A study of the TP53 Germline Mutation and Clinicopathologic Features in Thai Children with Adrenocortical Carcinoma

Objective: To determine the clinicopathologic features and germline tumor protein p53 (TP53) mutation in children with adrenocortical carcinoma (ACC). Material and Methods: This was a retrospective study. From 2009 to 2018, children with ACC from King Chulalongkorn Memorial Hospital and Phramongkutklao Hospital were enrolled into the study. Clinical presentations and hormonal profiles were recorded. Mutation analyses of the TP53 gene were acquired using the next-generation-sequencing method which was performed for germline samples of all patients and the parents of those who tested positive. Result: Two males and six females with ACC were enrolled into this study. The median age at diagnosis was 25.5 months (range 10 to 67 months). All participants had virilization, either virilization only (n=3) or associated with Cushing (n=5). All participants had had surgery to completely remove all of the tumor and three participants had received adjuvant chemotherapy consisting of etoposide, doxorubicin, and cyclophosphamide. All participants had three different known mutations in the TP53 gene: c.1010G>A (p.R337H), c.916C>T (p.R306*) and c.743G>A (p.R248Q). Two of the three participants with TP53 mutations had pulmonary metastasis. One participant had wild-type TP53 and pulmonary recurrence occurred one year after diagnosis. The median follow-up time was 31 months (range 10 to 168 months. As of this writing, seven participants survived without evidence of recurrence. No second malignancy was found in all participants. One participant who had c.916C>T died of pulmonary metastasis 10 months after diagnosis. Conclusion: We successfully identified three different germline TP53 mutations in patients with ACC. Progression to pulmonary metastasis and recurrence were higher among participants with TP53 mutations. The treatment outcome of childhood ACC was good when surgery and adjuvant chemotherapy were used

Germline <i>HAVCR2</i> mutations and their relation to the clinical spectrum of subcutaneous panniculitis-like T-cell lymphoma and hemophagocytic lymphohistiocytosis: results from a multicenter study and meta-analysis

Germline HAVCR2 mutations are frequently detected in subcutaneous panniculitis-like T-cell lymphoma (SPTCL) patients with/without hemophagocytic lymphohistiocytosis (HLH) but factors associated with variable manifestations remain undetermined. To evaluate clinical variations and associated factors in SPTCL and/or HLH with/without HAVCR2 mutations, we performed direct sequencing of HAVCR2 exon 2 using DNA from patients with SPTCL or idiopathic HLH/HLH-like systemic illnesses, defined by HLH alone without secondary causes. The systematic review and individual patient data (IPD) level meta-analysis which included the present and previously published studies reporting HAVCR2 mutations in SPTCL with/without HLH populations was subsequently conducted using random-effects meta-analysis and multivariate logistic regression. Among 34 patients enrolled, ten of 28 SPTCL patients developed HLH/HLH-like systemic illnesses. Six cases with HAVCR2Y82C mutation manifested with HLH without panniculitis. Male sex (P=0.03) and age <18 years (P=0.04) were associated with HLH, corresponding to the inverse correlation between age and HLH-2004 score (r=-0.40; P=0.02). Homozygous HAVCR2Y82C mutation was more common in the presence of HLH compared with the absence (75.0% vs. 44.4%; P=0.02). Using IPD from the present and the other three eligible cohorts (N=127), male sex, heterozygous and homozygous/compound heterozygous HAVCR2 mutations were associated with HLH by the adjusted odds ratio of 2.93 (95% confidence interval [CI]: 1.22-7.06), 4.77 (95% CI: 1.05-21.63) and 8.48 (95% CI: 2.98-24.10), respectively. Patients with male sex and/or germline HAVCR2 mutations showed an increased risk of developing HLH. Younger patients tended to manifest with HLH, while older patients typically presented with SPTCL with less frequent HLH/HLH-like systemic illnesses

Severe Dengue Is Associated with Consumption of von Willebrand Factor and Its Cleaving Enzyme ADAMTS-13

Severe dengue infections are characterized by thrombocytopenia, clinical bleeding and plasma leakage. Activation of the endothelium, the inner lining of blood vessels, leads to the secretion of storage granules called Weibel Palade bodies (WPBs). We demonstrated that severe dengue in Indonesian children is associated with a strong increase in plasma levels of the WPB constituents von Willebrand factor (VWF), VWF propeptide and osteoprotegerin (OPG). An increased amount of the hemostatic protein VWF was in a hyperreactive, platelet binding conformation, and this was most pronounced in the children who died. VWF levels at enrollment were lower than expected from concurrent VWF propeptide and OPG levels and VWF levels did not correlate well with markers of disease severity. Together, this suggests that VWF is being consumed during severe dengue. Circulating levels of the VWF-cleaving enzyme ADAMTS-13 were reduced. VWF is a multimeric protein and a subset of children had a decrease in large and intermediate VWF multimers at discharge. In conclusion, severe dengue is associated with exocytosis of WPBs with consumption of VWF and low ADAMTS-13 activity levels. This may contribute to the thrombocytopenia and complications of dengue

Severe Plasmodium falciparum Malaria Is Associated with Circulating Ultra-Large von Willebrand Multimers and ADAMTS13 Inhibition

Plasmodium falciparum infection results in adhesion of infected erythrocytes to blood vessel endothelium, and acute endothelial cell activation, together with sequestration of platelets and leucocytes. We have previously shown that patients with severe infection or fulminant cerebral malaria have significantly increased circulatory levels of the adhesive glycoprotein von Willebrand factor (VWF) and its propeptide, both of which are indices of endothelial cell activation. In this prospective study of patients from Ghana with severe (n = 20) and cerebral (n = 13) P. falciparum malaria, we demonstrate that increased plasma VWF antigen (VWF∶Ag) level is associated with disproportionately increased VWF function. VWF collagen binding (VWF∶CB) was significantly increased in patients with cerebral malaria and severe malaria (medians 7.6 and 7.0 IU/ml versus 1.9 IU/ml; p<0.005). This increased VWF∶CB correlated with the presence of abnormal ultra-large VWF multimers in patient rather than control plasmas. Concomitant with the increase in VWF∶Ag and VWF∶CB was a significant persistent reduction in the activity of the VWF-specific cleaving protease ADAMTS13 (∼55% of normal; p<0.005). Mixing studies were performed using P. falciparum patient plasma and normal pooled plasma, in the presence or absence of exogenous recombinant ADAMTS13. These studies demonstrated that in malarial plasma, ADAMTS13 function was persistently inhibited in a time-dependent manner. Furthermore, this inhibitory effect was not associated with the presence of known inhibitors of ADAMTS13 enzymatic function (interleukin-6, free haemoglobin, factor VIII or thrombospondin-1). These novel findings suggest that severe P. falciparum infection is associated with acute endothelial cell activation, abnormal circulating ULVWF multimers, and a significant reduction in plasma ADAMTS13 function which is mediated at least in part by an unidentified inhibitor

A New Cryogenic Apparatus to Search for the Neutron Electric Dipole Moment

A cryogenic apparatus is described that enables a new experiment, nEDM@SNS, with a major improvement in sensitivity compared to the existing limit in the search for a neutron Electric Dipole Moment (EDM). It uses superfluid $^4$He to produce a high density of Ultra-Cold Neutrons (UCN) which are contained in a suitably coated pair of measurement cells. The experiment, to be operated at the Spallation Neutron Source at Oak Ridge National Laboratory, uses polarized $^3$He from an Atomic Beam Source injected into the superfluid $^4$He and transported to the measurement cells as a co-magnetometer. The superfluid $^4$He is also used as an insulating medium allowing significantly higher electric fields, compared to previous experiments, to be maintained across the measurement cells. These features provide an ultimate statistical uncertainty for the EDM of $2-3\times 10^{-28}$ e-cm, with anticipated systematic uncertainties below this level

Imbalance of Angiopoietin-1 and Angiopoetin-2 in Severe Dengue and Relationship with Thrombocytopenia, Endothelial Activation, and Vascular Stability

Contains fulltext : 108223.pdf (publisher's version ) (Open Access)Abstract. The pathogenesis of plasma leakage during dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) is largely unknown. Angiopoietins are key regulators of vascular integrity: Angiopoietin-1 is stored in platelets and maintains vascular integrity, and endothelium-derived angiopoietin-2 promotes vascular leakage. We determined angiopoietin-1 and angiopoietin-2 levels in a cohort of children in Indonesia with DHF/DSS and related them to plasma leakage markers. Patients with DHF/DSS had reduced angiopoietin-1 and increased angiopoietin-2 plasma levels on the day of admission when compared with levels at discharge and in healthy controls. There was an inverse correlation between angiopoietin-1 and markers of plasma leakage and a positive correlation between angiopoietin-2 and markers of plasma leakage. Angiopoietin-1 levels followed the same trend as the soluble platelet activation marker P-selectin and correlated with platelet counts. Dengue-associated thrombocytopenia and endothelial activation are associated with an imbalance in angiopoietin-2: angiopoietin-1 plasma levels. This imbalance may contribute to the transient plasma leakage in DHF/DSS

Activation of Endothelial Cells, Coagulation and Fibrinolysis in Thai Children with Dengue Virus Infection.

Abstract Dengue virus is an arthropod borne flavivirus that most commonly causes a non-specific febrile illness: Dengue Fever (DF), and less frequently causes a life-threatening illness: Dengue Hemorrhagic Fever (DHF). The fatality rates of DHF are varying from 1 to 5% in tropical countries. Although severe bleeding remains the major cause of death, the pathogenesis of bleeding is poorly understood. This study was primarily designed to determine the extent of activation of endothelial cells, coagulation cascade and fibrinolysis in correlation with clinical severity and also to detect the best prognostic factor(s) for DHF during the febrile phase. We conducted a prospective cohort study in 42 children with Dengue infections (20 with DF and 22 with DHF); 38 age-matched normal Thai children served as the control group. Endothelial cell activation, coagulation and fibrinolysis parameters were measured in each patient during 3 phases of illness: febrile (Days 1–4), toxic (Days 5–6) and convalescent (Day 7–10). Bleeding scores (Buchanan G, et al, J Pediatr 2002) were also classified. Results: In DHF patients, during the febrile phase, plasma levels of von Willebrand Factor Antigen (VWF:Ag), tissue factor (TF), Thrombin-Antithrombin Complex (TAT), and D-dimer were significantly high, while platelet counts, fibrinogen, Protein C activity (PC:Ac) and thrombin activatable fibrinolysis inhibitor (TAFIa) were significantly low compared to the control group. During the toxic phase, activated Factor VII (FVIIa:c), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) were significantly increased compared to the control group. In DF patients, during the febrile phase, only plasma levels of VWF:Ag were significantly elevated, while platelet count, PC:Ac and TAFIa were significantly lowered compared to the control group. During the toxic phase, soluble thrombomodulin (sTM), FVIIa:c, and TAT were significantly raised compared to the control group. Compared to DF patients, DHF patients had significantly higher plasma concentrations of vWF:Ag (p=0.02), TF (p=0.01), and PAI-1 (p=0.038) during the febrile phase. Bleeding scores were positively correlated with plasma t-PA and clinical severity, and negatively correlated with plasma Factor VIII:C and TAFIa. Using logistic regression analysis, we found that an increase in plasma VWF:Ag was the only significant predictor (p=0.049) of progression to DHF. The endothelial cell injury and release of procoagulant components, activation of the coagulation cascade with thrombin generation, increased antifibrinolytic factors and consumption of natural anticoagulants each play important roles in the development of hemorrhage in Dengue patients. Plasma VWF:Ag is the best indicator of progression to DHF.</jats:p

Age-Related Reference Values of Molecular Markers in Endothelial Cell Activation, Coagulation and Fibrinolysis in Thai Children Versus Adults.

Abstract Introduction: The hemostatic system is a developing and changing process relative to age. Although advances in the knowledge of hemostatic mechanisms have led to the development of new methods for measuring peptides or enzyme-inhibitor complexes, there are very limited data concerning normal reference values for these in children. Objectives: The aim of this study is to distinguish the difference(s) in various endothelial cell activation and hemostatic parameters between children and adults, and to establish the normal range of these parameters in normal children in different age groups. Materials and Methods: Blood was obtained from 96 normal Thai children and adults whose screening coagulation tests were normal. All children were categorized into 3 age groups: 1–5 yrs, 6–10 yrs, and 11–18 yrs. Endothelial cell activation parameters: von Willebrand Factor Antigen and Activity (VWF:Ag and VWF:Ac) and soluble thrombomodulin (sTM); coagulation parameters: activated Factor VII (FVIIa:C), tissue factor (TF), and Thrombin-Antithrombin Complex (TAT); and fibrinolytic parameters: tissue plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1), D-dimer, thrombin activatable fibrinolysis inhibitor (TAFIa) and Protein C activity (PC:Ac) were measured. Tests Children’s Age Range Adults (Mean ± SD) 1–5 yr (n=19) 6–10 yr (n=26) 11–18 yr (n=25) (n=25) ANOVA *p ≤ 0.05 compared with adults; **p ≤ 0.001 compared with adults VWF:Ag (%) 93.3 ± 25.7 110.3 ± 27.1 98.4 ± 31.0 100.5 ± 25.2 VWF:Ac (%) 72.1 ± 18.9 91.1 ± 20.6 82.0 ± 24.2 84.7 ± 21.8 sTM (ng/mL) 3.9 ± 1.6** 3.1 ± 0.9 3.1 ± 1.3 2.4 ± 1.1 Fibrinogen (mg/dL) 360.7 ± 56.9 384.4 ± 78.9 410.1 ± 109.4 360.7 ± 51.0 FVIIa:C (%) 68.0 ± 23.8 69.9 ± 23.1 75.0 ± 22.9 75.6 ± 19.7 TF (pg/mL) 218.8 ± 57.8** 152.7 ± 38.2 144.5 ± 46.7 126.6 ± 42.2 TAT (ug/L) 4.5 ± 3.3 2.2 ± 0.7 2.4 ± 1.2 3.7 ± 4.9 t-PA (ng/mL) 1.32 ± 0.56 1.19 ± 0.48 1.16 ± 0.28 1.27 ± 0.32 PAI-1 (ng/mL) 22.8 ± 13.2 29.4 ± 14.2* 27.3 ± 18.3 18.8 ± 11.9 D-dimer (mg/L) 0.52 ± 0.42* 0.44 ± 0.46 0.28 ± 0.33 0.21 ± 0.09 TAFIa μg/mL) ( 40.6 ± 9.7 44.4 ± 6.9 46.1 ± 6.1 42.2 ± 5.7 Protein C:Ac (%) 87.2 ± 18.3* 101.7 ± 15.6 101.1 ± 22.5 102.4 ± 17.4 As seen in the Table, children in all age groups showed no significant difference in mean levels of VWF:Ag and Ac, fibrinogen, FVIIa:C, TAT, t-PA and TAFIa compared to adults. However, compared to adults, children aged 1–5 had significantly higher mean values of sTM (p=0.001), TF (p=&amp;lt; 0.001), and D-dimer (p = 0.015) whereas they had significantly lower mean levels of PC:Ac (p=0.023). The mean levels of PAI-1 in children of all groups were high, especially in children in the 6–10 yr age group (p = 0.032). These data indicate a physiologic difference in endothelial cell activation and hemostatic system between children and adults. Our data will serve as a useful reference guide in interpreting test results from children with suspected bleeding disorders.</jats:p