bluuebunny/crossref_metadata_2025_split huggingface dataset:
Renal Deafferentation Improves Renal Function and Blood Pressure, but not Cardiac Parameters in an experimental model chronic kidney disease.
Authors: Bergamaschi Cássia Toledo , Sanders-Silveira Sarah , Veiga Amanda Costa , Aires Regina Sousa , Milanez Maycon Igor , Campos Ruy Ribeiro , Nishi Erika Emy 
Published: April 2020

Chronic kidney disease is highly prevalent and its mortality is associated with cardiovascular disease. Experimental evidence shows that the renal nerves contribute to arterial hypertension and renal sympathetic hyperactivity worsening the disease; renal denervation has shown renoprotective and cardioprotective effects. However, renal sympathetic innervation plays a fundamental role in renal homeostasis and its removal is associated with impaired compensatory mechanisms during hemodynamic changes. Thus, the aim of this work was to investigate the effects of selective renal denervation of afferent fibers on renal and cardiovascular repercussions in chronic kidney disease (CKD) induced by 5/6 nephrectomy in Wistar rats. The use of animals was approved by the Federal University of São Paulo Animal Care and Use Committee and performed according to the National Institutes of Health guidelines for the conduct of experiments in animals. After five weeks of nephrectomy, denervation of renal afferent fibers with a periaxonal application of 33 mM capsaicin was performed. Cardiovascular and renal function parameters were evaluated three weeks after deafferentation.ResultsCKD induced increase in blood pressure (CTL 113±7.2 × CKD 185±14 mmHg), creatinine plasma levels (CTL 0.5±0.1 × CKD 1.2±0.5 mg/dl) and proteinuria (CTL 14±10 × 300±129 mg/24h). It was also observed an increase in cardiac hypertrophy, reduction in baroreflex sensitivity and in parasympathetic tone after autonomic double blockade. Afferent renal denervation reduced blood pressure (165±21 mmHg), improved creatinine plasma levels (0.8 ±0.2 mg/dl) and proteinuria (178±102 mg/24h). No alterations were observed after deafferentation over baroreflex sensitivity, cardiac hypertrophy and autonomic control to the heart. Together these results suggest that renal deafferentation improves blood pressure and renal function, without, however, improving the cardiac changes found in this CKD model. It is possible that the time expected after deafferentation or the stage of the disease in which it was performed was not sufficient to induce cardiac improvements. Further studies are necessary to elucidate this question.Support or Funding InformationCoordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001; by the São Paulo Research Foundation (FAPESP 18/02671‐3) and by the Brazilian National Research Council (CNPq)
Flow Cytometry Analysis of Anastasis using CellTrace Far Red (CTFR) cell tracker in Cultured Mammalian T Lymphocytes
Authors: Smith Perianne , Lauzon Robert 
Published: April 2020

Anastasis is a cellular recovery process that rescues apoptotic cells from the brink of death. Our lab has developed a model for studying anastasis using cultured mouse HT2 lymphocytes dependent on the cytokine interleukin 2 (IL2). Within 24 hours of IL2 deprivation, HT2 cells display characteristic changes underlying apoptosis, including cellular condensation, membrane blebbing and fragmentation into apoptotic bodies, phosphatidylserine externalization, caspase 3 activation as well as G1 cell cycle arrest. HT2 cells can subsequently be rescued by adding IL2 back into the culture medium. In this study, we have used quantitative flow cytometry in conjunction with CellTrace Far Red (CTFR) and SYTOX Green (live/dead indicator) to track the kinetics of anastasis and re‐entry of viable apoptotic cells into the cell cycle. HT2 lymphocytes continuously maintained in IL2 comprised a homogeneous, viable population of CTFR+ cells which decreased in fluorescence intensity with each subsequent cell cycle for up to 96 hours post‐CTFR labelling. To investigate apoptosis reversal, HT2 cells were labeled with CTFR, deprived of IL2 for 24 hours and then rescued with IL2 re‐addition for periods of 24, 48, and 72 hours. Our findings indicate that viable, CTFR+ HT2 cells gradually resumed cell cycle entry over the 72‐hour rescue period. As caspase activation represents a cellular marker of early‐mid stage apoptotic cells, we also investigated caspase 3/7 activity in viable CTFR+ cells using the Nucview‐488 fluorescent caspase 3 substrate. Three distinct populations of caspase‐high, medium and low, viable CTFR+ HT2 cells were observed at 24 hours following IL2 deprivation. IL2‐rescued cells subsequently segregated into caspase‐high and low populations, ultimately merging into a single, caspase‐low population by 72 hours post‐rescue. Collectively, our findings suggest that CTFR tracking can be used successfully as a tool to follow the kinetics of apoptosis reversal in IL2‐dependent mouse HT2 lymphocytes, and is a promising model for the study of the mechanistic basis of anastasis.Support or Funding InformationUnion College Faculty and Student Research Grants
Cardiovascular Responses During Graded Lower‐Body Negative Pressure in Heart Failure with Preserved Ejection Fraction
Authors: Alpenglow Jeremy , Bunsawat Kanokwan , Craig Jesse , Ryan John , Richardson Russel , Wray Walter 
Published: April 2020

Heart failure with preserved ejection fraction (HFpEF) represents approximately half of the known cases of heart failure in the United States, with equally poor outcomes and rising prevalence compared to heart failure with reduced ejection fraction (HFrEF). While changes in autonomic nervous system function is a well‐known feature of HFrEF pathophysiology, whether patients with HFpEF experience a similar decrement remains unknown. Therefore, we sought to evaluate the reflex cardiovascular responses to baroreflex unloading achieved via graded levels of lower‐body negative pressure (LBNP) in patients with HFpEF compared to healthy, age‐matched controls (CON). In six participants (three HFpEF (1 male, 70±3 yrs) and three CON (2 male, 74±2 yrs)), we assessed forearm blood flow (FBF; Doppler ultrasound), mean arterial blood pressure (MAP; Finapres), heart rate (HR; electrocardiogram) and forearm vascular conductance (FVC) at rest and throughout graded levels of LBNP (−10, −20, −30 and −40 mmHg). At rest, there were no significant differences between HFpEF and CON in FBF (109±32 vs. 85±29 ml/min), MAP (90±4 vs. 96±3 mmHg), HR (60±5 vs. 61±5 bpm), or FVC (1.19±0.33 vs. 0.89±0.28 ml/min/mmHg). The reflex tachycardic response to baroreflex unloading induced by LBNP was similar between patients with HFpEF and CON. Vasoconstriction in response to LBNP decreased FBF to a similar level in both HFpEF and CON (−10: 102±31 vs. 88±28 ml/min; −20: 95±30 vs. 76±25 ml/min; −30: 91±30 vs. 78±28 ml/min; −40: 91±33 vs. 71±21 ml/min, HFpEF vs. CON), with similar changes in FVC. Together, these preliminary data suggest that baroreflex‐mediated cardiac and vascular responses are not altered in patients with HFpEF, suggesting an absence of overt autonomic dysfunction is this patient group.Support or Funding InformationThis project is funded in part by the National Institutes of Health (HL118313, R56AG057584, T32HL139451) and the U.S. Department of Veterans Affairs (I01RX001311, I01RX001697, I01RX002323, I01RX000182).
Development of GPR183 Antagonists to Treat Neuropathic Pain
Authors: Latzo Nicholas George , Braden Kathryn , Giancotti Luigi , Chen Zhoumou , DeLeon Chelsea , Boehm Terri , D’Cunha Napoleon , Luongo Livio , Doyle Timothy , Kolar Grant , Walker John , Arnatt Christopher , Salvemini Daniela 
Published: April 2020

Chronic neuropathic pain affects millions of people in the United States today. Opioids are currently the frontline agent for the treatment of neuropathic pain; however, as their use continues to grow, serious side effects can arise including addiction and death. The need for a safer alternative to opioids is becoming increasingly more urgent. GPR183 (Ebstein‐Barr induced gene 2, EBI2) is a G‐protein coupled receptor (GPCR) that binds to 7a,25‐dihydroxycholesterol (7a,25‐OHC). A homology model for GPR183 was constructed and a pharmacophore hypothesis was built around the binding of other known ligands to the receptor. Over 5 million compounds were screened in silico for their similarity to our pharmacophore hypothesis and the top 20k hits were funneled into a high‐throughput docking protocol. Several compounds showed strong inhibition of 7a,25‐OHC calcium mobilization with nanomolar potency. Intrathecal injections of these GPR183 antagonists reversed CCI‐induced mechano‐allodynia in mice when given at peak neuropathic pain. These results identify a prominent and functional role for GPR183 in the spinal cord in the maintenance of neuropathic pain. A preliminary structure‐activity relationship study of one of these compounds is being performed to study the pharmacology and medicinal properties of this new class of compounds.Support or Funding InformationWe would like to acknowledge Dr. Dale Dorsett and the Saint Louis University Genomics Core Facility for their assistance with the RNA‐Sequencing. This research was supported by funds from Dr. Salvemini and Dr. Arnatt and T32 GM008306‐01 training grant (KB). Dr. Salvemini and Dr. Arnatt have filed a patent describing the intellectual property generated at SLU. All other authors claim no conflict of interest.
Isolation of the exon‐switching variant, CYP4F2b, from human hepatic carcinoma (HCC) and its role in modulating epithelial‐mesenchymal transition (EMT) through Twist2.
Authors: Kang Puneet , Rote Paul , Saleem Abraham , Danso Prince , Shirota Leticia , Hardwick James 
Published: April 2020

Cytochrome P450 IV (CYP4) are a family of omega‐hydroxylase enzymes. We have previously shown that these fatty acid omega‐hydroxylases function in the hydroxylation of various chain‐length saturated and unsaturated fatty acids in the endoplasmic reticulum. These dicarboxylate products then undergo peroxisomal β‐oxidation, yielding shorter chain fatty acids. In humans, the CYP4F family members hydroxylate long‐chain fatty acid (>C16) and the bioactive eicosanoids, arachidonic acid (HETE) and pro‐inflammatory leukotrienes B4 (LTB4). In the human liver, CYP4F2a metabolizes long‐chain fatty acids and arachidonic acid to the vasoactive 20‐HETE. We have identified a new variant, CYP4F2b from human hepatic carcinoma that is produced from the CYP4F2 gene by exon switching. We have compared the expression of CYP4 members in human HCC and matched healthy human liver. We observed increased expression of CYP4F2b in HCC compared to controls, as well as increased fatty acid omega‐hydroxylation, with associated elevations of acetyl‐CoA/acetate in HCC, and decreased activity and expression of the acetyl‐CoA thioesterase 12 (ACOT12). Increased acetyl‐CoA levels activate the basic helix‐loop‐helix (bHLH) transcription factor Twist2, which initiates an epithelial‐mesenchymal cell transition (EMT). We propose that increased omega‐hydroxylation of fatty acids by CYP4F2b and subsequent peroxisomal β‐oxidation increases cellular acetyl‐CoA levels in HCC, leading to activation of Twist2 and progression of EMT.Support or Funding InformationNEOMED foundation
Development of Craniofacial Biomineralization in Mice with Osteogenesis Imperfecta (OI)
Authors: Wright Tommy R. , McBride Alexandra H. , Organ Jason M. , Menegaz Rachel A. 
Published: April 2020

Osteogenesis imperfecta (OI), or “brittle bone disease,” is a rare autosomal dominant disorder characterized by mutations in the genes that encode the alpha 1 (COL1A1) and alpha 2 (COL1A2) chains of type I collagen, a significant component of bone. Misfolding of the mutated collagen polypeptides interfere with proper assembly of collagen chains, or the triple helical structures, leading to extreme skeletal fragility. OI manifests in a variety of types, with type I being the most mild and most common, and type III, the subject of this study, being more severe and more rare. Clinical manifestations of OI type III generally include a short stature, low bone mineral density (BMD), dental problems, hearing loss, and blue sclerae. In this study, we use a mouse model of OI type III to examine how mutations in type I collagen synthesis impact craniofacial biomineralization during postnatal growth.The homozygous recessive osteogenesis imperfecta murine (OIM) is a model of OI type III and is characterized by increased skeletal fractures, low postcranial BMD, osteopenia, progressive skeletal deformities, and small body size. Wild type mice (WT) and OIM littermates (n≥3/genotype) were micro‐CT scanned at weaning (4 weeks) and adulthood (16 weeks). BMD at three regions of interest (ROI) within the mouse skull was measured using Bruker CTAnalyzer software. ROIs were chosen based on the amount of load during mastication, and included the temporomandibular joint (TMJ), the parietal bone (1mm from the sagittal suture), and the anterior maxilla at the incisal alveolus. Mann‐Whitney U‐tests (α=0.05) were used to compare BMD values between the genotypes for each ROI.At weaning, OIM mice had significantly lower BMD values (g.cm−3 CaHA) than their wild type littermates at the TMJ (0.39 vs 0.58), parietal bone (0.35 vs 0.64), and anterior maxilla (0.53 vs 0.85). These statistical differences persist into adulthood. This decrease in BMD reflects decreased deposition of CaHA due to an abnormal collagen framework in the craniofacial skeleton of OIM mice. Ongoing efforts will expand this analysis to quantify the development of BMD at additional regions within the craniofacial skeleton.Support or Funding InformationThis research was supported by an Indiana University Collaborative Research Grant and the Ralph W. and Grace Showalter Trust.
An Investigation of Anki Flashcards as a Study Tool Among First Year Medical Students Learning Anatomy
Authors: Rana Tanvi , Laoteppitaks Chaiya , Zhang Guiyun , Troutman Gregory , Chandra Shruti 
Published: April 2020

PurposeAnatomy is a challenging course in medical school, in part because of the vast number of facts that must be memorized. Current literature suggests the value of spaced repetition in the learning of factual information, and online tools such as Anki flashcards have been developed to facilitate it. The purpose of this study is to investigate the role of curriculum‐specific Anki flashcards as a study tool for first year medical students learning anatomy.MethodsA set of Anki flashcards was created for the anatomy thread of Block 1 (back, pectoral, shoulder, vertebral regions) and Block 3A (cardiac & pulmonary regions) of the JeffMD curriculum at Sidney Kimmel Medical College. Upon completion of an initial survey, study volunteers were provided with the curriculum‐specific flashcards. Following the anatomy exam in each block, another survey was administered to gather data regarding participants’ usage and opinions of the flashcards.Results and Conclusions148 students completed the initial survey and were provided with the flashcards. Of the 78 students that used the curriculum‐specific flashcards and completed the post‐exam survey in Block 1, 20.5% found them extremely helpful, 29.5% very helpful, 33.3% moderately helpful, 15.4% slightly helpful, and 1.3% not at all helpful. Furthermore, 60.3% believed that the flashcards helped reduce anxiety related to the anatomy, while 18.0% did not believe that they reduced anxiety. Of the 60 students that used the curriculum‐specific flashcards and completed the post‐exam survey in Block 3A, 41.7% found them extremely helpful, 36.7% very helpful, 11.7% moderately helpful, and 10.0% slightly helpful. Moreover, 76.7% believed that the flashcards helped reduce anxiety related to the anatomy, while 15.0% did not believe that they reduced anxiety. These results suggest that spaced repetition of anatomy facts through curriculum‐specific Anki flashcards may have a positive impact on affective learning outcomes among first year medical students.Support or Funding InformationSidney Kimmel Medical College at Thomas Jefferson University
Larp6 Regulates Collagen mRNA by Targeted Unwinding of a Conserved Stem‐Loop
Authors: Baggs Eric , Warner Lisa 
Published: April 2020

Collagen is the most abundant protein in the human body and a key component of the extracellular matrix. Regulation of collagen production in key to the development of fibroproliferative disorders; everything from scarring to cardiovascular disease. La ribonucleoprotein 6 (Larp6) is an RNA binding protein which has recently been shown to regulate collagen production by binding to a conserved region at the translation start site of collagen mRNA. This interaction appears to be critical to directing RNA to the rough ER for translation initiation. Additionally, LARP6 interaction with a number of molecular chaperones appears to be important for regulating collagen production. We hypothesize that LARP6 utilizes conformational selection in the recognition and discrimination of collagen mRNAs and that tertiary interactions dictate mRNA fate. In vitro RNA chaperone assays, translation assays, RNAi, confocal microscopy, and NMR and SAX structural studies are employed to interrogate these interactions. New information regarding the structures and dynamics that determine the Larp6 interaction with collagen mRNA may inform novel therapeutic targets that can mitigate fibroproliferative disease.Support or Funding InformationP20GM109095
Role of Heme metabolism in the oxidative state of NO‐receptor soluble guanylyl cyclase (sGC) in aging mouse brain
Authors: Sharina Iraida , Martin Emil , Srinivasan Subhiksha 
Published: April 2020

IntroductionAging is an important risk factor for cardiovascular diseases and neurodegeneration. Heme synthesis and metabolism decline with age. Heme moiety is essential for proper function of the Nitric Oxide receptor soluble guanylyl cyclase (sGC). Evidence suggests the involvement of NO/cGMP signaling in human neurological disorders. sGC also an important role in maintenance of cerebral blood flow, vascular homeostasis and platelet function. NO binds to the ferrous heme moiety of sGC and activates cGMP‐forming activity hundred fold. Therefore, the integrity of reduced sGC heme moiety is essential for proper transduction of NO signal.Methods and ResultsWe investigated if the oxidative state and the relative content of sGC heme moiety changes with age in mouse brain tissue. Three groups of animals have been used in our studies: young, 3.5 months old, and mature, 15 months old, and old, 24 months, C57Bl6 mice. sGC‐dependent synthesis of cGMP in protein homogenates from mouse brains in response to sGC regulators was evaluated by quantifying the amount P32‐labeled cGMP using thin layer chromatography. To estimate the changes in the amount of heme‐deficient sGC, protoporphyrin IX (PPIX) was used as a heme‐independent sGC activator. NO‐independent activator BAY 58‐2667, which has a higher affinity for sGC with ferric heme, was used to evaluate the changes in the amount of sGC with oxidized heme. NO donor DEA‐NO was used to evaluate the level of sGC with ferrous heme. For each individual sample the ratios of sGC‐[HemeFe3+] vs sGC‐[HemeFe2+] activities (BAY 58‐2667/DEA‐NO) and heme‐deficient vs sGC‐[HemeFe2+] activities (PPIX/DEA‐NO) were determined. Our analysis demonstrated that the relative amount of heme‐deficient and heme‐oxidized sGC increased in both males and females of older age. Decline in sGC protein expression in oldest animal groups was detected by Western blot analysis. We also investigated if the accumulation of heme‐deficient sGC or sGC with ferric heme in ageing brain correlates with changes in the level of proteins involved in iron and heme metabolism. We found no difference in the level of HSP90 chaperone needed for sGC heme insertion and unchanged level in the iron chaperone PCBP2, which is essential for iron trafficking. The expression of divalent metal ion transporter (DMT1) was also not affected by age. However, we observe a decrease in the expression of Heme Oxygenase 1 (HO1) and Aconitase 2 proteins involved in maintenance of cellular oxidative balance in old animals of both genders.ConclusionOur data indicate that changes in heme homeostasis might contribute to the decline of NO/sGC in brain with aging.
Insight into the Metabolic Machinery of Deep‐Marine Microbes Grown with Light and Heavy Petroleum
Authors: Tejeda-Mora Andres Javier , Maturano Luis Donis , Macias-Contreras Marla I. , Guerrero Abraham , Gonzalez-Sanchez Ricardo , Hernandez-Lopez Edna , Vazquez-Duhalt Rafael , Licea-Navarro Alexei , Moreno-Ulloa Aldo 
Published: April 2020

Degradation of hydrocarbons by marine microbes is well studied and specific metabolic routes have been delineated. Deep‐marine sediments (>1000 m) harbor bacterial communities but remain poorly studied compared to those inhabiting shallow waters. While metagenomics studies have evidenced the metabolic potential of microbes from deep‐sea origins, more exploration through novel approaches of their metabolic machinery is warranted. In this study, we profiled the bacterial taxa (16S rRNA amplicon sequencing) and metabolome (Liquid chromatography coupled to high‐resolution mass spectrometry [LC‐HR‐MS]) of microbes from deep‐sea sediments of the Gulf of Mexico cultured with heavy and light petroleum (four distinct types, according to API classification). We utilized a comprehensive chemoinformatic analysis to detect the microbial chemistries using a repertoire of novel chemoinformatic software, and results were integrated with those obtained by the prediction of functional profiles using 16S rRNA data. We noted a better microbial growth in lighter vs. heavier petroleum after 28 days of culturing, wherein the genus Pseudomonas prevailed, to a similar extent, in all conditions. Hydrocarbon‐degradation pathways (e.g., benzoate, naphthalene, and toluene degradation) were represented by predicted functional profiles, but no differences were observed among petroleum types. PCA analysis of metabolic data showed differences among petroleum type. The number of metabolites and chemical diversity were reduced in heavier vs. lighter petroleum. The predominant chemical classes were coumarins and derivatives, prenol lipids, benzene and substituted derivatives (equal to prenol lipids), and prenol lipids for API 7–10, API 16–20, API 35, and API 40, respectively. Also, other chemical classes linked to hydrocarbon‐metabolism were detected. Manual inspection of selected metabolite clusters or groups of structurally‐related metabolites led to the putative identification of aminated, carboxylated, and hydroxylated chemistries linked to microbial metabolism of hydrocarbons. In conclusion, our study provides valuable microbiological and metabolomic information about one of the most understudied ecosystems in the world, the deep‐marine sediments.Support or Funding InformationThis research was funded by the National Council of Science and Technology of México – Mexican Ministry of Energy – Hydrocarbon Trust, project 201441. This is a contribution of the Gulf of México Research Consortium (CIGoM).