bluuebunny/crossref_metadata_2025_split huggingface dataset:
Validation of a Formaldehyde‐Free Embalming Method for Preparing High Fidelity Cadavers for Teaching Surgical and Procedural Skills
Authors: Goodwin Shannon J , Kibbee Richard , Ramnanan Christopher J , Jalali Alireza 
Published: June 2021

Cadavers embalmed using traditional formaldehyde‐based solutions are considered low‐fidelity in terms of representing the flexibility and texture of patients. Further, many current ‘soft’ embalming solutions are expensive, involve complicated procedures, and/or result in short‐term preservation periods. The objective of this study was to develop a low‐cost, formaldehyde‐free embalming solution that would produce high fidelity cadaveric models for teaching procedural or surgical skills in postgraduate medical education. The novel embalming formula was based on replacing formaldehyde with saturating levels of sodium chloride (357 grams/L) in combination with other conventionally used, low cost chemicals. Formaldehyde‐free cadavers (FFC) were compared with cadavers embalmed using a traditional formaldehyde‐based formula in terms of microbiological profiles, and in terms of feedback from residents and faculty who participated in cadaveric‐based workshops. Swabs from oral, nasal, and rectal cavities over a 100 day time course in formaldehyde‐free donors demonstrated a nearly complete reduction of microbes that was generally comparable to that observed in formaldehyde‐preserved cadavers. In terms of feedback, qualitative commentary was obtained from separate workshops for anesthesiologists, orthopaedic surgeons, plastics surgeons, ophthalmologists and neurosurgeons over a two year period (2016–2018). Across these different groups, clinicians generally perceived that FFC donors featured enhanced fidelity in terms of flexibility and texture, and produced more realistic and higher quality ultrasound images, relative to formaldehyde‐preserved donors. In particular, anaesthesiologists noted the potential utility of FFC specimens for practicing needle insertion techniques, while neurosurgeons and ophthalmologists had strong positive feedback regarding the texture and quality of brain specimens and eye musculature, respectively. FFC tissue remained viable at least two years post‐embalming for use in postgraduate workshops. In conclusion, the FFC model has the potential to generate low‐cost, high‐fidelity anatomical resources, with long‐term viability, for the purposes of surgical and procedural skills training.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Intergenerational Inheritance of the Effects of Exercise on Novel Object Recognition Memory and Pattern Separation Performance in Mice
Authors: McGreevy Kerry R. , Fontán‐Lozano Ángela , Tezanos Patri , Pallé Anna , Ferreiro‐Villar Iria , Trejo Jose Luis 
Published: June 2021

It is known that intergenerational inheritance can be induced by environmental changes in parents. These studies usually focus on negative stressors that affect the offspring adversely. There is increasing evidence that exercise improves learning and memory in humans and mice. These beneficial effects could be mediated by enhanced hippocampal neurogenesis. Whether these effects could be inherited by future generations remains to be seen. This study analysed if paternal exercise training could facilitate hippocampal‐dependent novel object recognition memory and pattern separation performance in offspring. Firstly, male progenitors were subjected to a forced moderate exercise protocol for eight weeks before mating. All male experimental animals underwent the following behavioural protocols: an activity cage and two different protocols of a novel object recognition memory test and a pattern separation test, both protocols differing in the level of difficulty. Regarding basal motor activity, no differences were found between experimental groups in a known environment. As for the novel object recognition and pattern separation tests, only male progenitors that had undergone the exercise protocol were able to discriminate the novel object and the change of pattern in the difficult protocols. No significant differences were found in postnatal behaviour. However, the behavioural outcomes in adulthood showed that only the offspring of male progenitors that had undergone physical exercise displayed facilitation in novel object recognition memory and pattern separation performance. In order to assess adult hippocampal neurogenesis, immunohistochemistry against different neurogenesis markers was performed. Furthermore, whole‐genome sequence analyses showed differential gene expression among experimental groups. These results suggest that beneficial effects of exercise training on learning and memory can be inherited.Support or Funding InformationWork was supported by project grants from Spanish Ministry of Economy and Competitiveness BFU2013‐48907‐R and BFU2016‐77162‐R (to J.L.T.). A.F. was funded by a JaE‐DOC CSIC grant, I.L.T. was funded by an FPI grant, and K.R.McG. was funded by a contract associated with the project grants above mentioned (to J.L.T.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Extracellular Matrix Protection Factor Increases Sulfated Proteoglycan Production in Serum‐free, Primary Cultures of Human Osteoarthritic Chondrocytes
Authors: Morris Gregory , Milton LaBraya , Popper Hannah , Buckley Andrea , Mattioli Patrisia , Behling Kathryn , Selim Abdulhafez , D'Angelo Marina 
Published: June 2021

Osteoarthritis (OA) is a common chronic condition which involves the loss of articular cartilage, the cushion between joints that provides a smooth surface for the bones to glide. Three compounds that make up most of the articular cartilage are type II collagen, proteoglycans, and water. In OA there is an increase in collagenase activity that degrades collagens and proteoglycans necessary for healthy cartilage. The composition of the extracellular matrix changes in OA as evidenced by production of type I collagen. Previous studies in our lab have shown that primary cultures of human osteoarthritic chondrocytes (HOACs) can be reared in serum‐free alginate culture but measurable extracellular matrix endpoints of the OA profile are dampened when chondrocytes isolated from sides of greater and least pathology in the joint are pooled together in culture. In this study, we obtained HOACs from patients undergoing total knee arthroplasty. Femoral condyles and the tibial plateau were labeled greater or least pathology, determined by gross anatomical observation, the cells were isolated and plated separately in three‐dimensional, twelve‐well, alginate cultures at a density of 1.8 × 106 cells per 0.5 mL. Samples were treated for 48 hours from day of plating with either 250nM Extracellular Matrix Protective Factor‐1 (ECPF‐1) or hyaluronic acid conjugated ECPF‐1 (HA‐ECPF‐1). Collagen I degradation and sulfated proteoglycan synthesis were measured in alginate‐associated matrix of the treated cultures. HOACs isolated from the side of greatest pathology responded to treatment by reducing collagen type I degradation (8.42ng/culture Control, 7.25ng/culture ECPF‐1 and 4.25ng/culture HA‐ECPF‐1) and increasing retention of sulfated proteoglycans in the extracellular matrix (0.36ug/culture Control versus 0.56ug/culture ECPF‐1 treated). These data confirm that 48 hour treatment in serum free HOAC culture is sufficient to measure the therapeutic efficacy of an OA drug intervention and that both formulations of ECPF‐1 can slow the progression of OA by altering production of sulfated proteoglycan and collagen type I.Support or Funding InformationThis work was supported in part by a grant from the New Jersey Health Foundation and Philadelphia College of Osteopathic Medicine's Center for Chronic Disorders of Aging and the Division of Research.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Interaction of V. parahaemolyticus Effector VopR and its Putative Chaperone VP1684
Authors: Lafrance Alexander , De Nisco Nicole , Jiang Catherine , Orth Kim 
Published: June 2021

Vibrio parahaemolyticus (V. para) is a Gram‐negative, halophilic bacteria that causes acute gastroenteritis in humans upon consumption of contaminated seafood. The V. para life cycle is facilitated by an array of virulence factors, including two type III secretion systems (T3SS). T3SS‐mediated translocation of effectors into host cells is dependent on delivery of each effector to the base of the T3SS needle by a chaperone. VopR is an effector secreted by the first V. para T3SS (T3SS1) that uses an N‐terminal PIP2 –binding domain (BPD) to localize to the plasma membrane and induces host cell rounding through an unknown catalytic mechanism. We hypothesize that VP1684, encoded by a gene directly upstream of vopR in the V. para genome, is the cognate chaperone of VopR, and that the BPD also constitutes the chaperone‐binding domain. Through affinity column purification coupled with FPLC filtration we demonstrate that VP1684 forms a homodimer in solution much like other T3SS effector chaperones, which corroborates bioinformatics analyses indicating that VP1684 shares conserved biochemical properties with T3SS effector chaperones. We also show that VP1684 forms a stable complex with VopR. Moreover, we reveal through confocal microscopy of HeLa cells infected with V. para that the VopR‐mediated cell rounding phenotype is VP1684‐dependent. Finally, we show via increases in host cell cAMP levels by translocated cyaA‐tagged effectors that the translocation of VopR into the host cell by T3SS1 also depends on the presence of VP1684. Taken together, these data suggest that VP1684 is acting as the previously unidentified cognate chaperone for VopR, and as a necessary factor in effector translocation plays a significant role in V. para pathogenicity.Support or Funding InformationThis work was funded by the HHMI, National Institutes of Health (NIH) grant R01‐AI056404, NIH grant 5T32AI007520‐20, the Welch Foundation grant I‐1561, and Once Upon a Time&[hellip]Foundation. STARS program funding provided by the state of Texas.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Mechanism of Protein Kinase A in Fibrolamellar Hepatocellular Carcinoma
Authors: faradonbeh mahsa shirani 
Published: June 2021

Fibrolamellar Hepatocellular Carcinoma (FL‐HCC) is a rare form of liver cancer that affects adolescents and young adults without a prior history of liver disease. Chemotherapy is ineffective in treating FL‐HCC, and surgical resection of the primary tumor and metastases is currently the only semi‐effective treatment. Reoccurrence rates in FL‐HCC are extremely high.A 400kB deletion in chromosome 19 results in the fusion of the first exon of a heat shock protein, DNAJB1, with the catalytic subunit of protein kinase A, PRKACA (1). The new chimeric protein, DNAJB1‐PRKACA, retains the full catalytic activity of protein kinase A. We have performed partial biophysical characterization of DNAJB1‐PRKACA in comparison to PRKACA. Our preliminary results indicate the structural stability of DNAJB1‐PRKACA may differ from that of PRKACA. We also report the DNAJB1‐PRKACA chimera forms a stable complex with its regulatory domains, and differences in structural stability may extend to the chimera/regulatory domain complex. Surface plasmon resonance (SPR) does not reveal any difference in the binding affinity of DNAJB1‐PRKACA and wild type PRKACA to their Regulatory subunit type Iβ.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Oxidative Metabolism and Comparative Analysis of Synthetic Cannabinoid N‐(1‐adamantyl)‐1‐(5‐fluoropentyl)indazole‐3‐carboxamide (5F‐AKB‐48) and the Unfluorinated Analog AKB‐48
Authors: Yarbrough Azure Leigh , Pinson Anna , Prather Paul L. , Fantegrossi William E. , Bush John M. , Fujiwara Ryoichi , Radominska‐Pandya Anna 
Published: June 2021

Recently, the rising frequency in the abuse of synthetic cannabinoids (SCBs) has resulted in the occurrence of numerous severe adverse effects. We have previously shown that several early generations of SCBs undergo extensive metabolism by cytochrome P450s (P450s) and that their metabolites retain biological activity at CB1 and CB2 receptors. In this study, we examined the oxidative metabolism of 5F‐AKB‐48, one of a new structural generation of SCBs. Activity towards 5F‐AKB‐48 was investigated using various human microsomes followed by studies utilizing multiple recombinant human P450s. Screening with these enzymes showed that only 4, CYP2D6, −2J2, −3A4, and −3A5, biosynthesized the 2 major products, mono‐ and dihydroxylated 5F‐AKB‐48, with activities in the range of 2–5 pmol/min/pmol CYP. CYP3A5 formed the most dihydroxylated metabolite, whereas CYP2J2, highly expressed in cardiovascular tissues, showed the biosynthesized the most monohydroxylated derivative. CYP2D6 produced only monohydroxylated derivatives whereas CYP3A4 produced mostly dihydroxylated ones. To elucidate the roles of CYP3A4 and CYP3A5, additional enzymatic reactions utilized genotyped HLMs containing normal activity for CYP3A4 but low or high activities of CYP3A5. Those experiments resulted in significantly different patterns of oxidized metabolites. An UPLC equipped with UV/Vis detector and/or MS/MS were used to analyze the reactions. The profiles of the products biosynthesized were significantly different between the fluorinated and unfluorinated analogs. As this structural difference is known to increase potency for CB1 receptors, understanding the involved mechanism is essential. In conclusion, extensive metabolic studies of SCBs, including investigation of the mechanism(s) of action leading to toxicity, could result in the development of life saving, efficacious treatments for SCB abuse.Support or Funding Information(INBRE 117‐1006096 2018 AP and NIH/NIDA DA039143 to ARP and PLP)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Do Lactic Acid Bacteria produce antifungal bacteriocins?
Authors: Turcotte Madison , Allen Mary E , Piefer Andrew J 
Published: June 2021

Bacteriocins are small antimicrobial peptides that are produced by bacteria and are known to inhibit the growth of closely related or non‐related bacterial strains without causing harm to the producer strain. Bacteriocins are categorized into several classes and lactic acid bacteria (LAB) are known to produce class IIa bacteriocins which are described as small, heat‐stable peptides. LAB produced bacteriocins could create a wide range of applications in food preservation, which may present a safer alternative than currently used chemical preservatives. The mold Fusarium graminearum frequently contaminates barley malt, which along with water, yeast, and hops form the ingredients of beer. Hydrophobic compounds produced by F. graminearum causes beer to retain high levels of carbon dioxide, which results in gushing, or excessive foaming when bottled. Gushing can damage the reputation of a brewery. Thus, malt producers are searching for a way to control this grain contaminant. Recently, several studies demonstrating potential antifungal properties of bacteriocins were reported. Using a sourdough starter, we were able to isolate environmental strains of LAB by growing them anaerobically on MRS agar medium. We will then test their potential antifungal properties against F. graminearum using traditional microbiological methods. If we can determine that bacteriocins produced by LAB are in fact antifungal, we will then use PCR to rapidly detect bacteriocin genes that might be present in these strains. Potential bacteriocins will be exposed to a variety of experimental conditions such as protease treatment, pH, and detergents for further classification and characterization.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐Fibrotic Effects of HDAC8 Inhibitor in Bleomycin‐Induced Pulmonary Fibrosis Mouse Model–Whole Transcriptome Analysis Using RNA‐seq
Authors: Saito Shigeki , Zhuang Yan , Lasky Joseph , Liu Yaozhong 
Published: June 2021

BackgroundIdiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease with high mortality. Current therapies for IPF have only limited efficacy, thus there is a critical need for more effective medications. We have previously reported that HDAC8 inhibition represses TGF‐β1‐induced fibroblast‐myofibroblast differentiation in lung fibroblasts and that an HDAC8‐selective inhibitor NCC170 ameliorates bleomycin‐induced pulmonary fibrosis in mice. Here we report our recent analyses of RNA‐seq data derived from the NCC170‐treated mice.MethodsOn Day 0, mice were given Bleomycin or PBS (=vehicle) via oropharyngeal aspiration. On Day 7–13, mice were given NCC170 or DMSO (=vehicle) daily via intraperitoneal injection. On Day 14, lungs were harvested and total RNA was isolated for RNA‐seq. Differentially expressed genes between the PBS+DMSO group (n=4) and the Bleomycin+DMSO group (n=4) as well as between the Bleomycin+DMSO group and the Bleomycin+NCC170 group (n=4) were identified using DESeq2. Differentially regulated pathways between these groups were identified using Generally Applicable Gene‐set Enrichment (GAGE) analysis. To confirm the findings from RNA‐seq and GAGE analyses, THP‐1 cells (human monocytic cells) were differentiated into macrophages with PMA, and then treated with HDAC8 siRNA or scramble siRNA. The effects of HDAC8 knockdown on expressions of pro‐fibrotic cytokines were assessed using immunoblots.ResultsThe comparison between the PBS+DMSO group and the Bleomycin+DMSO group revealed that the Bleomycin treatment upregulated 3330 genes (>1.5 fold change) and downregulated 2598 genes (<0.66 fold change) with FDR<0.05. The comparison between the Bleomycin+DMSO group and the Bleomycin+NCC170 group revealed that the NCC170 treatment upregulated 182 genes and downregulated 454 genes. GAGE analysis suggested that NCC170 repressed bleomycin‐induced activation of several pathways, including well expected pathways (e.g., extracellular matrix ‐ receptor interaction) as well as novel pathways (e.g., innate immunity). Our in vitro study confirmed that HDAC8 knockdown repressed pro‐fibrotic cytokines (e.g., IL‐6) in PMA‐treated THP‐1 cells.ConclusionOur RNA‐seq analysis provided novel insights into the mechanism through which HDAC8 inhibition exerts anti‐fibrotic effects in a bleomycin‐induced pulmonary fibrosis mouse model, a widely‐used animal model relevant to IPF. The role of HDAC8 in innate immunity during pulmonary fibrosis development should be investigated further.Support or Funding InformationWetmore FoundationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ERK Controls Myosin Phosphatase and Cell Migration through RSK Phosphorylation of MYPT1
Authors: Mendoza Michelle C , Samson Shiela , Elliott Andrew , Carney Keith 
Published: June 2021

Cell migration is essential to embryonic development, wound healing, and cancer cell dissemination. Cells move via leading edge protrusion, substrate adhesion, and retraction of the cell's rear. However, the signaling mechanisms by which extracellular cues control the actomyosin cytoskeleton to induce motility are poorly understood. The growth factor‐responsive and oncogenically activated ERK kinase promotes motility by signaling for actin polymerization‐mediated edge protrusion. ERK has previously been proposed to also control the contractile machinery through myosin light chain kinase (MLCK). Here, we show that ERK indeed signals to myosin, but rather through its substrate, p90 ribosomal S6 kinase (RSK). By probing the signaling and migration dynamics of a cell line panel, we found that RSK phosphorylates the myosin phosphatase targeting subunit (MYPT1) at Ser507, which promotes Rho kinase (ROCK) interaction with and inhibition of MYPT1. By inhibiting the myosin phosphatase, ERK and RSK promote the myosin II–mediated tension necessary for lamella expansion and protrusion and retraction persistence. This level of myosin activation promotes optimal edge dynamics for cell migration, rather than inducing a strong contractile response. In conclusion, our findings suggest that ERK activity can coordinately amplify both protrusive and contractile forces for optimal cell motility.Support or Funding InformationK01CA168850, ALA Research Grant, and American Cancer Society RSG CSM130435 to MCMThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of eIF2α in the Collagen Production in Hepatic Stellate Cell is Associated with Autophagic and Apoptotic Signaling
Authors: Qi‐Lin Liu , Teng Lin Hua , Liu Ze‐Feng , Li Fu‐Jian , Chen Li , Zhang Guo 
Published: June 2021

Background & AimsStudies have documented cellular apoptosis, autophagy and endoplasmic reticulum (ER) stress participate in the liver fibrogenesis. However, the specific mechanism remains unclear. This study was aimed to determine whether and how eIF2α protein play a role in the collagen generation in hepatic stellate cell (HSC) via autophagic signaling events.MethodsHSC‐LX2 cell line was activated by transforming growth factor beta1 (TGF‐β1) in the presence of eIF2α plasmid or vehicle control. Real‐time fluorescence quantitative PCR was then performed to compare the differential mRNA level of COL1, eIF2α, CHOP, Bcl‐2 and Bax. Western Blot was performed to analyze the protein expression of α‐SMA, COL1, LC3, P62, eIF2α, P‐eIF2α and Bcl‐2 in LX2 cells. Meanwhile, the numbers of apoptotic cells were measured by flow cytometry.ResultsCompare to vehicle control, the ER stress hallmarks of eIF2α, CHOP were at lower level in HSC‐LX2 cells in TGF‐β1 treated condition. While transfected eIF2α plasmid in HSC‐LX2 cells, it was found that CHOP and BAX significantly increased in these cells but substantial reduction in LC3, P62, Bcl‐2, α‐SMA and COL1 v.s. control(<0.05). Moreover, thus exogenously enhancing of eIF2α gene was found to induce apoptosis of HSC‐LX2 cells with a 50% increment (<0.05).ConclusionsOverexpression of eIF2α in HSCs may trigger autophagy inhibition and apoptosis promotion, thereby reduce collagen production in these cells, which provide a potential therapeutic strategy for hepatic fibrosis.Support or Funding InformationSupported by NSFC Grants 81360077, 81860654, 30630145, 81172260 and GXNSFC Grants 2015GXNSFDA139022 to Guo ZhangThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.