bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluating Nationwide Application of Minimally Invasive Surgery for Treatment of Small Bowel Neuroendocrine Neoplasms
Authors: Kaçmaz Enes , Klümpen Heinz‐Josef , Bemelman Willem A. , Nieveen van Dijkum Els J. M. , Engelsman Anton F. , Tanis Pieter J.
Published: March 2021

Aim

Open resection of small bowel neuroendocrine neoplasms (SB‐NEN) is still considered standard‐of‐care, mainly because of frequently encountered multifocality and central mesenteric masses. The aim of this study was to evaluate surgical approach for SB‐NEN at a national level and determine predictors for overall survival.

Methods

Patients with SB‐NEN who underwent resection between 2005 and 2015 were included from the Netherlands Cancer Registry. Patient and tumor characteristics were compared between laparoscopic and open approach. Overall survival was assessed by Kaplan–Meier and compared with the Log‐rank test. Independent predictors were determined by Cox proportional hazards model.

Results

In total, 482 patients were included, of whom 342 (71%) underwent open and 140 (29%) laparoscopic resection. The open resection group had significantly more multifocal tumors resected (24% vs. 14%), pN2 lymph nodes (18% vs. 7%) and stage IV disease (36% vs. 22%). Overall survival after open resection was significantly shorter compared to laparoscopic resection (3‐year: 81% vs. 89%, 5‐year: 71% vs. 84%, p = 0.004). In multivariable analysis, age above 60‐years (60–75, HR 3.38 (95% CI 1.84–6.23); > 75 years, HR 7.63 (95% CI 3.86–15.07)), stage IV disease (HR 1.86 (95% CI 1.18–2.94)) and a laparoscopic approach (HR 0.51 (95% CI 0.28–0.94)) were independently associated with overall survival, whereas multifocal primary tumor, grade and resection margin status were not.

Conclusion

Laparoscopic resection was the approach in 29% of SB‐NEN at a national level with selection of the more favorable patients. Laparoscopic resection remained independently associated with better overall survival besides age and stage, but residual confounding cannot be excluded.
Head/skull injury potential of empty 0.5-l beer glass bottles vs. 0.33-l Coke bottles
Authors: Nentwig C. , Steinhoff S. , Adamec J. , Kunz S. N.
Published: March 2021

The medical and biomechanical assessment of injuries from blows to the head is a common task in forensic medicine. In the context of a criminal justice process, the injury potential of different striking weapons is important. The article at hand compares the injury potential of assaults with a 0.5-l beer bottle and a 0.33-l Coke bottle, both made of glass. The research team hit 30 used empty 0.5-l beer bottles and 20 used empty 0.33-l Coke bottles manually on an aluminum dummy skull set on a force measuring plate, using acrylic and pork rind as a scalp surrogate. There was no significant difference in fracture threshold and energy transfer between the examined beer and Coke bottles. Both glass bottles are able to cause fractures to the facial bones while cranial bone fractures are primarily not to be expected. Blows with a 0.5-l beer bottle or with a 0.33-l Coke bottle to the head can transfer up to 1.255 N and thus are able to cause severe blunt as well as sharp trauma injuries.
Risk factors for mortality in infants with congenital diaphragmatic hernia: a single center experience
Authors: Brandt Jennifer Bettina , Werther Tobias , Groth Erika , Küng Erik , Golej Johann , Berger Angelika
Published: March 2021

Background

Despite current progress in research of congenital diaphragmatic hernia, its management remains challenging, requiring an interdisciplinary team for optimal treatment.

Objective

Aim of the present study was to evaluate potential risk factors for mortality of infants with congenital diaphragmatic hernia.

Methods

A single-center chart review of all patients treated with congenital diaphragmatic hernia over a period of 16 years, at the Medical University of Vienna, was performed. A comparison of medical parameters between survivors and non-survivors, as well as to published literature was conducted.

Results

During the observational period 66 patients were diagnosed with congenital diaphragmatic hernia. Overall survival was 84.6%. Left-sided hernia occurred in 51 patients (78.5%) with a mortality of 7.8%. In comparison, right-sided hernia occurred less frequently (n = 12) but showed a higher mortality (33.3%, p = 0.000). Critically instable patients were provided with venoarterial extracorporeal membrane oxygenation (ECMO, 32.3%, n = 21). Survival rate among these patients was 66.7%. Right-sided hernia, treatment with inhaled nitric oxide (iNO) over 15 days and the use of ECMO over 10 days were significant risk factors for mortality.

Conclusion

The survival rate in this cohort is comparable to the current literature. Parameters such as the side of the diaphragmatic defect, duration of ECMO and inhaled nitric oxide were assessed as mortality risk factors. This analysis of patients with congenital diaphragmatic hernia enhances understanding of risk factors for mortality, helping to improve management and enabling further evaluation in prospective clinical trials.
Dynamic contrast-enhanced magnetic resonance imaging for monitoring neovascularization during bone regeneration—a randomized in vivo study in rabbits
Authors: Righesso L. A. R. , Terekhov M. , Götz H. , Ackermann M. , Emrich T. , Schreiber L. M. , Müller W. E. G. , Jung J. , Rojas J. P. , Al-Nawas B.
Published: March 2021

Objectives

Micro-computed tomography (μ-CT) and histology, the current gold standard methods for assessing the formation of new bone and blood vessels, are invasive and/or destructive. With that in mind, a more conservative tool, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), was tested for its accuracy and reproducibility in monitoring neovascularization during bone regeneration. Additionally, the suitability of blood perfusion as a surrogate of the efficacy of osteoplastic materials was evaluated.

Materials and methods

Sixteen rabbits were used and equally divided into four groups, according to the time of euthanasia (2, 3, 4, and 6 weeks after surgery). The animals were submitted to two 8-mm craniotomies that were filled with blood or autogenous bone. Neovascularization was assessed in vivo through DCE-MRI, and bone regeneration, ex vivo, through μ-CT and histology.

Results

The defects could be consistently identified, and their blood perfusion measured through DCE-MRI, there being statistically significant differences within the blood clot group between 3 and 6 weeks (p= 0.029), and between the former and autogenous bone at six weeks (p= 0.017). Nonetheless, no significant correlations between DCE-MRI findings on neovascularization and μ-CT (r=−0.101, 95% CI [−0.445; 0.268]) or histology (r= 0.305, 95% CI [−0.133; 0.644]) findings on bone regeneration were observed.

Conclusions

These results support the hypothesis that DCE-MRI can be used to monitor neovascularization but contradict the premise that it could predict bone regeneration as well.
Dialectical Methodology of the Praxis of Biology
Authors: Gremmen Bart
Published: March 2021

Zwart uses Hegel’s dialectical method to develop a dialectical methodology for assessing biology as technoscience during the Anthropocene. In this paper I will evaluate this use of Hegelian dialectics in biology. I will first elaborate the meaning of Hegel’s method of “Dialectics”. This helps me to evaluate Zwart’s dialectical scientific methodology from the perspective of Hegel’s method of “Dialectics” and to evaluate Zwart’s dialectical scientific methodology from the perspective of the praxis of biology. Finally, I will oppose Zwart’s claim that the synthetic cell is an appropriate case study to demonstrate the relevance of dialectics for understanding contemporary biological research.
Real-to-virtual domain transfer-based depth estimation for real-time 3D annotation in transnasal surgery: a study of annotation accuracy and stability
Authors: Tong Hon-Sing , Ng Yui-Lun , Liu Zhiyu , Ho Justin D. L. , Chan Po-Ling , Chan Jason Y. K. , Kwok Ka-Wai
Published: March 2021

Purpose

Surgical annotation promotes effective communication between medical personnel during surgical procedures. However, existing approaches to 2D annotations are mostly static with respect to a display. In this work, we propose a method to achieve 3D annotations that anchor rigidly and stably to target structures upon camera movement in a transnasal endoscopic surgery setting.

Methods

This is accomplished through intra-operative endoscope tracking and monocular depth estimation. A virtual endoscopic environment is utilized to train a supervised depth estimation network. An adversarial network transfers the style from the real endoscopic view to a synthetic-like view for input into the depth estimation network, wherein framewise depth can be obtained in real time.

Results

(1) Accuracy: Framewise depth was predicted from images captured from within a nasal airway phantom and compared with ground truth, achieving a SSIM value of 0.8310 ± 0.0655. (2) Stability: mean absolute error (MAE) between reference and predicted depth of a target point was 1.1330 ± 0.9957 mm.

Conclusion

Both the accuracy and stability evaluations demonstrated the feasibility and practicality of our proposed method for achieving 3D annotations.
Von der Allgemeinen Erziehungswissenschaft zur Empirischen Bildungsforschung?
Authors: Westphal Andrea , Zawacki-Richter Olaf
Published: March 2021

Der vorliegende Beitrag beschäftigt sich mit den Publikationen, die in der Zeitschrift für Erziehungswissenschaft (ZfE) in den Jahren 1998–2017 veröffentlicht wurden. Angesichts der Veränderungen in der erziehungswissenschaftlichen Forschungslandschaft in der jüngeren Vergangenheit untersuchen wir, inwiefern sich eine veränderte Schwerpunktsetzung auch in den Beiträgen der ZfE nachweisen lassen. Dazu führen wir zunächst eine quantitative Textanalyse durch und identifizieren die häufigsten sowie die charakteristischen Bigramme (Zweiwortsequenzen) in vier aufeinanderfolgenden Fünfjahres-Abschnitten (1998–2002, 2003–2007, 2008–2012, 2013–2017). Zudem prüfen wir, inwiefern bestimmte Wortstämme (bspw. „erziehungswissenschaft“, „bildungsforsch“, „didakt“) über die Jahre hinweg häufiger auftreten. Schließlich erstellen wir mit dem Textmining Tool Leximancer™ concept maps, die Hinweise auf die semantische Struktur der Themengebiete und Schlüsselkonzepte geben. Die Ergebnisse deuten darauf hin, dass im gesamten Zeitraum mehrheitlich Beiträge mit empirischem Fokus publiziert wurden, ein inhaltlicher Fokus auf sozialen Aspekten von Bildung lag und die Beschäftigung mit der allgemeinen Erziehungswissenschaft abnahm.
Rape myth acceptance as a relevant psychological construct in a gender-unequal context: The Hungarian adaptation of the updated Illinois rape myths acceptance scale
Authors: Nyúl Boglárka , Kende Anna
Published: March 2021

The Updated Illinois Rape Myth Acceptance Scale (UIRMAS) has been widely used for measuring rape myth acceptance. The scale was created in the United States, however studies have shown that rape myth is a culturally and socially embedded phenomenon. Therefore, in order to measure rape myth acceptance in other parts of the world, the scale needs to be validated. Victim blaming and rape myths are both widespread in public reactions to rape in Hungary (i.e., in media reports and public opinion). Furthermore, Hungary can be characterized by a weak feminist movement and scoring low on gender equality measures. Nevertheless, we expected and found the reliability and validity of the Hungarian version of the Updated Illinois Rape myth acceptance Scale (UIRMAS). In Study 1 we conducted a confirmative factor analysis to assess the structural validity of the scale and identified the original factors of UIRMAS on a large convenience sample (N = 758, 25.4% men 74.6% women). In line with previous empirical evidence we also found that men, people with stronger just-world beliefs and higher sexism accepted rape myths more. In Study 2 we again found support for the original factor structure and construct validity of UIRMAS on a nationally representative sample (N = 1007, 49.2% men 50.8% women), and also tested its convergent and discriminant validity. The results suggest that UIRMAS is a valid and reliable scale in the Hungarian context that can, for example, be used for measuring impact assessment of interventions.
New Advances in Biostratigraphy of the Lower/Middle Norian Transition: Conodonts of the Dovško Section, Slovenia
Authors: Karádi Viktor , Kolar-Jurkovšek Tea , Gale Luka , Jurkovšek Bogdan
Published: March 2021

This study presents the results of the conodont biostratigraphy and microfacies analysis carried out on the pelagic limestones of the Upper Triassic Dovško Section in Slovenia, which represents the eastern part of the Slovenian Basin. The age of the section ranges from the Lacian 1 to the Alaunian 1. The Lacian part of the succession is predominantly characterized by the representatives of the genus Ancyrogondolella. Transitional morphologies towards Alaunian faunas first appear in the Lacian 3 and become common during the Lacian-Alaunian transition. This evolutionary development coincides with a shift in microfacies from a dominantly radiolarian-bearing mudstone-wackestone-packstone to a filament- dominated wackestone-packstone, and the formation of small neptunian dykes, which may reflect environmental perturbations and/or a change in basin geometry. The proliferation of the genera Epigondolella and Mockina is observed in the Alaunian part of the section, though the genus Ancyrogondolella is still present in this interval. Systematic description of the conodont taxa is provided, and seven new species and two new subspecies are established. The new advances will be of great value in further biostratigraphic studies, especially in areas without ammonoid faunas, and in the reconstruction of the paleogeography of the Slovenian Basin.
Dealing with Multicollinearity Problem in Analysis of Side Friction Characteristics Under Urban Heterogeneous Traffic Conditions
Authors: Singh Shishodiya Ghanshyam , Kumar S. Vasantha
Published: March 2021

Disturbance to the moving traffic, such as on-street parking and bus stop on carriageway normally called as side friction in the literature, is one of the major problems that impact the vehicular speed on urban arterials in developing countries like India. Regression model to study the mode-wise vehicular speeds using side friction and other influencing variables faces the problem of multicollinearity as there is a strong correlation between the independent variables which results in high R2 and high p values. Instead of dropping the highly correlated variables which is the conventional practice to deal with multicollinearity, we proposed an approach of taking a linear combination and ratio of independent variables. To test the applicability of the proposed approach, traffic data from five locations in Hyderabad and Vellore in India having a wide variety of geometric and traffic characteristics with one or more side frictions were collected and regression models were built. It was found that the proposed approach of taking linear combination and ratio worked very well as the models exhibited a high R2, low p values and low variance inflation factor. The regression models also forecast speeds with good and reasonable accuracy. For better understanding of the impact of side friction, the present study proposed an index called side friction index and it was found that the number of side friction elements and their impact are directly proportional. Also, the impact is not uniform across the different vehicle types. Though the present study focussed on side friction problem, the approach that we proposed in this paper to deal with multicollinearity can be applied in other fields as well.
Temporal network embedding using graph attention network
Authors: Mohan Anuraj , Pramod K V
Published: March 2021

Graph convolutional network (GCN) has made remarkable progress in learning good representations from graph-structured data. The layer-wise propagation rule of conventional GCN is designed in such a way that the feature aggregation at each node depends on the features of the one-hop neighbouring nodes. Adding an attention layer over the GCN can allow the network to provide different importance within various one-hop neighbours. These methods can capture the properties of static network, but is not well suited to capture the temporal patterns in time-varying networks. In this work, we propose a temporal graph attention network (TempGAN), where the aim is to learn representations from continuous-time temporal network by preserving the temporal proximity between nodes of the network. First, we perform a temporal walk over the network to generate a positive pointwise mutual information matrix (PPMI) which denote the temporal correlation between the nodes. Furthermore, we design a TempGAN architecture which uses both adjacency and PPMI information to generate node embeddings from temporal network. Finally, we conduct link prediction experiments by designing a TempGAN autoencoder to evaluate the quality of the embedding generated, and the results are compared with other state-of-the-art methods.
Red-headed ash borer Neoclytus acuminatus acuminatus (Fabricius) (Coleoptera: Cerambycidae): the global distribution, current spreading and the seasonal activity depending on its different habitats
Authors: Keszthelyi Sándor
Published: March 2021

Holarctic expansion of Neoclytus acumiantus acuminatus due to its polyphagous character has been a major concern for plant health, which raises several questions about its spreading directions. I have examined all bibliographical references, phytosanitary reports and authentic photographs from the online sources in the quest of the determination of the worldwide distribution and flight phenology of N. a. acuminatus. The expansion, including the western Palearctic regions as well as several Neotropical habitats, is proven and has become more intense in the last decades. It can be found mostly on the Holarctic region including 21 countries, which is supported by 51 publications. Its spreading mostly northward and eastward in the Palearctic is continuous due to anthropogenic effects, climate change as well as movement of infested materials. Flight phenology and population densities as a function of habitats were mapped. The detection in novel habitats of this pest due to imported wood materials and products is envisaged in the foreseeable future. The primary criterion for controlling the species adapted to the commercial trade and climatic change would be the elaboration of a monitoring system in affected and exposed areas.
30432 Novel insights from single-cell RNAseq analysis of the stromavascular fraction of human adipose tissue
Authors: Wang Bingyao , Smith Gregory , Ruf-Zamojski Frederique , Lee Gyu Ho , Sealfon Stuart C. , Albu Jeanine , Fried Susan K. , Karastergiou Kalypso
Published: March 2021

ABSTRACT IMPACT: Characterizing the cellular composition of human adipose tissue may contribute to the prevention and/or treatment of obesity-associated metabolic diseases. OBJECTIVES/GOALS: Our aims in this study were to use single-cell techniques 1. to characterize cell types within the stromavascular fraction of human adipose tissue, 2. to identify subsets of cells within each type (sub-clustering), 3. to identify gene sets and pathways that may provide information on the function and significance of each cell cluster. METHODS/STUDY POPULATION: Abdominal subcutaneous adipose tissue samples from n=6 healthy volunteers (1M, 5F, age 28-38 y, BMI 24.5-63.0 kg/m2) were collected by aspiration or during surgery. In 3 subjects, all females, paired femoral samples were also collected. After collagenase digestion approximately n=10,000 cells/sample were used for single-cell RNA sequencing using the 10X Genomics platform. After QC and downstream analysis, data were analyzed in Seurat v.3.1.5. We identified first different cell types and then subclusters in an unbiased fashion. Gene Set Enrichment Analysis (GSEA) was used for pathway analysis. RESULTS/ANTICIPATED RESULTS: Progenitor markers are related to extracellular matrix, eg DCN (logFC progenitors vs other cells types=3.17, expressed in 99.7% (pct.1) of progenitors, 26.1% (pct.2) of others). Endothelial and pericytes shared markers like RBP7 (logFC=1.67, pct.1 0.88, pct.2 0.39); pericytes also showed unique markers, eg RGS5 (logFC=2.29, pct.1 0.89, pct.2 0.17). Progenitors are further divided into 11 sub-clusters, one of which showed enrichment of CD36 (high proliferation potential), FABP4 (differentiation), and of the novel marker PALMD (logFC=7.13, pct.1 0.94, pct.2 0.48). All p<10E-5. GSEA analysis suggests that inflammatory pathways are downregulated in both adipose progenitors and endothelial/pericyte cells in the femoral compared to the abdominal depot. DISCUSSION/SIGNIFICANCE OF FINDINGS: Single cell RNA sequencing provides unique insights into the molecular profile of cell types and the identification of novel subsets of cells within the human adipose tissue. Such cellular heterogeneity may explain differences in adipose function between individuals and eventually in the risk of obesity-associated metabolic diseases.
12382 Circadian Disruption in Pancreatic Cancer Carcinogenesis
Authors: Schwartz Patrick B. , Walcheck Morgan T. , Matkowskyj Kristina A. , Bradfield Christopher A. , Ronnekleiv-Kelly Sean M.
Published: March 2021

ABSTRACT IMPACT: Circadian disruption is known to cause significant human pathology but has not been evaluated in pancreas cancer carcinogenesis; through understanding how disruption of circadian rhythms can lead to pancreas cancer development and spread, preventive and therapeutic strategies can be devised. OBJECTIVES/GOALS: Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer due to early spread and poor response to therapy. Identifying factors driving PDAC growth could lead to new therapeutic strategies. Thus, we evaluated the extent to which circadian rhythm disruption, a factor strongly associated with cancer formation, contributes to PDAC pathogenesis. METHODS/STUDY POPULATION: To achieve the objective, we evaluated mice with pancreas lineage Kras-mutation (KC mice), which are predisposed to develop the full spectrum of pancreas cancer precursor lesions (pancreatic intra-epithelial neoplasia or PANIN-1, 2, 3) and PDAC. We subjected KC mice to a light-dark phase shift protocol known to induce circadian disruption (KCCD, n = 18), and another group to standard lighting conditions (KCNC, n = 31), with equal numbers of males and females in each group. The mice were allowed access to food and water ad libitum until sacrifice at age 9 months. Histopathologic evaluation of the pancreas was then performed to assess for pancreatic inflammation, pancreatic precursor lesions (PANIN) and PDAC. Fisher’s Exact Test was used to evaluate differences in incidence. RESULTS/ANTICIPATED RESULTS: As expected, both groups of mice demonstrated 100% incidence of chronic pancreatitis and PANIN-1 (low-grade precursor lesion) at age 9 months. This is consistent with the KC phenotype. However, the KCCD mice demonstrated a significant increase in acute pancreatic inflammation (61.1% vs 19.4%, p = 0.005) compared to KCNC mice. Furthermore, intermediate grade precursor lesions (PANIN-2) were also significantly increase in the KCCD mice (38.9% vs 6.5%, p = 0.006). Incidence of high-grade precursor lesions (PANIN-3, or carcinoma in situ: 22.2% vs 9.7%) and PDAC (27% vs 19%) were also increased, but these were not statistically significant. These results are notable given the established progression from higher grade premalignant PANIN lesions (PANIN-2, PANIN-3) to PDAC. DISCUSSION/SIGNIFICANCE OF FINDINGS: Insight into how circadian disruption leads to increased PANIN-2 formation and increase in acute inflammation may be advantageous for understanding circadian disruption in PDAC carcinogenesis. The circadian clock is present in immune cells and disruption can induce immune dysregulation. This mechanism will be evaluated in follow up studies.
33886 Deregulation of the LANCL2 Pathway in Alzheimer's Disease
Authors: Birnbaumria Emily , Figueiredo-Pereira Maria , Rockwell Patrica , Henchcliffe Claire , Serrano Peter
Published: March 2021

ABSTRACT IMPACT: My work focuses on understanding the role of an immunomodulatory pathway in Alzheimer’s disease as a potential therapeutic target. OBJECTIVES/GOALS: My long-term goal is to develop effective drugs to halt the progression of Alzheimer’s disease (AD). The overall objective of this translational work is to study the deregulation of the Lanthionine Synthetase C-like 2 (LANCL2) immunomodulatory pathway in AD. I hypothesize that stimulation of the LANCL2 pathway will inhibit progression of AD. METHODS/STUDY POPULATION: These studies use a transgenic rat model of AD, and a LANCL2-binding drug, BT-11, that crosses the BBB. Tg-AD Fisher 344 rats express human APP with the Swedish mutation and human presenilin-1 with the ? exon 9 mutation. Tg-AD rats develop age-dependent AD-like pathology. Rodent chow containing BT-11 (10mg/Kg bw) was administered orally from 5 months of age (pre-pathology) to 11 months of age. Eight groups of rats (8 rats/group) were included: WT and Tg-AD x 2 sexes x 2 conditions (BT-11 treated and untreated). Rats are analyzed for: (1) Spatial learning and memory with an active place avoidance test. (2) AD pathology including amyloid plaques and activated microglia, with hippocampal immunohistochemistry. (3) LANCL2-signaling and other AD relevant pathways, with hippocampaI western blot and RNAseq analyses. RESULTS/ANTICIPATED RESULTS: I predict that BT-11 will prevent and/or mitigate some aspects of AD pathology displayed by the AD transgenic rats (Tg-AD). Tg-AD rats treated with BT-11 should perform better in the spatial learning and memory tasks, than the untreated Tg-AD rats. I also expect that the immunohistochemical analysis will reveal reduced pathological hallmarks, including amyloid plaques and reactive microglia, in hippocampal tissue of BT-11 treated versus untreated Tg-AD rats. Through western blot and RNAseq analyses, I will establish how BT-11 treatment affects the LANCL2 signaling cascade as well as other AD relevant pathways, in treated and untreated rats from both sexes and both genotypes. DISCUSSION/SIGNIFICANCE OF FINDINGS: My proposed translational research will address the potential of targeting the LANCL2 pathway to improve AD treatment. Discovering the effects of stimulating the LANCL2 pathway on AD pathology is a novel approach in AD drug development. I expect that my studies with BT-11 will positively influence therapeutic outcomes of this devastating condition.
37161 Triazole-based reversible inhibitors of spermine oxidase and implications for amelioration of neuronal injury
Authors: Furbish Amelia , Woster Patrick
Published: March 2021

ABSTRACT IMPACT: This project aims to investigate the impact of spermine oxidase inhibition on amelioration of neuronal injury. OBJECTIVES/GOALS: Our group has recently described a series of triazole-based reversible inhibitors of spermine oxidase (SMOX) (Holshouser et al. 2019). The purpose of the current project is to optimize our most promising inhibitors by structural modification, and to determine whether they can reduce oxidative damage in models of neuronal injury. METHODS/STUDY POPULATION: A small number of SMOX inhibitors have been described in the literature, however, currently available inhibitors lack selectivity for the enzyme and are associated with dose-limiting toxicity. For this project we used multiple medicinal chemistry techniques to synthesize novel triazole-based analogs of our most potent inhibitors as potential SMOX inhibitors. In addition, we plan to utilize virtual and physical screening methods to identify new potential scaffolds. Compounds with demonstrated activity against SMOX via enzymatic assay will then be evaluated in a cell-based model of neuronal injury. In a preliminary study, we investigated the ability of hydrogen peroxide to induce SMOX expression in an SH-SY5Y neuroblastoma cell line using western blot. RESULTS/ANTICIPATED RESULTS: We found that cellular SMOX protein increases in response to hydrogen peroxide exposure in a dose-dependent manner, indicating that this may be a viable cellular model for testing the efficacy of our experimental compounds. To extend these studies, we have developed a SMOX enzymatic assay that will be used for high-throughput screening of commercial libraries, as well as the South Carolina Compound Collection (SC3), which contains 100,000 proprietary, fully annotated analogs. As hits are identified, they will be synthesized and evaluated for potency and selectivity as SMOX inhibitors. The most potent and selective compounds will then be evaluated in our cellular model of neuronal injury. DISCUSSION/SIGNIFICANCE OF FINDINGS: Studies have linked the overexpression of SMOX and the production of associated toxic byproducts with increased susceptibility to excitotoxic stress and neuronal injury. Our objective is to develop potent and selective inhibitors for this enzyme that can serve as chemical probes for elucidating the role of this enzymatic pathway in neuronal injury.
45724 VC2 Oncolytic Virotherapy Induces Robust Systemic Anti-Tumor Immunity and Increases Survival in an Immunocompetent B16F10-derived Mouse Melanoma Model
Authors: Uche Ifeanyi Kingsley , Fowlkes Natalie , Vu Luan , Watanabe Tatiane , Carossino Mariano , Nabi Rafiq , Del Piero Fabio , Rudd Jared S. , Kousoulas Konstantin G. , Rider Paul J.F.
Published: March 2021

ABSTRACT IMPACT: Our data demonstrate that VC2 oncolytic virotherapy has significant clinical potential. OBJECTIVES/GOALS: Use our novel oncolytic herpes simplex virus type I (HSV-1), VC2, to understand how oncolytic virotherapy affects the immunosuppressive tumor microenvironment as a mechanism of efficacy. METHODS/STUDY POPULATION: We tested the efficacy of VC2 as an oncolytic virotherapy (OVT) in a syngeneic B16F10-derived mouse model of melanoma. We modified the B16F10 to express nectin-1 (B16F10n-1), the major receptor for HSV-1. Engrafted B16F10n-1 tumors were intratumorally treated with either phosphate-buffered saline (PBS) or 1x10^6 pfu VC2. At indicated time points, treated tumors were excised and processed for immunohistochemistry or flow cytometry analysis. For our experimental metastasis studies, mice were intravenously challenged with B16F10n-1 cells. For our depletion studies, CD4+ and CD8+ T cells were depleted in mice by treatment with mouse anti-CD4 and anti-CD8 monoclonal antibodies respectively, while the control mice were given Rat IgG2b isotype. RESULTS/ANTICIPATED RESULTS: We found that VC2 slowed tumor growth rates and significantly enhanced survival times over control treated mice. VC2-treated mice that survived initial tumor engraftment were able to reject a second tumor challenge and were also resistant to lung colonization (experimental metastasis) of tumor cells. Furthermore, VC2 treatment promoted increased intratumoral T cell infiltration and induced a strong antitumor effect that decreased growth rates of distant, untreated tumors. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our data demonstrate that VC2 OVT has significant clinical potential. Furthermore, due to the increased survival rates and CD8+ T cells dependence, our model will enable study of the immunological correlates of protection for VC2 OVT and OVT in general, as well as to inform the rational design of future OVs with improved therapeutic potentials.
48019 Create a mouse model of chronic sleep deprivation by specific-neuron targeted ablation
Authors: Imamura Toshihiro , Pack Allan I.
Published: March 2021

ABSTRACT IMPACT: A mouse model of minimally-invasive chronic sleep deprivation is essential for elucidating the impact of sleep deprivation on various health issues, and it would lead to the possibility of sleep as a therapeutic target. OBJECTIVES/GOALS: The lack of sleep has been associated with various health conditions. In mice, sleep deprivation has been achieved mainly by physical disturbances, which raises concern about confounding effects by stresses. Without physical disturbance, targeted neuron ablation can address this methodological flaw. METHODS/STUDY POPULATION: AdultVgat-IRES-cre mice undergo a stereotaxic injection of adeno-associated virus (AAV) vector containing mCherry-dtA to bilateral parafacial zone (PZ) to perform GABAergic neuron-specific cell ablation. Control mice receive an injection of AAV vector containing hSyn-DIO-mCherry. All mice are implanted with electroencephalogram and electromyogram (EEG/EMG) electrodes for sleep-wake analysis. After 7-10 days of the postoperative recovery period, mice are kept individually in a cage for sleep-wake state recording. EEG/EMG and video recording are used to measure total wake time, total sleep time, percent of rapid eye movement (REM) and non-REM sleep, and detailed characterization with spectral analysis. RESULTS/ANTICIPATED RESULTS: We anticipate that the ablation of GABAergic neurons in bilateral PZ decreases the fraction of sleep state in mice, especially non-REM sleep. In the Vgat-IRES-cre mice that received the injection of AAV vector containing mCherry-dtA, total sleep time is expected to be decreased constantly during the 8-week observation period. Sleep-wake staging by video activity recording is anticipated to be closely correlated with the gold standard staging by EEG/EMG. Possible stresses caused by the restriction of physical activity and handling of mice for EEG/EMG recording can be further minimized by the sleep-wake staging performed with the video activity recording. DISCUSSION/SIGNIFICANCE OF FINDINGS: The lack of sleep has been associated with negatively affecting overall health and is implicated in major health conditions including obesity, diabetes, and cardiovascular diseases. This chronic sleep deprivation mouse model can be used to understand the mechanisms of such detrimental effects on health, and would improve many health conditions.
49483 Evaluating the Role of IFNLR1 Receptor Dynamics and Plasticity in Regulating Cellular Response to Interferons
Authors: Evans Gray , Kappler Christiana S. , Liu Ray , Carten Juliana D. , Orr Cody M. , Stephenson Sarah , Traktman Paula , Duncan Stephen A. , Meissner Eric G.
Published: March 2021

ABSTRACT IMPACT: We hope to provide a more nuanced understanding of the type-III IFN system, thereby exploring its therapeutic potential in the realm of infectious diseases. OBJECTIVES/GOALS: The role of IFNLR1 receptor dynamics and plasticity in regulating the type-III IFN response is largely unknown. As a specific, powerful component of innate immunity, understanding how the type-III IFN system is regulated could lead to the development of novel therapeutic targets and strategies to face a multitude of viral illnesses. METHODS/STUDY POPULATION: To facilitate our investigation, we will generate doxycycline-inducible FLAG-tagged IFNLR1-expression plasmids representing all known transcriptional variants. These plasmids will allow us to: 1) Evaluate the effect of IFNLR1 surface abundance on the type-III IFN transcriptional profile and 2) Assess the extent of IFNLR1-FLAG co-localization with several notable intracellular structures using immunofluorescence, before and after stimulation with IFNL3. RESULTS/ANTICIPATED RESULTS: We have successfully generated three IFNLR1-FLAG transcriptional variants and confirmed inducible-expression and function in vitro. We are currently assessing the role of surface abundance, internalization, differential isoform expression, and trafficking. DISCUSSION/SIGNIFICANCE OF FINDINGS: By completing this study, we hope to provide a more nuanced understanding of the type-III IFN system, thereby exploring its therapeutic potential in the realm of infectious diseases.
68722 Role of ER calcium in beta cell senescence and diabetes pathophysiology
Authors: Weaver Staci A. , Kono Tatsuyoshi , Syed Farooq , Bone Robert , Evans-Molina Carmella
Published: March 2021

ABSTRACT IMPACT: The proposed study has the potential to inform new paradigms of type 1 diabetes prevention and therapy with the overall goal of improving β cell health during autoimmunity. OBJECTIVES/GOALS: Type 1 diabetes (T1D) results from immune-mediated destruction of pancreatic βcells. Recent data suggest that activation of senescence and acquisition of a senescence associated secretory phenotype (SASP) by βcells may contribute to T1D pathogenesis. However, the molecular mechanisms responsible for this phenotype are not well understood. METHODS/STUDY POPULATION: We hypothesize that loss of endoplasmic reticulum (ER) Ca2+ induces βcell senescence, SASP as well as mitochondrial dysfunction which drive T1D development. The current study utilizes SERCA2 KO INS-1 βcells (S2KO) exhibiting loss of ER Ca2+ and a SERCA2 haploinsufficient mice on a non-obese diabetic background (NOD-S2+/-) to test the role of ER Ca2+ loss during T1D development. Senescence associated βgalactosidase staining (SA-βgal), expression of senescence markers (RT-qPCR), mitochondrial function (Seahorse, TMRM) and mitochondrial copy number (qPCR) were all measured in S2KO versus WT βcells and are currently being measured in the NOD-S2+/- mouse model at 6, 8, 12, 14, and 16wks of age. RESULTS/ANTICIPATED RESULTS: RT-qPCR assays detecting senescence markers cdkn1a and cdkn2a and mitochondrial specific genes cox1 and nd1 were developed and validated in both INS-1 βcells and mouse islets. Mitochondrial function assay (Seahorse) was optimized for use in INS-1 βcells and is currently under development for use in intact mouse islets. S2KO βcells displayed increased SA- βgal staining as well as increased mitochondrial coupling efficiency (p=0.0146) and baseline mitochondrial copy number (p=0.0053) compared to WT βcells, suggesting a senescence phenotype and altered mitochondrial function. NOD-S2+/- mice exhibited increased expression of the senescence marker cdkn2a in the islet at 12wks (p=0.0117) compared to control mice, whereas cdkn1a remained unchanged across all timepoints tested. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our results suggest that loss of SERCA2 and reduced ER Ca2+ alter βcell mitochondrial function and are associated with features of senescence. Future studies will test whether SERCA2 activation and/or senolytic/senomorphic drugs are able to prevent or delay diabetes onset in NOD-S2+/- mice.
89723 CD105-targeted CAR T cells for the treatment of acute myeloid leukemia
Authors: Lontos Konstantinos , Wang Yiyang , Frisch Andrew , Colbert Mason , Lohmueller Jason , Delgoffe Greg M.
Published: March 2021

ABSTRACT IMPACT: Our work might lead to a new treatment for patients with acute myeloid leukemia OBJECTIVES/GOALS: Acute myeloid leukemia (AML) is a devastating hematologic malignancy, with dismal 5-year survival. Chimeric antigen receptor (CAR) T cells have been approved for B cell malignancies but not for AML. The goal of this study is to explore the safety and efficacy of CAR T cells targeting CD105 (endoglin) to treat AML. METHODS/STUDY POPULATION: We have constructed human and murine CAR T cells targeting CD105. The CARs were created by sequencing the V(D)J regions of hybridomas and designing single chain variable fragments that target CD105 which were subsequently introduced in a CAR backbone via Gibson assembly. The CAR T cells were produced via transduction using retrovirus or lentivirus. Leukemia cell lines were assessed for CD105 expression with flow cytometry. Killing assays were performed via measurement of luminescence of target cells after co-culture with CAR T cells. Activation assays were performed with co-culture of CAR T cells and target cells and measurement of activation markers with flow cytometry. To assess in vivo efficacy and safety, murine CAR T cells were infused into C57BL/6J mice carrying B16 melanoma after lymphodepletion. RESULTS/ANTICIPATED RESULTS: All human leukemia cell lines assessed (Nalm6, MOLM-14, MV4-11, Kasumi-1, THP-1) expressed some degree of endoglin apart from the T cell leukemia Jurkat. Human CD105 CAR T cells were activated by co-culture with leukemia cell lines and effectively killed leukemia cells in vitro in a CD105-specific manner. Murine CAR T cells killed efficiently both murine solid tumors (B16 melanoma) and murine leukemias (C1498) in vitro. Murine CAR T cells did not exhibit any toxicity when infused after low-dose lymphodepletion (cyclophosphamide 100mg/kg) but caused significant morbidity after higher doses (cyclophosphamide 200mg/kg). Murine CAR T cells delayed the growth of B16 melanoma in immunocompetent mice. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have constructed human and murine CD105 CAR T cells with excellent activity in vitro. The activity of human CD105 CAR T cells in xenografts and the biologic relevance of the toxicity of murine CD105 CAR T cells in humans needs to be further investigated. CD105 CAR T cells might prove an important therapeutic option for patients with AML.
34646 DEFICIENCY OF NOVEL ADIPOKINE TETRANECTIN INCREASES OBESITY AND INSULIN RESISTANCE IN FEMALES
Authors: Plasko George , He Sijia , Zhang Jingjing , Liu Fen , Bai Juli , Dong Lily , Liu Feng
Published: March 2021

ABSTRACT IMPACT: Novel adipokines like tetranectin help explain why some people progress from obesity to diseases like diabetes, atherosclerosis, and dislipidemia OBJECTIVES/GOALS: Obesity has an established association with diabetes, dyslipidemia, and atherosclerosis. Preventing progression from obesity to insulin resistance requires understanding of the regulatory mechanisms involved in the loss of insulin sensitivity. Adipose tissue is well known to function as an endocrine organ that produces many kinds of adipokines. METHODS/STUDY POPULATION: Blood sample analysis from human patients and mice was used to determine associations between tetranectin and obesity. Samples were tested with a monoclonal anti-tetranectin antibody for detection with western blot. A tetranectin mutant knock out mouse line was compared to wild type littermates on high fat diet for 4 months. Insulin tolerance tests and glucose tolerance were used to determine progression to insulin resistance and glucose intolerance. Histological analysis of metabolic tissue was used to demonstrate adipocyte hypertrophy and liver steatosis. RESULTS/ANTICIPATED RESULTS: In the current study, we report the identification and initial characterization of a novel adipokine tetranectin. Tetranectin, which is coded by the C-type lectin domain family 3 member B (CLEC3B) gene, is ubiquitously expressed in various mouse tissues, whereas it is highly enriched in white adipose tissue. We found that the serum level of tetranectin was much higher in both obese and diabetic patients. Knocking out the tetranectin gene in mice protected against glucose intolerance in males but reduced insulin and glucose tolerance in females, without effects on food intake and body weight for either sex. Mechanistically, tetranectin targets liver tissues and its deficiency increases lipid accumulation in hepatocytes in females. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have identified a novel adipokine which mediates a different metabolic crosstalk among tissues to maintain systemic glucose and lipid metabolism in different genders. Further investigation of tetranectin’s function could yield a new target for precise therapeutic treatment for obesity and its associated metabolic diseases in different genders
92180 Prevalence of Clostridioides difficile strains found in Texas soil
Authors: Zeidan Amina R. , Strey Kelsey , Vargas Michelle N. , Reveles Kelly R.
Published: March 2021

ABSTRACT IMPACT: This work investigates C. difficile strains in soil as a potential exposure for gut colonization and community-acquired infection of C. difficile. OBJECTIVES/GOALS: Identifying environmental sources of C. difficile can inform how non-hospital reservoirs can potentially contribute to C. difficile exposure and subsequent gastrointestinal colonization. The objective of the study was to identify C. difficile and toxin genes across various soil sources. METHODS/STUDY POPULATION: This was a cross-sectional study utilizing soil samples obtained throughout Texas, USA. All samples were collected between August and November of 2019 and 2020. Samples were taken from human and animal high contact areas, such as recreational parks. Samples were stored at -80oC until processing. DNA extractions were performed using the DNeasy Powersoil Pro Kit (Qiagen) per manufacturer’s instructions. Real-time PCR was also performed on extracted DNA using the Microbial DNA qPCR Multi-Assay Kit for Clostridium difficile Pathogenicity (Qiagen) for the identification of C. difficile, toxin A (TcdA), and toxin B (TcdB) genes. RESULTS/ANTICIPATED RESULTS: A total of 137 soil samples including dry dirt, sand, and wet soil near water sources were collected and processed for the presence of C. difficile. These included samples from parks and trails (42.3%), water sources (36.5%), and other public spaces (21.2%). C. difficile was identified in 59 (43.1%) soil samples: 6 (4.4%) with Toxin A and 2 (1.5%) with toxin B production. C. difficile was most prevalent among samples taken from parks and trails (50.0%), followed by other public spaces (48.3%), and water sources (32.0%). The median (IQR) Cq value for the C. difficile gene was 39.24 (33.45-40.47) among samples that tested positive. DISCUSSION/SIGNIFICANCE OF FINDINGS: We identified a high prevalence of Clostridioides difficile in soil samples, though toxin gene detection prevalence was low. Future studies will analyze other sources, including water and varying surface samples to obtain a comprehensive view of C. difficile in the environment.