excised from the SDS-PAGE gel is a fragment of perlecan.

<p>Mascot search results.</p>α<p>Ions score is −10*log(P), where P is the probability that the observed match is a random event. Individual ion scores >52 indicate identity of extensive homology (p<0.05).</p