Increased galactose expression and enhanced clearance in patients with low von Willebrand factor.

Glycan determinants on von Willebrand factor (VWF) play critical roles in regulating its susceptibility to proteolysis and clearance. Abnormal glycosylation has been shown to cause von Willebrand disease (VWD) in a number of different mouse models. However, because of the significant technical challenges associated with accurate assessment of VWF glycan composition, the importance of carbohydrates in human VWD pathogenesis remains largely unexplored. To address this, we developed a novel lectin-binding panel to enable human VWF glycan characterization. This methodology was then used to study glycan expression in a cohort of 110 patients with low VWF compared with O blood group-matched healthy controls. Interestingly, significant interindividual heterogeneity in VWF glycan expression was seen in the healthy control population. This variation included terminal sialylation and ABO(H) blood group expression on VWF. Importantly, we also observed evidence of aberrant glycosylation in a subgroup of patients with low VWF. In particular, terminal α(2-6)-linked sialylation was reduced in patients with low VWF, with a secondary increase in galactose (Gal) exposure. Furthermore, an inverse correlation between Gal exposure and estimated VWF half-life was observed in those patients with enhanced VWF clearance. Together, these findings support the hypothesis that loss of terminal sialylation contributes to the pathophysiology underpinning low VWF in at least a subgroup of patients by promoting enhanced clearance. In addition, alterations in VWF carbohydrate expression are likely to contribute to quantitative and qualitative variations in VWF levels in the normal population. This trial was registered at www.clinicaltrials.gov as #NCT03167320.

Novel insights into the clinical phenotype and pathophysiology underlying low VWF levels.

Critical clinical questions remain unanswered regarding diagnosis and management of patients with low von Willebrand factor (VWF) levels (30-50 IU/dL). To address these questions, the Low VWF Ireland Cohort (LoVIC) study investigated 126 patients registered with low VWF levels. Despite marginally reduced plasma VWF levels, International Society of Thrombosis and Haemostasis Bleeding Assessment Tool (ISTH BAT) confirmed significant bleeding phenotypes in the majority of LoVIC patients. Importantly, bleeding tendency did not correlate with plasma VWF levels within the 30 to 50 IU/dL range. Furthermore, bleeding phenotypes could not be explained by concurrent hemostatic defects. Plasma factor VIII to VWF antigen (VWF:Ag) ratios were significantly increased in LoVIC patients compared with controls (P < .0001). In contrast, VWF propeptide to VWF:Ag ratios >3 were observed in only 6% of the LoVIC cohort. Furthermore, platelet-VWF collagen binding activity levels were both significantly reduced compared with controls (P < .05). In response to 1-desamino-8-D-arginine vasopressin (DDAVP), peak VWF:Ag levels exceeded 100 IU/dL in 88% of patients and was sustained >100 IU/dL after 4 hours in 72% of subjects. In conclusion, our novel data suggest that low VWF levels can be associated with significant bleeding and are predominantly due to reductions in VWF synthesis and/or constitutive secretion. Although enhanced VWF clearance may contribute to the pathophysiology in some individuals, the absolute reduction in VWF plasma half-life is usually mild and not sufficient to significantly impact upon the duration of DDAVP-induced VWF response. This trial was registered at www.clinicaltrials.gov as #NCT03167320.

A novel role for von Willebrand factor in the pathogenesis of experimental cerebral malaria.

Plasmodium falciparum malaria infection is associated with an early marked increase in plasma von Willebrand factor (VWF) levels, together with a pathological accumulation of hyperreactive ultra-large VWF (UL-VWF) multimers. Given the established critical role of platelets in malaria pathogenesis, these increases in plasma VWF raise the intriguing possibility that VWF may play a direct role in modulating malaria pathogenesis. To address this hypothesis, we used an established murine model of experimental cerebral malaria (ECM), in which wild-type (WT) C57BL/6J mice were infected with Plasmodium berghei ANKA. In keeping with findings in children with P falciparum malaria, acute endothelial cell activation was an early and consistent feature in the murine model of cerebral malaria (CM), resulting in significantly increased plasma VWF levels. Despite the fact that murine plasma ADAMTS13 levels were not significantly reduced, pathological UL-VWF multimers were also observed in murine plasma following P berghei infection. To determine whether VWF plays a role in modulating the pathogenesis of CM in vivo, we further investigated P berghei infection in VWF(-/-) C57BL/6J mice. Clinical ECM progression was delayed, and overall survival was significantly prolonged in VWF(-/-) mice compared with WT controls. Despite this protection against ECM, no significant differences in platelet counts or blood parasitemia levels were observed between VWF(-/-) and WT mice. Interestingly, however, the degree of ECM-associated enhanced blood-brain barrier permeability was significantly attenuated in VWF(-/-) mice compared with WT controls. Given the significant morbidity and mortality associated with CM, these novel data may have direct translational significance.

N-linked glycans within the A2 domain of von Willebrand factor modulate macrophage-mediated clearance.

Enhanced von Willebrand factor (VWF) clearance is important in the etiology of von Willebrand disease. However, the molecular mechanisms underlying VWF clearance remain poorly understood. In this study, we investigated the role of VWF domains and specific glycan moieties in regulating in vivo clearance. Our findings demonstrate that the A1 domain of VWF contains a receptor-recognition site that plays a key role in regulating the interaction of VWF with macrophages. In A1-A2-A3 and full-length VWF, this macrophage-binding site is cryptic but becomes exposed following exposure to shear or ristocetin. Previous studies have demonstrated that the N-linked glycans within the A2 domain play an important role in modulating susceptibility to ADAMTS13 proteolysis. We further demonstrate that these glycans presented at N1515 and N1574 also play a critical role in protecting VWF against macrophage binding and clearance. Indeed, loss of the N-glycan at N1515 resulted in markedly enhanced VWF clearance that was significantly faster than that observed with any previously described VWF mutations. In addition, A1-A2-A3 fragments containing the N1515Q or N1574Q substitutions also demonstrated significantly enhanced clearance. Importantly, clodronate-induced macrophage depletion significantly attenuated the increased clearance observed with N1515Q and N1574Q in both full-length VWF and A1-A2-A3. Finally, we further demonstrate that loss of these N-linked glycans does not enhance clearance in VWF in the presence of a structurally constrained A2 domain. Collectively, these novel findings support the hypothesis that conformation of the VWF A domains plays a critical role in modulating macrophage-mediated clearance of VWF in vivo.

Physical Activity Surveillance in Adolescents with Type 1 Diabetes: A Pilot Mixed-Methods Investigation.

Type 1 diabetes (T1D) affects over 2,500 children in Ireland. Insulin replacement is the mainstay of treatment for T1D, and physical activity (PA) is an important, modifiable lifestyle factor for sustaining health. Surveillance of PA for both research and clinical purposes in paediatric T1D has been limited. This study deployed both quantitative (accelerometry) and qualitative (self-report) measures to assess habitual PA patterns in children with T1D. Twenty-one participants (9 females, 12 males) between 10 and 17 years (mean 13.7 ± 1.94 years) were recruited from an Outpatients Paediatric Diabetes Clinic. Total steps, standing time (minutes (mins)) and sitting time (mins) were recorded using the activPAL 3 microactivity monitor. Clinical parameters (HbA1c, insulin regimen, and weight centiles) were measured. A self-report diary was used to measure perceived activity levels. The findings of this study show that participant children with T1D are not achieving the required steps per day to sustain physical health (recommended minimum 11,500). Females (mean = 7,306 steps ± 5,468) achieved significantly less (p = 0.001) steps per day compared to males (10,806 steps ± 5,904). No significant differences were found between genders for sitting time or standing time. Overweight or obesity was identified in 44% of female participants and 15% of male participants. Mean HbA1c for both females 8.25% (67 mmol/mol) and males 7.97% (64 mmol/mol) was above the International Society for Pediatric and Adolescent Diabetes (ISPAD) recommended <7.0% (53 mmol/mol) for children. Further research is warranted to investigate PA promotion strategies in populations of children with paediatric T1D.

Parent and child perceptions of physical activity with type 1 diabetes.

INTRODUCTION: Type 1 diabetes (T1D) is a lifelong illness that affects over 2500 children in Ireland. Management involves complex daily regimens including frequent blood glucose monitoring, pharmacotherapy, dietary management, and physical activity (PA). PA is an important modifiable lifestyle factor. Unfortunately, children with T1D remain physically inactive. Children with T1D face disease-specific barriers and facilitators to PA engagement. All aspects of T1D management for children are supported or supervised by parents. Thus, the purpose of this study was to examine parents' and children's perceptions of barriers and facilitators to PA engagement. RESEARCH DESIGN AND METHODS: 43 parent and child dyads participated. Parents completed a self-report survey. Children completed a modified version of the Physical Activity Questionnaire for Children (PAQ-C) that explored habitual PA patterns, perceived facilitators and barriers to PA engagement. RESULTS: 21 females, 22 males and their parents (36 mothers, 7 fathers) participated. 69% of males and 90% of females reported that having diabetes did affect their PA participation. 54% of males and 48% of females were insufficiently active based on their total PAQ-C score (<2.9 and <2.7). 53% of parents reported that their children participated in school physical education. 21% of parents reported that their child did not participate in PA outside of the school setting. 23% of parents reported that they did not feel comfortable with their child participating in strenuous PA. A further 30% of parents reported that they only felt comfortable with their child participating in strenuous PA if supervised. 66% of parents reported their child should be more physically active. 83% of parents reported that having T1D did impact their child's PA level. CONCLUSIONS: This study demonstrates the potential influence of parents' perceptions on PA engagement in children with T1D. Additional education is needed to support the promotion of PA for children with T1D.

Thermoelectric properties of electrospun carbon nanofibres derived from lignin.

Developing sustainable and efficient thermoelectric materials is a challenge because the most common thermoelectric materials are based on rare elements such as bismuth and telluride. In this context, we have produced bio-based carbon nanofibres (CNFs) derived from mixtures of polyacrylonitrile and lignin using electrospinning. The addition of lignin (up to 70%) reduces the diameter of CNFs from 450 nm to 250 nm, increases sample flexibility, and promotes inter-fibre fusion. The crystalline structure of the CNFs was analysed by Raman spectroscopy. The electrical conductivity and the Seebeck coefficient were evaluated as function of the lignin content in the precursor and carbonised equivalents. Finally, a conversion of p-type to n-type semiconducting behaviour was achieved with a hydrazine vapour treatment. We observe a maximum p-type power factor of 9.27 µW cm-1 K-2 for CNFs carbonised at 900 °C with 70% lignin which is a 34.5-fold increase to the CNFs with 0% lignin. For the hydrazine treated samples, we observe a maximum n-type power factor of 10.2 µW cm-1 K-2 for the CNFs produced in the same way which is an 11.0-fold increase to the hydrazine-treated CNFs with 0% lignin.

Significant gynecological bleeding in women with low von Willebrand factor levels.

Gynecological bleeding is frequently reported in women with von Willebrand disease (VWD). Low von Willebrand factor (VWF) may be associated with significant bleeding phenotype despite only mild plasma VWF reductions. The contribution of gynecological bleeding to this phenotype has yet to be described. The optimal clinical bleeding assessment tool (BAT) to evaluate bleeding remains unclear. Using a standardized approach to phenotypic assessment, we evaluated gynecological bleeding and directly compared the Condensed Molecular and Clinical Markers for the Diagnosis and Management of type 1 VWD (Condensed MCMDM-1 VWD) and International Society on Thrombosis and Haemostasis (ISTH) BAT scores in 120 women enrolled in the Low von Willebrand in Ireland Cohort study. Heavy menstrual bleeding (HMB) was reported in 89% of female participants; 45.8% developed iron deficiency. Using identical data, Condensed MCMDM-1 VWD menorrhagia domain scores were significantly lower than ISTH BAT scores (2 vs 3; P < .0001), the discrepant results related to 40% of women not seeking medical consultation for HMB, reducing the sensitivity of the Condensed score. For those who reported HMB to physicians, the low VWF diagnosis was not expedited (age at diagnosis 34.2 vs 33.4 years in women failing to present; P = .7). Postpartum hemorrhage (PPH) was self-reported in 63.5% of parous women (n = 74); 21.6% required transfusion, critical care, radiological, or surgical intervention. Our data demonstrate that gynecological bleeding is frequently reported in women with low VWF; despite pregnancy-related increases in plasma VWF levels, these women may experience PPH. Defining the optimal management approach for these patients requires further research. This trial was registered at www.clinicaltrials.gov as #NCT03167320.