bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adiposity of the B6 Mouse Lung
Authors: Meyerholz David K. , Beck Amanda P. 
Published: April 2020

In biomedical research, mice are studied using a variety of techniques that require a baseline knowledge of microscopic anatomy, these include histochemistry, immunohistochemistry and immunofluorescence. Many biomedical personnel lack this background in their training. The lung is primarily composed of two zones: the conducting zone (airways) and the respiratory zone (alveoli). Morphologic differentials for alveolar structures are few in the lung, but in our experience adipose tissues in the lung can be a source of confusion and misinterpretation. In this study, we examined lungs from adult B6 mice (>3 months, n=16, female) that were obtained from de‐identified archival control tissues (UI Comparative Pathology Laboratory) to define lung adiposity. White adipose tissues (WAT) were detected in all lung samples (100%, 16/16). WAT was often seen as small aggregates of 5–15+ adipocytes embedded in loose connective tissue adjacent to large airways/vessels. In sites of smaller airway branching, WAT was rarely detected and usually only as a few solitary adipocytes. WAT adipocytes were distinguished from alveolar structures by their location, significantly reduced cellularity, and residence often in a collagenous matrix. Brown adipose tissues (BAT) were not observed in this survey. We have characterized the localization and appearance of adipose tissue in the lungs of female B6 mice. Awareness that WAT is present in lung tissues along with knowledge of its distinguishing features versus alveoli can help investigators make more clear and accurate interpretations of the lung data.Support or Funding InformationNIH
Cosolvent Molecular Dynamics Simulation‐Based Discovery of Potential Allosteric Sites on Regulator of G Protein Signaling 4
Authors: Chan Wallace , Carlson Heather , Traynor John 
Published: April 2020

Regulator of G Protein Signaling 4 (RGS4) is member of a family of intracellular proteins that are characterized by the presence of a 125 amino‐acid RGS homology domain (RH) which binds to the Ga subunit of heterotrimeric G proteins. By doing so RGS4 acts as a GTPase‐accelerating protein or GAP to expedite the conversion of the GTP‐bound active form of Gα subunit of heterotrimeric G proteins to the GDP‐bound inactive form, thereby terminating signaling downstream of G protein coupled receptors (GPCRs). RGS4 is expressed widely in both the periphery and central nervous system and has been implicated in pain, schizophrenia, and the control of cardiac contractility. Inhibitors of RGS4 have been developed, but these all bind covalently to a cysteine (Cys95) in the RH domain. Therefore, we sought to identify alternative druggable sites on this protein using cosolvent molecular dynamics simulations. This technique employs a low concentration of organic probes (pyrimidine, acetonitrile, isopropyl alcohol) to find druggable “hot spots” on a protein based on the binding preference of the probes. This methodology has the advantage of utilizing protein dynamics to find druggable pockets that would be otherwise inaccessible with a single snapshot crystal structure. The top predicted sites were located in all areas of the RH domain of RGS4. Subsequently, statistical coupling analysis, a technique that identifies co‐evolving residues from analysis of multiple sequence alignments, was used to map out a functionally‐significant sector on the surface of RGS4 which overlapped with the identified hot spots, allowing for prioritization. The first site identified was expectedly the Gα subunit binding site. The second was located in a cleft in the proximity of lysine residues thought to be the binding site for phosphatidylinositol (3,4,5)‐trisphosphate (PIP3) binding. Since PIP3 is a known inhibitor of the GAP activity of RGS4, future directions entail the investigation of PIP3‐bound conformations of RGS4 by molecular dynamics simulation to better understand the mechanism of deactivation. Of other predicted sites, one is in close proximity to the purported allosteric site that contains Cys95, while the others are putative novel sites for allosteric control of RGS4.Support or Funding InformationSupported by R01 DA‐035316.
Attenuation of Scopolamine‐Induced Learning and Memory Impairment by Ceriporia lacerata Mycelial Culture in C57BL/6 Mouse Model
Authors: Lee Sujin , Yun Hyun Seok , Oh Jisun , Kim Jong-Sang 
Published: April 2020

Ceriporia lacerata (provided from FUGENBIO Co., Ltd., Seoul, S. Korea) is one of white‐rot fungi. The submerged culture and its extract of the fungal mycelia have been reported to contain various bioactive substances, including extracellular polysaccharide and β‐glucan, and to exert the anti‐diabetic effect in mice. Our in vitro study showed that the ethanol extract of C. lacerata mycelial culture powder protected HT22 mouse hippocampal neuronal cells from glutamate‐induced cytotoxicity. The present study aimed at examining whether oral supplementation of the cultured C. lacerata mycelia assists the restoration of learning and memory capability diminished by scopolamine injection in C57BL/6 mice. Results exhibited that the treatment with C. lacerata mycelial culture powder (□) improved spatial learning and memory impairment in scopolamine‐injected mice as tested by behavioral tests (Y‐maze task, Passive avoidance task, and Morris water maze task), (□) reduced histological damage in the hippocampal CA1 region, (□) increased the expression level of cytoplasmic heme oxygenase‐1 (HO‐1) in hippocampal tissue homogenates, and (□) decreased the levels of oxidative stress markers in the plasma, in comparison to mice treated with scopolamine alone. These findings suggest that oral supplementation of C. lacerata mycelial culture powder could prevent scopolamine‐induced learning and memory deficit possibly by protecting hippocampal damage from oxidative stress.Support or Funding InformationThis work was supported by the “Food Functionality Evaluation Program” with the Ministry of Agriculture, Food, and Rural Affairs (2019).
Using NMR Derived Structural Information Complemented by Molecular Docking and MM/GBSA Rescoring to Mitigate CYP3A4 Time‐Dependent Inhibition
Authors: Wilderman Ross , Halpert James 
Published: April 2020

Time‐dependent inhibition (TDI) of human CYP3A4 is a significant challenge in drug development and is the cause of multiple clinically relevant drug‐drug interactions. The vasodilator diltiazem causes TDI when metabolized by CYP3A4. We hypothesized that introduction of steric bulk into the CYP3A4 active site by site‐directed mutagenesis will disrupt active site contacts, alter diltiazem orientation in the CYP3A4 active site, and decrease TDI, as previously seen with midazolam. CYP3A4‐ligand interactions were monitored using spectral binding titrations. In parallel, we performed molecular docking of diltiazem and the N‐demethylation products into CYP3A4 models of previously tested mutants that introduce steric bulk into the enzyme active site, followed by molecular mechanics generalized Born surface area (MM‐GBSA) rescoring. An NMR‐derived model of diltiazem in the CYP3A4 active site revealed that the dimethylamino group that undergoes CYP3A4‐mediated N‐demethylation is oriented toward the heme iron. Diltiazem caused a Type II spin shift upon binding to CYP3A4, indicating interaction of the amine with the heme iron. Docking of diltiazem, N‐desmethyldiltiazem, or N, N‐didesmethyldiltiazem into wild‐type CYP3A4 resulted in the amino group of each compound oriented toward the heme iron. In contrast, docking these three compounds into CYP3A4 I120W resulted in the amino group orienting away from the heme. Furthermore, T309Y docking results showed reorientation of diltiazem, which would prevent the initial N‐demethylation event, and I301W and A370F resulted in N‐desmethyldiltiazem reorientation. MM‐GBSA rescoring of docking experiments confirmed that all compounds used orient with the amino group toward the heme in wild‐type CYP3A4, all amino groups away from the heme in I120W, and reorientation of N‐desmethyldiltiazem in I301W and A370F. Furthermore, in rescoring experiments, N‐desmethyldiltiazem was reoriented in the Y307W active site, and N, N‐didesmethyldiltiazem was reoriented in the T309Y active site in addition to reorientation of diltiazem. Substitution of I120 and T309 with a larger residue decreased CYP3A4 TDI. In summary, a hybrid NMR‐computational approach was used to rapidly generate a model of diltiazem in complex with CYP3A4. These results indicate that NMR‐derived models of protein‐ligand interactions provide a structure‐based approach for decreasing metabolic liabilities of CYP enzyme substrates.Support or Funding InformationThis work was supported by internal funding from the University of Connecticut to James Halpert.
Applicability of Capnography in the Adult ICU During Physical Therapy: A Systematic Review
Authors: Smith Karen R. , Hubbert Doug R. , Murphree Lisa K. 
Published: April 2020

PurposeThe purpose of this review was three‐fold: first, evaluate the strength and quality of the current research evidence on usage of capnography, second, determine whether current evidence regarding the use capnography during physical therapy should be considered when making decisions regarding patient’s response to physical therapy; and third, identify weaknesses in the current evidence and area for continued research.Background/SignificanceCapnography is typically used in emergency departments and postoperatively but is not commonly used during physical therapy. Pulse oximetry is typically used to monitor oxygen desaturation and hypoxemia during physical therapy but does not predict or monitor respiratory depression. Capnography monitoring during exercise and mobility could have benefits for physical therapists monitoring patients during therapy as well as provide prognostic value of patient progress.Methods and MaterialsA literature search was completed using: Science Direct, IOP Science, PLOS one, Google scholar, Wiley, EBSCO Host, CINAHL Complete, Cochrane Library, and PubMed. Keywords: capnometry, capnography accuracy, physical therapy, application of capnography, mobility, monitoring. Articles selected for review were those of original research or systematic review published since 2009 and evaluated for quality using the Oxford Centre for Evidence‐Based Medicine 2011 levels of evidence system.ResultsTwenty‐seven articles are included for this study. Six articles were included in the introduction. Seven articles examined the current practices for use of capnography; six studies looked at the use of capnography in the intensive care unit. Three articles on pulse oximetry and capnography with mobility, two articles on the use of monitoring ventilation and oximetry during exercise, and three studies on biologically‐based assessment tools for physical therapy.ConclusionsEvidence suggests that monitoring partial pressure end‐tidal carbon dioxide (PETCO2) levels in patients during rest and exercise may provide more prognostic value of possible adverse events in patients with conditions such as heart failure. Monitoring ventilatory responses in patients is valuable in physical therapy as it indicates tolerance to activity. Evidence suggests there is a significant correlation between PETCO2 and cardiac output. Recent research also suggests regardless of mask or cannula used on patients, diagnostic accuracy of measuring PETCO2 is similar during normal and hyper‐ventilatory states. With increasing application of capnography in intensive care settings, studies involving usage in physical therapy are warranted.
Cancer Attenuates Nutrient Activation of Skeletal Muscle mTORC1 in Female Mice
Authors: Counts Brittany , Halle Jessica , Carson James 
Published: April 2020

Cancer‐induced metabolic disruptions can alter tissue specific energy demands and has the potential to alter muscle responses to feeding. Mechanistic target of rapamycin complex 1 (mTORC1) signaling axis is activated by nutrient stimuli, and cancer cachexia can suppress basal muscle mTORC1 signaling in mice. However, gaps still remain in our understanding of how the cancer environment effects skeletal muscle nutrient regulation of mTORC1 signaling.PurposeWe examined the effect of feeding on skeletal muscle mTORC1 signaling in female tumor bearing mice.MethodsFemale C57BL/6 (B6) and ApcMin/+ (MIN) mice were fasted for 12‐hrs during the light cycle. Following the 12‐hr fast, mice were given access to a food pellet for 1‐hr (B6 FED:N=7, MIN FED:N=8) or fasted for another hour (B6 FAST:N=6, MIN FAST:N=7). Circulating blood glucose, and plasma IL‐6 and insulin were taken prior to sacrifice. Gastrocnemius muscle was homogenized for protein analysis. Statistical significance was set at p£0.05.ResultsThe MIN FED and MIN FAST groups were not different in percent body weight change, polyp number, plasma IL‐6, and hindlimb muscle mass. 1‐hour food intake was not different between B6 FED and MIN FED mice. Both MIN FED and B6 FED increased stomach mass, blood glucose, and circulating insulin levels. Compared to FAST mice, FED mice increased p70S6K and rpS6 phosphorylation, and was positively associated to insulin in both B6 and MIN mice. 4E‐BP1 phosphorylation was increased in the B6 FED compared to B6 FAST, but this response was blunted in the MIN FED. Additionally, circulating insulin concentration was associated to 4E‐BP1 phosphorylation in the B6 FED, but not in the MIN FED.ConclusionThe cancer environment can suppress the nutrient‐induced phosphorylation of skeletal muscle 4E‐BP1. Future studies are warranted to determine if the combination of exercise or muscle contraction can improve skeletal muscle mTORC1’s response to nutrients in the female cancer environmentAcknowledgementsNCI R01‐CA121249
Evaluating Data to Determine when Biotherapeutic Vitamin A Supplementation can be Scaled Back in Children <5 Years Old
Authors: Tanumihardjo Sherry A. 
Published: April 2020

The World Health Organization recommends that high dose vitamin A supplements be administered to children under the age of 5 years to be used as a biotherapeutic to prevent morbidity and mortality. Vitamin A is essential for vision, immunity, reproduction, and growth. In addition to high dose supplements, many countries add preformed vitamin A to staple foods during processing. The Global Alliance for Vitamin A (GAVA) has set up a decision framework to follow to determine if a country can scale back high dose vitamin A supplements in preschool children. The objective of this work was to evaluate data available in African countries to determine if vitamin A supplements can be scaled back. The framework includes evaluating demographic information, dietary intake data, and biomarkers of vitamin A status. All of these data were available for one province in South Africa, one district in Zambia, and a subgroup of children enrolled in a demographic health survey in Ghana. In South Africa, it was determined that there is enough evidence to scale back high dose supplements in one province, but not enough data to scale back supplementation in the whole country. A similar finding was determined in one area of Zambia where dietary intake is adequate. In Ghana, the evidence is mounting that status has improved, but there are still pockets of vitamin A deficiency that likely benefit from high dose supplementation. In conclusion, overlapping interventions in some countries likely leads to adequate vitamin A intakes and in some cases excess. Vitamin A supplementation is used as a therapy to save children’s lives, but in some areas it may no longer be necessary. The repercussions of vitamin A deficiency are devastating and the world has the means to eradicate it; however, vitamin A excess to the point of toxicity also has health concerns. Therefore, balancing needs with intakes is important for overall health. The GAVA framework can be used in any country that has all of the needed data to evaluate their situation or to determine where more data may be needed.Support or Funding InformationSupport for these studies was obtained from the International Atomic Energy Agency (South Africa), HarvestPlus (Zambia), and GroundWork (Ghana). Evaluation of the data was done independently from the funding agencies on published data.
A cis‐regulatory Element in Intron 1 Cooperates with the Core Promoter during Salinity Induction of Tilapia Glutamine Synthetase
Authors: Kim Chanhee , Kültz Dietmar 
Published: April 2020

Mozambique tilapia (O. mossambicus) is a euryhaline fish that tolerates salinity ranging from fresh water (FW) to 120 g/kg (> 3x seawater, SW). Hyperosmotic stress associated with salinity transfer from FW to SW strongly induces glutamine synthetase (GS) transcription in tilapia, including a brain cell line model (OmB cells). In this study we cloned the 5′ regulatory sequence (RS) (including 5′‐flanking region and 5′‐UTR) of the O. mossambicus GS gene to identify osmotic response elements that mediate transcriptional induction. Using enhancer trapping, we discovered a novel osmosensitive mechanism that utilizes an unusual enhancer cassette in intron 1 of the GS gene. This cassette includes a single copy of a cis‐regulatory element (CRE), the osmolality/salinity‐responsive element 1 (OSRE1). We show that intronic OSRE1 is necessary but not sufficient for intron 1‐mediated hyperosmotic GS induction. Furthermore, intron 1 and its interaction with the GS core promoter complex are also necessary for hyperosmotic GS induction. We propose a novel model for hyperosmotic intron 1‐mediated induction of the O. mossambicus GS gene that is based on the interaction of an OSRE1 enhancer, intron 1, and the core GS promotor.Support or Funding InformationThis study was funded by a grant from NSF (IOS‐1656371).
Lectin Staining Biomarkers for Gene Therapy in GNE Myopathy
Authors: Crowe Kelly E. 
Published: April 2020

GNE myopathy (GNEM) is an autosomal recessive disease characterized by muscle wasting and weakness. This disease is caused by mutations in the GNE (UDP‐N‐acetylglucosamine (GlcNAc) 2‐epimerase/N‐acetylmannosamine (ManNAc) kinase) gene, which encodes a bifunctional enzyme that catalyzes the committed step of sialic acid (SA) biosynthesis. As such, GNE mutations found in GNEM impair enzymatic function and generally result in a decrease of SA. Efforts to treat GNEM with gene therapy are currently underway, but given the slowly progressing muscle pathology of GNEM, it becomes important to establish molecular biomarkers to assess such a therapy during the relatively brief timescale of a clinical trial. To do so, we have compared SA abundance in converted myoblasts from GNEM and otherwise normal patients using fluorescently labeled lectins, which can reflect cellular SA levels. Here, we describe changes in SA abundance in GNE myopathy that could serve as surrogate biomarkers for a GNE gene therapy clinical trial.Support or Funding InformationThis work was supported by a grant from the Neuromuscular Disease Foundation (NDF).
Class III PI3‐Kinase in Angiotensin II‐Induced Cardiac Hypertrophy
Authors: Sun Chengwen , Zhong Tiecheng , Guo Ang , O’Rourke Stephen T. , Guo Lirong 
Published: April 2020

Pathological cardiac hypertrophy could lead to heart failure and sudden cardiac death. Our previous study demonstrated that Class I PI3‐kinase (PI3KC1) contributes to the development of cardiac hypertrophy by elevating intracellular reactive oxygen species (ROS) levels. In the current study, we evaluated the role of Class III PI3‐kinase (PI3KC3) in Ang II‐induced cardiac hypertrophy. Treatment of cultured cardiomyocytes with Ang II (10−6 M) significantly increased the cellular surface area, which was measured under a fluorescence microscope after staining with anti‐alpha‐sarcomeric actin antibodies. In cardiomyocytes, Ang II (10−9‐10−5 M) induced a dose‐dependent activation of PI3KC3, detected using an ELISA kit. More interestingly, Ang II‐induced cardiac hypertrophy was dramatically exacerbated by co‐treatment with 3‐Methyladenine (3‐MA, 10−6 M), a PI3KC3 inhibitor, suggesting that activation of PI3KC3 may play a protective role in Ang II‐induced cardiac hypertrophy. To identify the intracellular signaling pathway involved in the PI3KC3‐mediated protective effect, mitochondrial ROS was measured using MitoSOX in cardiomyocytes treated with control, Ang II (10−6 M), Ang II plus 3‐MA (10−6 M), and 3‐MA alone. The results demonstrated that Ang II treatment significantly increased mitochondrial ROS production and that Ang II‐induced increases in ROS generation were potentiated by co‐treatment with 3‐MA. It is well known that the major intracellular source of ROS is the damaged mitochondria, which is scavenged by autophagy (also called mitophagy) in cardiomyocytes. Thus, we measured autophagy activity by detection of autophagosomes using staining method with MAP‐LC3 antibodies in cardiomyocytes treated with Ang II. Treatment of cardiomyocytes with Ang II (10−9 to 10−5 M) significantly increased autophagy activity with two phases (increased activity at low doses, decreased activity at high doses, peaked at 10−7 M). More interestingly, Ang II (10−6 M)‐induced increases in autophagy were dramatically attenuated by 3‐MA (10−6 M). Taken together, these data suggest that activation of PI3KCIII has a protective effect on Ang II‐induced hypertrophy by enhancing autophagy activity and decreasing mitochondrial ROS generation in cardiomyocytes.Support or Funding InformationNIH HL143519