Venezuelan equine encephalitis (VEE) complex alphaviruses are important re-emerging arboviruses that cause life-threatening disease in equids during epizootics as well as spillover human infections. We conducted a comprehensive analysis of VEE complex alphaviruses by sequencing the genomes of 94 strains and performing phylogenetic analyses of 130 isolates using complete open reading frames for the nonstructural and structural polyproteins. Our analyses confirmed purifying selection as a major mechanism influencing the evolution of these viruses as well as a confounding factor in molecular clock dating of ancestors. Times to most recent common ancestors (tMRCAs) could be robustly estimated only for the more recently diverged subtypes; the tMRCA of the ID/IAB/IC/II and IE clades of VEE virus (VEEV) were estimated at ca. 149-973 years ago. Evolution of the IE subtype has been characterized by a significant evolutionary shift from the rest of the VEEV complex, with an increase in structural protein substitutions that are unique to this group, possibly reflecting adaptation to its unique enzootic mosquito vector Culex (Melanoconion) taeniopus. Our inferred tree topologies suggest that VEEV is maintained primarily in situ, with only occasional spread to neighboring countries, probably reflecting the limited mobility of rodent hosts and mosquito vectors.

Mycotic aortic aneurysms are infrequent but challenging cases. We present a 68-year-old man with evolving infrarenal aortic and right common iliac artery aneurysms from an infection with Yersinia enterocolitica. This is a rare but virulent cause of aortitis. The patient underwent open resection and debridement with anatomic reconstruction using an aortic homograft. He recovered well and quickly returned to normal functional status. This represents the first successful anatomic aortic repair using homograft with this organism. Here, we review the literature and outcomes associated with this unusual pathogen. With favorable anatomy and expedient operative management, good results can be obtained.

Background. Enthusiasm for endovascular therapies has led to the use of stent graft repairs for popliteal artery aneurysms. In this report, we review our experience with this technique.Methods. A retrospective review was performed of all endovascular popliteal artery aneurysm repairs (EVPARs) performed by the vascular surgery group at a tertiary care medical center. Patient demographic data, operative details, and outcomes were examined.Results. We performed 33 EVPARs in 28 patients. All patients were male with a mean age of 76 years (range, 60-91). Mean aneurysm diameter was 3.2 cm (range, 1.5-6.3). All repairs were performed using a self-expanding covered stent graft. Among the patients, 18% were symptomatic at the time of repair. The median number of stents used was 2 (range, 1-4). Median duration of stay was 1 day (range, 0-12). The 1-year and 2-year patency were 87% and 81%, respectively, with a mean follow-up of 23 months. Loss of patency was associated with both poor distal runoff (P = .007) and increasing number of stents used (P = .03). Early complications were seen in 4 patients including: slent oversizing leading to in-folding, perforation of a tibial artery, access site hematoma, and access vessel dissection.Conclusion. As experience with EVPAR continues to grow, caution must be applied in its use. Careful patient selection, proper operative technique, and correct vessel sizing are required for good outcomes. Poor distal runoff and use of numerous stents leads to diminished patency rates.

Background The objective of this study was to identify the self-reported barriers to and facilitators of prescribing low-molecular-weight heparin (LMWH) thromboprophylaxis in the intensive care unit (ICU). Methods We conducted an interviewer-administered survey of 4 individuals per ICU (the ICU director, a bedside pharmacist, a thromboprophylaxis research coordinator, and physician site investigator) regarding LMWH thromboprophylaxis for medical-surgical patients in 27 ICUs in Canada and the United States. Items were generated by the research team and adapted from previous surveys, audits, qualitative studies, and quality improvement research. Respondents rated the barriers to LMWH use, facilitators (effectiveness, affordability, and acceptability thereof), and perceptions regarding LMWH use. Results Respondents had 14.5 (SD, 7.7) years of ICU experience (response rate, 99%). The 5 most common barriers in descending order were as follows: drug acquisition cost, fear of bleeding, lack of resident education, concern about bioaccumulation in renal failure, and habit. The top 5 rated facilitators were preprinted orders, education, daily reminders, audit and feedback, and local quality improvement committee endorsement. Centers using preprinted orders (mean difference [P<.01]) and computerized physician order entry (P<.01) compared with those centers not using those tools reported higher affordability for these 2 facilitators. Compared with physicians and pharmacists, research coordinators considered ICU-specific audit and feedback of thromboprophylaxis rates to be a more effective, acceptable, and affordable facilitator (odds ratio, 6.67; 95% confidence interval, 1.97-22.53; P<.01). Facilitator acceptability ratings were similar within centers but differed across centers (P≤.01). Conclusions This multicenter survey found several barriers to use of LMWH including cost, concern about bleeding, and lack of resident knowledge of effectiveness. The diversity of reported facilitators suggests that large scale programs may address generic barriers but also need site-specific interprofessional knowledge translation activities.

Venous thromboembolism (VTE) is a common postsurgical complication, the incidence of which can be reduced with the use of various prophylactic measures. Proper use of these measures requires an understanding of each individual patient's risk of VTE. Risk assessment models have been developed to aid clinicians in quantifying the likelihood of an individual VTE formation. In this review, we discuss several models of risk assessment for general and vascular surgery patients with a focus on both sensitivity and prospective validation by external sources. In addition, strategies to improve proper implementation of prophylactic measures are highlighted.

Recent studies have demonstrated the capability of second generation sequencing (SGS) to provide coverage of short tandem repeats (STRs) found within the human genome. However, there are relatively few bioinformatic software packages capable of detecting these markers in the raw sequence data. The extant STR-calling tools are sophisticated, but are not always applicable to the analysis of the STR loci commonly used in forensic analyses. STRait Razor is a newly developed Perl-based software tool that runs on the Linux/Unix operating system and is designed to detect forensically-relevant STR alleles in FASTQ sequence data, based on allelic length. It is capable of analyzing STR loci with repeat motifs ranging from simple to complex without the need for extensive allelic sequence data. STRait Razor is designed to interpret both single-end and paired-end data and relies on intelligent parallel processing to reduce analysis time. Users are presented with a number of customization options, including variable mismatch detection parameters, as well as the ability to easily allow for the detection of alleles at new loci. In its current state, the software detects alleles for 44 autosomal and Y-chromosome STR loci. The study described herein demonstrates that STRait Razor is capable of detecting STR alleles in data generated by multiple library preparation methods and two Illumina(®) sequencing instruments, with 100% concordance. The data also reveal noteworthy concepts related to the effect of different preparation chemistries and sequencing parameters on the bioinformatic detection of STR alleles.

Microbial pathogens of humans, animals, and plants can serve as potential agents of biowarfare, bioterrorism, and biocrime. Previously, the Microbial Rosetta Stone (MRS) Central database, an easily accessible informational resource tool, was developed to assist law enforcement personnel in the event of a disease investigation by providing key information on pathogens of concern. Although the database already contained information on a few high-profile plant pathogens, the coverage was insufficient considering the large number of plant pathogens that pose a threat, not only to agricultural production but also to natural plant resources such as forests and rangelands. In this project, 100 plant pathogens of high consequence were selected for study, existing literature on these agents was reviewed, and both the sources and key pathogen information provided therein were curated in the new Agricultural Database (AgDB), an accessory to the existing MRS Central Database. Chosen for inclusion in the MRS Central AgDB were plant pathogens having significant potential for damage to U.S. agricultural and natural ecosystems. The selection process included review of several previously developed plant-pathogen threat lists and recommendations from experts within the U.S. plant biosecurity community. Pathogen information was collected by searching a number of relevant literature databases, sites on the World Wide Web, and other resources. For inclusion in the MRS, the information was curated into categories: pathogen taxonomy, nomenclature synonyms, disease symptoms and geographic distribution, plant hosts, insect vectors, detection and diagnostic methods, laboratory and field protocols, sample collection, and epidemiology. The resulting AgDB enhances the MRS Central Database by summarizing and linking key information on high-threat plant diseases and their causal agents to relevant scientific literature and internet resources. The AgDB contains critical, key information on high-consequence plant pathogens, curated in a format that is readily accessible and easily searched. The resource enhances the existing MRS Central Database and provides law enforcement, forensic, and investigative personnel with an additional tool with which to respond to microbial emergencies, particularly those affecting the agricultural and environmental sectors.

High-risk neuroblastoma (NB) represents a problematic tumor phenotype associated with a dreary outlook. Modern molecular achievements over the last decade have seen the increase and implementation of 'omics technologies in oncology that promises to provide for a deeper comprehension of complex tumor pathways. The emerging concept of analyzing NB-specific 'omics profiles to better understand and define the behavior of advanced-stage tumors along with providing direct and targeted therapy may ultimately translate into improved outcomes for high-risk NB. Knowledge of NB proteomics has gradually become available, but the challenge remains to integrate data obtained from different levels of biological organization. In this review, we provide an overview of the proteomics-based techniques that can be used to advance and accelerate the discovery of novel molecular biomarkers for NB. By citing specific examples, we discuss how proteomics has contributed to the early detection of advanced-stage NB and minimal residual disease. We end by contemplating the emerging technologies that are likely to have a high impact on the field of NB in the near future.

Fetal imaging has dramatically impacted neonatal care by providing an advanced warning of many different congenital anomalies. The advancements and widespread use of fetal imaging has, however, increased the identification of various incidental findings that is creating new challenges for neonatal diagnosis and treatment. We report such a case where a fetal pancreatic neuroblastoma (NB) was incidentally detected by computed tomographic scan of the maternal abdomen. Primary pancreatic NB is a very uncommon childhood neoplasm that, to our knowledge, has never been previously reported in the English language presenting in either the prenatal or neonatal periods. A 21-year-old woman complaining of acute abdominal pain and carrying a 35 weeks' gestation fetus was referred for computed tomographic scan because of the concern of maternal appendicitis. That scan was inconclusive for appendicitis but did incidentally detect a fetal mass in the general region of the pancreas. At 36 weeks' gestation, the fetus developed signs of distress, which prompted a cesarean delivery. Neonatal workup confirmed the presence of an abdominal mass in the region of the pancreas, but precise anatomic localization was not possible. Also noted on neonatal workup were elevated urinary catecholamines consistent with a hormonal active tumor. These findings prompted an abdominal exploration of this neonate, which revealed a solid tumor contained in the distal pancreas. The mass was managed by an uncomplicated distal pancreatectomy. The neonate fully recovered, and histologic diagnosis revealed NB, whereas the postoperative urine catecholamines normalized. This case underscores the unintended clinical challenges created by widespread fetal imaging, while presenting the first prenatally diagnosed case in the English language medical literature and earliest treated patient with pancreatic NB.

Background. Solid pseudopapillary neoplasms of the pancreas (SPN) account for less than 1% of all pancreatic tumors. The goal of this study was to better understand the nature of these rare tumors through analysis of patients' clinical presentations and outcomes following surgical resection.Methods. A multi-institutional retrospective review was conducted of all patients who underwent surgical resection from 1994 to 2008.Results. Twenty-one patients were identified with SPN. Twenty patients were female. Median age at presentation was 34y. The most common presenting symptom was abdominal pain (67%). All patients underwent resection: distal pancreatectomy (9), pancreaticoduodenectomy (5), central pancreatectomy (6), and laparoscopic excision/enucleation (1). A R-0 resection was obtained in all patients. Median tumor size was 5.5cm. AJCC stages were stage I (18), stage II (1), stage III (2), and stage IV (0). Postsurgical complications occurred in 52% of patients, with pancreatic fistulae being the most common (29%). The median follow-up time was 55 mo. All patients remain alive without evidence of recurrence.Conclusion. Solid pseudopapillary neoplasms of the pancreas are atypical pancreatic tumors. SPN usually occur in young women who present with abdominal pain. Oncologic outcomes in patients who undergo surgical resection are excellent. (C) 2009 Elsevier Inc. All rights reserved.

Neuroblastoma (NB) is one of the most common solid tumors of childhood and displays a remarkable diversity in both biologic characteristics and clinical outcomes. Availability of high-throughput 'omics technologies and their subsequent application towards oncology has provided insight into the complex pathways of tumor formation and progression. Investigation of NB 'omics profiles may better define tumor behavior and provide targeted therapy with the goal of improving outcomes in patients with high-risk disease. Utilization of these technologies in NB has already led to advances in classification and risk stratification. The gradual emergence of NB-directed proteomics adds a layer of intricacy to the analysis of biologic organization but may ultimately provide a better comprehension of this complex disease. In this review, we cite specific examples of how NB-directed proteomics has provided information regarding novel biomarkers and possible therapeutic targets. We finish by examining the impact of high-throughput 'omics in the field of NB and speculate on how these emerging technologies may further be incorporated into the discipline.

Background. Children with advanced-stage neuroblastoma (NB) traditionally experience poor outcomes. Because early detection of advanced-stage disease may impact survival, finding new targets for early diagnosis is crucial. Evidence suggests the tumor microenvironment may have profound effects on cancer progression.Methods. As little is known concerning the NB-host microenvironment, this study applied proteomic techniques, two-dimensional polyacrylamide gel electrophoresis (21) PAGE) combined with matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry to determine protein differences between cell cultured NB and tumors grown in mice for 2, 4, and 5 wk.Results. We found an increase in proteins in cultured NB compared with implanted mouse tumors during tumor progression. Additionally, analyzing in vivo tumors to cultured NB, we observed less expressed proteins. However, 16 out of 19 proteins were of mouse origin, thus inferring host-derived factors contributing to tumor growth.Conclusion. We show that the dynamic relationship between NB and host microenvironment is important for tumor growth and better understanding of this milieu maybe relevant towards finding unique approaches for identifying advanced-stage disease. Published by Elsevier Inc. All rights reserved.

Background Neuroblastoma (NB) is the most common extracranial solid tumor in children. At the time of diagnosis, the tumor has metastasized in as many as 7 of 10 cases, and survival in high-risk patients remains poor. Accurate classification of high-risk patients is very important since this determines treatment plan, and although a consensus risk classification system has been established for NB, it contains few specific molecular markers that account for aggressive nature and metastatic potential of the tumor. Bin1 expression is reduced in breast, NB, and other cancer types and the reduction correlates with high-risk clinical features. Here we hypothesize that Bin1 has an inhibitory role in metastasis, and therefore decrease in its expression may be a marker of high-risk NB. Procedure. Initially, breast cancer and NB cell lines derived from metastasis were examined for Bin1 expression. Then, a stable Bin1-overexpressing NB cell line was created and evaluated for in vitro metastatic behaviors using anoikis, invasion, and migration assays, and chemoresponsiveness using MTT assay. Results. Reduced Bin1 was detected in all cancer cell lines examined, and forced Bin1 overexpression increased NB cell anoikis and enhanced the cell killing by doxorubicin. However, Bin1 overexpression did not significantly affect cell invasion, motility, or proliferation. Conclusions. Bin1 appears to function as a metastasis suppressor and chemosensitizer in NB, and resistance to anoikis may be an important metastatic mechanism. Thus, Bin1 expression status could serve as a marker for metastatic potential and chemosensitivity thereby allowing for more accurate classifications of high-risk NB patients. Pediatr Blood Cancer 2009;53:332-337. (C) 2009 Wiley-Liss, Inc.

I would like to thank the authors for their comments regarding our recent article “Impact of omphalocele size on associated conditions” [ [1] Kumar H.R. Jester A.L. Ladd A.P. Impact of omphalocele size on associated conditions. J Pediatr Surg. 2008; 43: 2216-2219 Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar ]. As they very eloquently point out and defend with the published literature, our profession has yet to offer scientifically based definitions to either designation of small or large omphaloceles. With this lack of standardization in the segregation of children with this anomaly, the corresponding published literature does not lend itself to straightforward conclusions that can be generalized to treatment or expected outcomes. In the end, the clinical designation may very well be based on the suitability of the defect for primary closure or not, yet the successful use of prosthetics in closure may confound this naive simplification.

Introduction Solid tumors, such as neuroblastoma (NB), are associated with a heterogeneous cell environment. Multicellular tumor spheroid (MCTS) cultures have been shown to better mimic growth characteristics of in vivo solid tumors. Because tumor spheroid growth patterns may be quite different from standard two-dimensional culture systems, we sought to compare the protein expression profiles of two- and three-dimensional in vitro NB cultures, i.e., monolayers and MCTS.Materials and methods Human NB cells were grown as both monolayers and spheres. Nuclear and cytosolic proteins were analyzed for differentially secreted proteins by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and selected polypeptides were identified by mass spectrometry (LC-MS/MS).Results Several metabolic (transketolase, triosephosphate isomerase, pyruvate kinase M1/M2, alpha enolase, and phosphoglycerate mutase-1), cell stress response (heat shock proteins (HSP) 90, 70, and 60; antioxidant, thioredoxin), cell structure (septin 2, adenyl cyclase-associated protein-1), tubulin beta-2 chain, actin, translationally controlled tumor protein and cofilin), signal transduction (peptidyl prolyl cis/trans isomerase A), biosynthetic (phosphoserine aminotransferase) and transport (cellular retinoic acid binding protein 1) polypeptides were overexpressed in spheroids. Several protein groups were differentially expressed between NB monolayers and spheroids.Conclusion The altered proteins among NB spheroids may represent an important link between monolayer cell cultures and in vivo experiments and thus a more ideal in vitro culture system for determining the precise threedimensional microenvironment of NB.

Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults. Median survival from the time of diagnosis is less than a year, with less than 5% of patients surviving 5 years. These tumors are thought to arise through two different pathways. Primary GBMs represent de novo tumors, while secondary GBMs represent the malignant progression of lower-grade astrocytomas. Moreover, despite improvements in deciphering the complex biology of these tumors, the overall prognosis has not changed in the past three decades. The hope for improving the outlook for these glial-based malignancies is centered on the successful clinical application of current high-throughput technologies. For example, the complete sequencing of the human genome has brought both genomics and proteomics to the forefront of cancer research as a powerful approach to systematically identify large volumes of data that can be utilized to study the molecular and cellular basis of oncology. The organization of these data into a comprehensive view of tumor growth and progression translates into a unique opportunity to diagnose and treat cancer patients. In this review, we summarize current genomic and proteomic alterations associated with GBM and how these modalities may ultimately impact treatment and survival.

A35 Bin1 protein displays multiple functions such as suppression of oncogenic transformation and promoting apoptosis. Loss or reduction of Bin1 expression has been evident in many human cancers including breast, prostate, and neuroblastoma (NB), and correlated with the poor outcome of patients. NB originates from precursor cells of the peripheral nervous system and is the most common extracranial solid tumor in children. The tumor is metastatic in 70% of NB patients at the time of diagnosis and the overall prognosis is particularly poor despite aggressive therapies (10-40% survival in high-risk cases). Here we show that Bin1 is frequently reduced or lost in cell lines derived from the bone marrow metastasis or abdominal mass of NB patients. To establish the direct functional link between Bin1 expression and NB, we created NB cell lines that stably overexpress Bin1. Because Bin1 has a role in apoptosis and because survival of cancer cells in the systemic circulation plays a crucial role in the multistep metastatic process, we examined the role of Bin1 in cell survival under an anchorage disrupted condition. Forced Bin1 overexpression in the stable cells increased cell death when cultured in Poly-Hema coated plates. This type of cell death is believed to be a specialized apoptosis (anoikis) . Additionally, many chemotherapeutic agents kill tumor cells via apoptosis, and we also evaluated the influence of Bin1 on chemosensitivity. The results showed that Bin1 overexpression enhances cell killing by Doxorubicin, a commonly used anticancer agent. Proteomic analyses of cell lysates from Bin1 transfectants versus the vector controls revealed differentially expressed proteins. This indicates that Bin1 may modify the metastatic behaviors and chemoresponsiveness in NB via regulating the expression levels of other cellular proteins. Our results establish Bin1 as an important factor for NB metastasis and chemosensitivity. Thus, loss of Bin1 could serve as a prognostic marker for metastatic potential and therapeutic response, and targeting Bin1 and Bin1-regulated pathways could be a rational strategy for the treatment of high-risk NB patients.