bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of a Novel Hydroalcoholic Extract of Lens Culinaria Against the Dexamethasone‐Induced bone loss and Chemotherapy Cytotoxicity in Murine and Human Cells
Authors: Tricarico Domenico , Antonacci Marina , Di Benedetto Jacopo R. , Maqoud Fatima , Scala Rosa , Centoducati Gerado , Colonna Nicola , Leonetti Francesco 
Published: May 2022

Lens culinaria showed nutritional health benefit in humans. Here, we evaluated the effects of a novel hydroalcoholic extract (70% ethanol, 1E030521DIPFARMTDA) from Lens culinaria (Terre di Altamura SRL, Altamura BA) to prevent cytotoxic damage from chemotherapeutic agents such as staurosporine, a non‐selective inhibitor of tyrosine kinases, cisplatin the antineoplastic alkylating agent, doxorubicin as antimitotic, and the glucocorticoid dexamethasone targeting osteoblast.On renal Hek293, staurosporine caused a significant reduction from ‐20% to ‐60% (p <0.05) in cell proliferation and the extract was significantly effective in bringing these values between 70% and 100% (p <0.05) with comparable activity to that of diazoxide. Also, in the clonogenic assay, the extract was effective to prevent the reduction of colonies numbers caused by staurosporine.Cisplatin caused a slight reduction of ‐10% in Hek293 cell proliferation and a significant reduction of ‐30% in SHSY5Y cell proliferation respectively after 48 and 72 hours of incubation time. In both cases, the extract showed a significant cytoprotective effect. On the primary culture of murine osteoblasts, doxorubicin reduced survival by ‐30%, dexamethasone reduced it by ‐15%. The extract was significantly effective in preventing cell count‐reduction induced by dexamethasone but not by doxorubicin.A UV/VIS spectroscopy analysis was carried out. We obtained three peaks at the wavelength of 350, 260 and 190 nm which correspond to families of compounds such as flavonoids, proanthocyanins and salicylates.Therefore, this novel extract prevented the cytotoxic damage induced by cisplatin and staurosporine on the renal cell line, and by cisplatin also on the neuronal cell line by clonogenic assay and DNA staining. The extract was significantly effective in preventing the reduction of cell proliferation induced by dexamethasone in murine osteoblasts suggesting of potential use in bone loss diseases. It is not yet clear which of these families molecules composing the extract is responsible for these effects or if there is a synergistic action.
Reverting Colitis Symptoms by Oral Administration of L. reuteri Expressing CXCL12 Acting Through Novel Pathway Demonstrated in Two Experimental Animal Models
Authors: Öhnstedt Emelie , Donas Cuadra Cristian , Lofton Tomenius Hava , Frank Peter , Vågesjö Evelina , Roos Stefan , Phillipson Mia 
Published: May 2022

IntroductionIn USA and Europe, it is estimated that more than 3 million people are suffering from inflammatory bowel disease (IBD) and another 1.7 million cancer patients treated with checkpoint inhibitors (ICI) are diagnosed with manifest ICI‐induced colitis. The current treatments with steroids and available biologics act systemically and possess limitations with loss of response and severe adverse events and may even limit the effect of the ICI therapy. Thus, new local‐acting therapies are needed to serve the full unmet need amongst these patients.We have recently published that oral administration of L. reuteriR2LC to mice increases IgA production in Peyer’s patches, resulting in modified gut microbiota, altered physiology and immune cell phenotype in the colon and prophylactic administration reduced inflammation in the DSS‐induced colitis.Methods, the novel drug candidate, ILP100, which is a genetically modified lactic acid bacteria that express the chemokine CXCL12 were generated and evaluated as a therapeutic in two different models of colitis, the DSS‐model and a model of checkpoint inhibitor induced colitis (using aPD1 and aPDL1).Results, Peroral administration of ILP100 to mice with overt colitis interrupts disease progression and improves symptoms assessed as disease activity index (DAI) and colon shortening. Further analysis showed that treatment with ILP100 increased infiltrating immune cells expressing TGF‐ß and IL‐10 as well as other tolerogenic immune cell populations including Treg and Tr1 in the colonic lamina propria. There were no changes in the small intestinal lamina propria or mesenteric lymph nodes and there was no systemic exposure detected. Further, efficacy comparison studies demonstrate a reduced severity and a faster recovery of the ILP100 treated mice compared to TNF‐α inhibitor and α4β7‐inhibitor treated mice. The findings have been confirmed over a large dose range. Even though the ICI‐induced colitis did not manifest in symptoms such as body weight loss or blood in feces, fibrosis was developed measured as a reduction of the mean colon length of 12‐18 % in the ICI‐treated groups compared to control, with a tendency to a reduced colon shortening in the ICI + ILP100‐treated groups. Histological analysis of intestines samples from the checkpoint inhibitor is currently ongoing.In conclusion, we demonstrate that peroral administration of ILP100 efficiently ameliorate overt colitis in the DSS‐model by inducing healing through a local shift into an anti‐inflammatory and tolerogenic immune signature in the colon mucosa. Further, our data suggest that peroral administration of ILP100 ameliorate colitis in an ICI model.
The diagnostic and prognostic value of COL5A2 high expression and hypomethylation in recurrent low‐grade glioma
Authors: Duan Hubin , Hao Chunyan , Wang zhuangzhuang , Xue Wen , Li Mingli , Qiao Binjun , Zhang Jinbao , Jing Zhijie , Chen Xianghui , Zheng Juan , Qiao Xiaohong 
Published: May 2022

ObjectionTo investigate the diagnostic and prognostic value of COL5A2 high expression and hypomethylation in recurrent low‐grade glioma.MethodsFour independent databases, Chinese Glioma Genome Atlas(CGGA), The Cancer Genome Atlas(TCGA), Gravendeel and Rembrandt, were used to analyze the differential expression and prognostic of COL5A2 in different grades and types (primary and recurrent) of LGG. And further analyzed the differential expression and prognosis of COL5A2 and its methylation between primary and recurrent LGGs (including their pathological subtypes), and correlation between COL5A2 expression and methylation in low‐grade gliomas and their clinical phenotypes. SPSS and GraphPad Prism statistical software were used to assess the prognostic and diagnostic value of COL5A2 expression and its methylation in recurrent LGG by binary Logistic regression, Cox regression and Receiver Operating Characteristic (ROC) curve. The correlation between COL5A2 expression and immune cell infiltration in TCGA for LGG was analyzed using TIMER and TISIDB database. Gene Set Cancer Analysis (GSEA) was used to investigate the signaling pathway of COL5A2.ResultsCOL5A2 is highly expressed in different grades of gliomas and recurrent LGG, and overexpression of COL5A2 is detrimental to the prognosis of LGG. The expression level of COL5A2 was negatively correlated with its methylation level, and the hypomethylation of COL5A2 was not conducive to the prognosis of LGG. COL5A2 may promote the recurrence and progression of LGG through post‐ transcriptional methylation modifications and immunoinfiltration, and may provide a reference for the diagnostic threshold of recurrent LGG.ConclusionsOur study identified COL5A2 overexpression and hypomethylation as independent risk factors and prognostic factors for different grades of glioma and recurrent LGG, and has some reference value for the diagnosis of recurrent LGG.
Contribution of stress granules and spliceosomal components to the formation of chemoresistance in ovarian cancer cells
Authors: Shnaider Polina , Malyants Irina , Ivanova Olga , Arapidi Georgij , Lagarkova Maria , Govorun Vadim , Shender Victoria 
Published: May 2022

The emergence of chemoresistance in cancer cells is one of the main reasons for the high mortality from oncological diseases. We have shown that signaling molecules from dying cancer cells contribute to a more aggressive therapy‐resistant phenotype. However, the molecular mechanisms underlying this effect remain elusive.Here we aimed to find out which proteins secreted by dying tumor cells get into recipient cells and to determine the molecular cascades triggered in response to incubation with therapy‐induced secretomes. We modified the SILAC (Stable Isotope Labeling by Amino acids in Cell culture) technology. A heavy‐labeled primary culture of ovarian cancer cells was treated or untreated with cisplatin for 48 h. Then using ultrafiltration cartridges, a fraction of extracellular vesicles (EV) were isolated from culture media. The obtained EV were resuspended in a medium containing Medium‐labeled amino acids and added to the recipient chemonaive unlabeled tumor cells for 24 h. During incubation, recipient cells absorbed EV with Heavy‐labeled proteins from donor cells and synthesized new proteins with Medium label. Then, we performed proteomic analysis of recipient cells. In total, we identified 4224 proteins. Interestingly, dying tumor cells secrete and recipient cells absorb a large number of spliceosomal and ribosomal proteins along with translation initiation complex. These proteins are often presented in stress granules. In response to EV from dying tumor cells, recipient cells upregulated the translation of genes responsible for the regulation of the cell cycle and oxidative phosphorylation.To our knowledge, this is the first report about the secretion of stress granules and spliceosomal components into the extracellular space and their internalization in other cells. To confirm these data we performed transfection of donor cells with constructs encoding the fluorescent proteins TIA1 (protein of stress granules) and SRSF4 (protein of the spliceosome). We showed that both proteins are re‐located together from the nucleus into the cytoplasm under the cisplatin treatment. Then, we demostrated the absorption of fluorescent‐labeled proteins TIA1 and SRSF4 by recipient cells after incubation with therapy‐induced EV. Our findings demonstrate that in response to cisplatin treatment tumor cells secrete spliceosomal components into the extracellular space (presumably as a part of stress granules), then these components are absorbed by recipient chemonaive cells and trigger a restructuring of metabolism and cell cycle.
Anti‐Epileptic Drug and Teratogen Valproic Acid Induces Microglial Cell Death in a Valproic Acid Murine Model of Autism
Authors: Loyola Allison , Bhushan Alok , Awale Prabha S. 
Published: May 2022

Autism Spectrum Disorder (ASD) is a neurodevelopmental condition that affects approximately 1 in 44 children in North America, and its association with neuronal connectivity is an area of intense research. Valproic acid (VPA) is a multi‐target drug widely used to treat epilepsy. It is classified as a teratogen and as a histone deacetylase inhibitor (HDACi) and fetal exposure to VPA increases the risk of ASD. The VPA model has been well characterized for behavioral and neuronal deficits, including hyperconnectivity. The cause of hyperconnectivity is poorly understood however, it is speculated that lack of pruning may be partly responsible for this anomaly. Microglia, the principal immune cells of CNS regulate dendrite and synapse formation during early brain development. The effect of VPA on microglia during early development has not been well characterized and may provide potential hints regarding the etiology of this disorder. Preliminary studies from our lab indicate that VPA reduces microglial number during peak periods of postnatal synaptogenesis in the mouse brain primary cortex. Nevertheless, the cause of this reduction is not known. Therefore, in this study, we determined the mechanism for reduced microglial numbers in the VPA model of autism.Pregnant BALBc mice were administered 600 mg/kg of VPA by subcutaneous injection on day 13.5 of gestation. We harvested the mice brains at embryonic (E) day 14, E17, and postnatal day 1 (PD1). Brain tissue was double immunolabeled for microglia (IBA1) and cell death marker (TUNEL). We found a gradual increase in TUNEL staining beginning E14 to PD1 compared to age‐matched controls. The mechanism for reduced microglial number may be due to cell death. Thus, our findings possibly have pathophysiological/pathogenic implications in autism and may, ultimately, lead to the discovery of new therapeutic target(s) for autism.
Anatomy Education‐Paradigm Shift from Passive to Active Learning‐Effects on Student Engagement, Comprehension and Retention A Review of Literature from 2012 to 2022
Authors: Gamo Joel A. 
Published: May 2022

IntroductionThere has been a dramatic paradigm shift from passive to active teaching and learning methods. Passive teaching is faculty‐centric, while active teaching is student‐centric which many academic scholars believe will lead to better student engagement, comprehension, and knowledge retention. Instead of just pouring information into their brains, students must be responsible for their learning, and teachers only act as facilitators rather than the sage on the stage. There is a need to formulate curriculum map designs and lesson plans that leads to higher‐order learning using Bloom's taxonomy in teaching.ObjectiveThe objective is to review literature that compares the passive and active teaching strategies on their advantages and disadvantages regarding student engagement, comprehension, and knowledge retention.MethodologyA review of the literature was conducted on passive and active learning methods in anatomy education from 2012 to 2022.ResultsThe faculty‐centric passive teaching with lectures and videos make students disengaged. On the other hand, in active learning, students are more engaged, have better comprehension, retention and are more responsible for their learning. Active teaching utilizes discussion fora, collaborative and peer teaching, flipped classrooms, blended classes, games, concept maps, chunking, polling, and augmented reality.The pyramid of retention from the National Training Laboratories in Bethel, Maine, showed that passive teaching led to 30 % retention while active teaching had superior retention from 50 to 90 %.ConclusionActive learning methods have more advantages regarding student engagement, comprehension and, retention than passive teaching. In addition, the faculty may combine both strategies to maximize learning among the students.
Analysis of the lake trout heart, blood, liver, and brain proteome using evolutionary proteomics
Authors: Alwine Shelby E. , Dupree Emmalyn , Crimmins Bernard , Holsen Thomas , Darie Costel 
Published: May 2022

IntroductionSalvelinus namaycush (lake trout) is a top‐predator fish in the Great Lakes region. The Great Lakes Fish Monitoring and Surveillance Program (GLFMSP) use lake trout as a bioindicator of chemical stress by monitoring the concentrations of persistent, bioaccumulative and toxic (PBT) chemicals in their bodies. Studies have shown that elevated concentrations of PBTs can cause changes in transcribed genes, translated mRNAs, proteins produced, and post‐translational modifications in aquatic species. Though lake trout is used to monitor chemical concentrations for over 50 years, there is very little information available on the proteome of this species. A well‐developed protein database for the lake trout would help elucidate the effects of PBT chemicals on this species providing a direct health metric for the Great Lakes ecosystem.MethodsIn this study, heart, blood, brain, and liver samples from lake trout were analyzed by SDS‐PAGE, followed by in‐gel trypsin digestion and analysis by nanoLC‐MS/MS. The raw data was searched against different NCBI and UniProtKB databases in Mascot Daemon and the output was analyzed by Scaffold 4.3 software. Databases used include Actinopterygii, Salmonidae, Salvelinus, as well as the highly studied species Oncorhynchus mykissand Danio rerio.Preliminary DataPreviously, our group has reported a large number of proteins for lake trout liver, heart and blood (Dupree et al., Proteomics, 2019, PMID: 31578773 and Dupree et al., Proteomics, 2021, PMID: 34676671). In lake trout liver we identified 4194 proteins using the NCBI databases and 3811 potential protein hits in the UniProtKB databases. In the blood and heart we identified 838 and 580 proteins, respectively, using NCBI databases and 1180 and 561 potential protein hits for the heart and blood, respectively in BR2. A lake trout brain proteomics analysis completed using an in‐house SDS‐PAGE, followed by in gel digestion and nanoLC‐MS/MS analysis. The Mascot database search is underway. Once protein hits are cataloged, BLAST and Scaffold‐based comparisons of the liver, heart, blood and brain will be performed. Additional search of the data against the other fish relatives will also be completed. Through this process, evolutionary relationships for the lake trout species will also be explored. The current study will add to our understanding of the lake trout proteome that will be used to understand the biochemical/physiological effects of legacy PBT chemicals in the Great Lakes ecosystem.Novel AspectUse of four types of lake trout tissue together to develop a protein database for the species.
Engineering Spinal Interneurons for Repair of the Injured Cervical Spinal Cord
Authors: Fortino Tara A. , Randelman Margo L. , Ponna Anish , Schardien Kayla A. , Niceforo Alessia , Hall Adam A. , Zholudeva Lyandysha , Lane Michael A. 
Published: May 2022

Cervical spinal cord injuries (SCI), the most common SCI, compromise respiratory networks leading to life‐threatening impairments in breathing. Following a cervical injury, extensive damage occurs to the phrenic motor network (controlling the diaphragm) consisting of phrenic motoneurons and interneurons. Despite these devastating consequences, there is now significant evidence for plasticity and limited spontaneous recovery. Our ongoing work has identified spinal interneurons (SpINs), in particular the excitatory V2a subtype, as key therapeutic targets for promoting plasticity and functional recovery of the phrenic motor network. Efforts to enhance this plasticity have shown some promise, but it is likely that treatments capable of promoting repair (e.g., cell transplantation) will provide the best therapeutic outcome.Building on a long history of transplanting spinal neural precursor cells (NPCs) to promote spinal cord repair, the present work harnesses advances in stem cell engineering to enrich donor NPCs with specific subsets of stem cell derived SpINs. We hypothesize that NPCs enriched for V2a SpINs will have enhanced therapeutic efficacy over NPCs alone and promote phrenic motor recovery. We will test this hypothesis using a clinically relevant model of a cervical contusion in the adult rat, and test the functional contribution of donor V2a SpINs using inhibitory chemogenetics (hM4Di).Adult female Sprague‐Dawley rats received mid‐cervical contusion injury (Infinite Horizon impactor, intended impact force 200 kilodyne). One‐week post‐injury, aggregates of NPCs with mouse embryonic stem cell derived V2a SpINs were injected into the lesion cavity. One month post‐transplantation, animals were re‐anesthetised and traced with a retrograde, transneuronal tracer, pseudorabies virus, applied to the diaphragm muscle to assess donor‐host connectivity with the phrenic network. Three days later, animals underwent terminal diaphragm electrophysiology, at which time clozapine was injected intraspinally into the transplant to silence donor SpINs. Preliminary data revealed that clozapine‐induced suppression of donor cell activity reduced diaphragm activity ipsilateral to injury, with no measurable effect on the contralateral side. Consistent with this, immunohistochemical assessment revealed donor cells survived and synaptically integrated with the injured host phrenic network. This ongoing research offers the first in‐depth assessment of how these donor SpIN populations contribute to phrenic recovery.
Assembly‐line catalysis in bifunctional terpene synthases: (+)‐Copalyl diphosphate synthase from fungal Penicillium species
Authors: Ronnebaum Trey A. , Eaton Samuel A. , Brackhahn Emily A. E. , Faylo Jacque L. , Gupta Kushol , Christianson David W. 
Published: May 2022

The magnificent chemodiversity of terpenoid natural products is largely rooted in two types of terpene synthases: prenyltransferases and cyclases. The unusual diterpene (C20) synthase, (+)‐copalyl diphosphate synthase from fungal Penicilliumspecies (PvCPS, PfCPS),is the first bifunctional terpene synthase identified with both prenyltransferase and class II cyclase activities in a single polypeptide chain. The C‐terminal prenyltransferase α‐domain generates the C20 linear isoprenoid, geranylgeranyl diphosphate, which is then cyclized to (+)‐copalyl diphosphate at the interface of the N‐terminal βγ‐domains. We, first, establish that PvCPS exists as a hexamer – a unique quaternary structure for known αβγ terpene synthases.1 Hexamer assembly is corroborated by 1) crystal structures of the prenyltransferase α‐domain obtained from limited proteolysis of full‐length PvCPS, 2) the ab initio modeling of full‐length PvCPS derived from small‐angle X‐ray scattering data, and 3) preliminary 3D reconstructions of PfCPS by cryo‐EM. Interestingly, biophysical experiments with PvCPS suggest that oligomer formation is dynamic since the hexamer dissociates into lower‐order species at lower concentrations. However, enzyme concentration does not affect prenyltransferase activity in vitro.Assembly‐line catalysis provides an efficient carbon management system for generating high‐value terpenoid natural products. With the goal of generating different C5n isoprenoid products, we next explored the structure‐function relationship of the prenyltransferase domain within the assembly‐line platform of PvCPS. Steady‐state kinetics, product analysis, and crystal structures of various structure guided site‐directed variants afforded a functional C15 prenyltransferase chimera.2 This variant will be used in future engineering experiments to create bifunctional sesquiterpene synthases.References1. Ronnebaum, T.A.; Gupta, K.; Christianson, D.W., J. Struct. Biol.2020, 210 (1), 107463.2. Ronnebaum, T.A.; Eaton, S.A.; Brackhahn, E.A.E.; Christianson, D.W., Biochem. 2021, 60 (42), 3162‐3272.