Activated protein C ß-glycoform promotes enhanced noncanonical PAR1 proteolysis and superior resistance to ischemic injury.

Activated protein C (APC) is an anticoagulant protease that initiates cell signaling via protease-activated receptor 1 (PAR1) to regulate vascular integrity and inflammatory response. In this study, a recombinant APC variant (APC(N329Q)) mimicking the naturally occurring APC-ß plasma glycoform was found to exhibit superior PAR1 proteolysis at a cleavage site that selectively mediates cytoprotective signaling. APC(N329Q) also enhanced integrin αMß2-dependent PAR1 proteolysis to exert significantly improved antiinflammatory activity on macrophages compared with wild-type APC. Recent therapeutic applications of recombinant APC in ischemic stroke models have used APC variants with limited anticoagulant activity to negate potential bleeding side effects. Using a mouse model of ischemic stroke and late t-PA intervention, the neuroprotective activity of a murine APC variant with limited anticoagulant activity (mAPC(PS)) was compared with an identical APC variant except for the absence of glycosylation at the APC-ß sequon (mAPC(PS/N329Q)). Remarkably, mAPC(PS/N329Q) limited cerebral ischemic injury and reduced brain lesion volume significantly more effectively than mAPC(PS). Collectively, this study reveals the importance of APC glycosylation in controlling the efficacy of PAR1 proteolysis by APC and demonstrates the potential of novel APC variants with superior cytoprotective signaling function as enhanced therapeutic agents for the treatment of ischemic stroke.

Serum Immune System Biomarkers Neopterin and Interleukin-10 Are Strongly Related to Tryptophan Metabolism in Healthy Young Adults.

BACKGROUND: Changes in tryptophan metabolism through the vitamin B-6-dependent kynurenine pathway have been linked to activation of the immune system. OBJECTIVE: We hypothesized that blood concentrations of tryptophan and its catabolites were associated with biomarkers relevant to inflammatory processes in healthy noninflamed subjects. METHODS: Healthy young adults (n = 737) aged 18-28 y without any known diseases or clinical evidence of inflammation provided blood samples for analysis of serum tryptophan/kynurenine metabolites, neopterin, C-reactive protein (CRP), and plasma pyridoxal 5'-phosphate (PLP) with LC-tandem mass spectrometry methodologies. A panel of cytokines was measured in serum by using high-sensitivity ELISA assays. Anthropometric and lifestyle data were collected by questionnaire. Multiple linear regression analysis to determine the effect of measured serum cytokine concentrations as predictors of tryptophan metabolites was performed on inverse normal-rank transformations of the data, adjusted for sex, body mass index, smoking, alcohol intake, and contraceptive use in women. RESULTS: Median serum CRP and neopterin concentrations were well below established clinical cutoffs for inflammation. We observed significant positive associations between serum interleukin-10 (IL-10) and serum kynurenine (P = 0.0002), the kynurenine-to-tryptophan ratio (KTR) (P = 0.003), 3-hydroxykynurenine (P = 0.01), and 3-hydroxyanthranilic acid (P = 0.04). Serum neopterin was positively associated with kynurenine, the KTR (both P < 0.0001), and anthranilic acid (P = 0.004), and was negatively associated with serum tryptophan (P = 0.01) and PLP (P < 0.0001). Serum tumor necrosis factor α was also negatively associated with tryptophan (P < 0.001). CONCLUSIONS: In healthy young adults with no apparent inflammatory conditions, serum tryptophan metabolites are significantly associated with key immune system biomarkers. The observed association between IL-10 and kynurenine is unexpected and suggests that kynurenine-linked mechanisms promoting negative regulation of inflammatory responses are associated with normal immune homeostasis.

The effect of percutaneous coronary intervention on habitual physical activity in older patients.

BACKGROUND: Given the ongoing burden of cardiovascular disease and an ageing population, physical activity in patients with coronary artery disease needs to be emphasized. This study assessed whether sedentary behaviour and physical activity levels differed among older patients (≥75 years) following percutaneous coronary intervention (PCI) for acute coronary syndrome (ACS) consisting of ST-segment elevation myocardial infarction (STEMI) and non STEMI (NSTEMI) versus an elective admission control group of stable angina patients. METHODS: Sedentary behaviour and physical activity were assessed over a 7-day period using wrist-worn triaxial accelerometers (GENEActiv, Activinsights Ltd, UK) in 58 patients following PCI for, STEMI (n = 20) NSTEMI (n = 18) and stable angina (n = 20) upon discharge from a tertiary centre. Mean ± Standard deviation age was 79 ± 4 years (31% female). RESULTS: STEMI and NSTEMI patients spent more time in the low acceleration category (0-40 mg) reflecting sedentary time versus stable angina patients (1298 ± 59 and 1305 ± 66 vs. 1240 ± 92 min/day, p < 0.05). STEMI and NSTEMI patients spent less time in the 40-80 mg acceleration category reflecting low physical activity versus stable angina patients (95 ± 35 and 94 ± 41 vs. 132 ± 50 min/day, p < 0.05). Stable angina patients spent more time in the higher acceleration categories (80-120 and 120-160 mg) and moderate-to-vigorous physical activity (defined as 1 and 5 min/day bouts) versus NSTEMI patients (p < 0.05). For acceleration categories ≥160 mg, no differences were observed. CONCLUSIONS: Patients presenting with ACS and undergoing PCI spent more time in sedentary behaviour compared with stable angina patients.

Engineering activated protein C to maximize therapeutic efficacy.

The anticoagulant-activated protein C (APC) acts not solely as a crucial regulator of thrombus formation following vascular injury, but also as a potent signalling enzyme with important functions in the control of both acute and chronic inflammatory disease. These properties have been exploited to therapeutic effect in diverse animal models of inflammatory disease, wherein recombinant APC administration has proven to effectively limit disease progression. Subsequent clinical trials led to the use of recombinant APC (Xigris) for the treatment of severe sepsis. Although originally deemed successful, Xigris was ultimately withdrawn due to lack of efficacy and an unacceptable bleeding risk. Despite this apparent failure, the problems that beset Xigris usage may be tractable using protein engineering approaches. In this review, we detail the protein engineering approaches that have been utilized to improve the therapeutic characteristics of recombinant APC, from early studies in which the distinct anti-coagulant and signalling activities of APC were separated to reduce bleeding risk, to current attempts to enhance APC cytoprotective signalling output for increased therapeutic efficacy at lower APC dosage. These novel engineered variants represent the next stage in the development of safer, more efficacious APC therapy in disease settings in which APC plays a protective role.

Teacher-Implemented Response Interruption and Redirection: Training, Evaluation, and Descriptive Analysis of Treatment Integrity.

Response interruption and redirection (RIRD) is an effective intervention for decreasing stereotypy. During RIRD, contingent on occurrences of stereotypy, therapists interrupt the behavior and prompt the participant to complete an alternative response. Although RIRD has been implemented by teachers in classrooms, it requires continuous monitoring of participants to be implemented with fidelity and may be difficult for teachers to manage. The present study evaluated the effectiveness of RIRD when implemented in classrooms. In addition, we evaluated if novice teaching assistants could be trained to implement RIRD. Finally, a descriptive analysis of treatment integrity errors during RIRD was conducted. Three children and teaching assistants participated. Following a written instructions baseline, the teaching assistants were trained to implement RIRD using modeling, rehearsal, and feedback. The training increased the accuracy of RIRD implementation for all participants. Incorrectly initiating and terminating RIRD were the most common treatment integrity errors observed.

A follow-up study of chronic fatigue syndrome in children and adolescents: symptom persistence and school absenteeism.

This is a follow-up study of 28 young people aged between 7 and 17 meeting the Oxford criteria for the diagnosis of chronic fatigue syndrome treated in a specialist paediatric/psychiatric service. Retrospective case note analysis revealed a wide range and duration of symptoms together with high levels of school absenteeism prior to the diagnosis. The mean follow-up interval after discharge from the specialist service was 3 years and although most of the young people regarded themselves as fully recovered by this time, improvement was variable and about one third were still experiencing disabling symptoms. The illness had impacted on the education or career plans of all the young people to some extent with 15 experiencing difficulty returning to school. This article highlights the need for early recognition and diagnosis of chronic fatigue syndrome in young people and the importance of continuing paediatric support to reduce symptom persistence in the sensitive recovery period. Maintaining school attendance by close liaison between health and education services both before and after diagnosis and treatment is also vital if long-term morbidity is to be reduced.

Physical activity in the management of patients with coronary artery disease: a review.

Cardiovascular diseases remain the leading cause of death in the Western world despite advances in therapeutics and interventions. The prescription of physical activity is a key component of cardiac rehabilitation following myocardial infarction. This review aims to outline the impact of physical activity in particular patient cohorts with coronary artery disease. The current understanding of the mechanisms involved in atherosclerosis, plaque rupture and thrombosis, and how these can be modified by physical activity, are also discussed. There is the potential for future research to investigate the clinical and mechanistic effects of different exercise types, intensities, duration, and frequencies in patients hospitalized for coronary artery disease.

IFNL cytokines do not modulate human or murine NK cell functions.

The interferon-lambda (IFNL) cytokines have been shown to be important in HCV infection with SNPs in the IFNL3 gene associated with both natural and treatment induced viral clearance. We have recently shown that rs1299860 (an IFNL3 associated SNP) and an NK cell gene, KIR2DS3, synergised to increase the odds of chronic infection in a homogenous cohort of Irish women infected with HCV. To characterise a biological basis for the genetic synergy, we investigated for any evidence that IFNL cytokines regulate NK cell functions. Using a range of functional responses, we did not find any evidence of NK cell activation by IFNL3, IFNL1 or IFNL2 cytokines. Similar results were found using human and murine NK cells. In addition, and in contrast to our preliminary study, we did not find any evidence that IFNL cytokines inhibited NK cell cytokine production; thus, the biological basis for the genetic synergy remains to be discovered.