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Competing Interests: The authors report no conflicts of interest.

Oncogenic fusions involving neurotrophic receptor tyrosine kinase (NTRK) genes are being increasingly identified in a range of mesenchymal tumours unrelated to infantile fibrosarcoma or cellular congenital mesoblastic nephroma, where the canonical ETV6-NTRK3 fusion was first described more than two decades ago. Recognition of these NTRK-rearranged tumours poses a diagnostic challenge to surgical pathologists due to their non-specific clinical and pathological features. However, their recognition is of heightened importance, particularly in locally advanced and metastatic sarcomas, due to the recent availability of selective and highly effective targeted therapy. Herein, we present an Australian multi-institutional series of six of these rare NTRK-rearranged mesenchymal neoplasms to share the local experience and diagnostic challenges as well as to highlight key morphological patterns and immunoprofiles that provide the most helpful clues in routine practice. We also propose a diagnostic algorithm for the detection of these fusions, drawing attention to the limitations of ancillary studies including immunohistochemistry against tropomyosin receptor kinase (TRK) protein, fluorescence in situ hybridisation (FISH) and next generation sequencing., (Copyright © 2020 Royal College of Pathologists of Australasia. All rights reserved.)

Objective: Oxidized phospholipids (OxPL), such as the oxidized derivatives of 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine, 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine, and 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, have been shown to be the principal biologically active components of minimally oxidized LDL (low-density lipoprotein). The role of OxPL in cardiovascular diseases is well recognized, including activation of inflammation within vascular cells. Atherosclerotic Apoe mice fed a high-fat diet develop antibodies to OxPL, and hybridoma B-cell lines producing natural anti-OxPL autoantibodies have been successfully generated and characterized. However, as yet, no studies have been reported demonstrating that treatment with OxPL neutralizing antibodies can be used to prevent or reverse advanced atherosclerosis. Approach and Results: Here, using a screening against 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine/1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, we generated a novel IgM autoantibody, 10C12, from the spleens of -/- mice fed a high-fat diet develop antibodies to OxPL, and hybridoma B-cell lines producing natural anti-OxPL autoantibodies have been successfully generated and characterized. However, as yet, no studies have been reported demonstrating that treatment with OxPL neutralizing antibodies can be used to prevent or reverse advanced atherosclerosis. Approach and Results: Here, using a screening against 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine/1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, we generated a novel IgM autoantibody, 10C12, from the spleens of Apoe -/- mice fed a long-term Western diet, that demonstrated potent OxPL neutralizing activity in vitro and the ability to inhibit macrophage accumulation within arteries of Apoe -/- mice fed a Western diet for 4 weeks. Of interest, 10C12 failed to inhibit atherosclerosis progression in Apoe -/- mice treated between 18 and 26 weeks of Western diet feeding likely due at least in part to high levels of endogenous anti-OxPL antibodies. However, 10C12 treatment caused a 40% decrease in lipid accumulation within aortas of secreted IgM deficient, sIgM -/- Apoe -/- , mice fed a low-fat diet, when the antibody was administrated between 32-40 weeks of age., Conclusions: Taken together, these results provide direct evidence showing that treatment with a single autoimmune anti-OxPL IgM antibody during advanced disease stages can have an atheroprotective outcome.

Adiponectin (APN) and its receptors have been reported to be associated with metabolic phenotypes. To better understand the effects of APN levels and its receptors on diabetic retinopathy (DR), we investigated the association of the plasma APN level and variations in APN-related genes with DR, individually and in combination.Patients with type 2 diabetes (T2D; N = 1604), above 20 years of age from the Taiwanese population participated in the study. Demographic information, blood pressure, and serological markers were recorded at enrollment. Genomic DNA was isolated and genotyped. The plasma APN levels were measured by enzyme-linked immunosorbent assay.T2D patients with DR (N = 632) had diabetes for a longer duration, and had higher HbA1c, and systolic and diastolic blood pressure compared to those without DR (N = 972) (P < .001, for all the parameters). Overall, 10 single nucleotide polymorphisms (SNPs) in ADIPOQ and CDH13 susceptibility loci were associated with DR. Gene risk score (GRS) was calculated based on 10 SNPs for each subject and the cumulative effect of genes was observed. Among the subjects with plasma APN level (N = 518), natural logarithm (LN) of APN (LN [APN]; odds ratio [OR] = 1.63, 95% confidence interval [CI] = 1.19-2.25) and GRS (OR = 1.90, 95% CI = 1.11-3.26 for middle range of GRS, and OR = 2.61, 95% CI = 1.48-4.59 for high range of GRS) were independent risk factors for DR after adjustment for other parameters.In conclusion, the plasma APN level and the genetic variations in adiponectin receptors were associated with DR.

Background: The vascular complications of diabetes mellitus are the major causes of morbidity and mortality among people with diabetes. Circular RNAs are a class of endogenous noncoding RNAs that regulate gene expression in eukaryotes. In this study, we investigated the role of circular RNA in retinal vascular dysfunction induced by diabetes mellitus., Methods: Quantitative polymerase chain reactions, Sanger sequencing, and Northern blots were conducted to detect circular HIPK3 (circHIPK3) expression pattern on diabetes mellitus-related stresses. MTT (3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assays, EdU (5-ethynyl-2'-deoxyuridine) incorporation assays, Transwell migration assays, and Matrigel assays were conducted to detect the role of circHIPK3 in retinal endothelial cell function in vitro. Retinal trypsin digestion, vascular permeability assays, and ELISA assays were conducted to detect the role of circHIPK3 in retinal vascular dysfunction in vivo. Bioinformatics analysis, luciferase activity assays, RNA pull-down assays, and in vitro studies were conducted to reveal the mechanism of circHIPK3-mediated retinal vascular dysfunction., Results: circHIPK3 expression was significantly upregulated in diabetic retinas and retinal endothelial cells following stressors related to diabetes mellitus. circHIPK3 silencing or overexpressing circHIPK3 changed retinal endothelial cell viability, proliferation, migration, and tube formation in vitro. circHIPK3 silencing in vivo alleviated retinal vascular dysfunction, as shown by decreased retinal acellular capillaries, vascular leakage, and inflammation. circHIPK3 acted as an endogenous miR-30a-3p sponge to sequester and inhibit miR-30a-3p activity, which led to increased vascular endothelial growth factor-C, FZD4, and WNT2 expression. Ectopic expression of miR-30a-3p mimicked the effect of circHIPK3 silencing on vascular endothelial phenotypes in vivo and in vitro., Conclusions: The circular RNA circHIPK3 plays a role in diabetic retinopathy by blocking miR-30a function, leading to increased endothelial proliferation and vascular dysfunction. These data suggest that circular RNA is a potential target to control diabetic proliferative retinopathy., (© 2017 American Heart Association, Inc.)

Background: Unrecognized hemorrhage and unguided resuscitation is associated with increased perioperative morbidity and mortality. The authors investigated peripheral venous waveform analysis (PIVA) as a method for quantitating hemorrhage as well as iatrogenic fluid overload during resuscitation., Methods: The authors conducted a prospective study on Yorkshire Pigs (n = 8) undergoing hemorrhage, autologous blood return, and administration of balanced crystalloid solution beyond euvolemia. Intra-arterial blood pressure, electrocardiogram, and pulse oximetry were applied to each subject. Peripheral venous pressure was measured continuously through an upper extremity standard peripheral IV catheter and analyzed with LabChart. The primary outcome was comparison of change in the first fundamental frequency (f1) of PIVA with standard and invasive monitoring and shock index (SI)., Results: Hemorrhage, return to euvolemia, and iatrogenic fluid overload resulted in significantly non-zero slopes of f1 amplitude. There were no significant differences in heart rate or mean arterial pressure, and a late change in SI. For the detection of hypovolemia the PIVA f1 amplitude change generated an receiver operator curves (ROC) curve with an area under the curve (AUC) of 0.93; heart rate AUC = 0.61; mean arterial pressure AUC = 0.48, and SI AUC = 0.72. For hypervolemia the f1 amplitude generated an ROC curve with an AUC of 0.85, heart rate AUC = 0.62, mean arterial pressure AUC = 0.63, and SI AUC = 0.65., Conclusions: In this study, PIVA demonstrated a greater sensitivity for detecting acute hemorrhage, return to euvolemia, and iatrogenic fluid overload compared with standard monitoring and SI. PIVA may provide a low-cost, minimally invasive monitoring solution for monitoring and resuscitating patients with perioperative hemorrhage.

Background and Purpose: A role of neural networks in the development of poststroke dementia has not been clearly established. We hypothesized that stroke-mediated disruption of subcortical cholinergic pathway or large-scale neural networks contributes to poststroke dementia., Methods: A matched case-control study was conducted in a predetermined cohort with acute ischemic stroke. Cases were defined as newly developed dementia diagnosed >3 months after stroke using the Korean Vascular Cognitive Impairment Harmonization Standards. Each case was matched to 2 controls for age, education, and initial stroke severity. The Cholinergic Pathways HyperIntensities Scale was applied with some modifications to characterize disruption of cholinergic pathways by acute stroke lesions. Involvement of major cortical hub locations of the default mode network, central executive network, and salience network was also investigated., Results: After matching, 38 cases and 66 matched controls were included. Cholinergic Pathways HyperIntensities Scale scores were significantly higher in cases than in controls (2.2±2.9 versus 0.9±1.4). Acute ischemic lesions affecting the default mode and central executive networks were more frequently observed in cases compared with controls (36.8% versus 7.6% and 26.3% versus 6.1%, respectively). These findings remained significant in the multiple logistic regression models adjusted for various sets of potential confounders. Lesion location analysis revealed that cases were more likely to have acute lesions in the left corona radiata, hippocampal formation, and posterior parietal cortex., Conclusions: Disruption of cholinergic pathways and major hubs of large-scale neural networks might contribute to newly developed dementia after acute ischemic stroke.

Background: Color flow Doppler ultrasonography has been used to confirm caudal epidural injection, but its ability to detect accidental intrathecal injection is unknown. We hypothesized that, when using color flow Doppler, the injection of fluid into the epidural space would result in turbulent flow which would appear as a burst of color while intrathecal injection would show an absence of a color flow Doppler signal., Methods: Two groups of pediatric patients (up to 6 years of age) were prospectively enrolled for this observational study during a 2-month period. One group (group E) consisted of patients suitable for elective surgery using caudal epidural analgesia, and the other (group I) included patients receiving lumbar puncture for intrathecal chemotherapeutic injection. After induction of general anesthesia and placement of the patient in the lateral position, an 8 MHz curved array probe (Sonosite TITAN, Bothell, WA) was applied to obtain a transverse image of the lumbar region (L1-L3). Real-time images using color flow Doppler were obtained and recorded during initial injections of 2 consecutive (20 seconds apart) aliquots of 0.1 mL/kg medication of local anesthetic (0.25% bupivacaine) or chemotherapy drugs (mixture of methotrexate, cytarabine, and hydrocortisone) at a rate of 0.5 to 1.0 mL/s. After obtaining the study images, the rest of the medication was injected in standard fashion. A blinded anesthesiologist later evaluated the recorded images to determine a positive or negative result (positive = presence of turbulence as illustrated by a medley of color; negative = no turbulence or color). Sensitivity, specificity, and positive and negative predictive values were calculated for those patients who had successful analgesia (group E) and intrathecal (group I) injections., Results: Forty recorded images from 41 patients (group E, n = 21; group I, n = 20) were included in the analysis. The observed sensitivity, specificity, positive predictive value, and negative predictive values were all 100%. The lower 95% confidence limits were 0.832., Conclusion: In the context of this study, color flow Doppler could differentiate epidural from intrathecal injection into the caudal space of children up to 6 years of age using a 0.1 mL/kg injection volume and injection rate of 0.5 to 1.0 mL/s.

Objectives: To describe acute injury characteristics in children and youth soccer players and to identify the characteristics of patients who required hospital admission., Methods: The analysis of the study was based on the Canadian Hospitals Injury Reporting and Prevention Program. A total of 32,149 patients (aged 5-19 years) with soccer-related injuries presenting to 16 participating hospital emergency departments from 1994 to 2004 were included in the analysis., Results: Males had the highest proportion of injuries (62%). The leading injuries were sprains/strains (38%), followed by fractures/dislocations (31%) and superficial injuries (23%). A total of 896 cases (3%) required hospital admission. Based on logistic regression analysis, being a male, playing unorganized soccer, having multiple body injuries, playing soccer outside school premises, and playing during the summer/fall increased the likelihood of hospital admission. Moreover, having a head/face/neck injury (Odds ratio [OR], 1.3; 95% confidence interval [95% CI], 1.1-1.7) and trunk injury (OR, 1.7; 95% CI, 1.2-2.4) as compared with an upper extremity injury and having injuries from contact with structures/surfaces (OR, 3.1; 95% CI, 2.2-4.3) and with other players (OR, 2.5; 95% CI, 1.8-3.5) as compared with ball contact had the highest odds of hospital admission., Conclusions: Soccer accounted for a significant proportion of injuries presented to Canadian Hospitals Injury Reporting and Prevention Program emergency departments during 1994-2004. Further studies investigating potential interventional programs and techniques among this population are highly warranted.

Background: Roughly, 10% of elderly patients develop postoperative cognitive dysfunction. General anesthesia impairs spatial memory in aged rats, but the mechanism is not known. Hippocampal neurogenesis affects spatial learning and memory in rats, and isoflurane affects neurogenesis in neonatal and young adult rats. We tested the hypothesis that isoflurane impairs neurogenesis and hippocampal function in aged rats., Methods: Isoflurane was administered to 16-month-old rats at one minimum alveolar concentration for 4 h. FluoroJade staining was performed to assess brain cell death 16 h after isoflurane administration. Dentate gyrus progenitor proliferation was assessed by bromodeoxyuridine injection 4 days after anesthesia and quantification of bromodeoxyuridine+ cells 12 h later. Neuronal differentiation was studied by determining colocalization of bromodeoxyuridine with the immature neuronal marker NeuroD 5 days after anesthesia. New neuronal survival was assessed by quantifying cells coexpressing bromodeoxyuridine and the mature neuronal marker NeuN 5 weeks after anesthesia. Four months after anesthesia, associative learning was assessed by fear conditioning. Spatial reference memory acquisition and retention was tested in the Morris Water Maze., Results: Cell death was sporadic and not different between groups. We did not detect any differences in hippocampal progenitor proliferation, neuronal differentiation, new neuronal survival, or in any of the tests of long-term hippocampal function., Conclusion: In aged rats, isoflurane does not affect brain cell death, hippocampal neurogenesis, or long-term neurocognitive outcome.

The use of ultrasound guidance has provided an opportunity to perform many peripheral nerve blocks that would have been difficult to perform in children based on pure landmark techniques due to the potential for injection into contiguous sensitive vascular areas. This review article provides the readers with techniques on ultrasound-guided peripheral nerve blocks of the extremities and trunk with currently available literature to substantiate the available evidence for the use of these techniques. Ultrasound images of the blocks with corresponding line diagrams to demonstrate the placement of the ultrasound probe have been provided for all the relevant nerve blocks in children. The authors hope that this review will stimulate further research into ultrasound-guided regional anesthesia in infants, children, and adolescents and stimulate more randomized controlled trials to provide a greater understanding of the anatomy and physiology of regional anesthesia in pediatrics.

Background: Anesthetic agents cause cell death in the developing rodent brain and long-term, mostly hippocampal-dependent, neurocognitive dysfunction. However, a causal link between these findings has not been shown. Postnatal hippocampal neurogenesis affects hippocampal function into adulthood; therefore, the authors tested the hypothesis that isoflurane affects long-term neurocognitive function via an effect on dentate gyrus neurogenesis., Methods: The S-phase marker 5-bromodeoxyuridine was administered at various times before, during, and after 4 h of isoflurane given to postnatal day (P)60 and P7 rats to assess dentate gyrus progenitor proliferation, early neuronal lineage selection, and long-term survival of new granule cell neurons. Fear conditioning and spatial reference memory was tested at various intervals from 2 weeks until 8 months after anesthesia., Results: In P60 rats, isoflurane increased early neuronal differentiation as assessed by BrdU/NeuroD costaining, decreased progenitor proliferation for 1 day, and subsequently increased progenitor proliferation 5-10 days after anesthesia. In P7 rats, isoflurane did not induce neuronal lineage selection but decreased progenitor proliferation until at least 5 days after anesthesia. Isoflurane improved spatial reference memory of P60 rats long-term, but it caused a delayed-onset, progressive, persistent hippocampal deficit in P7 rats in fear conditioning and spatial reference memory tasks., Conclusion: The authors conclude that isoflurane differentially affects both neurogenesis and long-term neurocognitive function in P60 and P7 rats. Neurogenesis might mediate the long-term neurocognitive outcome after isoflurane at different ages.

Background: Millions of neonates undergo anesthesia each year. Certain anesthetic agents cause brain cell death and long-term neurocognitive dysfunction in postnatal day (P)7 rats. Despite its intuitive appeal, a causal link between cell death and neurocognitive decline after anesthesia has not been established. If one existed, the degree of cell death would be expected to correlate with the degree of neurocognitive dysfunction caused by anesthesia. The authors therefore tested if cell death caused by various durations of isoflurane at 1 minimum alveolar concentration causes duration-dependent long-term neurocognitive dysfunction., Methods: Isoflurane was administered to P7 rats at 1 minimum alveolar concentration for 0, 1, 2, or 4 h. To control for the respiratory depressant effects of anesthesia, a group of rats was treated with 4 h of carbon dioxide. Cell death was assessed by FluoroJade staining 12 h after the end of each intervention, and neurocognitive outcome was assessed 8 weeks later by using fear conditioning, spatial reference memory, and spatial working memory tasks., Results: Widespread brain cell death was caused by 2 h and 4 h of isoflurane and by 4 h of carbon dioxide. The degree and distribution of thalamic cell death was similar in 4 h isoflurane-treated and 4-h carbon dioxide-treated rats. Only 4 h of isoflurane caused a long-term neurocognitive deficit affecting both spatial reference memory and spatial working memory. Working memory was improved in carbon dioxide-treated rats., Conclusion: Isoflurane-induced brain cell death may be partly caused by hypercarbia. The inconsistencies between cell death and neurocognitive outcome suggest that additional or alternative mechanisms may mediate anesthesia-induced long-term neurocognitive dysfunction.

Objectives: 201Tl SPECT is used successfully in the diagnosis of recurrent supratentorial glioma and in the evaluation of its response to chemotherapy. However, different methods are used to measure relative tracer uptake in tumour and background. The objective of this study was to assess the interobserver variability of such methods, and, if possible, to provide nomograms for data conversion., Methods: Using baseline and follow-up SPECT scans from 20 patients with recurrent glioma treated with chemotherapy, three observers applied manual and semi-automatic ROI techniques to define activity in tumour (manual, semi-automatic) as well as in reference tissue (scalp, mirror, hemisphere)., Results: All tumour ROI techniques had intra-class correlation coefficients (ICC) > or = 0.80 indicating almost perfect agreement. The main source of variation with the manual techniques was the tumour intensity; with the semi-automatic method, observer agreement was independent of the level of tumour activity. Agreement for background ROIs was also adequate, but the mirror technique tended to perform poorer at follow-up SPECT scans (ICC 0.68). Measurement of fractional change during treatment revealed no significant differences between observers for any of the investigated ROI methodology variants. Conversion of quantitative methods to measure fractional change was possible using linear regression analysis., Conclusion: 201Tl SPECT in recurrent glioma appears to be a robust method with acceptable interobserver variability. The clinical field in neuro-oncology should consider including 201Tl SPECT parameters in monitoring response to chemotherapy.

Background: This is the first blinded, randomized, placebo-controlled clinical trial to evaluate the efficacy of poly-N-acetyl glucosamine (p-GLcNAc) in improving hemostasis in patients undergoing cardiac catheterization., Methods: Patients were randomly assigned to have either a placebo-treated (n = 17) or a p-GlcNAc-treated (n = 16) 3 x 3-cm patch topically placed at the femoral insertion site at the completion of their catheterization procedure with a mechanical pressure clamp applied over it. The amount of pressure was measured., Results: Although the placebo group had slightly higher clamp pressure applied to the femoral arterial puncture site at the end of the catheterization procedure (189 +/- 47 vs. 149 +/- 49 mm Hg, p = 0.042), the time to effective hemostasis (16 +/- 7 vs. 10 +/- 3 minutes, p = 0.01) was decreased in the p-GlcNAc group by 37%., Conclusion: The application of p-GlcNAc patches improved hemostasis at the arterial puncture site in patients undergoing cardiac catheterization.