bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Mouse Model for Cerebro‐Costo‐Mandibular Syndrome (CCMS) with an Intronic Deletion in SnrpB
Authors: Alam Sabrina , Majewski Jacek , Majewska Loydie 
Published: April 2020

SNRPB, an essential effector of the spliceosomal complex, is implicated in the splicing of mature mRNA and is mutated in a rare craniofacial disorder known as Cerebro‐costo‐mandibular syndrome (CCMS). Most CCMS patients carry point mutations in the highly conserved alternative exon 2 (AE2) of SNRPB, leading to increased inclusion of a pre‐termination codon (PTC) and reduced levels of the protein‐coding transcript. Given that SNRPB is implicated in all steps of splicing, it was thought that its requirement would be equivalent in all cells and tissues. However, CCMS patients predominantly present with micrognathia and rib defects with variable penetrance. We hypothesize that a subset of transcripts is highly sensitive to the levels of SNRPB during development and is tissue specific. Here, we are generating a mouse model for CCMS to uncover those transcripts that play an essential role in craniofacial development. Using the CRISPR/Cas9 gene‐editing system, we developed a mutant mouse line with a 61 base‐pair (bp) intronic deletion upstream of the AE2. Interestingly, a subset of the heterozygous and homozygous embryos and pups for this deletion show morphological abnormalities in the face, head, ribs, and limbs. Moreover, a significant percentage of the mutants with the 61‐bp deletion die in their first year of life. A subgroup of the heterozygous and homozygous mutant embryos show increased inclusion of the PTC containing AE2 and reduced SNRPB protein levels. We presume that this predominant inclusion of the AE2 induces non‐sense mediated decay resulting in decreased levels of SNRPB. As our data suggest that the 61 bp intronic sequence plays a critical role in regulating SNRPB levels in mice, we are currently investigating if similar intronic sequences in intron 2 of human SNRPB are involved in the preferential splicing of AE2. In addition, we generated a SnrpB conditional knockout mouse model, which will be used in parallel with the deletion model to investigate the cell‐type‐specific roles of SNRPB during craniofacial development. Our novel model for CCMS will further our understanding of the role of SNRPB in craniofacial development.Support or Funding InformationCanadian Institutes of Health Research (CIHR)
Changes in HAS3 and CaSR Expression Following Daily Neonatal CPAP in Mice
Authors: Mayer Catherine A. , Martin Richard J. , Pabelick Christina B. , Prakash Y.S. , MacFarlane Peter M. 
Published: April 2020

Continuous positive airway pressure (CPAP) is a common mode of respiratory support for preterm infants. Recent studies from our lab, however, have shown that CPAP may contribute to the pathophysiology of airway hyperreactivity (AHR) associated with wheezing disorders in former preterm infants. The long‐term effects of CPAP on lung‐development, however, still remain largely unknown. Both hyaluronan synthase 3 (HAS3) and the calcium‐sensitive receptor (CaSR) have been associated with various respiratory morbidities. In the present study, we used a neonatal mouse model of daily CPAP treatment to investigate whether the long‐term effects on AHR are associated with changes in airway HAS3 and CaSR expression. Neonatal mice were exposed to CPAP (6cm H2O) for 3 hours per day for the first 7 postnatal days. Control animals experienced the same conditions as experimental animals only with no CPAP. On postnatal day 21, the lungs from these animals were harvested for either real time rt‐PCR or immunohistochemistry to assess changes in HAS3 and CaSR expression. We observed increased expression of multiple genes involved in hylauraonan synthesis, most notably has3, as well as increased CaSR expression, in the lungs of mice two weeks after CPAP ended when compared to control mice. The increased gene expression of has3 and CaSR was also reflected in increased protein expression as confirmed by immunohistochemistry. HAS3 was expressed predominantly in airway epithelium and the increased CaSR expression correlated with increased αSM actin. To further investigate the role of CaSR in CPAP‐induced AHR, CaSR KO mice (CaSR(−/−)) were exposed to CPAP and airway reactivity to methacholine challenge was assessed two weeks later (P21 days). Compared to control mice, AHR was not expressed in CaSR(−/−) mice suggesting resistance to CPAP treatment. These data suggest increased airway HAS3 and CaSR may be important modulators of CPAP‐induced AHR and remodeling.Support or Funding InformationHLR01138402HLR01056470
Significant role for intestinal urate transporters in host defense
Authors: Kazi Mirajul Hoque , Kuhns Victoria Halperin , Dixon Eryn , Woodward Owen M. 
Published: April 2020

Humans, because of the loss of the uricase gene, have higher levels of serum urate (SUA) than most other mammals, prompting speculation of potential advantages of higher SUA levels. Others have proposed the role of urate as a chemical danger signal, and gout research has clearly shown urate crystal deposition stimulates a profound inflammatory response. Recently, we created a hyperuricemia mouse model through the insertion of a point mutation (Q140K) into the critical urate transporter ABCG2. Though these animals are hyperuricemic, they display no evidence of crystal deposition. In addition, we observed reduction in ABCG2 mediated intestinal secretion was key in the animal’s urate homeostatic mechanisms and the cause of the hyperuricemia. Thus, we hypothesized gastrointestinal infection may lead to dramatic changes in serum urate levels. In a mouse model of shigella infection we found 2‐fold increases in serum urate in infected mice. Because the intestines are a critical source of cytokines and other immune system signal, we next sought to better understand the role for urate. Is it a byproduct of infection or a signal that helps coordinate an immune response and host defense? To answer this question, we challenged the two mice genotypes, WT and the hyperuricemic Q140K, with intraperitoneally injected 100μg lipopolysaccharide (LPS, E. coli 055: B5). At baseline, pre‐LPS, a comparison between WT and the Q140K mice revealed no significant differences in our panel of cytokines. However, twenty‐four hours after LPS challenge showed significantly greater increases in the Th2 cytokines: IL‐5, ‐10, and IL‐13 (100%, 168%, and 100% respectively) in the hyperuricemic Q140K mice. We observed no differences in IL‐4 production, Th1 cytokines IL‐2, and TNF, or chemokines Ccl5, and Cxcl10 between the two genotypes. These results suggest the increased Th2 cytokine production in Q140K mice is due to hyperuricemia. In addition to the Th2 cytokines, vascular endothelial growth factor (VEGF) levels were increased by 2.3‐fold in Q140K mice as compared to WT in response to LPS challenge, potentially influencing the Th2 cytokine production and moderating immunopathology. We conclude hyperuricemia pre‐conditions the immune system, specifically Th2 cytokine, resulting in far greater responses to potential infectious pathogens and ultimately inhibiting disease progression.Support or Funding InformationNIH, NIDDK
Direct Pathway For Bacillus anthracis tRNA Asparaginylation
Authors: Hernandez Jose Giron , Sheppard Kelly 
Published: April 2020

Protein synthesis is essential for life and requires the correct pairing of amino acids to their cognate transfer tRNA (aminoacylation). To date, only two routes exist to attach asparagine (Asn), to it cognate tRNAAsn: the direct and the indirect pathways. The direct path uses asparaginyl‐tRNA synthetase to attach Asn to tRNAAsn when free Asn is present. The Asn is synthesized by asparagine synthetase A. The second pathway involves synthesizing Asn on the tRNA itself. The causative agent for anthrax, Bacillus anthracis, appears to encode both routes for Asn‐tRNAAsn formation. We would like to understand why B. anthraciscodes for both routes. This semester we have focused on the direct pathway. We report the successful purification of the asparagine synthetase A and asparaginyl‐tRNA synthetase along with activity measurements of asparagine synthetase A in order to characterize the direct pathway under different physiologically relevant conditions.Support or Funding InformationNational Science Foundation (MCB‐1615770)
Building Bridges Back to Back: 5 years of an outreach initiative in Anatomical Sciences and Medical Education between Brazil and US
Authors: Curcio Daniella , Laitman Jeffrey T. 
Published: April 2020

The Building Bridges program has fostered collaborations and exchange experiences in Anatomical Sciences among students, researchers, and educators through lectures, colloquia, and student‐faculty exchange sessions. These have taken place in Sao Paulo (Santa Casa School of Medical Sciences), Ribeirao Preto (University of Sao Paulo School of Medicine, USP‐RP) and Rio de Janeiro (Federal University of Rio de Janeiro, UFRJ; Federal University of the State of Rio de Janeiro, UNIRIO; and the Brazilian National Academy of Medicine), since 2015. In August 2019, our group represented the American Association of Anatomists in London, at the 19th Congress of the International Federation of Associations of Anatomists (IFAA), contributing in a workshop addressing the topic International Connections of Anatomists in Global Health. Here, we comment on the outcomes of this workshop and report our upcoming endeavor in New York: the mini symposium Building Bridges Back to Back that will be hosted at the Icahn School of Medicine at Mount Sinai, December 2019. The meeting celebrates the 5th year anniversary of our international outreach initiative. It will bring together leaders from Medical Education from Brazil and connect other participants through videoconference channels. In the Medical Education session, the program includes the Dean of the Santa Casa School of Medical Sciences of Sao Paulo—Dr. Carlos Alberto Malheiros, as keynote lecturer. In the academic publishing session, the keynote lecturers will be the Director of the Graduate Program and Chief of the Surgery Department of the Medical Course at the Federal University of the State of Rio de Janeiro (UNIRIO), member of the Brazilian Academy of Medicine—Dr. Rossano Fiorelli; and the Editor in Chief of the Anatomical Record—Dr. Kurt Albertine. Within the two main sessions, Medical Education and Academic Publishing, the meeting will also cover new advances in Brazilian science, bringing leaders in education and research from different states in Brazil and the US. The event marks the launch of an international exchange program at the Icahn School of Medicine at Mount Sinai and Brazilian medical schools. Our goal is to offer opportunities for Brazilian students to experience medical learning in New York with the intent to help forge new leaderships and role models in medicine and health sciences in Brazil. We expect that this meeting will build the foundation to this exchange project and enhance the cooperation with the health care, medical and anatomical communities of the two countries.Support or Funding InformationMeeting support funded in part by The Anatomical Record; Research reported funded in part by CNPq grant PDE #249582/2013–9
FGF1 receptor antagonist decreases biliary proliferation, fibrosis, and senescence in a mouse model of chronic cholestasis
Authors: O’Brien April , White Tori , Francis Heather , Meng Fanyin , Alpini Gianfranco , Glaser Shannon 
Published: April 2020

BackgroundAcute or chronic cholestasis is a hallmark for a wide variety of liver diseases such as primary sclerosing cholangitis (PSC), alcoholic liver disease (ALD) and cholestasis of pregnancy. Available treatments for cholestasis are few and the subsequent scarring often necessitates the patients undergo a costly and risky liver transplantion. Fibroblastic growth factor 1 (FGF1) is involved in a wide range of cellular activities and has been previously shown to act in both a protective and detrimental manner as it participates in the fibrotic process of various tissue systems. Therefore, we aimed to evaluate the role of FGF1 in the progression of senescence, inflammation and fibrosis in an MDR−/− mouse model of PSC.MethodsFVB and MDR−/− mice were treated with 100ng/kg/day for 7‐days with the FGF receptor inhibitor, AZD4547. Liver, serum, and pure cholangiocytes were obtained. Hepatic fibrosis was evaluated by H&E and Sirius Red staining, and qPCR for collagen‐1a. The inflammatory response was evaluated by qPCR in pure cells for IL1‐b, IL‐6, and IL‐33 and immunofluorescent staining of F4/80. Biliary proliferation was determined through IHC CK‐19 expression and qPCR for Ki67 and PCNA. Senescence was evaluated by qPCR for p53, p18 and p21. FGFR expression was determined by qPCR. FGF1 serum expression was measured using a commercially available ELISA kit and in OCT liver sections by immunofluorescence (IF). In vitro, human intrahepatic biliary epithelial cells (HIBepiC) were treated with 150ng/ml of either FGF1 or AZD4547 with or without the presence of 100ng/ml of LPS for 24 hr before evaluating inflammatory markers, biliary proliferation and markers of fibrosis.ResultsAs expected, MDR−/− mice had a significant increase in biliary proliferation and fibrosis when compared to the FVB control. MDR−/− animals treated with AZD4547 experienced a significant reduction in fibrosis, with a subsequent decrease in collagen 1a1 gene expression. Treatment with AZD4547 also decreased senescence and inflammatory markers in MDR−/− mice evaluated by IHC and IF. Serum analysis showed a dramatic increase in FGF1 levels that were significantly reduced upon treatment. In vitro studies also had significant reductions in inflammatory and fibrotic markers.ConclusionFGF1 receptor antagonist decreased biliary proliferation, senescence, inflammation and fibrosis in cholangiocytes and surrounding liver in a mouse model of PSC. Targeting the FGFR pathway may provide a therapeutic option for patients suffering from various forms of chronic cholestasis such as PSC.Support or Funding InformationNIH RO1 DK110035R01 DK115184
Exploring the scope of high throughput micro‐permethylation based glycomics.
Authors: Shajahan Asif , Supekar Nitin T. , Heiss Christian , Azadi Parastoo 
Published: April 2020

The glycosylation plays important roles in various molecular and cellular functions such as cell signaling, adhesion, development, immune recognition, lubrication, cellular defense etc. The protein glycosylation also contributes towards the stability, serum half‐life, immunogenicity and more importantly, biological activity of therapeutic glycoproteins, including antibodies, vaccines, biomarkers etc. This justifies the increasing demand for the qualitative and quantitative characterization and validation of glycosylation profile on glycoproteins. However, profiling of glycosylation is challenging owing to the glycosylation microheterogeneity, non‐template driven synthesis and their lower abundances. We have recently developed a high throughput N‐ and O‐glycomics strategy through a microscale permethylation based glycan derivatization followed by mass spectrometry (MS). Through this strategy facile but detailed quantitative structural profiling of glycans in a high throughput fashion can be attained. Here, we are reporting the wide applicability of our high throughput glycomics on glycoconjugates from various sources, including human serum, mammalian cells, milk oligosaccharides and purified glycoproteins via an automated tandem ESI‐MSn acquisition platform. The released or free glycans are permethylated in microscale in a 96‐well plate or microcentrifuge tubes and analyzed by an automated ESI‐MSn system after a C18 tip‐based cleanup. An internal standard based method was also incorporated for the absolute quantitation of glycan species. We have employed an in‐depth automated structural characterization MS program for N/O‐glycans through an automated ESI‐MSn acquisition up to MS5 level. Through this automated process, detailed glycan structural topology can be identified including glycan termini, branching, isomers, sialic acid linkages, and bisecting GlcNAc on the glycans. This approach can facilitate profiling of glycosylation on large sample sets, including clinical glycan biomarkers, milk oligosaccharides, and other glycoprotein therapeutics.Support or Funding InformationNational Institutes of Health grant (No. 1S10OD018530) to Dr. Parastoo Azadi
Identifying Variants and Prevalence of the Superficial Brachial Artery – A Case Series
Authors: Bolden Alexys M. , Schmitt Emily A. , Alsharaf Fatemah , Zdilla Matthew J. , Lambert H. Wayne 
Published: April 2020

Superficial brachial artery (SBA) is the term used for when the brachial artery (BA) itself or an early branch from the BA travels superficial to the roots of the median nerve (MN) or the MN itself. The purpose of this study was to determine the prevalence of the aforementioned SBA variations, to identify early branching variants of the SBA, and to explore the appropriateness of the anatomical terminology of these SBA variants due to ambiguity in the nomenclature. Assessment of 87 cadavers (174 upper extremities) led to the discovery of nine (5.2%) SBA variations which were observed in a total of seven (8.0%) cadavers (five females, two males). Three different SBA variation types were observed: Variation 1 was identified when the entire BA ran superficial to the median nerve roots, present in 1 of 174 limbs (0.57%); Variation 2 was noted when the BA bifurcated proximal to median nerve roots, resulting in superficial brachial and brachial arteries, seen in 6 of 174 limbs (3.4%); and Variation 3, wherein the BA bifurcated distal to the median nerve roots at the location of an additional connection between the musculocutaneous and median nerves, also resulting in superficial brachial and brachial arteries in 2 of 174 limbs (1.1%). The branches of the BA were followed to discern their branching patterns distally. In the six examples described as Variation 2, four (67%) of the SBAs traveled distally to become the radial artery only (Variation 2A), and two (33%) traveled distally to become both the radial and ulnar arteries (Variation 2B). For the two examples identified as Variation 3, both of the SBA vessels turned into solely the radial arteries distally. Therefore, Variations 2A and 3 both travel distally to become solely the radial artery; however, there is confusion in the literature as to whether these SBA variants can also be called high origin radial arteries (HORAs). Therefore, clarification of the current criteria for naming these brachial artery variants will also be discussed. Due to the unusual locations of SBAs, these variant arteries may cause confusion during surgery and in the interpretation of imaging studies.Support or Funding InformationWV Research Challenge Fund [HEPC.dsr.17.06]
Exercise‐enriched extracellular vesicles benefit brain health by decreasing protein aggregation
Authors: Geiger Paige C. , Deng Fengyan , Miller Joshua , Morris Jill , Vidoni Eric , Burns Jeffrey 
Published: April 2020

Aerobic exercise results in improved brain health during both healthy aging and dementia. However, the precise biological mechanisms that drive exercise‐induced benefits for brain health remain unknown. One possible mechanism by which exercise may benefit the brain is by facilitating the delivery of molecular mediators within extracellular vesicles (EVs; exosomes and microvesicles). EVs are bilayer‐phospholipid enclosed vesicles that carry protein and microRNA cargo and are released into the blood from many organs and cells. EVs have been shown to regulate nerve regeneration, synaptic function and behavior. Importantly, when directly injected into rodent brains, EVs can effectively eliminate protein aggregates like amyloid‐β‐peptide (Aβ), which are closely linked to Alzheimer’s Disease (AD) pathology. We hypothesize that EVs isolated from human serum post‐exercise could effectively clear Aβ aggregates in neuronal cells. To test this hypothesis, we isolated EVs from cognitively healthy subjects and early onset AD subjects (8 non‐dementia subjects, ND and 4 early AD subjects) 15 minutes after maximal aerobic exercise (EX). EV size did not change with exercise in either ND or AD subjects, while EV concentration increased by 20% post‐EX in both groups. EV function was tested by incubating serum EVs with human neuronal SY5Y cells over expressing β‐amyloid precursor protein (APPswe). We found that post‐EX‐derived EVs were more effective at alleviating Aβ protein aggregation (40% decrease in aggregation compared to pre‐EX‐derived EVs). Heat Shock Proteins (HSPs), molecular chaperones that play a critical role in protein folding/misfolding, were increased in post‐EX EVs. These findings demonstrate a novel way in which exercise could benefit the brain, through the cell‐to‐cell signaling network of extracellular vesicle communication.Support or Funding InformationResearch support by the University of Kansas Alzheimer’s Disease Center.
Endogenous Promoters enable efficient CRISPR/Cas9 in a Tilapia Cell Line Model
Authors: Hamar Jens , Kültz Dietmar 
Published: April 2020

Cell cultures are effective supplemental models to study specific biochemical pathways used for environmental adaption in animals. They enable isolation from system influence and facilitate control of the extracellular environment. The recent rise of CRISPR/Cas9 gene editing has provided an additional powerful tool to aid in these studies. For work focusing on fish species many fish cell lines now exist. However, conventional tools and methods for implementing CRISPR/Cas9 gene targeting in these cells doesn’t always provide sufficient results as seen in other animal cell models. A tilapia brain cell line (OmB) developed in our lab has proven useful for gene expression studies. Previous work with multiple conventional methods attempting to apply CRISPR/Cas9 failed to indicate genomic alteration at the targeted sites. Here we present a plasmid vector based system utilizing both Polymerase II and III tilapia endogenous promoters that enables proficient gene targeting. This system demonstrated efficient cleavage in most target sites attempted with efficiency as high as 80%. These tools will allow generation of knockout cell lines for gene targeting studies in tilapia and other phylogenetically related fish species.Support or Funding InformationThis study was funded by a grant from NSF (IOS‐1656371) and BARD (IS‐48000‐ISR).