ProteinSeq: High-Performance Proteomic Analyses by Proximity Ligation and Next Generation Sequencing

Despite intense interest, methods that provide enhanced sensitivity and specificity in parallel measurements of candidate protein biomarkers in numerous samples have been lacking. We present herein a multiplex proximity ligation assay with readout via realtime PCR or DNA sequencing (ProteinSeq). We demonstrate improved sensitivity over conventional sandwich assays for simultaneous analysis of sets of 35 proteins in 5 µl of blood plasma. Importantly, we observe a minimal tendency to increased background with multiplexing, compared to a sandwich assay, suggesting that higher levels of multiplexing are possible. We used ProteinSeq to analyze proteins in plasma samples from cardiovascular disease (CVD) patient cohorts and matched controls. Three proteins, namely P-selectin, Cystatin-B and Kallikrein-6, were identified as putative diagnostic biomarkers for CVD. The latter two have not been previously reported in the literature and their potential roles must be validated in larger patient cohorts. We conclude that ProteinSeq is promising for screening large numbers of proteins and samples while the technology can provide a much-needed platform for validation of diagnostic markers in biobank samples and in clinical use

Obsessive-compulsive disorder in children and adolescents: Parental understanding, accommodation, coping and distress

Parental accommodation of pediatric OCD is common and is associated with negative affect in parents. Qualitative accounts of caring for a child with OCD are limited and no studies have assessed differences between mothers and fathers in accommodation, coping and distress. The current study used a mixed methods approach to understand parental accommodation, negative affect and coping. Forty-one mothers and 29 fathers of 43 children with OCD were asked to write narratives about their understanding and management of OCD and to complete measures of accommodation, coping, and distress. Symptom accommodation was high with almost half of the parents watching the child complete rituals or waiting for the child on a daily basis. Analysis of parental narratives indicated a distressing struggle between engaging in and resisting accommodation in order to manage their own and their child’s anger and distress. T-tests and correlation analysis indicated that accommodation did not differ significantly between mothers and fathers but was more strongly associated with negative affect in mothers. Analyses indicated that mothers reported using all types of coping strategy more often than fathers, particularly escape-avoidance, taking responsibility and using social support. Escape-avoidance coping was positively correlated with accommodation and negative affect in both mothers and fathers. Interventions that target parental constructions of OCD and their behavioural and emotional responses to it may assist in reducing the occurrence of accommodation, avoidant coping and parental distress