bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis on the Construction of School-enterprise Cooperation Base for Transportation Majors
Authors: Lu Mingyu , Feng Pengfei
Published: June 2021

The transportation specialty cannot be combined with reality in the construction. Traffic engineering and transportation are the combination of my country’s existing planning, and the profe ssional potential is huge. The main mission of applied universities is to cultivate high-quality applie d talents. Serving local economic and social development, the school-enterprise cooperation practice education base is the most important way to achieve the above goals. It is of great significance to i mprove the quality of application-oriented talent training and promote the technological transformati on and upgrading of enterprises. The construction is guided by the educational concept of “enterpri se-oriented, project-driven, ability training and overall development” to build a teaching platform for production, study and research. Establish a curriculum system that can optimize the corresponding curriculum, strengthen practical teaching, Better promote the training of talents towards a good situat ion and continue to develop steadily. Transportation majors are under construction.
Research on the Innovation of Supply Chain Financial Operation mode in the Pharmaceutical Circulation Field---Taking the Accounts Receivable Securitization as an Example
Authors: Kong Yan , Wang Yanna
Published: June 2021

In recent years, the scale of China’s pharmaceutical circulation market has continued to expand, and the government has issued relevant policies to regulate the market. For example, the implementation of two-vote system has led to the increase of accounts receivable of pharmaceutical circulation enterprises year by year with a decrease of accounts receivable turnover rate and an increasing pressure of capital turnover. The enterprise accounts receivable securitization is an important way for enterprises to broaden financing channels, reduce financing costs, revitalize existing assets and improve the efficiency of asset use, while the securitization of statement type assets is the only standardized financial tool to reduce the leverage ratio of enterprises and realize financial optimization. Asset securitization can relieve the pressure of capital turnover and reduce the cost of financing. The case analysis part of this paper analyzes the factors that promote the financial innovation of enterprise’s supply chain from the macro and micro perspectives. And also from the perspective of the capital demand of upstream and downstream enterprises, this part analyzes the motivation of developing supply chain financial innovation. the innovation of supply chain financial operation mode.
Research on the regional public brand of agricultural products based on block chain technology—Exploration of digital economy innovation
Authors: Meng Weiwei
Published: June 2021

Structural oversupply pushes fresh agricultural products to the buyer’s market, and the market’s demand for regional public brands of agricultural products is increasing day by day. Based on block chain technology, this paper analyzes the countermeasures for the construction of regional public brand of agricultural products, which is also an effective measure to promote digital economy innovation and accelerate the promotion of digital industrialization..
Research on outsourcing level measurement and influencing factors of China’s agricultural listed companies
Authors: Ren Jianhua , Lei Hongzhen
Published: June 2021

Outsourcing is beneficial for enterprises to integrate production factors, strengthen core forces and realize value chain appreciation. Based on the data of 42 agricultural listed companies from 2,015 to 2,019, the outsourcing level of agricultural enterprises was measured, and the influencing factors of agricultural enterprise outsourcing were analyzed. The results show that the level of outsourcing of agricultural enterprises in 2015 to 2,019 showed a slow downward trend, the outsourcing level of agricultural enterprises has policy continuity, and the outsourcing level of the current period is affected by the outsourcing level of the previous period; the larger the market scale, the higher the proportion of business outsourcing; and the transaction costs significantly inhibit the outsourcing decision of agricultural enterprises. Finally, the paper puts forward the policy suggestions to promote the healthy development of the agricultural outsourcing market.
An Efficient and Practical Chemoenzymatic Route to (3R,3aR,6R,6aR)-Hexahydrofuro[3,2-b]furan-6-amino-3-ol (6-Aminoisomannide) from Renewable Sources
Authors: Zullo Valerio , Iuliano Anna , Petri Antonella
Published: June 2021

The synthesis of 6-aminoisomannide is easily achieved starting from the renewable, inexpensive, and commercially available isosorbide in 66% overall yield. A biocatalyzed highly regioselective acetylation of the 3-endo hydroxyl group of isosorbide was followed by the stereospecific interconversion of the 6-exo hydroxyl group into an azido group, through reaction with trifluoromethanesulfonic anhydride, followed by nucleophilic displacement of the triflate group by sodium azide. Finally, reduction of the azido group and deacetylation of the 3-hydroxy group were performed in one pot using LiAlH4.
Orbital angular momentum communications based on standard multi-mode fiber (invited paper)
Authors: Wang Jian , Chen Shi , Liu Jun
Published: June 2021

Orbital angular momentum (OAM) modes, having unique properties of a helical phase structure and doughnut intensity profile, have been widely studied in fiber-optic communications, in terms of OAM modulation and OAM multiplexing. In general, different types of specialty fibers with a ring-shape structure are preferred for more stable OAM transmission, which, however, may face greater manufacturing challenge and larger fiber loss compared to standard multi-mode fibers (MMFs). Therefore, the widely deployed and commercially available standard MMFs that can support hundreds of OAM modes have recently attracted great attention. In this paper, we review recent research progress in OAM communications based on standard MMFs. First, the basic concept of OAM and different types of specially designed OAM fibers are briefly introduced. Then, the OAM mode properties in MMFs and recent works, including OAM mode modulation, multiple-input multiple-output (MIMO)-free OAM mode group multiplexing, small-scale partial MIMO assisted OAM mode multiplexing, and OAM-based heterogeneous fiber-optic networks, are presented. The OAM communications using other widely deployed standard single-mode fibers are also briefly introduced as supplementary. Finally, key challenges and perspectives of OAM communications based on standard MMF are discussed and summarized.
Time-domain spectroscopy of methane excited by resonant high-energy mid-IR pulses
Authors: Crippa G , Faccialà D , Prasannan Geetha P , Pusala A , Musheghyan M , Assion A , Bonanomi M , Cinquanta E , Ciriolo A G , Devetta M , Fazzi D , Gatto L , De Silvestri S , Vozzi C , Stagira S
Published: June 2021

We describe the implementation of nonlinear time-domain spectroscopy of rotovibrational IR-active modes in methane through broadband Four-Wave Mixing driven by resonant high-energy mid infrared laser pulses. At high driving pulse intensities we observe an efficient vibrational ladder climbing triggered in the molecules. This study opens the possibility to impulsively and selectively excite molecules of biological interest to high-lying vibrational states and to characterize their dynamics.
Breathing New Life into the German Utility Model System in the Industry 4.0 Era
Authors: Takenaka Toshiko
Published: June 2021

Key players of Industry 4.0, high-tech firms and individual inventors, do not use patents to exclude others due to the open source philosophy and the risk of a counter-infringement assertion. These key players have developed new uses of patents: (1) defensive use to avoid litigation and (2) proactive use to promote open innovation. They want to be rewarded with the freedom to operate instead of monopoly profits through exclusivity. To better serve the key players in light of these new uses, the German utility model system should be reformed by incorporating a statutory license to implement the open source philosophy and adopting a simplified claiming and disclosure format. The current German translation requirement should be removed to globalize the system.
Alpha‐synuclein binds to neuron‐specific glycans
Authors: Rhoades Elizabeth , Birol Melissa
Published: June 2021

Fibrillar deposits of the intrinsically disordered protein α‐Synuclein (αS) are the defining pathological feature of Parkinson's disease, the second most common neurodegenerative disorder. αS is abundant in the brain, where its localization to presynaptic termini and ability to stabilize curved membranes support a role in synaptic vesicle exocytosis/endocytosis, although its precise function is not known. N‐terminal acetylation is universally present on αS in vivo, in samples derived from both healthy persons and Parkinson's disease patients. Prior work from our group and others have demonstrated that N‐terminal acetylation (αSacetyl) impacts the fundamental biophysical properties of αS. Here, using cell‐derived proteolipid vesicles, we find that αSacetyl binds specifically to neuronal‐lineage cell membranes. Cleavage of complex N‐linked glycans dramatically decreases binding by αSacetyl while having a negligible impact on binding by the unmodified protein, αSunmod. αSacetyl selectively binds to glycans cleaved from cultured neuronal cells and removal of cell‐surface glycans dramatically reduces its cellular uptake. Based on these findings we propose that N‐terminal acetylation confers glycan recognition and binding to αS and that it is a critical modification for functional interactions with cellular protein binding partners. Moreover, understanding the basis of this interaction may have critical implications in obtaining a clearer understanding in the function of αSacetyl.

Support or Funding Information

NIH NINDS NS079955

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using lipidomics pathway analysis to identify potential therapeutic targets
Authors: Wakelam Michael , Nguyen An , Zhang Qifeng
Published: June 2021

Advances in LC‐MS/MS methodologies have resulted in an appreciation of the complexity of the lipid composition of cells and tissues. Theoretically mammalian cells can contain many thousands of individual lipid molecular species and whilst it is unlikely that all are present in a single cell, lipidomic experiments have demonstrated the presence of more than a thousand species. The breadth of this species diversity is a particular consequence of the range of fatty acid species and this emphasises the importance of fatty acid synthases, desaturases and elongases in regulating cell function. The integrated regulation of changes in lipid species that occur during and that can regulate cell functions, including signalling and metabolism, highlights the need for bioinformatics analysis to fully interpret lipidomics data.

We have adopted two experimental systems: colorectal tumour tissue and rhinovirus‐infected human bronchial epithelial cells, to develop this analysis. Lipidomic analysis that compared colorectal tumour tissue of defined stage with matched normal tissue showed changes in more than 700 lipid species, this also demonstrated considerable changes in acyl chain saturation and chain length accompanying tumour progression. In the rhinovirus‐infected cells there was considerable temporal‐associated changes in a number of lipids including PIP, cardiolipin, PC and DG, there were also clear changes in the acyl chains of a number of neutral and phospholipids.

Pathway analysis highlighted key changes in sphingolipid metabolising enzymes in both experimental systems including sphingosine kinase and ceramide synthase activities. Further, adopting the network optimising Prize‐collecting Steiner tree problem methodology, we were able to identify fatty acid synthases, acyl transferases and ceramidases as potential therapeutic targets. The importance of a number of these activities in viral infection was confirmed by observing inhibitory effects of selective inhibitors upon infection of human bronchial epithelial cells by rhinoviruses.

The lipidomics‐determined changes also point to important roles for remodelling of membrane lipid acyl chains suggesting that changes in membrane fluidity, protein binding or curvature may have important roles in infection and disease progression.

Support or Funding Information

Supported by grants from the Medical Research Council (MR/M004821/1) and the Biotechnology and Biological Sciences Research Council (BBS/E/B/000C0432; BB/P013384/1).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A systems‐level understanding of glycosylation signaling networks
Authors: Hsieh‐Wilson Linda C.
Published: June 2021

The dynamic posttranslational modification of proteins by O‐linked‐β‐N‐acetylglucosamine (O‐GlcNAc glycosylation or O‐GlcNAcylation) plays many diverse roles in cellular physiology and disease. We will describe a systems‐level approach for studying the function, specificity, and dynamic regulation of O‐GlcNAcylation. Elucidation of the signaling networks associated with the O‐GlcNAc modification provides new insights into its central functions and may reveal new therapeutic approaches to enable its selective modulation.

Support or Funding Information

NIH (R01 GM084724)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Transcriptomics profiling of breast cancer cells, MDA‐MB‐231, exposed to TNF‐α: focus on pro‐survival and pro‐inflammatory regulated pathways.
Authors: Bauer David F. , Mazzio Elizabeth A. , Soliman Karam F.
Published: June 2021

Triple‐negative breast cancer (TNBC) is characterized by the absence of estrogen, progesterone and HER2 receptors. TNBC is considered to have a poor prognosis due to the limited treatment options with the exclusion of hormone receptor‐based chemotherapies. Aggressive TNBC growth is accompanied by infiltration of tumor‐associated macrophages which can establish a perpetual state of inflammation rendering sustained release of proinflammatory cytokines such as tumor necrosis factor (TNF)‐α, which can lead to immunological evasion. In this work, we evaluated the effects of TNF‐α on the changes to the whole transcriptome: including mRNAs, long intergenic non‐coding RNA transcripts (lincRNA) in MDA‐MB‐231 cells. The data show enormous upregulation (60‐fold change FC, p<.0001) in CXCL8 chemokine (C‐X‐C motif) ligand 8, with a statistically significant elevation of other transcripts including IL‐6 (+10.47), TNFRSF9 (9.58), TNFAIP3 (+7.52) and CCL2 (+3.5). On the other side, there was a reduction in numerous non‐coding microRNA including MIR29A (−8.8), MIRLET7A2 (−8.7), MIR222 (−4.91), MIR100 (−4.91), MIR27B (−4.68) and MIR181B1 (−7.95) in addition to Glutathione S – transferase (GSTT2) (−5.59). Pathway analysis shows that the most relevant effects of TNF‐α are on driving NF‐kB related survival, cytokines, and inflammatory pathway elements and TNF‐α related pathways.

Support or Funding Information

Supported by grants from the National Institute on Minority Health and Health Disparities RCMI grant G12MD007582 and COE grant P20 MD006738.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Excessive UBE3A dosage impairs retinoic acid signaling and synaptic plasticity in autism spectrum disorders
Authors: Hu Ronggui
Published: June 2021

The autism spectrum disorders (ASDs) are a collection of human neurological disorders with heterogeneous etiologies. Hyperactivity of E3 ubiquitin (Ub) ligase UBE3A, stemming from 15q11‐q13 copy number variations, accounts for 1%–3% of ASD cases worldwide, but the underlying mechanisms remain incompletely characterized. Here we report that the functionality of ALDH1A2, the rate‐limiting enzyme of retinoic acid (RA) synthesis, is negatively regulated by UBE3A in a ubiquitylation‐dependent manner. Excessive UBE3A dosage was found to impair RA‐mediated neuronal homeostatic synaptic plasticity. ASD‐like symptoms were recapitulated in mice by overexpressing UBE3A in the prefrontal cortex or by administration of an ALDH1A antagonist, whereas RA supplements significantly alleviated excessive UBE3A dosage‐induced ASD‐like phenotypes. By identifying reduced RA signaling as an underlying mechanism in ASD phenotypes linked to UBE3A hyperactivities, our findings introduce a new vista of ASD etiology and facilitate a mode of therapeutic development against this increasingly prevalent disease.

Support or Funding Information

This work was founded by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000 and XDA12040323), the Natural Science Foundation of China (31470770 and 81525019 to RH), the Ministry of Science and Technology of China (2013CB910900 to RH).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Attenuating Effects of 1,2,3,4,6 Penta‐O‐Galloyl‐β‐D‐Glucose (PGG) on the Expression of Proteins Involved in Alzheimer's Disease in LPS/IFNγ Activated BV‐2 Microglial Cells.
Authors: Mendonca Patricia , Taka Equar , Soliman Karam F.
Published: June 2021

Alzheimer's disease (AD) is a chronic and progressive neurodegenerative disorder associated with memory and cognition impairment. The main pathological hallmarks are abnormal intraneuronal cytoskeletal changes, known as neurofibrillary tangles, and the extracellular protein deposits of amyloid beta plaques (Aβ). Although the currently available drugs for AD do not prevent or reverse the formation of Aβ deposits, medicinal plants have been reported for possible anti‐AD activity and are promising therapeutic agents for neurodegenerative disorders. 1,2,3,4,6‐Penta‐O‐galloyl‐β‐D‐glucose (PGG), which is a naturally occurring polyphenolic compound, has shown to exhibit anti‐aggregation activity on Alzheimer's Aβ proteins. Its oral intake reduced Aβ plaque burden and Aβ peptide content in brain tissue of transgenic mice. In this study, we investigated the effect of PGG on the expression of proteins involved in neurodegenerative diseases in LPS/IFNγ activated BV‐2 microglial cells. Proteomic results identified 17 proteins whose expression level was significantly downregulated by PGG, including septin‐7, ataxin‐2, and adenylosuccinate synthetase isozyme 2 (ADSS). These proteins were previously described as being expressed in neurodegenerative diseases and/or involved in the Wnt signaling pathway, which is associated with the development of neuronal circuits and control of AD pathogenesis. PGG inhibitory effect on ataxin‐2, septin‐7, and ADSS was confirmed at the protein and transcription level, and the proteins were classified according to biological processes and molecular functions using NCBI and PANTHER databases. Since these selected proteins are involved in synapse impairment induced by Aβ protein, they are interesting candidates for possible involvement in the underlying mechanisms of synaptic dysfunction in neurodegenerative diseases. Therefore, these results suggest that PGG may have a role in halting neuropathologic effects of AD by targeting ataxin‐2, septin‐7, and ADSS.

Support or Funding Information

Supported by NIH Grants G12 MD007582 and P20 MD 006738

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elp1‐dependent Shp1 Phosphatase Regulation and its Essential Role in Familial Dysautonomia Pathogenesis
Authors: Tourtellotte Warren G.
Published: June 2021

Familial Dysautonomia (FD; Riley‐Day Syndrome; HSAN3) is a rare heritable disease characterized by debilitating sensory and sympathetic neuropathy. It is caused by a germline mutation of the Elp1 gene that leads to exon mis‐splicing, nonsense‐mediated truncation of the Elp1 protein and loss of Elp1 protein primarily in sympathetic and nociceptive sensory neurons. How Elp1 functions in FD disease‐vulnerable neurons is very poorly understood.

Using Elp1 conditional knockout mice that we generated to recapitulate the molecular and physiologic phenotypes associated with human FD, we identified abnormalities in sympathetic neurons that explain the pathophysiologic basis for disease. Sympathetic neurons isolated from Elp1 conditional knockout mice have abnormalities in nerve growth factor signaling which is essential for their survival. NGF is normally acquired from peripheral tissues by innervating axons and NGF retrograde signal transduction to the neuron cell body is essential for their normal survival and differentiation. We found that sympathetic neurons lacking Elp1 have abnormalities in retrograde NGF signaling and as a consequence they have impaired survival and differentiation in response to NGF. After binding to NGF, the terminal axon TrkA (NTRK1) receptors are internalized and phosphorylated to activate downstream signaling pathways essential for sympathetic neuron survival and differentiation. We found that Elp1‐ deficient neurons normally bind NGF and they normally internalize and activate (phosphorylate) TrkA receptors in response to NGF. However in the absence of Elp1, TrkA receptor phosphorylation, which is essential for activation and downstream signaling, was markedly diminished. Shp1 phosphatase, which normally binds to TrkA receptors and terminates signaling by dephosphorylation, was found to be hyperactivated in the absence of Elp1. Shp1 hyperactivation results in precocious TrkA receptor dephosphorylation and attenuation of retrograde signaling. Pharmacological treatment with either Shp1 phosphatase inhibitors or molecular inhibition of Shp1 phosphatase activity resulted in complete rescue of TrkA dephosphorylation and restoration of normal NGF‐dependent retrograde neuron survival.

These results demonstrate that sympathetic neuron death in patients with FD is due to loss for normal retrograde NGF signaling mediated by Shp1 phosphatase hyperactivity and precocious attenuation of TrkA signaling. Inhibition of Shp1 phosphatase activity may provide a novel therapy for sympathetic neuron loss in FD. Future studies are focused on understanding how Elp1 regulates Shp1 phosphatase activity and whether it may regulate Shp1 phosphatase activity in other signal transduction pathways involved in immune function and oncogenesis.

Support or Funding Information

Support: NIH: NICHD, NINDS and NOD.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Search for Hidden Germline Mutations in Bilateral Retinoblastoma
Authors: Okonkwo Oluchukwu
Published: June 2021

Retinoblastoma (RB) is a cancer in the retina that develops in children usually under the age of five and can cause blindness, loss of the affected eye, or death by metastasis, if not diagnosed and treated early. It arises as a result of bi‐allelic inactivation of the RB1 gene in a retinal cell that prevents the production of the pRB protein, causing uncontrollable cell proliferation in the retina. The Genetic Diagnostic Laboratory in the Perelman School of Medicine at the University of Pennsylvania has used Sanger sequencing to identify disease‐causing mutations in the RB1 gene in thousands of individuals diagnosed with bilateral retinoblastoma. Current testing focuses on coding regions and splice site consensus sequences, and various mutations have been found in 97% of individuals tested. This study aimed to identify pathogenic exonic or deep intronic mutations in the germline of six individuals which belong to the 3% of mutation‐negative cases. For three of these individuals, RNA isolated from the lymphocytes of the whole blood was converted into cDNA, RB1 transcript amplified with gene specific primers and sequenced by Sanger sequencing. The cDNA of RB1 had indicated one sample had a heterozygous deletion of exon 10, but the cause of the exonic deletion was unknown. To investigate this, several primer pairs were designed to amplify and sequence regions flanking the deleted region in the genomic DNA, and an inversion between exons 10 and 11 was suspected. For the other two patients, who are sisters, an exonic deletion of exon 2 was found but proven to be benign. For the other three individuals, previous Sanger sequencing of exons had not indicated the presence of any pathogenic mutations. An AmpliSeq panel of three hundred and four primer pairs was designed to amplify and sequence both coding and non‐coding regions spanning the RB1 gene. Amplicons were prepared using genomic DNA isolated from these three individuals and next generation sequencing on the Ion Torrent PGM platform was performed. Upon analysis, three pathogenic mutations present at low allele frequencies were identified in each individual. These were low frequency mosaic mutations that may not be detected through Sanger sequencing. Therefore, the AmpliSeq design panel can provide information about intronic mutations and low frequency mosaic mutations, and it can serve as an assay complimentary to Sanger sequencing when investigating missing mutations in RB1 gene.

Support or Funding Information

Jennifer Richards‐Yutz, Tappan Ganguly, Eric Toorens, Jennifer Rosado, Richard Grant, Kimberly Moran, Arupa Ganguly. Genetic Diagnostic Laboratory at the University of Pennsylvania, Philadelphia, PA

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of High‐fidelity Patient Simulation Training on Medical Student Self‐efficacy and Competence
Authors: Klein Barbie
Published: June 2021

High‐fidelity patient simulation (HFPS) is an instructional adjunct that incorporates realistic physical environments with interactive computerized patient manikins for training and assessment. This research compared the self‐efficacy ratings and competence, as measured by Objective Structured Clinical Examination (OSCE) scores, between medical students exposed to HFPS (intervention) and medical students with little to no exposure with this pedagogy (control). The groups that were examined included: first‐year (MS1), second‐year (MS2), and third‐year (MS3) medical students. A questionnaire was developed and validated to assess clinical self‐efficacy of patient care. Compared to the control groups, average OSCE composite scores were higher for the MS1 (93.49, SD = 4.24) and MS3 intervention groups (75.27, SD = 5.39), but differences were not significant (MS1: t(27) = 1.090, p = .285; MS3: t(7) = 2.263, p = .058). Pearson's correlations between average self‐efficacy ratings and OSCE scores exhibited negative correlations for all control groups, ranging from weak to strong. Weak to moderate positive correlations were observed for the MS1 and MS3 intervention groups. Both MS2 intervention and control groups had moderate negative correlations, suggesting the MS2 intervention group and all control groups inaccurately assessed their actual ability to successfully provide patient care. Although statistical significance was not achieved, the general trends observed are consistent with the literature and warrant further investigation into the dynamic interactions of HFPS exposure on medical student self‐efficacy and external measures of competence.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of a novel anatomy education tool for teaching dermatomes, cutaneous nerve maps and surface anatomy concepts: The Dry‐Erase Anatomy Mannequin (DrEAM)
Authors: Maini Arjun K. O. , Farias Anna
Published: June 2021

Dermatomes are areas of skin whose sensory innervation can be traced back to a single spinal nerve. Knowledge of dermatome patterns is fundamental to the study of human surface anatomy and is diagnostically useful for a range of neurologic pathologies and injuries. Dermatomes and other surface anatomy concepts are often visually depicted in lectures and textbooks through illustrations. Such visual depictions are two‐dimensional and typically only show the anterior and posterior surface maps of the body side‐by‐side. However, dermatomes and other surface projections, such as cutaneous nerve maps, are 3D shapes that often pass continuously from anterior to posterior. Thus, learning these concepts from such limited visual depictions can provide perceptual challenges to anatomy students who must eventually apply 2D mental schema onto their 3D patients. Additionally, there is a relative lack of emphasis placed on teaching surface anatomy, both within textbooks and as part of formal anatomy curricula. Various instructional methods and low‐fidelity models have been developed by others attempting to convey surface anatomy concepts including body painting exercises and drawing on cloth‐covered anatomy models, to name a few.

Here, we report the development of a new educational tool: The Dry‐Erase Anatomy Mannequin (DrEAM). Our novel low‐fidelity learning tool consists of common retail clothing mannequins coated in a commercially available dry‐erase finish and allows students to mark, erase and remark 3D surface anatomy shapes and contours, such as dermatomes, using dry‐erase markers. We also document our recent deployment of the tool as part of our “Dermatome Day” ‐ an informal, interactive extracurricular anatomy teaching session for undergraduate medical students. Other potential use‐cases for the DrEAM in teaching surface anatomy concepts within a range of health professions curricula are further proposed. Lastly, we discuss the formal evaluation of the DrEAM to determine its efficacy in improving student learning outcomes related to surface anatomy concepts as compared to other traditionally used methods. We hypothesize that its use will promote increased understanding of surface anatomy concepts amongst medical students and lead to their improved performance on related test questions when compared to more traditional teaching methods.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel Strategies and Approaches to Improve the Efficacy of Targeted Therapy in Gastric Cancer
Authors: Kitahara Shuji , Duda Dan , Matsui Aya , Shigeta Kohei , Aoki Shuichi , Ezaki Taichi
Published: June 2021

Gastric cancer (GC) is the third most common cancer and the second most deadly cancer worldwide. The vast majority of GCs are adenocarcinomas, which can be further subdivided into intestinal and diffuse types according to the Lauren classification. In all subtypes, a tumor microenvironment (TME) characterized by active angiogenesis, fibrosis and inflammation is critical in the local and metastatic progression of these tumors. This is in part due to increased hypoxia and suppression of the immune system's ability to fight the cancer. VEGF is a key factor for this abnormal structure and function of the TME of GC. Using surgical samples, we found that tumor angiogenesis is increased with clinical stage progression. In addition, we found that a substantial fraction of GC cells expressed receptor (VEGFR)‐2. Blocking VEGFR2 pathway is a standard of care in GC. However, clinical responses are transient and the survival benefits are limited in GC patients. One of the cues to drive evasive resistance is hypoxia, which can be further increased after treatment with potent anti‐VEGF drugs. Interestingly, using low does anti‐VEGFR‐2 antibodies could transiently normalize tumor vasculature and convert the immunosuppressive TME into an immunostimulatory one. This effect could improve the outcome of various immunotherapies by increasing flow and oxygenation in other cancers. To examine the role of VEGFR‐2 in GC, we blocked VEGFR‐2 in tumor cells versus endothelial cells using species specific monoclonal antibodies and orthotopic models of human GC xenografts in immunodeficient mice. In these experiments, blocking VEGFR2 in endothelial cells normalized tumor blood vessels and reduced hypoxia and fibrosis, which was not seen when blocking VEGFR2 in GC cell. Moreover, a growth delay was seen only when blocking VEGFR2 in endothelial cells. These data are important because anti‐angiogenic therapy might potentially shift the TME towards promotion of anti‐tumor response to other therapies. These studies might lead to rapid implementation of the new combination therapy with anti angiogenic agent into clinical trials through our collaborations.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Catalase and glutathione reductase co‐localize with insulin in pancreatic beta cells of normal and diabetic rats
Authors: Adeghate Ernest , Al Darmaki Reem , Baniyas May , D'Souza Crystal , Lotfy Mohamed , Tariq Saeed
Published: June 2021

Background

Long‐term diabetes mellitus (DM) is associated with hyperglycemia. Hyperglycemia causes oxidative stress which may subsequently reduce the level of endogenous antioxidants. Hyperglycemia‐induced tissue stress has been implicated in the formation of advanced glycated end products and other factors responsible for the development of short and long‐term complications of DM.

Aims

Our study aims to determine whether catalase (CAT)‐ and glutathione reductase (GR) are present in pancreatic islet cells and if yes, which of the islet cells contain CAT and or GR?

Methods

DM was induced in adult male Wistar rats with streptozotocin (60 mg/kg body wt., intraperitoneal). Four weeks after the induction of DM, the rats were anesthetized and part of the pancreas was removed, incubated overnight in Zamboni's fixative and processed for double‐labelled immunofluorescence using antibodies against insulin, glucagon, somatostatin, pancreatic polypeptide, CAT and GR. A portion of the fresh pancreatic was also processed for immunoelectron microscopy using different gold particle sizes as probes for either pancreatic hormone or antioxidant.

Results

The number of insulin‐, CAT‐ and GR‐immunoreactive cells was significantly (p < 0.05) reduced after the onset of DM. In contrast, the percentage distribution of glucagon‐, somatostatin‐, and pancreatic polypeptide‐positive cells increased in pancreatic islets of diabetic rats, when compared to non‐diabetic controls. CAT and GR were observed in insulin‐containing pancreatic beta cells. A decrease in the number of insulin‐secreting cells was associated with a concomitant reduction in either CAT or GR. Immunoelectron microscopy showed that CAT‐ and GR‐labelled immunogold particles are present in association with insulin in secretory granules of pancreatic beta cell. The number of insulin‐, CAT‐ and GR‐coated gold particles was markedly (p < 0.05) reduced in pancreatic beta cells of diabetic rats.

Conclusion

Co‐localization of CAT and GR with insulin suggests that these antioxidants may play role in the maintenance of pancreatic beta cell structure and function.

Support or Funding Information

Supported by research grants from the CMHS (Grant # NP‐14‐32) NP and UAEU (Grant # ZCHS‐7‐2014)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemically Altered Epigenetic Landscape: Transcriptional Reprogramming at Precise Space and Time
Authors: islam kabirul
Published: June 2021

Epigenetics is a set of DNA sequence‐independent biochemical processes which dictate the transcriptional potential of a gene and are essential to the phenotypic diversity of multicellular organisms. Cells employ a range of epigenetic mechanisms, most prominent being covalent chemical modifications, such as DNA and histone methylation. A wide range of proteins partakes in establishing (writers), recognizing (readers) and removing (erasers) these chemical entities in the chromatin for gene regulation to occur. We have engineered the protein‐ligand and protein‐protein interfaces of these biochemical pairs by employing the bump‐hole approach to develop orthogonal epigenetic regulators. These engineered epigenetic machines are employed to construct a non‐native chromatin landscape suitable for reprogramming gene expression with high temporal control in response to the cues administered by the ‘bumped’ ligands and inhibitors.

Support or Funding Information

University of Pittsburgh and the National Institute of Health

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rift Valley Fever Virus Nucleocapsid Protein Binds to C/D Box Motifs of snoRNA
Authors: Hayashi Miyuki , Hornak Katherine E. , Lachy Jean‐Marc , Lodmell J. Stephen
Published: June 2021

Rift Valley fever virus (RVFV) is a mosquito‐borne RNA virus that infects livestock and humans. RVFV is widespread in Africa and the Arabian Peninsula, however, no approved vaccines or treatments are available. Nucleocapsid (N) protein in RVFV is RNA binding protein and recognizes its binding target possibly by interacting with a secondary structure or specific sequence of RNA. N protein encapsidates viral RNA in virions to protect its genome. Early in the infection cycle N protein expression is very high, and is more abundant than would be required to interact only with viral RNA. Therefore, it is possible that N protein not only encapsidates its genome but has other functions during infection. In fact we showed, using systematic evolution of ligands by exponential enrichment (SELEX) and cross‐linking, immunoprecipitation and deep sequencing (CLIP‐seq) that N protein is able to bind host RNAs as well as RVFV genomic RNAs. In this study, we investigated characteristics of N protein binding partners. Our data show that N protein preferentially binds to RNAs that contain CUGA/UGA motifs. These motifs are also highly represented in the C and D boxes in small nucleolar RNAs (snoRNAs), whose usual function is to direct 2′‐O‐methylation of specific nucleotides in rRNA. In addition, our CLIP‐seq data, showed that N protein prefers binding to box C/D class snoRNAs over other classes of stable RNAs. Together, these data strongly suggest a non‐canonical function for N protein during RVFV infection that involves binding of small nucleolar RNAs, although the downstream effect of this binding interaction is still incompletely understood and is under investigation.

Support or Funding Information

This project was supported by NIH grants R15AI105737 to JSL and NIGMS P20GM103546 (S. Sprang, PI)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying Regulatory Targets of the Small RNA MtlS in Vibrio cholerae
Authors: Mendez‐Contreras Sabrina Isabel , Tsao‐Wu Maya , Liu Jane
Published: June 2021

In the facultative pathogen Vibrio cholerae, MtlS is a small RNA (sRNA) transcribed antisense to the 5′ untranslated region of mtlA, which codes for the transporter of the sugar alcohol mannitol. MtlS works in cis as a translational repressor of mtlA and it is postulated that the sRNA allows the bacteria to respond rapidly to changes in environmental carbon sources. Because bacterial sRNAs can target multiple genes, we hypothesized that MtlS may regulate other genes in addition to mtlA. Previous work in the lab used RNA target‐prediction programs and mass spectrometry‐based proteomics to identify additional MtlS targets. The results identified 6 potential targets: VCA1045 (mtlA), VCA0820, VCA1046, VC1560, VC1899, and VC2683. The objective of this study was to determine if the putative targets were post‐transcriptionally regulated by MtlS. We constructed gfp translational reporter fusions of the candidate genes in a V. cholerae ΔmtlS strain carrying either a control vector or a plasmid ectopically expressing mtlS. An mtlA‐GFP fusion served as a positive control and exhibited the expected down‐regulation of gfp expression when mtlS was expressed. We have tested reporter fusions to all the candidate genes for regulation by MtlS by monitoring total cell fluorescence and western blot analysis. In an experiment done in duplicate in the presence of MtlS, during stationary phase growth we observed 20% down‐regulation of VC1899, encoding a protein with homology to nucleic acid‐binding proteins. In an experiment done in duplicate, we have also observed 30% logarithmic and stationary phase downregulation of VCA1046, encoding the mannitol dehydrogenase mtlD, in the presence of MtlS. Results based on another experiment conducted in duplicate indicate 20% logarithmic and stationary phase down‐regulation of VC1560, encoding a protein with homology to a catalase‐peroxidase, in the presence of MtlS. Tests with VC2683, encoding a putative cystathionine gamma‐synthase, have been inconclusive, as both down‐regulation and up‐regulation by MtlS has been observed; further experiments will need to be conducted with the VC2683‐gfp fusion. The VCA0820‐gfp fusion does not exhibit any fluorescence, in the presence or absence of MtlS. This work positively contributes to the notion that a single sRNA can have multiple regulatory targets. Ultimately, this work will expand our knowledge on MtlS's role in helping V. cholerae adapt quickly to changes in environmental carbon sources and on sRNAs in general.

Support or Funding Information

NIH R15 AI090606

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression and purification of the antigen binding fragment of the germline‐encoded precursor to the murine anti‐S1P metalloantibody in the baculovirus‐insect cell system
Authors: Farokhi Elinaz
Published: June 2021

Sphinosine‐1‐phosphate (S1P) is a biologically active lipid that plays a signaling role in angiogenesis and in tissue remodeling after cardiac ischemia. In recent years, antibodies that target biologically active lipids have been studied as promising therapeutic agents. We determined the 1.9 Å x‐ray crystal structure of a humanized version of the murine anti‐S1P antibody, and reported the novel finding that it employs two bridging calcium ions in binding to its lipid antigen. In order to study the chemical properties of this novel metalloantibody, we are developing methods for recombinant expression and purification. Previously, germline‐encoded light and heavy immunoglobulin genes that served as precursors to the mature anti‐S1P antibody have been engineered into a plasmid vector for recombinant expression in E. coli. Recovery of soluble protein required that both light and heavy genes be directed to the periplasm of the Gram‐negative bacteria through implementation of PelB signal peptides on both genes in a coexpression plasmid. Unfortunately, the yield for the recombinant antibody was low and insufficient for further analyses. Curiously, the affinity chromatography purified protein partitions into two well resolved pools upon further purification by size exclusion chromatography. Sf9 insect cells infected with recombinant baculoviruses have been successfully used to express high yield secreted proteins. Selection of appropriate promoters and signal peptides has been shown to significantly affect protein yield. We employed the pFastBac Dual vector system in which one gene is expressed under the control of the AcNPV polyhedrin promoter while the other is under the control of the AcNPV P10 promoter and the gp64 signal peptide to express the histidine‐tagged heavy (VH and CH1 domains) and light chains of the metalloantibody precursor. The protein concentration is 12 mg/ml in 500 ml suspension insect cells. As part of our efforts to determine whether the incorporation of metals arises during affinity maturation of the antibody or if it is inherent to the germline encoded sequence and as a starting point to study the chemical potential of metalloantibodies, we used ICP‐MS to determine the Ca concentration in samples treated with EDTA. The concentration of Ca in the Fab antibody was significantly higher than the negative control antibody and the buffers. Our next step is to mutate residues that were previously shown to coordinate metals in the anti‐S1P antibody to introduce novel metal binding potential into an antibody scaffold.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Demonstration of the contributions of pulmonary CYPs to naphthalene‐induced airway toxicity using lung‐Cpr‐null mice
Authors: Kovalchuk Nataliia , Zhang Qing‐Yu , Van Winkle Laura , Ding Xinxin
Published: June 2021

Naphthalene (NA) is an omnipresent air pollutant and respiratory toxicant. NA induces cytotoxicity in airways following bioactivation by CYP (P450) enzymes. Previous studies suggested that liver and lung are both capable of bioactivating NA in vitro and in vivo. However, direct evidence for a specific role of pulmonary P450 enzymes in mediating NA‐induced airway toxicity, particularly upon exposure to NA via the systemic route, is still lacking.

The aim of this study was to examine the specific contribution of pulmonary CYPs to airway toxicity of NA using a lung‐Cpr‐null mouse. We predicted that lung epithelial cells that lack Cpr expression, which would have little microsomal P450 activity, would be resistant to NA‐induced cytotoxicity. Cre‐mediated deletion of Cpr in the lung‐Cpr‐null mouse occurred in a substantial proportion of the Club cells, which, before undergoing Cpr deletion, had high ability to bioactivate NA. Under the conditions used, doxycycline‐induced deletion of Cpr occurred in 66% of Club cells in proximal airways and 86% of Club cells in distal airways, based on the results of a dual immunofluorescence study. Levels of NA and NA‐GSH (a biomarker of NA bioactivation) were similar in the plasma of lung‐Cpr‐null mice and their control littermates after administration of a single bolus dose of NA (200 mg/kg). Histological examination of airways revealed that NA‐induced damage of epithelial cells is focal and mild in lung‐Cpr‐null mice compared to severe exfoliation of epithelial cells in most airways in NA‐treated control mice. Moreover, the number of proliferating (BrdU‐positive) airway epithelial cells was 4‐fold lower in NA‐treated lung‐Cpr‐null mice than in NA‐treated control littermates.

These results confirm that CYPs in the lung can mediate NA‐induced airway epithelial damage in vivo.

Support or Funding Information

supported in part by NIH grant ES020867

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.