Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children

Bilateral near-infrared spectroscopy for detecting traumatic vascular injury

Extremity wounds account for most battlefield injuries. Clinical examination may be unreliable by medics or first responders, and continuous assessment by experienced practitioners may not be possible on the frontline or during transport. Near-infrared spectroscopy (NIRS) provides continuous, noninvasive monitoring of tissue oxygen saturation (StO2), but its use is limited by inter-patient and intra-patient variability. We tested the hypothesis that bilateral NIRS partially addresses the variability problem and can reliably identify vascular injury after extremity trauma. This prospective study consisted of 30 subjects: 20 trauma patients with extremity injury and 10 healthy volunteers. Bilateral StO2 tissue sensors were placed on the thenar eminence or medial plantar surface. Injured and non-injured extremities within the same patient (ΔStO2) were compared using Wilcoxon signed ranks test. Receiver operating characteristic curves were generated and areas under the curve (AUCs) were calculated for ΔStO2 of 6, 10, and 15. Values are expressed as median (interquartile range). Trauma patients were age 31 y (23 y), 85% male, with injury severity score of 9 (5). There were seven arterial and three venous injuries. Most involved the lower extremity (n = 16; 80%) and resulted from a penetrating mechanism (n = 14; 70%). ΔStO2 between limbs was 20.4 (10.4) versus 2.4 (3.0) (P < 0.001) for all patients with vascular injury versus patients and volunteers with no vascular injury. ΔStO2 reliably identified any vascular injury (AUC, 0.975; P < 0.001), whereas pulse examination alone or in combination with Doppler exam could detect only arterial injury. A ΔStO2 of 6 had the greatest sensitivity and specificity (AUC, 0.900; P < 0.001). Continuous monitoring of bilateral limbs with NIRS detects changes in perfusion resulting from arterial or venous injury and may offer advantages over serial manual measurements of pulses or Doppler signals. This technique may be most relevant in military and disaster scenarios or during transport, in which the ability to monitor limb perfusion is difficult or experienced clinical judgment is unavailable

Initial hematocrit in trauma: a paradigm shift?

After severe trauma and hemorrhage, it is generally assumed that the rate of fluid shift from the interstitial space into the vasculature is relatively slow and that initial hematocrit (Hct) does not reflect estimated blood loss. This study challenges that idea and tests the hypothesis that initial Hct correlates with signs of shock and hemorrhage in trauma patients. Data were retrospectively reviewed from 198 trauma patients requiring emergency surgery at a Level I center from July 2009 to April 2010. Patients were divided into quartiles based on the initial Hct measured within 10 minutes of arrival. Categorical data were compared using χ(2) test or Fisher's exact test, as appropriate. Normally distributed data were compared using Student's t test or analysis of variance. Nonparametric data were compared with a Mann-Whitney U test or Kruskal-Wallis test. Post hoc analysis was conducted using the Bonferroni correction or paired Mann-Whitney U tests. The study population was 83% male, aged 35 ± 1 years (mean ± SE), with 71% penetrating injuries. Lower initial Hct correlated with hypotension (p < 0.001), acidosis (p = 0.003), altered mental status (p < 0.001), Injury Severity Score (p < 0.001), Revised Trauma Score (p < 0.001), estimated blood loss (p < 0.001), and usage of packed red blood cells (p < 0.001), fresh frozen plasma (p = 0.003), crystalloid (p = 0.021), and vasopressors (p < 0.001). Admission Hct correlates with signs of shock and hemorrhage in trauma patients requiring emergency surgery because fluid shifts rapidly from the interstitial space into the vasculature. This finding of a rapid Hct change contradicts the current teaching in most trauma textbooks

Persistence of hypercoagulable state after resection of intra-abdominal malignancies

The hypercoagulable state associated with cancer imparts considerable risk for venous thromboembolism. Surgical resection of malignancies should theoretically reverse tumor-induced hypercoagulability. However, coagulation changes in cancer patients postresection have not been described thoroughly. Conventional coagulation tests are unable to detect hypercoagulable states. In contrast, rotational thromboelastography (ROTEM) can detect hypo- or hypercoagulable conditions. We hypothesized that the cancer-induced hypercoagulable state would improve after surgical resection. After informed consent, blood samples of patients undergoing surgical resection for curative intent were analyzed with serial ROTEM. Thirty-five patients (mean ± SD age 66 ± 17 years; 67% male) had cancers involving the pancreas (n = 12 [34%]), esophagus (n = 10 [29%]), stomach (n = 7 [20%]), bile ducts (n = 3 [9%]), and duodenum (n = 3 [9%]). Preoperative ROTEM identified 14 (40%) who were hypercoagulable. After surgical resection, patients became progressively hypercoagulable with more rapid clot formation time (low clot formation time, high alpha) and higher maximum clot firmness. By week one, 86% (n = 30) had abnormal ROTEM values, including 17 of 21 (81%) who had normal coagulation profiles preoperatively. Most (n = 30 [86%]) remained hypercoagulable at 3 to 4 weeks. Rotational thromboelastography identifies baseline hypercoagulability in more than one third of patients with intra-abdominal malignancies. This is among the first studies to demonstrate progressive hypercoagulability that persists for at least 1 month after resection. These data support postdischarge thromboprophylaxis regimens in high-risk cancer patients

Pre-existing hypercoagulability in patients undergoing potentially curative cancer resection

Rotational thromboelastometry (ROTEM) is a new point-of-care test that allows a rapid and comprehensive evaluation of coagulation. We were among the first to show that ROTEM identifies baseline hypercoagulability in 40% of patients with intra-abdominal malignancies and that hypercoagulability persists for ≥1 month after resection. The purpose of this follow-up study was to confirm and extend these observations to a larger population in outpatient preoperative clinics. The hypothesis is that pre-existing hypercoagulability is present in patients undergoing surgery for malignant disease and that coagulation status varies by tumor type. After informed consent, preoperative blood samples were drawn from patients undergoing exploratory laparotomies for intra-abdominal malignancies and analyzed with ROTEM. Eighty-two patients were enrolled, including 72 with a confirmed pathologic diagnosis and 10 age-matched controls with benign disease. The most common cancers involved the pancreas (n = 23; 32%), esophagus (n = 19; 26%), liver (n = 12; 17%), stomach (n = 7; 10%), and bile ducts (n = 5; 7%). Preoperative hypercoagulability was detected in 31% (n = 22); these patients were more likely to have lymphovascular invasion (88% vs 50%; P = .011), perineural invasion (77% vs 36%; P = .007), and stage III/IV disease (80% vs 62%; P = .039). More patients with pancreatic tumors (9/23, 39%) were hypercoagulable than with esophageal (3/19, 16%) or liver (2/13, 15%, P = .034) tumors. When only resectable malignancies were considered, clot formation was more rapid (low clot formation time, high alpha) with enhanced maximum clot strength (high maximum clot firmness) in pancreatic versus esophageal or liver cancers and in all cancers versus those with benign disease. Preoperative hypercoagulability can be identified with ROTEM and is associated with lymphovascular/perineural invasion and advanced-staged disease in cancer. Compared with other tumor types, pancreatic adenocarcinomas have the greatest risk for hypercoagulability

Insertion of central venous catheters induces a hypercoagulable state

Central venous catheters (CVCs) increase the risk of venous thromboembolism. We have previously demonstrated that pulmonary artery catheters are associated with a hypercoagulable state in an animal model and in patients. The purpose of this study is to determine whether the insertion of a CVC is associated with a similar response. 7F femoral artery catheters were placed in healthy anesthetized swine (N = 16). Serial arterial blood samples were drawn immediately before and after an 8.5F jugular vein CVC and then for 3 hours after CVC removal. Samples were analyzed using kaolin-activated thromboelastography (TEG) at precisely 2 minutes. Human: An institutional review board-approved prospective observational trial was conducted, with informed consent, in patients with critical illness (N = 8) at a Level I trauma center. Blood was drawn from indwelling arterial catheters immediately before and 60 minutes after CVC insertion. Samples were stored in sodium citrate for 15 minutes before TEG. Routine and special coagulation tests were performed on stored samples in the hospital pathology laboratory. Insertion of a CVC decreased TEG clotting time (R) by 55% in swine and by 29% in humans (p < 0.001 and 0.019, respectively). Initial clot formation time (K) was reduced by 41% in swine and by 36% in humans (p = 0.003 and 0.019). Fibrin cross-linking (α) was accelerated by 28% in swine and by 17% in humans (p = 0.007 and 0.896), but overall clot strength (maximum amplitude) was not affected. There was no change in routine or special coagulation factors, including von Willebrand factor, antithrombin III, prothrombin time, international normalized ratio, or activated partial thromboplastin time. In animals, the hypercoagulable TEG response was persistent for 3 hours after CVC removal and was prevented by pretreatment with enoxaparin (n = 4) but not heparin (n = 2). In healthy swine and patients with critical illness, a systemic hypercoagulable state occurred after CVC insertion, and this may partially account for an increased risk of venous thromboembolism. However, because the sample size was small and not powered to detect changes in coagulation proteins, no inferences can be made about the mechanism for the hypercoagulable response

Change in Hematocrit during Trauma Assessment Predicts Bleeding Even with Ongoing Fluid Resuscitation

This study tests the hypothesis that a change in hematocrit (ΔHct) during initial trauma work-up is as reliable as conventional vital signs for detecting bleeding, even with ongoing fluid resuscitation. Consecutive trauma patients admitted to a Level I trauma center receiving two Hct measurements during initial resuscitation between January 2010 and January 2011 were stratified based on estimated blood loss greater than 250 mL (bleeding) or nonbleeding. Sensitivity, specificity, and receiver operating characteristic curves were calculated for systolic blood pressure (SBP), heart rate, base deficit, and ΔHct. In 168 (72%) nonbleeding versus 64 (28%) bleeding patients, age and gender were similar. Arrival SBP was highly specific (90 to 99%) but poorly sensitive (13 to 31%) for detecting bleeding. Combinations of vital signs increased specificity, albeit at the expense of sensitivity. For bleeding versus nonbleeding patients (all receiving resuscitation fluid), ΔHct was 9.0 versus 1.8, ΔHct/liter was 4.8 versus 1.5, and ΔHct/liter/hour was 2.8 vs 0.6 (all P < 0.001). Only SBP (area under the curve [AUC] 0.608 to 0.695) and ΔHct (AUC 0.731 to 0.921) were significant for identifying bleeding with ΔHct 6 or greater being the best predictor (sensitivity 89%, specificity 95%, AUC 0.921). During ongoing fluid resuscitation of a trauma victim, ΔHct is the single most reliable indicator of continuing blood loss. A ΔHct 6 or greater during initial resuscitation is highly suspicious for ongoing blood loss, but even lesser changes have predictive value. Altogether, these results support the idea that fluid shifts are rapid after hemorrhage and Hct can be valuable during initial trauma assessment

Hypercoagulability and other risk factors in trauma intensive care unit patients with venous thromboembolism

Thromboelastography (TEG) on hospital admission can identify hypercoagulable trauma patients at risk for venous thromboembolism (VTE), but the value of TEGs obtained after multiple interventions, including tranexamic acid (TXA), has not been defined. We test the following hypotheses. (1) TEG on intensive care unit (ICU) admission can help stratify patients screened with Greenfield's risk assessment profile (RAP) for VTE. (2) TXA is a VTE risk factor, and its effect on fibrinolysis can be identified with TEG. Trauma patients who survived to the ICU with RAP ≥ 10 received serial venous duplex ultrasound examinations and blood samples for coagulation analysis at admission to the ICU and weekly thereafter. Six hundred seventy-eight patients were screened and 121 were enrolled; 76% blunt injury, Injury Severity Score (ISS) 27, 13% mortality. Thromboprophylaxis was administered to 90% of the patients and was started a median of 2 days after hospital admission. VTE was detected in 28% (n = 34) of the patients (27 deep vein thrombosis and 7 pulmonary emboli) and occurred a median 10 days after admission. Twenty-nine percent (n = 10) of VTE occurred within 2 days of admission. Most variables were similar between those with and without VTE, but the VTE group received more operations (3 (2) vs. 2 (2), p = 0.044), had increased ICU days (25 (34) days vs. 15 (18) days, p = 0.004), and was more likely to have abdominal injury with Abbreviated Injury Scale (AIS) score > 2 (59% vs. 39%, p = 0.050). Upon ICU admission, standard coagulation markers were within normal limits, while TEG demonstrated hypercoagulability, but neither was associated with VTE. Repeat TEG one week after admission (n = 58) remained hypercoagulable but transitioned to a different pattern with increased clot strength. TXA was associated with reduced fibrinolysis on initial TEG (p < 0.05) but was not associated with VTE. Trauma ICU patients with RAP ≥ 10 are hypercoagulable at admission to ICU and remain so during recovery. They have a ≥ 25% rate of VTE, despite thromboprophylaxis. TXA is associated with reduced fibrinolysis but does not increase VTE rates. Neither TEG nor standard coagulation markers (measured on ICU admission) stratify high-risk patients who develop VTE from those who do not. Prognostic study, level III