bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell‐based hormone therapy prevents diastolic dysfunction after estrogen loss in the Spontaneously Hypertensive Rat (SHR)
Authors: Groban Leanne , Sun Xuming , Sivanandane Sittadjody , Wang Hao , Hodge Hunter S. , Basu Swati , Kim-Shapiro Daniel G. , Opara Emmanuel C. 
Published: April 2020

Introduction. Preclinical left ventricular diastolic dysfunction (LVDD) is a forerunner to heart failure with preserved ejection fraction, which is seen more often in older women than men of the same age. While hormone therapy plays an important role in the management of vasomotor and genitourinary symptoms experienced by many women during the menopausal transition, its efficacy with regard to maintenance of cardiac structure and function and prevention of LVDD disease progression with aging has not been tested due to the concern for off‐target health risks (e.g., cancer, blood clots). We developed a cell‐based hormone therapy (cHT) approach that recapitulates native cell–cell interactions between ovarian granulosa and theca cells in a 3D bioengineered construct to mimic the dynamic release of sex hormones into the circulation. Our first report in ovariectomized rats shows that ovarian constructs are able to ameliorate various somatic adverse effects of hormone deficiency (e.g. bone loss). To extend these findings to cardiac health, we sought to determine the efficacy of cHT in preserving cardiac diastolic function after estrogen loss using a hypertensive rodent model.Methods14 SHR females underwent bilateral ovariectomy (OVX) while 5 SHRs received sham surgery at 12 weeks of age; 8 weeks post‐surgery, OVX rats were randomized (n=4–5/group) to vehicle, cHT, or pharmacologic hormone therapy (pHT: E2 (10mg/kg/day) and P4 (2mg/kg/day) subcutaneous pellets) and compared to gonad‐intact rats treated with vehicle (VEH). Body weights and tail cuff systolic blood pressure (SBP) were measured biweekly. Following 10 weeks of treatment, rats were lightly anesthetized (isoflurane) for Doppler measures of diastolic function. Data were analyzed by one‐way ANOVA. P<0.05 was significant.ResultsOVX rats, irrespective of treatment, had higher body weights when compared to sham (P<0.001). Uterine atrophy by OVX was minimized by pHT and cHT, reflecting modest E2‐related effects (Figure
A). SBP increased progressively over time (P<0.01), without overt differences across treatment groups. Even so, both cHT and pHT prevented adverse OVX‐related effects on myocardial relaxation (e') and Doppler‐derived filling pressures (E/e') when compared to vehicle (Figures
B, C).ConclusionThese data show that novel cell‐based hormone therapy is effective in preventing diastolic dysfunction from surgically‐induced menopause in the SHR. Additional studies are underway to determine cHT’s long‐term (6 months) treatment efficacy in middle‐to‐advanced aged OVX rats.Support or Funding InformationNIH AG061588Figure 1
Dynamic Phosphorylation of Yeast eIF4A Drives Changes in Gene Expression that Couple Protein Synthesis with Cell Division
Authors: Sahoo Ansuman , Ashraf Marium , Walker Sarah E. 
Published: April 2020

The eukaryotic translation initiation factor 4A (eIF4A) supports mRNA recruitment to the ribosomal preinitiation complex through activity of its conserved DEAD‐box RNA helicase motif. Interestingly, in plants eIF4A is differentially phosphorylated near the DEAD motif during various phases of the cell cycle by CDKA, lowering translation activity during mitosis (
Bush et al., 2016). The same Threonine residue of yeast eIF4A (T146) is present in a consensus CDK1/CDKA/Cdc28 motif, and is highly phosphorylated in large‐scale studies (
Soulard et al., 2010). To further dissect the molecular function of eIF4A, we are analyzing specific in vitro and in vivo changes that take place due to phosphorylation of eIF4A under normal conditions and in response to stress. We have mutated eIF4A residue T146 to phosphodeficient (Ala) and phosphomimetic (Glu, Asp) forms, and analyzed the effects of these mutations on growth and translation. Our data indicate that the phosphorylation status of eIF4A sensitizes yeast cells to membrane stressors in a manner affected by eIF4A•eIF4B coupling, and that constitutive phosphorylation of this residue is lethal due to arrest prior to cell division. Mechanistically, phosphorylation disrupts RNA binding by eIF4A. Together our results suggest that dynamic phosphorylation of eIF4A modulates translation changes that promote cell division and resistance to external stressors.Support or Funding InformationThis work was supported by NIH grant R00GM119173 and startup funds from SUNY at Buffalo‐College of Arts and Sciences
Digital radiological integration into the first year medical anatomy course: A systematic literature review
Authors: Williams Sarah M. 
Published: April 2020

As diagnostic digital radiology is becoming the standard in medicine, numerus medical schools have incorporated digital radiology (i.e. CTs, MRIs, Xrays and Ultrasound) into their preclinical curriculum. There are a lack of resources that provide a comprehensive insight into the most common methods of digital radiological integration into the first year medical anatomy course. The purpose of this study is to perform a meta‐analysis on most common methods of digital radiological integration and to determine the overall trend in the usefulness of this technique as a teaching method. Utilizing the PICO (Population, Intervention, Comparison, Outcomes) reporting system, two of the following questions are proposed: (i) What is the most common method of integration of digital radiology in the first year medical anatomy course? (ii) What are the learner outcomes for assessment of anatomical knowledge using digital radiological imaging? The systematic literature review is conducted in accordance with Preferred Reporting Items for Systematic reviews and Meta‐Analysis (PRISMA) guidelines. Identification of citations (n=1688) were obtained searching keywords “radiology” AND “anatomy” AND “medical student” in the following four databases: PubMed, Embase, Web of Science, and Scopus (Table 1). Duplicates were excluded using the “find duplicates” tool in Medeley citation software and articles published prior to 2009 were excluded (n=276). A reviewer team will be assembled to analyze the degree of inclusion of remaining articles screened (n=1412). Results indicate that there are diverse ways to incorporate radiological images into a gross anatomy course and the method of incorporation may affect the successfulness of the method.A summary of the screening process. Out of 1688 articles, 1412 advanced to be analyzed by the reviewer team based on the eligibility inclusion criteria.Figure 1
Feline Chondroblastic Osteosarcoma: Comparative Analysis by Pathology Techniques
Authors: Takahashi Tamy Verony , de Albuquerque Lima Mobricci Luciana , Hyodo Vivian , Duarte Janaína 
Published: April 2020

Even though primary bone tumors are considered uncommon in cats, osteosarcoma is the most common, with a prevalence of 70% to 80% of all primary bone tumors reported in this species. However, the number of cases reported are still scarce when compared to those reported in dogs. The goal of this study was to diagnose, through necropsy and histopathology, a case of feline chondroblastic osteosarcoma presented to a private practice in Brazil, describing the macroscopic and microscopic findings. The necropsy was performed at the Pathology Laboratory of Paulista University, located in São José dos Campos, Brazil, using an adaptation of the Ghon method, developed by Professor Janaína Duarte, DVM, PhD. Histopathology was performed by a veterinary diagnostic laboratory located in São Paulo, Brazil.A female, spayed, 12 years old, domestic short hair cat, was presented to an ophthalmology specialist practice with a complaint of suspected glaucoma, which a skull radiograph later showed to be, in fact, exophthalmus caused by a mass in the left eyeball topography, with loss of definition of the zygomatic bone’s rostral aspect and the adjacent maxilla. Euthanasia was performed due to worsening of the general state and economic limitations to follow further diagnose and treatment. Necropsy revealed a lobulated, semi‐hard, pale red mass, measuring approximately 4.0 × 3.0 × 1.0 cm, adhered to the left mandible vertical ramus and pressing against the left eyeball. Transverse cutting showed a translucent, whitish, partially mineralized surface. Histopathology exhibited proliferation of spindle shaped cells, with marked degeneration, cytoplasmic retraction and acidophilic staining, arranged in disorganized bundles, permeated by immature and mineralized osteoid matrix. The nuclei were rounded, hyper‐chromatic with occasional nucleoli evidence. Figures of mitoses were not identifiable due to cellular degeneration. It was also seen that between the cells there were multiple focuses of marked chondroid tissue proliferation, highly suggestive of chondroblastic osteosarcoma.Macroscopic findings during necropsy showed a mass with malignant and invasive biological behavior, affecting adjacent structures such as the eyeball and the zygomatic arch, which is consistent with those of histopathology that showed proliferation and marked degeneration of atypical spindle shaped cells with infiltration of the chondroid matrix by osteoid tissue.Location of the mass, filling the left eye orbit (arrow).Figure 1Proliferation of spindle snapped cells permeated by osteoid and chondroid matrix.Figure 2
Coordinated Gene Expression of Na+/Cl−cotransporter 1 (ncc1) and Clc family Cl−channel 2 (clcn2c) in Kidney and Urinary Bladder of Euryhaline Tilapia
Authors: Nelson Nastasia , Petro-Sakuma Cody , Celino-Brady Fritzie , Seale Andre , Breves Jason 
Published: April 2020

Euryhaline fishes maintain hydromineral balance in a broad range of environmental salinities through the functional coordination of multiple tissues, including the gill, intestine, kidney, and urinary bladder. Fishes inhabiting freshwater (FW) environments are faced with the diffusive loss of ions and the osmotic gain of water. In turn, both kidney and urinary bladder support the reabsorption of Na+ and Cl− and the excretion of excess water. While functional responses by these two organs to FW conditions have been previously described, the subcellular pathways for Na+ and Cl− reabsorption are not well understood. We hypothesized that euryhaline tilapia (Oreochromis mossambicus), employ Na+/Cl− cotransporter 1 (Ncc1) and Clc family Cl− channel 2 (Clcn2c) for the active reabsorption of Na+ and Cl− and thus their respective gene transcripts are expressed under FW conditions. In the current study, we report ncc1 and clcn2c gene expression patterns in both steady‐state animals and animals abruptly transferred from seawater (SW) to FW. In kidney and urinary bladder, ncc1 and clcn2c gene expression was markedly enhanced (>30‐fold) in FW‐ versus SW‐acclimated tilapia. By 6 hours after transfer from SW to FW, ncc1 and clcn2c in both kidney and urinary bladder were coordinately elevated compared with controls continuously maintained in SW. These patterns in the kidney and urinary bladder occurred in concert with branchial processes supportive of Na+ and Cl− absorption from the external environment. Collectively, our results suggest that both Ncc1 and Clcn2c contribute to the functional plasticity of kidney and urinary bladder that underlies acclimation to variable environmental salinities.Support or Funding InformationSupported by the National Science Foundation (IOS‐1755131 and ‐1755016).
Autophagy Inhibition Sensitizes Liver Kinase B1 (LKB1)‐Deficient Kras‐Driven Lung Tumors to MEK Inhibitor Trametinib
Authors: Bhatt Vrushank Dharmesh , Khayati Khoosheh , Kong Jerry , Raju Akash , Su Xiaoyang , Guo Jessie Yanxiang 
Published: April 2020

Tumor suppressor liver kinase B1 (LKB1) activates 5’‐adenosine monophosphate protein kinase (AMPK) and plays an essential role in maintaining energy homeostasis. LKB1 mutations are the third most frequent mutation detected in non‐small cell lung cancer (NSCLC). Compared to p53 mutation, co‐mutations in LKB1 with KRAS causes aggressive tumor growth and resistance to chemotherapy as well as immunotherapy. Thus, identifying a novel target to improve LKB1‐deficient Kras‐driven NSCLC treatment is urgently needed. Our previous work demonstrates that autophagy inhibition is synthetically lethal in KrasG12D/+;Lkb1−/− (KL) mediated tumorigenesis; in contrast to intact autophagy KL lung tumors, loss of an autophagy‐essential gene Atg7 dramatically impaired both tumor initiation and tumor growth. This is in sharp contrast to wild‐type (WT) LKB1 (KrasG12D/+;p53−/−(KP)) tumors that are less sensitive to autophagy gene ablation. These observations indicate that LKB1 mutations predispose KRAS NSCLC to autophagy inhibition and that LKB1 mutations could be explored as a predictive biomarker for precision lung cancer therapy. To further value our discoveries in clinical translational ability, we treated mouse lung tumor derived cell lines (TDCLs) with FDA approved autophagy inhibitor hydroxychloroquine (HCQ) and found that the KL TDCLs were much more sensitive to HCQ‐induced cell death compared with Lkb1‐WT KP TDCLs. Furthermore, a combination treatment of HCQ with mitogen‐activated protein kinase kinase (MAPKK or MEK) inhibitor Trametinib showed synergistic anti‐proliferative effects in KL TDCLs, but not in KP TDCLs. To elucidate the underlying mechanism of increased sensitivity of KL TDCLs to Trametinib by autophagy ablation, we performed metabolomics profiling of KL TDCLs with Trametinib, HCQ alone or in combination treatment. We found that the levels of several glycolytic and TCA cycle intermediates, as well as amino acids were significantly upregulated upon treatment with Trametinib, which were significantly reduced by the combined treatment with HCQ and trametinib. Moreover, Trametinib treatment led to an increased level of ATP, which was decreased by the combination of HCQ and Trametinib. Similarly, we observed anti‐tumor synergistic effects of the combination treatment in KL allograft tumor growth. Our observations suggest that KL TDCLs resist to Trametinib treatment by upregulating autophagy to maintain energy homeostasis for cancer cell survival. Therefore, a combination therapy of autophagy and MEK inhibition could be a novel therapeutic strategy to specifically treat LKB1‐deficient NSCLC.Support or Funding InformationThis research has been supported by the research funds from NIH K22 CA190521 grant, American Cancer Society Early Investigator Pilot Award, Lung Cancer Research Foundation grant and startup funding from Rutgers Cancer Institute of New Jersey.
A Three‐Pronged Synergistic Approach to Increase the Biofeedstock Viability of Chlorella vulgaris
Authors: Slone Danielle Renee , Smythers Amanda Louise , Kolling Derrick 
Published: April 2020

Over the last several decades, the global temperature has increased at an alarming rate, increasing the need for efficient renewable energy sources.
1 Microalgae are a promising source of biodiesel due to their high rate of lipid accumulation, but are not efficient enough to produce biofuel on a scale matching current fuel needs.
2Chlorella vulgaris is a fast‐growing unicellular green microalga with a high percent of lipids per biomass, making it a promising candidate for biodiesel production. Previous research has used two approaches to increase the accumulation of lipids and photosynthetic activity: the addition of exogenous glycerol or the addition of excess Mn2+. When used separately, the addition of glycerol increases neutral lipid production in C. vulgaris while the addition of Mn2+ increases photosynthetic capacity.
3 Glycerol was found to be directly incorporated into the triacylglycerol (TAG) backbone, which allowed for an increased flux toward TAG synthesis without significantly changing the energy exerted. Manganese was shown to increase the rate of electron transfer through photosystem II, increasing the overall photosynthetic output. While it was assumed that combining these conditions would further optimize the cultures for lipid production, this was not the case: when used simultaneously, they decrease TAG production. To improve lipid productivity, a three‐pronged synergistic approach was taken combining Mn2+‐replete conditions, nitrogen deprivation, and exogenous glycerol. Nitrogen was deprived from the cells, because under nitrogen deplete conditions cells shift away from protein synthesis and towards energy storage compounds that can be used to produce TAGs. Cultures were then monitored for turbidity, chlorophyll content, lipid productivity, biomass, and TAG content. Cells grown using this three‐pronged approach showed an increase in the amount of TAGs produced over time as the glycerol concentrations were increased. This novel approach can be used to increase lipid synthesis to increase biofuel feasibility.Support or Funding Information#NNX15AI01H.D.S. was supported from the Marshall University Department of Chemistry alumni donors to the Marshall University Department of Chemistry’s Summer Undergraduate Research Fellowship.D.R.J.K. and A.L.S. were supported by the National Science Foundation under Cooperative Agreement No. OIA‐1458952. A.L.S. was also funded by the NASA West Virginia Space Grant Consortium, Grant #NNX15AK74A and Training Grant
Stiffness Induces NAFLD‐like Metabolic Dysfunction in Primary Hepatocytes
Authors: Kidambi Srivatsan 
Published: April 2020

BackgroundNon‐alcoholic fatty liver disease (NAFLD), the liver manifestation of the metabolic syndrome linked to obesity and insulin resistance, affects 25% of the general population both in western and developing countries. The development and progression of NAFLD is mediated by increase in lipid accumulation and triglycerides levels, excessive oxidative stress and inflammation that in turn exacerbates the diseased condition and impairs metabolic parameters. NAFLD progression is also characterized by liver stiffness due to extensive remodeling of the extracellular matrix. The role of matrix stiffness as related to subtle but pivotal changes in hepatocytes physiology and metabolic dysfunction is under explored. The overall goal of our study is the development and implementation of a platform that enables the convergence of engineered cell microenvironments with the phenotypic and functional analysis of hepatocytes. Using our innovative biomimetic liver fibrosis model that allows modulation of substrate stiffness, we investigated the role of liver matrix stiffness in modulating hepatocytes metabolic function in fibrotic‐like microenvironment.Materials and MethodsPrimary hepatocytes were cultured on our novel polymer film coated polydimethylsiloxane (PDMS) gels with 2 kPa, 9 kPa 25 kPa and 55 kPa elastic modulus mimicking healthy, early fibrotic, fibrotic and extremely fibrotic substrates. A wide range of functional/metabolic analysis were performed to determine: lipogenic and oxidation genes, glutathione expression, dynamics by Western Blot; Respirometry by Seahorse XFe24 Analyzer.ResultsWe demonstrated that stiffness impedes hepatic urea and albumin production, expression of drug transporter gene and epithelial cell phenotype marker, hepatocyte nuclear factor 4 alpha (HNF4a). We also demonstrated that hepatocytes cultured on NAFLD‐like stiffness showed an induction of lipogenic genes along with lowered‐oxidation genes expression, mitochondrial respiration, and glycolytic capacity, 2) increased ROS production, and 3) disruption of the mitochondrial fusion process and dynamics. We observed a significant increase in oxidized glutathione (GSSG) and reduced glutathione (GSH) in hepatocytes cultured on NAFLD‐like stiffness compared to healthy liver stiffness. Similar effect was observed in hepatocytes isolated from NAFLD rat models indicating correlation to physiological conditions. These data suggest a plausible mechanism that increased stiffness modulates hepatocyte function causing liver functional failure.ConclusionsTogether, all these data demonstrates the plausible role of stiffness in regulating hepatocytes function and contribute to metabolic dysregulation in NAFLD. Understanding the impact of stiffness on hepatocytes biology will provide significantly more nuanced data to aid drug development for NAFLD.Support or Funding InformationNIH R01AA027189‐01A1; Nebraska Research Initiative; NIH 1P20GM113126Project SummaryFigure 1
High Fat Feeding Primes Mice to Binge Alcohol‐induced Liver Injury: Critical Role of Cathelicidin as a Chemoattractant
Authors: Li Fengyuan , Liu Yunhuan , Gu Zelin , Jiang Mengwei , Zhang Lihua , McClain Craig J. , Feng Wenke 
Published: April 2020

BackgroundAlcohol consumption and obesity are known synergistic risk factors of steatohepatitis. Combination of high fat diet (HFD) and acute alcohol synergistically induced acute liver inflammation via enhanced hepatic neutrophil infiltration. CRAMP (Cathelicidin‐related antimicrobial peptide), the murine ortholog of LL‐37, which is the only known member of human cathelicidin antimicrobial peptide family, possess both anti‐ and pro‐inflammatory activity. Our previous studies showed that CRMAP deficiency exacerbated binge‐on‐chronic alcohol‐induced liver injury. Here we aimed to investigate the role of CRAMP in alcohol‐induced liver injury in HFD fed mice, and to explore the underlying mechanism.MethodsCRAMP KO mice (Camp−/−) and wild type (WT) mice were fed control diet (CD) or HFD for 10 weeks, a bolus of alcohol was gavaged 9 hours before sample harvesting.ResultsTen‐week HFD + acute alcohol markedly induced liver steatosis and injury in WT mice, reflected by significantly increased liver macrosteatosis and serum levels of AST and ALT. Immunofluorescent assay showed a significantly stronger staining of Ly6G in the livers of mice treated with HFD + acute alcohol compared to CD feeding + acute alcohol, indicating an increased neutrophil infiltration. Interestingly, HFD+acute alcohol treatment significantly increased hepatic protein levels of CRAMP, which co‐localized with Ly6G‐positive neutrophils. In addition, hepatic mRNA expression of chemokines, Mcp‐1 and Cxcl‐2, was markedly increased. Depletion of Camp gene (Camp−/−) markedly reduced HFD + acute alcohol‐induced liver steatosis, injury and neutrophil infiltration, along with reduced expression of Mcp‐1. Importantly, feeding Camp−/− mice with HFD + acute alcohol significantly reduced the mRNA expression level of Cxcr‐2, a known CRAMP receptor. In vitro study showed that synthetic CRAMP peptide increased neutrophil migration, which was dependent on Cxcr‐2 expression upon treatment by fatty acid and ethanol. In addition, significantly decreased mRNA levels of hepatic Tgf‐β and Col‐1a1 were found in Camp−/− mice compared to WT mice, suggesting that CRAMP may play a role in hepatic stellate cells activation.ConclusionsHFD + acute alcohol treatment synergistically induces liver injury through systemic upregulation of chemoattractant CRAMP and subsequent enhancement of hepatic neutrophil infiltration. CRAMP deficiency protects against acute alcohol‐induced steatohepatitis in the HFD primed mice.Support or Funding InformationSupported by grants from NIHSchematic hypothesis of CRAMP‐mediated neutrophils infiltration in HFD and alcohol‐induced liver injury in miceFigure 1