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68 articles published in JoVE
Adapting Gastrointestinal Organoids for Pathogen Infection and Single Cell Sequencing under Biosafety Level 3 (BSL-3) Conditions Megan L. Stanifer1,2, Steeve Boulant2,3,4 1Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 2Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, 3Department of Infectious Diseases, Virology, Heidelberg University Hospital, 4Research Group "Cellular Polarity and Viral Infection", German Cancer Research Center (DKFZ) This protocol describes how to infect human intestinal organoids from either their apical or basolateral side to characterize host/pathogen interactions at the single-cell level using single-cell RNA sequencing (scRNAseq) technology.
Retrograde Parotid Gland Infusion through Stensen's Duct in a Non-Human Primate for Vectored Gene Delivery Guy El Helou1, Joseph F. Goodman2, Maria Blevins3, David L. Caudell4, Todd A. Ponzio3, John W. Sanders3,5 1Department of Medicine, Division of Infectious Diseases and Global Medicine, University of Florida, 2Department of Otolaryngology, George Washington University School of Medicine, 3Department of Medicine, Section on Infectious Diseases, Wake Forest Baptist Medical Center, 4Department of Pathology, Wake Forest School of Medicine, 5Department of Medicine, Section of Infectious Diseases, Hefner Veterans Affairs Medical Center Salivary glands have been proposed as a tissue target site for gene therapy, especially in the area of vaccination by gene transfer. We demonstrate gene delivery in a non-human primate model utilizing retrograde parotid infusion.
A Preclinical Model of Exertional Heat Stroke in Mice Michelle A. King1, Jamal M. Alzahrani1, Thomas L. Clanton1, Orlando Laitano1,2 1Department of Applied Physiology and Kinesiology, College of Health and Human Performance, University of Florida, 2Department of Nutrition and Integrative Physiology, College of Health and Human Sciences, Florida State University The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.
A Magnetic-Bead-Based Mosquito DNA Extraction Protocol for Next-Generation Sequencing Tse-Yu Chen1, Adam E. Vorsino2, Kyle J. Kosinski1, Ana L. Romero-Weaver1, Eva A. Buckner1, Joanna C. Chiu3, Yoosook Lee1 1Florida Medical Entomology Laboratory, Department of Entomology and Nematology, Institute of Food and Agricultural Sciences, University of Florida, 2U. S. Fish and Wildlife Service, Pacific Islands Fish and Wildlife Office, 3Department of Entomology and Nematology, University of California Davis Described here is a DNA extraction protocol using magnetic beads to produce high quality DNA extractions from mosquitoes. These extractions are suitable for a downstream next-generation sequencing approach.
Field-Based Thermal Physiology Assay: Cold Shock Recovery under Ambient Conditions Emily S. Khazan1 1School of Natural Resources and Environment, University of Florida Here, a low-cost, accessible protocol is described to evaluate cold shock recovery of butterflies under ambient environmental conditions.
Activity of Posterior Lateral Line Afferent Neurons during Swimming in Zebrafish Elias T. Lunsford1, James C. Liao1 1Department of Biology, University of Florida, The Whitney Laboratory for Marine Bioscience We describe a protocol to monitor changes in the afferent neuron activity during motor commands in a model vertebrate hair cell system.
Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification Owen Hudson1, Sumyya Waliullah1, Justin Hand2, Romina Gazis-Seregina3, Fulya Baysal-Gurel4, Md Emran Ali1 1Department of Plant Pathology, Coastal Plain Experiment Station, University of Georgia, 2University of Georgia Cooperative Extension, 3Department of Plant Pathology, Tropical Research & Education Center, University of Florida, 4Department of Agricultural and Environmental Sciences, Otis L. Floyd Nursery Research Center, Tennessee State University We developed a method to detect Phytophthora capsici zoospores in water sources using a filter paper DNA extraction method coupled with a loop-mediated isothermal amplification (LAMP) assay that can be analyzed in the field or in the lab.
TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis Silveli Suzuki-Hatano1, Ang-Chen Tsai1, Audrey Daugherty1, Christina A. Pacak1 1Department of Pediatrics, University of Florida We present an optimized tandem mass tag (TMT) labeling protocol that includes detailed information for each of the following steps: protein extraction, quantification, precipitation, digestion, labeling, submission to a proteomics facility, and data analyses.
Quantifying Human Norovirus Virus-like Particles Binding to Commensal Bacteria Using Flow Cytometry Jasmine L. Madrigal1, Melissa K. Jones1 1Microbiology and Cell Science Department, University of Florida The goal of this protocol is to quantify binding of the eukaryotic pathogen human norovirus to bacteria. After performing an initial virus-bacterium attachment assay, flow cytometry is used to detect virally-bound bacteria within the population.
At-Risk Butterfly Captive Propagation Programs to Enhance Life History Knowledge and Effective Ex Situ Conservation Techniques Jaret C. Daniels1,2, Geena M. Hill2,3, Kristin A. Rossetti2, Stephanie J. Sanchez2, Jacob A. Hornfeldt1 1Department of Entomology and Nematology, University of Florida, 2Florida Museum of Natural History, University of Florida, 3School of Natural Resources and Environment, University of Florida Here, we present protocols for 1) the laboratory captive propagation of the federally endangered Miami blue butterfly (Cyclargus thomasi bethunebakeri), and 2) assessing basic life history information such as immature development time and number of larval stadia. Both methods can be adapted for use with other ex situ conservation programs.
Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation Helen Borges1, Alexandra Dufau1,2, Bhaskar Paneri1, Adam J. Woods3, Helena Knotkova4,5, Marom Bikson1 1Department of Biomedical Engineering, The City College of New York, CUNY, 2Department of Clinical and Health Psychology, Center for Cognitive Aging and Memory, 3McKnight Brain Institute, University of Florida, 4MJHS Institute for Innovation in Palliative Care, 5Department of Family and Social Medicine, Albert Einstein College of Medicine When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).
Rapid Collection of Floral Fragrance Volatiles using a Headspace Volatile Collection Technique for GC-MS Thermal Desorption Sampling Haleigh A. Ray1, Charles J. Stuhl2, Jennifer L. Gillett-Kaufman1 1Entomology and Nematology Department, University of Florida, 2Center for Medical, Agricultural and Veterinary Entomology, Agricultural Research Service U.S. Department of Agriculture Here, we present a protocol for collecting the floral fragrance volatiles from blooming flowers, using a non-destructive sampling procedure.
Porcine Model of Infrarenal Abdominal Aortic Aneurysm Alexander H. Shannon1, J. Michael Cullen1, Jolian J. Dahl1, Erik J. Scott1, Zachary Tyerman1, Michael D. Spinosa1, William G. Montgomery1, W. Forrest Johnston2, Guanyi Lu3, Morgan Salmon*1, Gorav Ailawadi*1,4, Gilbert R. Upchurch Jr.*3 1Department of Surgery, University of Virginia School of Medicine, 2Department of Surgery, Ochsner Medical Center, 3Department of Surgery, University of Florida, 4The Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine This novel model creates robust infrarenal abdominal aortic aneurysms in swine using a combination of balloon angioplasty, elastase/collagenase perfusion, topical elastase application, and oral compound β-aminopropionitrile administration, which interferes with collagen cross-linking.
Murine Surgical Model of Topical Elastase Induced Descending Thoracic Aortic Aneurysm Zachary Tyerman1, Jolian Dahl1, Alexander Shannon1, W. Forrest Johnston2, Nicholas H. Pope1, Guanyi Lu3, Gilbert R. Upchurch Jr.3, Gorav Ailawadi1,4, Morgan Salmon1 1Department of Surgery, University of Virginia School of Medicine, 2Department of Surgery, Ochsner Medical Center, 3Department of Surgery, University of Florida, 4The Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine We describe a surgical protocol to consistently induce robust descending thoracic aortic aneurysms in mice. The procedure involves left thoracotomy, thoracic aorta exposure, and placement of a sponge soaked in porcine pancreatic elastase on the aortic wall.
Sampling for Estimating Frankliniella Species Flower Thrips and Orius Species Predators in Field Experiments Joseph Funderburk1, Xavier Martini1, Josh Freeman1, Iris Strzyzewski1, Edward Traczyk1, Thomas Skarlinsky2, Scott Adkins3 1North Florida Research and Education Center, University of Florida, 2Animal and Plant Health Inspection Services, Plant Protection Quarantine, Miami Plant Inspection Station, United States Department of Agriculture, 3Agricultural Research Service, US Horticultural Research Laboratory, United States Department of Agriculture Presented here is a protocol to determine the number of thrips and minute pirate bug predators in crops over multiple dates in field experiments. Also illustrated is how to determine the efficacy of management tactics against thrips and evaluate the benefits of predation by minute pirate bugs.
Manipulation of Color Patterns in Jumping Spiders for Use in Behavioral Experiments Malika Ihle1, Lisa Anne Taylor1,2 1Department of Entomology and Nematology, University of Florida, 2Florida Museum of Natural History, University of Florida The goal of this protocol is to manipulate the color patterns of jumping spiders and other very small arthropods with paint in order to study questions related to sexual selection, sexual cannibalism, predation, aposematism, or any other field of animal coloration.
HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries Huacheng Luo1, Amin Sobh2, Christopher D. Vulpe2, Edmond Brewer1, Sinisa Dovat1, Yi Qiu3, Suming Huang1 1Department of Pediatrics, Pennsylvania State University College of Medicine, 2Department of Physiological Sciences, University of Florida, 3Department of Anatomy and Cell Biology, University of Florida A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.
Construction and Use of an Electrical Stimulation Chamber for Enhancing Osteogenic Differentiation in Mesenchymal Stem/Stromal Cells In Vitro Liudmila Leppik1, Mit B. Bhavsar1, Karla M.C. Oliveira1, Maria Eischen-Loges1, Sahba Mobini1,2, John H. Barker1 1Frankfurt Initiative for Regenerative Medicine, Johann Wolfgang Goethe-University, 2J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida Here we present a protocol for the construction of a cell culture chamber designed to expose cells to various types of electrical stimulation, and its use in treating mesenchymal stem cells to enhance osteogenic differentiation.
A Controlled Mouse Model for Neonatal Polymicrobial Sepsis Byron Brook*1, Nelly Amenyogbe*1, Rym Ben-Othman2, Bing Cai2, Danny Harbeson1, Freddy Francis1, Aaron C. Liu1, Natallia Varankovich2, James Wynn3,4, Tobias R. Kollmann1,2 1Department of Experimental Medicine, University of British Columbia, 2Department of Pediatrics, Division of Infectious Diseases, University of British Columbia, 3Department of Pediatrics, College of Medicine, University of Florida, 4Department of Pathology, Immunology, and Laboratory Medicine, University of Florida This protocol provides the necessary steps to establish and evaluate neonatal sepsis in 7-day-old mice.
Reverse Dissection and DiceCT Reveal Otherwise Hidden Data in the Evolution of the Primate Face Anne M. Burrows1,2, Kailey M Omstead1, Ashley R Deutsch3,5, Justin T. Gladman4, Adam Hartstone-Rose3 1Department of Physical Therapy, Duquesne University, 2Department of Anthropology, University of Pittsburgh, 3College of Sciences, North Carolina State University, 4Shared Materials Instrumentation Facility, Duke University, 5Department of Anthropology, University of Florida Facial expressions are a mode of visual communication produced by mimetic muscles. Here, we present protocols for the novel techniques of reverse dissection and DiceCT to fully visualize and assess mimetic muscles. These combined techniques can examine both morphological and physiological aspects of mimetic musculature to determine functional aspects.
Electrophysiological Recording of The Central Nervous System Activity of Third-Instar Drosophila Melanogaster Daniel R. Swale1, Aaron D. Gross2, Quentin R. R. Coquerel3, Jeffrey R. Bloomquist3 1Department of Entomology, Louisiana State University AgCenter, 2Department of Entomology, Virginia Tech, 3Department of Entomology and Nematology, Emerging Pathogens Institute, University of Florida This protocol describes a method to record the descending electrical activity of the Drosophila melanogaster central nervous system to enable the cost-efficient and convenient testing of pharmacological agents, genetic mutations of neural proteins, and/or the role of unexplored physiological pathways.
A Direct Force Probe for Measuring Mechanical Integration Between the Nucleus and the Cytoskeleton Qiao Zhang1, Andrew C. Tamashunas1, Tanmay P. Lele1 1Department of Chemical Engineering, University of Florida In this protocol, we describe a micropipette method to directly apply a controlled force to the nucleus in a living cell. This assay allows interrogation of nuclear mechanical properties in the living, adherent cell.
Stomata Tape-Peel: An Improved Method for Guard Cell Sample Preparation Sheldon Lawrence II1,2,3, Quiying Pang2,3,4, Wenwen Kong2,3, Sixue Chen1,2,3,5 1Plant Molecular and Cellular Biology Program, University of Florida, 2Department of Biology, University of Florida, 3Genetics Institute (UFGI), University of Florida, 4Alkali Soil Natural Environmental Science Center, Northeast Forestry University, 5Interdisciplinary Center for Biotechnology Research (ICBR), University of Florida This protocol describes a method of preparing enriched stomatal guard cells that is useful for physiological and other biological studies.
Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1 Ozlem Yaren1, Barry W. Alto2, Kevin M. Bradley3, Patricia Moussatche3, Lyudmyla Glushakova3, Steven A. Benner1,3 1Foundation for Applied Molecular Evolution (FfAME), 2Florida Medical Entomology Laboratory, University of Florida, 3Firebird Biomolecular Sciences LLC Current multiplexed diagnostics to detect Zika, chikungunya, and dengue viruses require complex sample preparation and expensive instrumentation, and are difficult to use in low resource environments. We show a diagnostic that uses isothermal amplification with target-specific strand displaceable probes to detect and differentiate these viruses with high sensitivity and specificity.
High Resolution 3D Imaging of the Human Pancreas Neuro-insular Network Elizabeth Butterworth1, Wesley Dickerson2, Vindhya Vijay3, Kristina Weitzel1, Julia Cooper1, Eric W. Atkinson4, Jason E. Coleman5, Kevin J. Otto4, Martha Campbell-Thompson1 1Department of Pathology, Immunology and Experimental Medicine, University of Florida, 2Heller School for Social Policy and Management, Brandeis University, 3Department of Medicine, College of Medicine, University of Florida, 4Department of Biomedical Engineering, College of Engineering, University of Florida, 5Department of Pediatrics, College of Medicine, University of Florida Here, we present a protocol to image human pancreas sections in three dimensions (3D) using optimized passive clearing methods. This manuscript demonstrates these procedures for passive optical clearing followed by multiple immunofluorescence staining to identify key elements of the autonomic and sensory neural networks innervating human islets.
Capturing Flow-weighted Water and Suspended Particulates from Agricultural Canals During Drainage Events Jehangir H. Bhadha1, Anne Sexton1, Timothy A. Lang2, Samira H. Daroub1 1Soil and Water Sciences Department, University of Florida, 2Everglades Research and Education Center, University of Florida Nutrients present in particulate form can contribute significantly to the overall loads in agricultural drainage waters. This study describes a novel method to capture flow-weighted water and suspended particulates from farm canal drainage over the entire duration of the drainage event.
Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition Jeremy J. Flint1,2, Kannan Menon2,3, Brian Hansen4, John Forder2,3,5, Stephen J. Blackband1,2,6 1Department of Neuroscience, University of Florida, 2McKnight Brain Institute, University of Florida, 3Department of Biomedical Engineering, University of Florida, 4Center for Functionally Integrative Neuroscience, Aarhus University, 5Department of Radiology, University of Florida, 6National High Magnetic Field Laboratory, Florida State University The current protocol describes a method by which users can maintain viability of acute hippocampal and cortical slice preparations during the collection of magnetic resonance microscopy data.
Controlled-release of Chlorine Dioxide in a Perforated Packaging System to Extend the Storage Life and Improve the Safety of Grape Tomatoes Xiuxiu Sun1,2, Elizabeth Baldwin1, Anne Plotto1, Jan Narciso1, Christopher Ference1, Mark Ritenour2, Ken Harrison3, Joseph Gangemi3, Jinhe Bai1 1USDA, ARS, USHRL, 2Indian River Research and Education Center, University of Florida, 3Worrell Water Technologies, LLC Here, we describe a protocol for the application of a novel, slow-release ClO2 product that reduces spoilage and extends the shelf life of fresh fruit. The slow-release ClO2 product was added to standard commercial grape tomato packaging and tested against Escherichia coli and Alternaria alternata.
A Method for Evaluating Insecticide Efficacy against Bed Bug, Cimex lectularius, Eggs and First Instars Brittany E. Campbell1, Dini M. Miller2 1Department of Entomology and Nematology, University of Florida, 2Department of Entomology, Virginia Tech Insecticide evaluations are often targeted against adult insects, rather than immature stages. Here, we present a protocol for evaluating insecticides against bed bug eggs with a comparison to the first nymphal bed bug stage. These protocols could be adjusted for other insects to evaluate insecticide efficacy in non-adult life stages.
Endobronchial Ultrasound-guided Intratumoral Injection of Cisplatin for the Treatment of Isolated Mediastinal Recurrence of Lung Cancer Hiren J. Mehta1, Michael A. Jantz1 1Division of Pulmonary/Critical care/Sleep Medicine, University of Florida College of Medicine The management of isolated recurrent lung cancer in a previously-irradiated field is challenging. Here, we describe an endobronchial ultrasound (EBUS)-guided cisplatin injection for the management of patients with localized lung cancer recurrence in a previously-radiated field.
Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays Alvine C. Mehinto1, B. Sumith Jayasinghe2, Darcy R. Vandervort1, Nancy D. Denslow2, Keith A. Maruya1 1Southern California Coastal Water Research Project Authority, 2Department of Physiological Sciences, University of Florida A protocol to screen for endocrine activity in organic extracts of water samples, including treated wastewater effluent and surface (receiving) water, was adapted using commercially available division-arrested ("freeze and thaw") in vitro transactivation bioassays.
A Repetitive Concussive Head Injury Model in Mice Zhihui Yang1,2, Fan Lin1,2, Amanda S. Weissman1,2, Emily Jaalouk1,2, Qing-shan Xue3, Kevin K.W. Wang1,2 1Program for Neurotrauma, Neuroproteomics & Biomarkers Research, University of Florida, 2Department of Psychiatry, University of Florida, 3Department of Neuroscience, University of Florida Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.
A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees Caroline A. Efstathion1, William H. Kern, Jr.1 1Department of Entomology and Nematology, University of Florida Preventing colonization of bird nest boxes by invasive Africanized honey bees is important for conservation efforts of nest-site limited birds. We provide an integrated pest management approach to "push" bees away from nest boxes with a repellant insecticide, permethrin, and "pull" them toward pheromone baited swarm traps.
Murine Model of Intestinal Ischemia-reperfusion Injury Ekaterina O. Gubernatorova1,2, Ernesto Perez-Chanona3, Ekaterina P. Koroleva1, Christian Jobin3, Alexei V. Tumanov1,2 1Trudeau Institute, 2Engelhardt Institute of Molecular Biology, 3Departments of Medicine and Infectious Diseases and Pathology, University of Florida Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.
Delivery of In Vivo Acute Intermittent Hypoxia in Neonatal Rodents to Prime Subventricular Zone-derived Neural Progenitor Cell Cultures Heather H. Ross1, Milap S. Sandhu1, Sharareh Sharififar1, David D. Fuller1 1Department of Physical Therapy, University of Florida This article describes the methodology for administering short periods of intermittent hypoxia to postnatal day 1-8 mouse or rat pups. This approach effectively elicits a robust tissue level "priming effect" on cultured neural progenitor cells that are harvested within 30 min of hypoxia exposure.
Isolation and Propagation of Circulating Tumor Cells from a Mouse Cancer Model Dibash K. Das1,2, Michelle K. Naidoo1, Adeodat Ilboudo1, Pascal DuBois1, Victoria Durojaiye1, Chen Liu3, Olorunseun O. Ogunwobi1,2 1Department of Biological Sciences, Hunter College of The City University of New York, 2Departments of Biology and Biochemistry, The Graduate Center of The City University of New York, 3Department of Pathology, Immunology, and Laboratory Medicine, University of Florida Circulating tumor cells (CTCs) have been shown to play an important role in tumor metastasis. Here, a method for the isolation and propagation of CTCs from the whole blood of a syngeneic mouse tumor model of hepatocellular carcinoma (HCC) metastasis is described.
Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms Eric M Deshaies1,2, Mark R Villwock1, Amit Singla3, Gentian Toshkezi4, David J Padalino1 1Neurovascular and Stroke Center, Crouse Neuroscience Institute, 2Department of Biology, Syracuse University, 3Department of Neurosurgery, University of Florida, 4Department of Neurosurgery, SUNY Upstate Medical University Minimally invasive thumb-sized pterional craniotomy for aneurysm clipping has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.
An In Vitro Adult Mouse Muscle-nerve Preparation for Studying the Firing Properties of Muscle Afferents Joy A. Franco1, Heidi E. Kloefkorn2, Shawn Hochman3, Katherine A. Wilkinson4 1Department of Mechanical Engineering, San José State University, 2J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, 3Department of Physiology, Emory University School of Medicine, 4Department of Biological Sciences, San José State University Muscle sensory neurons are involved in proprioceptor signaling and also report on metabolic state and injury related events. We describe an adult mouse in vitro muscle-nerve preparation for studies on stretch-activated muscle afferents.
Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction Ji-Yoen Kim*1, Stacy D. Grunke*1, Yona Levites2, Todd E. Golde2, Joanna L. Jankowsky1,3 1Department of Neuroscience, Baylor College of Medicine, 2Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, 3Department of Neurology and Department of Neurosurgery, Baylor College of Medicine Here we demonstrate a technique for widespread neuronal transduction by intraventricular injection of adeno-associated virus into the neonatal mouse brain. This method provides a rapid and easy way to attain lifelong expression of virally-delivered transgenes.
Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water Bledar Bisha1, Jaclyn A. Adkins2, Jana C. Jokerst3, Jeffrey C. Chandler1, Alma Pérez-Méndez4, Shannon M. Coleman4, Adrian O. Sbodio5, Trevor V. Suslow5, Michelle D. Danyluk6, Charles S. Henry2, Lawrence D. Goodridge7 1Department of Animal Science, University of Wyoming, 2Department of Chemistry, Colorado State University, 3Department of Environmental and Radiological Health Sciences, Colorado State University, 4Department of Animal Sciences, Colorado State University, 5Department of Plant Sciences, University of California, Davis, 6Department of Food Science and Human Nutrition, Citrus Research and Education Center, University of Florida, 7Department of Food Science and Agricultural Chemistry, McGill University A protocol involving integrated concentration, enrichment, and end-point colorimetric detection of foodborne pathogens in large volumes of agricultural water is presented here. Water is filtered through Modified Moore Swabs (MMS), enriched with selective or non-selective media, and detection is performed using paper-based analytical devices (µPAD) imbedded with bacterial-indicative colorimetric substrates.
Transmitting Plant Viruses Using Whiteflies Jane E. Polston1, H. Capobianco1 1Deptartment of Plant Pathology, University of Florida Improving our understanding of and our ability to manage many of the insect-transmitted plant viruses requires the use of the vector. Insect transmission of plant viruses is a tritrophic interaction, and as such requires the manipulation of insects, virus, and plant. Vectors must be reared in large numbers and manipulated in such a way as to insure high rates of transmission to test plants. The basics of rearing and manipulating the whitefly, Bemisa tabaci (Hemiptera: Aleyrodidae), a vector of plant viruses belonging to several emerging plant virus genera, representing a large number of economically significant viruses is presented.
Protocol for Relative Hydrodynamic Assessment of Tri-leaflet Polymer Valves Sharan Ramaswamy1, Manuel Salinas1, Rob Carrol2, Karla Landaburo1, Xavier Ryans1, Cynthia Crespo1, Ailyn Rivero1, Faris Al-Mousily3,4, Curt DeGroff3, Mark Bleiweis3, Hitomi Yamaguchi2 1Tissue Engineered Mechanics, Imaging and Materials Laboratory, Department of Biomedical Engineering, Florida International University, 2Department of Mechanical and Aerospace Engineering, University of Florida, 3College of Medicine, University of Florida, 4King Faisal Specialty Hospital and Research Center, Jeddah, Saudi Arabia There has been renewed interest in developing polymer valves. Here, the objectives are to demonstrate the feasibility of modifying a commercial pulse duplicator to accommodate tri-leaflet geometries and to define a protocol to present polymer valve hydrodynamic data in comparison to native and prosthetic valve data collected under near-identical conditions.
Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes Simone K. Bedoya*1, Tenisha D. Wilson*1, Erin L. Collins1, Kenneth Lau1, Joseph Larkin III1 1Department of Microbiology and Cell Science, The University of Florida With effector functions distinct from other T cell subsets, Th17 cells have been centrally implicated in inflammatory autoimmunity. This in vitro Th17 differentiation protocol provides a means to determine whether naïve CD4+ T lymphocytes can differentiate into Th17 cells, and to further examine their role in autoimmunity and host response.
Assessing Forelimb Function after Unilateral Cervical SCI using Novel Tasks: Limb Step-alternation, Postural Instability and Pasta Handling Zin Z. Khaing*1, Sydney A. Geissler*1, Timothy Schallert2, Christine E. Schmidt1,3 1Department of Biomedical Engineering, The University of Texas at Austin, 2Department of Psychology, The University of Texas at Austin, 3The J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida Three new behavioral tests (forelimb step-alternation, postural instability test, pasta handling test) for evaluating forelimb function after cervical spinal cord injury in rodents are described.
Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior Ethan M. Anderson1,2, Richard Mills3, Todd A. Nolan4, Alan C. Jenkins4, Golam Mustafa1, Chris Lloyd3, Robert M. Caudle1,2, John K. Neubert4 1Department of Oral and Maxillofacial Surgery, University of Florida College of Dentistry, 2Department of Neuroscience, McKnight Brain Institute, University of Florida College of Medicine, 3Stoelting Co., 4Department of Orthodontics, University of Florida We present a user-friendly, high-throughput operant system for the evaluation of pain behaviors in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) can assess pain through a reward/conflict paradigm thus providing a more humane way of testing. This protocol will yield more clinically relevant and translational data from rodents.
Using High Resolution Computed Tomography to Visualize the Three Dimensional Structure and Function of Plant Vasculature Andrew J. McElrone1,2, Brendan Choat3, Dilworth Y. Parkinson4, Alastair A. MacDowell4, Craig R. Brodersen5 1U.S. Department of Agriculture, 2Department of Viticulture and Enology, University of California - Davis, 3Hawkesbury Institute for the Environment, University of Western Sydney, 4Advanced Light Source, Lawrence Berkeley National Lab, 5Citrus Research & Education Center, University of Florida High resolution x-ray computed tomography (HRCT) is a non-destructive diagnostic imaging technique that can be used to study the structure and function of plant vasculature in 3D. We demonstrate how HRCT facilitates exploration of xylem networks across a wide range of plant tissues and species.
Purification of Transcripts and Metabolites from Drosophila Heads Kurt Jensen1, Jonatan Sanchez-Garcia1, Caroline Williams2, Swati Khare1, Krishanu Mathur1, Rita M. Graze3, Daniel A. Hahn2, Lauren M. McIntyre3, Diego E. Rincon-Limas1,4, Pedro Fernandez-Funez1,4 1Department of Neurology, McKnight Brain Institute, University of Florida, 2Department of Entomology and Nematology, University of Florida, 3Genetics Institute, Department of Molecular Genetics and Microbiology, University of Florida, 4McKnight Brain Institute, Department of Neuroscience, Genetics Institute, Center for Translational Research on Neurodegenerative Diseases, and Center for Movement Disorders and Neurorestoration, University of Florida We describe here the procedures for the extraction and purification of mRNA and metabolites from Drosophila heads. We are applying these techniques to better understand the cellular perturbations underlying neuronal degeneration. These methodologies can be easily scaled and adapted for other "omic" projects.
Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion Robert W. Regenhardt1, Saeed Ansari2, Hassan Azari3, Kenneth J. Caldwell2, Adam P. Mecca1 1Department of Physiology and Functional Genomics, University of Florida, 2Department of Neurosurgery, McKnight Brain Institute, University of Florida, 3Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.
Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat Saeed Ansari1, Hassan Azari2, Kenneth J. Caldwell1, Robert W. Regenhardt3, Vishnumurthy S. Hedna4, Micheal F. Waters4, Brian L. Hoh1, Adam P. Mecca3 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 3Department of Physiology and Functional Genomics, University of Florida, 4Department of Neurology, University of Florida Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.
Vertical T-maze Choice Assay for Arthropod Response to Odorants Lukasz Stelinski1, Siddharth Tiwari1 1Entomology and Nematology Department, University of Florida A vertical, T-maze olfactometer is described for assaying the behavioral response of arthropods. The olfactometer allows the experimenter to measure choices performed by test subjects when subjected to two potential odor fields. Both attraction to and repulsion from odorants can be measured with this device.
Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry Kaustuv Saha1, Jarod Swant1, Habibeh Khoshbouei1,2 1Department of Neuroscience, University of Florida, 2Department of Psychiatry, University of Florida The amperometric technique measures dopamine release from a single cell by detecting the oxidative current produced by spontaneous dopamine oxidization. Simultaneous voltage clamp and amperometry methodology reveal the mechanistic relationship between the overall "activity" of dopamine transporter and the regulatory role of this activity on the reverse transport of dopamine.
Staining Protocols for Human Pancreatic Islets Martha L. Campbell-Thompson1, Tiffany Heiple1, Emily Montgomery1, Li Zhang1, Lynda Schneider1 1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.
Collection Protocol for Human Pancreas Martha L. Campbell-Thompson1, Emily L. Montgomery1, Robin M. Foss1, Kerwin M. Kolheffer1, Gerald Phipps1, Lynda Schneider1, Mark A. Atkinson1 1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.
GC-based Detection of Aldononitrile Acetate Derivatized Glucosamine and Muramic Acid for Microbial Residue Determination in Soil Chao Liang1,2, Harry W. Read2, Teri C. Balser2,3 1DOE-Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, 2Department of Soil Science, University of Wisconsin, Madison, 3Department of Soil and Water Science, University of Florida We describe a method protocol for the GC-based analysis of the aldonitrile acetate derivatives of glucosamine and muramic acid extracted from soil. For elucidation of the chemical mechanism, we also present a strategy to confirm the structure of the derivative and the ion fragments formed upon electron ionization.
Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE) Loic P. Deleyrolle1, Mark R. Rohaus1, Jeff M. Fortin1, Brent A. Reynolds1, Hassan Azari1,2 1Department of Neurosurgery, The University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This video protocol demonstrates the application of the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) for the identification and separation of different sub-populations of cells in human glioblastoma based on frequency of cell division.
The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytometry Hassan Azari1,2, Sharareh Sharififar1, Jeff M. Fortin1, Brent A. Reynolds1 1Department of Neurosurgery, The University of Florida, 2Laboratory for Stem Cell Research, Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This video protocol demonstrates a novel method for the generation and subsequent purification of neuronal progenitor cells from a renewable source of neural stem cells (NSCs) based on their physical (size and internal granularity) and fluorescent properties using flow cytometry technology.
Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry Jennifer Parker1, Ning Zhu2, Mengmeng Zhu2, Sixue Chen1,2,3,4 1Plant Molecular and Cellular Biology Program, University of Florida, 2Department of Biology, University of Florida, 3Interdisciplinary Center for Biotechnology Research, University of Florida, 4Genetics Institute, University of Florida Reactive oxygen species level is elevated when cells encounter stress conditions. Here we show the example of 3'-3' diaminobenzidine staining as well as cysTMT labeling and mass spectrometry to profile the redox proteome in Pseudomonas syringae treated tomato leaves.
Pouring and Running a Protein Gel by reusing Commercial Cassettes Alexander C. Hwang1, Paris H. Grey1, Katrina Cuddy1, David G. Oppenheimer1,2,3 1Department of Biology, University of Florida, 2UF Genetics Institute, University of Florida, 3Plant Molecular & Cellular Biology Program, University of Florida Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences This video protocol demonstrates the isolation and expansion of stem like cells from surgically resected human glioblastoma mutliforme (GBM) tumor tissue using the neurosphere assay culture method.
Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes Jing Chen1, Scott Grieshaber1, Clayton E. Mathews1 1Departments of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida Cell-mediated lymphocytotoxicity (CML) assays can be used to test autoreactive responses and study mechanisms of cell death in vitro. However, using live-cell confocal microscopic imaging techniques with fluorescent dyes, the type and kinetics of cell death as well as the pathways utilized can be studied in greater detail.
High-throughput Screening and Biosensing with Fluorescent C. elegans Strains Chi K. Leung1, Andrew Deonarine1, Kevin Strange2, Keith P. Choe1 1Department of Biology, University of Florida, 2Mount Desert Island Biological Laboratory A procedure for liquid-based culturing and dispensing of C. elegans strains expressing fluorescent reporter proteins is described that does not require expensive sorting equipment. This approach can be applied to numerous inducible C. elegans genes for drug discovery or biosensing of contaminants.
Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice Saeed Ansari*1, Hassan Azari*1,2, Douglas J. McConnell1, Aqeela Afzal1, J Mocco1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.
High-Efficiency Transduction of Liver Cancer Cells by Recombinant Adeno-Associated Virus Serotype 3 Vectors Chen Ling1, Yuan Lu1, Binbin Cheng1, Katherine E. McGoogan1, Samantha W.Y. Gee1, Wenqin Ma1, Baozheng Li1, George V. Aslanidi1, Arun Srivastava1 1Department of Pediatrics, Division of Cellular and Molecular Therapy, University of Florida In this article, we describe the identification of the adeno-associated virus serotype 3 (AAV3) as the most efficient vector for targeting human liver cancer cells.
Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells Hassan Azari1,2, Sharon A. Louis3, Sharareh Sharififar1, Vinata Vedam-Mai1, Brent A. Reynolds1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, 3STEMCELL Technologies, Inc. This video protocol demonstrates how to discriminate and enumerate bona fide neural stem cells in a mixed population of neural precursor cells using the neural colony-forming cell assay.
Establishing Embryonic Mouse Neural Stem Cell Culture Using the Neurosphere Assay Hassan Azari1,2, Sharareh Sharififar2, Maryam Rahman2, Saeed Ansari2, Brent A. Reynolds2 1Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 2Department of Neurosurgery, The University of Florida This video protocol demonstrates the application of the neurosphere assay for the isolation and expansion of neural stem cells from the ganglionic eminences of embryonic day 14-mouse brain.
Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay Hassan Azari1,2, Maryam Rahman2, Sharareh Sharififar2, Brent A. Reynolds2 1 Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 2Department of Neurosurgery, University of Florida This video protocol demonstrates the neurosphere assay method to generate and expand neural stem cells from the adult mouse periventricular region, and provides technical insights to ensure one can achieve reproducible neurosphere cultures.
Cargo Loading onto Kinesin Powered Molecular Shuttles Yolaine Jeune-Smith1, Ashutosh Agarwal2, Henry Hess2 1Department of Materials Science and Engineering, University of Florida, 2Department of Biomedical Engineering, Columbia University Molecular shuttles consisting of functionalized microtubules gliding on surface-adhered kinesin motor proteins can serve as a nanoscale transport system. Here, the assembly of a typical shuttle system is described.
The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice Kelly A. Tennant1, Aaron L. Asay2, Rachel P. Allred3, Angela R. Ozburn4, Jeffrey A. Kleim5, Theresa A. Jones1,2 1Institute for Neuroscience, University of Texas at Austin, 2Department of Psychology, University of Texas at Austin, 3Department of Neurology, University of Florida, 4Department of Psychiatry, University of Texas Southwestern Medical Center, 5Department of Neuroscience, McKnight Brain Institute, University of Florida The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.


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Outing – publiczne ujawnienie czyjejś przynależności do społeczności LGBT wbrew woli lub oczekiwaniom albo bez wiedzy tej osoby. Outing może dotyczyć osób publicznych, których przynależność do LGBT jest ujawniana do wiadomości społeczeństwa za pośrednictwem środków masowego przekazu.

Outingu nie należy mylić z coming outem, polegającym na samodzielnym i dobrowolnym ujawnieniu faktu bycia gejem, lesbijką, osobą biseksualną czy transpłciową.

Opinie
Zdaniem większości osób, outing uderza w godność i prywatność osoby, której dotyczy. W zależności od poziomu uprzedzeń danego środowiska, może stanowić istotną przeszkodę w jej rozwoju zawodowym.

Niektórzy publicyści twierdzą, że outing jest dopuszczalną "bronią ostateczną" przeciwko osobom homoseksualnym, których "zachowanie leży w rażącej sprzeczności nie tylko z głoszonymi poglądami, ale też wyrządza szkodę ludziom homoseksualnym". Chodzi tu zazwyczaj o osoby homoseksualne, które na forum publicznym (zwykle społecznym, religijnym lub politycznym) otwarcie i umyślnie działają na szkodę środowiska gejowskiego. Tak rozumiany outing jest formą walki o równouprawnienie mniejszości seksualnych. Najczęściej podawanym przykładem tego typu sytuacji jest dyskusja na temat potrzeby outingu homoseksualnych polityków polskiej prawicy, która jest nieprzychylna wobec środowisk LGBT. Polscy działacze na rzecz praw LGBT, Robert Biedroń, Szymon Niemiec oraz Jacek Kochanowski, wypowiadali się jednakże przeciw outingowi. Kochanowski odciął się od fundacji LGBT, której rzecznik zagroził politykom prawicy ukrywającym swój homoseksualizm ujawnieniem ich nazwisk, a Biedroń zapowiedział, że Kampania Przeciw Homofobii nie zamierza wykorzystywać w swej działalności tak kontrowersyjnych metod.

Jednocześnie wiele osób popiera dokonywanie outingu, także w odniesieniu do osób publicznych, których działalność nie stoi w sprzeczności do interesów społeczności LGBT. Michael Musto, amerykański dziennikarz dokonujący outingu celebrytów, twierdzi: "wyoutowałem Rosie i Ellen i dziś nawet trudno sobie wyobrazić, że kiedykolwiek były w szafie". Tarja Halonen, działaczka na rzecz osób LGBT i prezydent Finlandii, w swojej książce krytykuje osoby ukrywające swój homo- lub biseksualizm. Krytyka osób LGBT ukrywających swoją orientację seksualną, często pociągająca za sobą poparcie dla outingu, wynika z przekonania, że widoczność osób nieheteroseksualnych w przestrzeni publicznej jest niezbędna dla tworzenia społeczeństwa tolerancyjnego wobec LGBT.

Opinia publiczna
W 2008 roku amerykański serwis "PlanetOut.com" przeprowadził badanie m.in. na temat outingu osób publicznych wśród grupy reprezentującej całą populację (nie różnicowaną ze względu na orientację seksualną) oraz wśród grupy gejów i lesbijek:

Zobacz też
 coming out

Przypisy

Linki zewnętrzne
 Kogo wydają geje?

LGBT
Psychologia społeczna

redpajama_set_name: RedPajamaWikipedia


Zizzy's interest in beef began as a young man when he went into the cow/calf business with some friends. This early experience whet his appetite for raising cattle. Throughout his years of owning and running a general construction business with his brother Pete, Zizzy never lost his interest in beef.
The genetics of raising cattle fascinate me and I like the work of continually refining and improving the herd.
Today, when Zizzy is not in his construction office, you'll find him on the farm. While most of the daily operations are handled by his Farm Manager, Daniel Stewart, Zizzy takes an active role in managing his herd. His current focus is to be able to strip graze intensively enough to be able to eliminate the need for mowing and baling hay.


You might be used to being lectured by your dentist when you go in for your appointments, and you could be wondering what you can do to make your dentist happy next time that you head to an appointment. A few ways that you can make your dentist happy have been listed here.
First of all, even though things might pop up from time to time, it is important to make an effort to always be on time for your appointments. After all, your dentist is probably a busy person who has a lot of patients to take care of, and if you're late for your appointment, then you can mess up his or her schedule. Plus, it can affect you if you're late for your appointments, too; after all, if you show up late, your dentist might have to rush through your appointment. Showing up on time will help you make sure that you get your dentist's full attention and care at your appointment, it'll help make sure that the patients behind you don't have to wait for their own appointments, and it will help your dentist avoid a stressful and hectic day.
One of the best things that you can do both to make your dentist happy and to prevent dental problems is to make sure that you take good care of your teeth in between appointments. Eating a healthy diet, such as a diet rich in calcium, can help you make sure that your teeth are strong. Additionally, avoiding foods that are filled with sugar or that are brightly colored can damage or stain your teeth. Keeping up with regular brushing and flossing is also essential if you want to keep your dental care professional happy.
If you know that you have a dental appointment coming up in the next few hours, it's a good idea to avoid eating foods that might give you bad breath, such as dishes with lots of garlic or raw onions. Even though your dentist might not say anything to you, the odor might be a little unpleasant when your dentist has to perform work or an exam of the inside of your mouth. It's not a necessity, but it's nice to keep your dentist -- as well as the dental hygienist -- in mind when choosing your diet in the hours before your appointment, and brushing your teeth well before you go to your appointment can help, too.
For more information, check out a website like https://www.belgradedental.com/.


The Massillon Museum staff have produced catalogs in conjunction with exhibits and major anniversaries since the early 2000s. These include scholarly essays, archival information, objects and archives from our permanent collection and other institutions. These publications may be viewed in the Velma B. Erwin Research Room, purchased through our online shop or in person at our OhRegionalities shop in the Museum lobby.


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Jenna Bush Hager Launches Book Club, Announces First Pick
"I think in a world that can feel very, sort of, technologically-obsessed, it's kind of fun to go back to the old-school book clubs"
By Courtney Gisriel | TODAY	• Published at 2:55 pm on March 7, 2019
When anyone on the TODAY team's looking for a book recommendation, there's only one person to turn to: Jenna Bush Hager.
The daughter of a librarian and our resident bookworm, Jenna's a wealth of knowledge when it comes to the best new reads. Now, she wants to share her passion with TODAY fans.
That's why she's kicking off a new book club for fellow bibliophiles!
"I love book recommendations from my family and friends," Jenna shared. "So to have that same community with the TODAY show viewers I think is going to be really fun."
"I think in a world that can feel very, sort of, technologically-obsessed, it's kind of fun to go back to the old-school book clubs," she continued. "My mom was in one. My grandma was in one. I'm in one. And now we get to do it. Even though it is through tech, we get to be connected and have conversations."
Every great book club has to start with a great page turner, so without further ado, the first pick for #ReadWithJenna is (drumroll please...) "The Last Romantics," by Tara Conklin!
Described as a "sweeping yet intimate epic about one American family," it's racked up plenty of praise since its release in early February. It follows the family of poet Fiona Skinner over two decades as they're forced to confront the life choices they've made.
"I'd read a review about it and I was like, 'OK, I'm dying to read that. Who's going to read it with me,'" Jenna said of this month's book. "The author wrote another book that I loved, 'The House Girl,' ... so I was excited about this."
So, how can you get involved? It's easy! As you read along, we'll be posting updates on the TODAY Instagram page with thought-provoking conversation starters and hope you'll engage with the rest of the #ReadWithJenna community to make this book club your own.
So, what are you waiting for! Here's an excerpt from the first chapter of "The Last Romantics," to get you started:
The day of our father's funeral was dank, mid-March. Ronald Reagan was president, the Cold War dragged on, Star Wars had made us all believe in forces we could not see. At that time Bexley was a town where people greeted each other by name at the post office or the bank and no one cared who had money and who did not. The doctor and the mill worker both visited my father for root canals, and both drank beer at the same drafty tavern. The dark Punnel River meandered along the east side of town and gave us something to do on summer days. This was still the era when a ninety-minute commute to New York City seemed absurd, and so the people who lived in Bexley, for the most part, worked in Bexley.
It was no surprise when the whole town turned out for our father's funeral. Hundreds, it seemed to me. Thousands. Noni led us through that awful day with an iron grip on two of our eight hands. She alternated, she did not play favorites. She had four children, and we all needed to feel the warmth of her palm.
To discover more book club content, check out the Read More TODAY page and follow along with #ReadWithJenna on Instagram and Twitter.
This story first appeared on today.com. Get more at TODAY:
This Blockbuster in Oregon is now the last one in the world
Tiffani Thiessen recalls how Luke Perry made her feel she 'belonged' on '90210'
How to help Alabama tornado victims and impacted communities
Copyright Today Digital Originals

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The Lexus Warranty: Is The Automaker's Coverage Worth It?
If you're in the market for a new Lexus, check out the warranty conditions before making a purchase
Linsay Thomas
All-new Lexus vehicles come with a standard factory warranty that aims to keep you covered if you have problems with your vehicle during the first few years and/or several thousand miles on the road. The automaker provides a decent factory warranty compared to some of its competitors and one that's more robust than average. If you're looking to buy a new Lexus, it's important that you're aware of what the warranty will provide as well as its limitations.
Written by Linsay Thomas
Lexus Warranty At-A-Glance
or 50,000 miles
Bumper-to-Bumper
or 100,000 miles
Long powertrain warranty duration
Additional coverage for hybrid vehicles
Parts coverage for one year, except for batteries
A shorter bumper-to-bumper warranty period compared to some other manufacturer warranties
Wheel alignment and balancing are only covered for one year or 12,000 miles
EXTENDED WARRANTY COVERAGE MORE … FOR LESS
Save $350 on an Endurance warranty plan with coupon code: DRIVE350
Going with a factory extended warranty is a good option for basic protection, but you can get more and save more with Endurance, ranked the top extended warranty provider by Consumer Advocates, Consumer Affairs, and Retirement Living.
Save $350 with coupon code: DRIVE350
CarShield — USA's #1 Auto Protection Provider
CarShield pays claims directly to a US or Canadian ASE-Certified® repair facility of your choice. Our contracts have low or no deductibles to prevent stress to your wallet at a critical time. Never worry about being stranded somewhere after a breakdown. All of our vehicle repair contracts include towing services to transport you and your vehicle to a safe repair facility.
Nationwide coverage for cars new and old
CARCHEX has something for everyone. Have an old car? No problem. Live in California? Not an issue. CARCHEX 's plethora of warranty plans includes options for cars up to twenty years old with up to 250,000 miles, regardless of any existing or expired warranties. Click below or call 1-877-207-3304 for a free quote.
Warranty In-Depth
The basic warranty is intended to cover nearly everything from the body to the electrical systems. However, tires are covered by a separate warranty provided by the tire manufacturer.
Lexus is the luxury vehicle division of Toyota, and its bumper-to-bumper and powertrain warranties are slightly better than its parent company regarding the coverage periods.
Bumper-to-Bumper Coverage
The basic warranty coverage protects a new Lexus vehicle for up to four years or 50,000 miles, whichever occurs first. This warranty covers repairs and adjustments to correct defects in materials or workmanship of any part supplied by Lexus.
The exceptions are certain situations such as fire, accidents, thefts, water contamination, and normal wear and tear maintenance services (i.e., replacement of fluids, filters, spark plugs, fuses, worn wiper blades, brake pads, etc.).
The basic warranty limits the scope of the coverage compared to most extended and third-party warranty options.
Powertrain Coverage
The 70,000-mile powertrain limited warranty is slightly better than some competitors in its offerings, covering most of the engine and transmission components for six years.
Specifically, you will get coverage for engine parts, which includes the cylinder block and head and all internal parts, timing belt and cover, flywheel, oil pan, water pump, fuel pump, engine mounts, engine control computer, seals and gaskets.
The powertrain warranty also covers transmission/transaxle parts (case and all internal parts, torque converter, clutch cover, mounts, engine control computer, seals and gaskets), and the front-wheel-drive system or rear-wheel drive system.
Lexus provides hybrid vehicles with an eight-year or 100,000-mile hybrid system warranty. It covers the hybrid control module, hybrid battery control module, hybrid battery, and inverter with converter.
The roadside assistance includes battery jump-starts, tire changes, lockouts, and fuel delivery and towing. The automaker provides emergency service 24 hours a day, 365 days per year for four years on a new vehicle and two years or unlimited miles on an L/Certified vehicle based on the vehicle's original date of purchase.
The company also provides a Service Part Warranty. It's a one-year warranty, regardless of mileage. Any repair or replacement resulting from a defect in the service part is covered, with the exception of batteries, from the date the part was installed or the remainder of any applicable new vehicle warranty, whichever provides greater coverage.
The Corrosion Perforation Warranty covers repair or replacement of any original body panel that rusts, but it's subject to the exceptions listed in the Basic Warranty. Coverage is for six years, regardless of mileage.
Lexus also has an optional Accessory Limited Warranty for genuine parts and accessories to replace or repair any accessory that is defective. For accessories purchased at the same time as the new vehicle, the Lexus Accessory Warranty coverage is four years or 50,000 miles from the vehicle's in-service date. When accessories are purchased following a new vehicle purchase, coverage is one year for parts and labor, regardless of mileage.
Lexus provides a slightly better-than-standard factory warranty for its brand-new vehicles
You aren't alone. Warranties have many benefits that aren't always clear. If you're new to the industry, check out our Factory Warranty Guide that helps explain some key fundamentals.
The factory car warranty is fairly comprehensive, making it a decent deal for new Lexus owners. Lexuses are some of the most dependable cars on the road, so they likely won't experience any major problems during the first few years that you drive them. However, it's still good to have a comprehensive warranty in case something goes awry.
Since the warranty is transferable, it's a bonus for potential buyers if you plan on quickly reselling your Lexus. Also, if you're a hybrid owner, you'll have peace of mind in case the system fails prematurely. In addition, the factory warranty offers extras such as roadside assistance, towing, and parts coverage.
The main problem with the Lexus factory warranty is its limitations, which is common across the board with most factory warranties. Even though the basic warranty is comprehensive in the parts that it covers, the circumstances it covers them in are limited. For example, the list of exclusions includes the following:
Major modifications
Periodic maintenance costs
General wear and tear
While the bumper-to-bumper warranty is slightly better than the average factory warranty offered by other automakers, it's still not as good as some of its competitors that offer warranties that cover more years and mileage.
In addition, if you are a frequent driver or commute long distances, you may exceed the mileage limitation within a couple of years. The roadside assistance limits are also short compared to some competitor warranties.
Common Repair Costs
Some of the most common issues and repair costs with Lexus vehicles:
Lexus GS300: Sludge build up in the engine ($134-$149 for an oil change)
Lexus GX470: Clunking noise from the rear of the vehicle when stopping (diagnostic and testing $88-$111)
Lexus ES300: Check engine light due to failed oxygen sensor (diagnostic and testing $88-$111)
Lexus IS250: Clicking noise from rear suspension (diagnostic and testing $88-$111)
Q: Do all-new Lexus vehicles come with a factory warranty?
A: Yes, they have both bumper-to-bumper and powertrain warranties.
Q: Do use Lexus vehicles come with a factory warranty?
A: Lexus provides a two-year, unlimited miles warranty only on an L/Certified vehicle.
Q: Does the factory warranty automatically transfer to a new owner?
A: No. The transfer has to be done by the new owner by who must provide proof of ownership.
Q. Can Toyota do warranty work on a Lexus?
A. No. While you can service your Lexus vehicle at a Toyota dealership because they share many of the same mechanical components, warranty services must be carried out at a Lexus dealership.
Q. Should I buy an extended Lexus warranty?
A. Many Lexus owners keep their vehicles for 10 years or more than 100,000 miles, so an extended warranty is not a bad idea. Lexus-branded extended protection plans aren't the only option for Lexus owners looking to extend coverage after their factory warranties have expired. You can forego Lexus's own plan in favor of what is known as a third-party, independent Vehicle Service Contract.
Q: Are Lexuses expensive to fix?
A: Many parts on a Lexus come from Toyota, which is one of the world's largest automakers. As a result, many of the parts are common and readily available. Statistically, Lexus is one of the lowest-maintenance cars during the first 10 years of its life.
Q: Does Lexus include free maintenance?
A: Every new Lexus includes complementary first and second vehicle service agreements They are provided at six months or 5,000 miles and one year or 10,000 miles.
Is Lexus' Warranty Worth It?
The Lexus factory warranty is a pretty decent offering from the manufacturer for any new vehicle.
The coverage limits of both the bumper-to-bumper and powertrain warranties are better than average, and the coverage is long enough to protect a new vehicle during the most common period for factory defects.
If you have your eyes on a new Lexus, you should feel pretty confident with the included factory warranty.
Here are a few more resources you can use:
Lexus phone number: 1-800-255-3987
Lexus website
Lexus recalls
How Much Does an Extended Car Warranty Cost?
Don't get surprised by the real costs many extended car warranties have
Who Has the Best Car Warranty?
It's time to look at major automakers and the factory warranties they can offer to you
A Review of CarShield's Extended Warranty Plans
Check out what you may get with a CarShield extended warranty
Other 'Factory Warranties' Reviews You Might Like:
VW CPO Warranty Lincoln CPO Warranty Rolls Royce Warranty Mitsubishi Warranty Porsche Warranty MINI Cooper Warranty Suzuki's Limited Warranty Land Rover Warranty Honda Accord Warranty Kia Warranty Bmw Warranty Honda Warranty Mazda Warranty Toyota Warranty Hyundai Warranty Subaru Warranty Ford Warranty Nissan Warranty


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From Paris To Berlin (And Most Other Western European Capitals)
Just like John in the post before this one, I too watched Stage 5 of the Tour de France. However, unlike John, my motivation was not to watch the cycling but rather the route in southern France that finished in the small town of Privas.
Most people (French people that I have met included) have never heard of Privas. With a population of less than 10,000 people it is easy to understand why. However, the town is a commune of France, and capital of the Ardèche department, the smallest administrative centre of any department in France. The reason I know is Privas is because I was lucky enough to visit the area 5 times during 2010-2013 while completing my PhD. It was fantastic to see the sprint finish into the town, along it's narrow streets, a far cry from the Champs-Élysées where the tour will finish.
And turning to Paris, where the Tour will end on the 20th of September, one can imagine that the Champions League loss is still fresh in the memory of Parisians. The end of the Tour might be a welcome distraction for many. The club's first appearance in the showpiece European game did not work out as expected. And while the champion of the Tour de France will always be crowned in Paris, the Champions League trophy has never been to the French capital.
And Paris is not alone in this regard.
One my second trip to Privas in 2010 I spent the entirety of my TGV journey from Paris Charles de Gaulle to Valence (south of Lyon) reading the wonderful Soccernomics by Kuper and Szymanski. I couldn't put it down. It was one of those rare books that I had to stop myself from reading, so as to have it for just a little longer. One of the earlier chapters recalls the success of club football teams in the European Cup. It is somewhat remarkable how so few clubs from Western capital cities have won the competition. PSG and Paris remain in this list.
The Table below lists the number of times the European Cup/Champions League trophy has been won by capital cities and regional cities in Western Europe. Just two other countries have clubs teams that have won the cup - Romania and the Former Yugoslavia. In both cases the teams were from the capitals (Steaua București (1986) and Red Star Belgrade (1991). Both were under dictatorship communists government at the time of their success.
And this is what make the list to the left even more interesting. The obvious outlier is Spain. Real Madrid have won a remarkable 13 European titles. Adding in Lisbon (Benfica), the two national capitals on the Iberian peninsula have won 15 European Cups/Champions Leagues. It is worth noting however that both cities had authoritarian governments until the 1970s. Benefica won both titles (1961 and 1962) under this system and Real Madrid their first six.
Western European capitals that have been fully democratic since 1945 have a pretty dismal record at winning the tournament. Ajax of Amsterdam (if you agree this is the capital and not The Hague) is the only bright spot. In fact, even London with the might of Chelsea, Arsenal and Tottenham had to wait until 2012 to win the competition for the first time.
Berlin, Edinburgh, Rome and Paris are all devoid of success. This is not to mention Athens, Bern, Brussels, Copenhagen, etc. who will probably never boast the winner. Instead the power lies in the regional cities. Places like Munich, Milan, Liverpool, Manchester, Barcelona and Turin. And historically in cities like Birmingham, Nottingham, Hamburg, Rotterdam, Eindhoven, Glasgow, Porto and Dortmund.
Kuper and Szymanski hypothesize why this might be the case, and why capitals like Paris could emerge as winners in the not too distance future. But for now, Neymar and co. will have to wait.


The lightweight and versatile women's Air Max Thea joins the latest SNKRS of the Week selection. The sneaker features a breathable textile upper with leather overlays topping an injected Phylon sole. Spring tones comes to life courtesy of a white, blue lagoon and ghost green colorway.


Shopping for a Tesla Appears Fairly Straightforward. However There Are a Few Issues to Know. - Information by Automobilnews.eu - Automobile News and Trends - You must read this !
Shopping for a Tesla Appears Fairly Straightforward. However There Are a Few Issues to Know.
When Tesla stated on Thursday night time that probably the most primary, $35,000 model of its Mannequin Three would now not be supplied on-line, my colleagues and I had been confused.
Isn't a Silicon Valley firm speculated to be all concerning the web? Why do I want to make use of a telephone or, worse, go to one in all Tesla's showrooms and work together with a gross sales consultant in actual life to order a high-tech automotive?
I needed to attempt the method for myself to see if Tesla's new coverage made issues unnecessarily difficult.
Although I ended wanting putting an order, it turned out to be fairly straightforward to stroll by the acquisition course of for a Mannequin 3, each on-line and over the telephone.
Regardless of some conflicting info, my telephone conversations with individuals at three Tesla shops within the New York space about shopping for a Mannequin Three had been environment friendly and matter-of-fact. On-line, it took me lower than two minutes to pick a automotive and attain the bank card info web page.
However you'll be able to't essentially get the model you're after, even from a retailer. And I did be taught a couple of issues any potential Tesla purchaser ought to know.
Now that Tesla isn't promoting the bare-bones Mannequin Three on-line, the least costly automotive you should buy on its web site goes for $39,500.
However Tesla lists the automotive's value as $28,950. That determine is later described as "after financial savings," and the corporate even shows the per-month price of a lease in these phrases.
Because it seems, Tesla is utilizing some inventive math to get to that quantity. Financial savings embrace a $3,750 tax credit score presently obtainable to Tesla consumers, however additionally they embrace cash that you'd in any other case spend on fuel for those who purchased a regular automotive.
Utilizing a instrument on the Tesla web site, you'll be able to modify that determine by altering the variety of miles per 30 days you may drive or the price of gasoline.
The corporate explains what it's doing, however this nonetheless struck me as a questionable gross sales tactic, particularly because the value "after financial savings" is the primary value you see on the web site.
What does 'customary' get me?
For consumers in search of the $35,000 base mannequin of the Mannequin 3, the one option to get one with out trekking to a retailer is to make a telephone name.
When it introduced the gross sales change, Tesla stated the "customary" Mannequin Three was really a software-limited model of the dearer automotive you should buy on-line. Which means the vary of the automotive on a full cost is restricted, for instance, not by battery physics however by software program that goals to maintain it that means.
A Tesla gross sales consultant on Lengthy Island defined this, too, and stated a purchaser might pay the distinction for a extra succesful model at a later date and the automotive could be up to date over the airwaves.
For me, the most important level of confusion needed to do with take a look at drives. Tesla says on its web site that you could drive a automotive for per week, or lower than 1,000 miles, and nonetheless return it. A gross sales consultant in Westchester County repeated this.
He additionally stated that if I did a take a look at drive earlier than shopping for I'd have solely in the future to return the automotive, so I reasoned it was most likely higher to purchase first, drive later.
What I used to be advised, nevertheless, conflicted with the coverage as Elon Musk, Tesla's chief government, not too long ago acknowledged it on Twitter: that returns had been potential inside one week no matter a take a look at drive.
A Tesla spokeswoman stated this was a latest change. Phrase could not have reached all of the shops.
One other level on which the shops appear out of step with the corporate's get together line is in estimating the time it might take to get a automotive. In response to the web site, a automotive ought to be obtainable in two weeks, however the Westchester retailer stated it'd take as much as 4, particularly if I actually needed a less-expensive model of the Mannequin 3. The shop in Manhattan stated I couldn't get a $35,000 Mannequin Three there in any respect. The most cost effective model it had obtainable would price $45,500.


Starring Kris Demeanor, Siobhan Cooney, Natalie Marshall, and Tori Darr.
Based on the short story "The Weather" from Deborah Willis' book Vanishing and Other Stories (finalist for the Governor General's Literary Award), this short film follows the influence that a city girl has on the lives of an aging cowboy, Braden, and his daughter, Edith. Braden's wife Nina left him and their rural Albertan home after a tornado wreaked havoc on their farm. Heartbroken, Edith and Braden attempt to continue their lives as usual until Edith befriends a new student: Rae. Rae's parents moved her out to the country to get her away from some bad habits she picked up in the city. Braden and Edith welcome Rae into their home despite Braden's concerns about the influence Rae will have on his daughter. He neglects, however, to recognize the influence Rae has on him.
This short film is currently in post-production and will be submitted to film festivals in winter 2018.


Collective favorites + inspiration from the studio.
These are some particular works that inspire us.
George Pratt, I particularly like this landscape because of how "moody" it is.
Its really dark but has enough warmth in it to evoke a pleasant emotion.
this one in person at the Cleveland Museum of Art.
think otherwise I defy you to give it a shot!
I seem to gravitate to the more abstract expressionistic landscapes from the 40's and 50's.
minimum, and really use color and simple forms in an almost naïve way to convey their image.
are uncannily similar, and I love the painterly quality of their simple forms and planes.
To me, they are almost reminiscent of the works of Hans Hoffman.
have always stayed with me as an artist and awaken me each time I see them.
Mark English has been one of the leading illustrators in the US for decades.
1). "Young Corn" by Grant Wood, 1931.
the image still radiates a warm feeling.
2). "Castrovalva" by M. C. Escher, 1930.
viewpoint and contrast do give it an austere quality.
3). "Relayer" by Roger Dean, 1974.
paintings a wonderful, distinctive look.
I think my favorite landscape of all time is this Twachtman. On the flip side, I also enjoy landscapes pushed into abstraction. A perfect example for me would be Diebenkorn's early works in New Mexico / circa 50'-60's. Another Painter that had a big influence on me was my professor at RISD Thomas Sgouros. His early watercolors were amazing and to me are at the same level as Andrew Wyeth. The Cleveland Museum has one of his pieces from the 50's which I have yet to see. Now 83, his eyesight is failing only seeing blurred shapes and color. He now paints "The Remembered Landscapes" large scale oils.
in style at that time.
washes and reflected light and colors.


Thu, 28 March 2019 – Ana Petrache (IRH-ICUB Fellow), What can someone achieve through a fake eschatology?
Thu 21 June 2018 – Matthew Dentith (New Europe College / IRH-ICUB Alumnus), What's fake about fake news?
Thu 22 February 2018 – Adina Camelia Bleotu (IRH-ICUB Grant), Scalar implicatures with quantifiers in Romanian. Are children more logical than adults?
Thu 11 January 2018 – Viktor Ilievski (IRH-ICUB Fellow) – Aristotelian causes in Plato's Timaeus?
Thu 06 April 2017 – Liviu Giosan (MIT/ Woods Hole Oceanographic Institution/ IRH-ICUB Visiting Professor), Anthropocene: A Chance to Bridge the Gap between the 'Two Cultures'?
Thu 28 April and Thu 5 May 2016 – Easter holyday.
Thu 24 March 2016 – Peter Anstey (Visiting Professor at the IRH-ICUB/ University of Sydney), Experimental moral philosophy?
Thu 3 December 2015 – Ionut Tudorie (IRH-ICUB) – May they not decay after death: In Search of the Uncorrupt Body of Byzantine Emperor Michael VIII Palaiologos (1258-1282).
Thu 29 October 2015 – Ciprian Jeler (IRH-ICUB) – What kind of environment is required for natural selection?


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TrailLink, powered by Rails-to-Trail Conservancy, is the ultimate trail-finder app for all outdoor enthusiasts. Download TrailLink today to enjoy full-length trail descriptions, reviews, photos, and detailed trail maps not found anywhere else! TrailLink is powered by Rails-to-Trails Conservancy (RTC) expert trails database which offers unique trail-finder functionality for more than 30,000 miles of multi-use trails.
The TrailLink app also offers in-app purchases for RTC's detailed offline trail maps which can now be downloaded in the TrailLink app to fully empower your on-the-trail adventures! Take TrailLink with you and let Rails-to-Trails Conservancy be your expert guide out on the trail. Try it out and register today to get your first map for free!
TrailLink offers in-app purchase map bundles which allow you to download RTC's detailed trail maps for offline use at 3 maps for $1.99, 7 maps for $3.99, or 15 maps for $7.99 to fully empower your on-the-trail adventures! Or, if you prefer, you can purchase the Unlimited subscription in the app directly through your iTunes Account for $29.99 for 12 months of unlimited offline map downloads. The TrailLink Unlimited subscription will automatically renew for $29.99 after 12 months unless you cancel your subscription at least 24hrs before the end of the current subscription period. You can cancel at any time and manage your subscription, including turning off auto-renewal, in your Account Settings.
Plus, your purchase helps support to the mission of Rails-to-Trails Conservancy!
Rails-to-Trails Conservancy's GIS mapping initiatives have produced more than 30,000 miles of highly accurate trail maps which include tens of thousands of amenity waypoints such as parking lots, trailheads, tunnels, restrooms, drinking fountains & much more — all available for download in the TrailLink app!
•Support the mission of Rails-to-Trails Conservancy!
Download TrailLink today and let RTC guide your way!
I love this app. We just got into biking this year and have enjoyed trying out new routes. I prefer biking on trails more than roads and have downloaded a dozen maps so far. They are very useful since most trials don't have adequate signage and therefore we can use gps with the app.
My only complaint is this. We prefer to ride a minimum of 20 miles at a time. If I see a trail that is only 3 miles long I will skip it. Not worth the drive. But I've realized that a lot of these continue where another trail ended. It's really hard to see this on the app. Would be nice to have a filter on the app that would show all the trails in an area.
All in all I love this app. Can't wait to go out and explore more trails.
Since I travel a lot, I find this app useful for zeroing in on trails that I can enjoy while in new places. The app saves me time by providing local information & helpful hints to maximize my experience. Thanks!!
Also, I cannot understand how anybody could ever complain about paying a bit for work that others do, to make your life more enjoyable. Seriously, the app creator/manager seems to be communicative w users & wants to improve the app ongoing.
Thanks for your support, Staeces! I think folks sometimes forget RTC is a non-profit trails organization and our primary mission is to help folks get out and enjoy trails! It's not an easy feat to create a model that sustainable but we try our best to keep improving our website and apps! Happy Trails!
The iPad version is simply a scaled up version of the iPhone version. Very annoying. I tried to write a review for the extension of the Bruce Freeman Rail Trail in Acton, MA, and I was limited to about 3 lines of text without a scrolling capability. I was unable to complete my review because I could not see what I was typing. Too bad, because I was attempting to post a 5 star rating for the trail but the app let me down. The app also only works in portrait mode 😕 which is VERY annoying. Perhaps these issues are why I had removed the app from my iPad (I needed to re-download the app to post a review). Please fix it!


text:
Generation Climate Kid
Cynical millennials are partying while the planet burns. The teen idealists of Gen Z are revolting. But who's right and who's the most woke? Four pairs of siblings challenge each other on who deserves the moral top bunk.
Introduction: Johanna Kinnock
Move over millennials, there's a new generation in town and they don't compromise on the climate. The world is burning – a fact that we millennials were aware of our whole lives – so why didn't we do anything? Even though the gap in years between teens and twentysomethings is small, the gap in activism is clear. Gen Z'ers have taken to the streets in school strikes across the world, following the lead of The Almighty Greta and many other young climate heroes. When we see this up-for-it agit-youth, it brings a tear of pride to our eye. But it also makes us feel like that scene in 21 Jump Street where Channing Tatum and Jonah Hill go back to high school and realise it's now cool to be vegan and wear your ruck-sack "two-strap": old.
To test the freakonomics of Generation Climate Kid – and to mark the start of Extinction Rebellion's fortnight of protests –we checked for variables by choosing pairs of siblings from different sides of the generational divide. On the agenda: the impending apocalypse, to protest or not to protest, and whether it's possible to sort all that out during family discussions, drunken or otherwise. And, as we investigated Gen Z's all-empowering wokeness, we learned how to efficiently tease your brother with an anti-nuclear power tote bag. Because, ultimately, annoying your sibling will always be the most sustainable source of pleasure, right?
Bella and Neve years before they discovered oat milk hot chocolates
Neve, 17, in London studying A‑levels
I've recently started Year 13 but I've spent the last few months complaining that I'm done with education. I don't think university is the path for me because all I seem to care for is shouting about how badly we've treated this planet.
The question, "Are you an activist?" bothers me. We're at a point, a sixth mass extinction point, in which you can't just not care.
My big sister dresses and prides herself as the "alternative" type: floaty, tie-dyed dresses, wears no shoes to be at one with the ground. You can picture it. But I do ask myself how much Bella is actually doing for the actual ground. Since joining the nine-to-five work system I have seen nor heard little action from her. She's given 20 holiday days a year, but so far she has spent every one jetting around the world to different festivals.
Meanwhile, I miss school stomping through Westminster with a placard and do my best to flood the family group chat with petitions, articles and photos of me climbing trucks at a Fridays for Future strike.
I'm not claiming to be Neve Thunberg. I've given up milk, meat and fish, but still struggle to turn down the last bite of a friend's chicken burrito. I justify this by saying I've not actually funded the meat industry and I'm reducing food waste in our society. But, yes, I need to practice what I preach a bit harder.
Growing up entirely with social media has played a massive role in this difference of outlook. Clips, solemnly narrated by celebrities, of starving polar bears and raging forest fires fill my screen time. A constantly updating reel of the "current best available science", as Greta calls it, is frustrating to read if no action follows. My micro-generation is angry. Maybe it's because our supposed role models are doing so little. Or because we are so frequently denied a voice in issues that will most certainly affect us for decades longer than the babies in suits that are currently leading our world into the flames.
If we're lucky, we'll get those decades. And I'm not planning on wasting them. As the planet and its youth become increasingly fired up, I hope my sister puts on some shoes and joins the march.
Bella, 24, in London working in TV production
My precocious little sister's probably said I'm a fake hippie. That I flounce around in my blown-out trousers and my Indian silk and I've long since located my chakras. But I'm a fraud, right? What am I really doing to protect the earth I love, other than dance around on it barefoot every summer?
Definitely not enough. But I promise, I do try. I've been a vegetarian for a long time – since before it was cool – but I can't bear to lose the joy of cheese just yet. I've had a bamboo toothbrush before, but hell, they can get expensive. And, as we've all been reminded, cocaine isn't vegan, guys.
To get serious for a moment – at a time when I know we must – I'll admit it: the older I get, the easier it is to be pacified by these bite-size chunks of eco-goodness. I'm becoming increasingly complacent, one oat-milk latte at a time.
The sad truth of it all is that adult life in the city moves too fast to stop and think about every change I could be making. I barely have time to look after my houseplants, let alone the planet. That doesn't mean I've stopped trying altogether, just that maybe 17-year-olds have a bit more time on their hands to fuck around with multi-coloured Sharpies? And when little sis starts paying rent in London, then we can talk about buying shampoo in glass bottles.
It's not like I've never marched for something I believe in. Were you even alive for the Iraq war march, Neve? I marched against Brexit (on a Saturday, I'll admit). I marched against tuition fees. I remember how fun it is to be kettled (let's not pretend). But it's a lot easier to walk out of school than the office.
It's soul-destroying to realise that each of these tragic political decisions have steamrolled through without my generation's voice making much of a difference. Which makes it incredible – miraculous, even – that the generation coming up behind us remains so unflinchingly optimistic.
With that in mind, if it forces me to up my eco-game, maybe a little sibling rivalry is a good thing. I'm glad this fierce little imp is on my shoulder, poking my hippie-ocrisy. After all, one day our own kids will be climbing trees together. And it would be good if there were still some left to climb.
Theo learning to gag Hannah so he can bang on about the benefits of nuclear energy
Theo, 17, in Copenhagen studying Sound Design
I've been into climate stuff ever since my 7th grade (Year 8) physics teacher taught us about climate change and alternative energy sources – including nuclear power. I do my bit for the climate, like not eating meat and wearing second-hand clothes. But mainly I'm pro-nuclear power as a sustainable energy source. Even the idea of it is so taboo here – my parents are of that Seventies generation who'd grown up hearing that anything nuclear was just straight-up bad and dangerous. They thought I was a total radical just for bringing it up, but I think they're coming around to it now. Especially since I talk about it at any opportunity.
And then there's my sister… Hannah is supposedly against nuclear power, but I kind of think she wears her pro nuclear-power tote just to bug me. Whenever we end up discussing the climate at home it always ends with bad vibes; it can literally ruin an evening. But how am I meant to be sure of my opinions if I can't use them to nuance my family's views? They think I'm chatting bollocks about stuff I don't know about, but the fact is: I've got the facts. I've got an arsenal of facts. If you want I can send you some links.
Hannah, 23, in Copenhagen studying Anthropology
I grew up knowing all about climate change but it was never something I seriously acted on: I knew it existed but I didn't know it in my bones.
All families have their little roles and I used to just put my younger brother into this box as "the stubborn one" because he insisted on not flying or eating red meat. I figured he was just being a typically difficult teenager. The other day we were all having dinner and he was talking about a trip he's taking to Norway. As the helpful big sis that I am, I told him I knew a website with cheap flight tickets. Theo just rolled his eyes: "Obviously we aren't flying."
I get the sense he thinks my parents and I are hypocrites who romanticise climate-friendly behaviour like buying organic food when it benefits our self-image as a progressive family. Basically, he thinks we're uninformed and a bit ridiculous. For example, I wear this "nuclear power, no thanks" tote, which I think looks cool. Theo literally refuses to look at it when he comes over. What I'm impressed about in him and his generation is that they are dealing with this deep-rooted climate anxiety in the calm and rational way that we never did. Him and his friends are all so… earnest about it that they might actually manage to turn this thing around. God, it's annoying though.
Neena and Seb: making eco-activism trendy via their fashion and haircut choices since 2002
Neena, 21, in Cambridge, studying French
Climate change was always a secondary consideration when we'd talk about social justice stuff in school. I've been into feminism for ages and that seemed much more pressing at the time. Climate change was abstract and even sort of tame. I think my little brother Seb's generation came of age when politics as usual got disrupted, when Trump – a literal climate-denier – got elected.
That's why Gen Z is less intimidated to be political: if none of the politicians are making sense, you don't have to have super eloquent arguments before you speak out, either. When I was a teenager I don't think we even saw it as an option to go out and protest, and it wasn't necessarily cool to be politically active. It sounds bad but when I was 14, posting something environmentalist online was perceived as a bit lame. Now I think it's normal, and even trendy.
And Seb? Seb goes about climate change in his very Seb way – more fact than feeling.
Seb, 17, in Brussels doing A‑levels
My sister and I are very different. It sounds weird but she's much more emotionally driven. She really cares about people's feelings and that's what drives her politics. She'd get really affected by testimonials of people having to evacuate their houses, whereas I worry about the numbers and science behind it all.
I think because there is such a substantial movement now it feels less intimidating. And we're more pragmatic – for sure Greta has helped rally the youth, but, honestly, she just said what we were all thinking. Millennials aren't old enough that I can be mad at them, but Gen X – don't get me started on Gen X. They knew for so long and didn't do anything because they were more interested in short-term gains.
At school the climate is basically the main thing. People talk about it loads. The other day I asked the people who organise the school bus to stop printing out 250 pages of admin every day. Little things like that.
Social media got us here. But the fact that being engaged is considered cool also means that you now have the types of people that go to the marches to post it on social media – and then Uber home whilst eating a plastic-wrapped sandwich.
But that's OK: their presence helps the cause either way. It's funny when people think we are this boring generation who don't go out and just sit at home typing up resolutions for the UN the whole time. We still go out, we still enjoy being teenagers, but we also enjoy going to protests and doing something about the climate. You can thank us later.
Cam and Iona travelling on a zero-emissions vehicle
Cam, 23, in London training as a solicitor
I remember being introduced to an "inconvenient truth" early on but I never thought it would affect me, really. Climate change was a bit like ISIS: far away, very real, but ultimately not part of my life. I did become a veggie seven years ago but from the POV of animal rights – Paul McCartney saying "if slaughterhouses had glass windows" and all that.
The environmentalist focus on meat-consumption was later. That's how my little sister was introduced to it; she just turned veggie recently. I'm really glad she's finding a political cause to get into. When I was her age I got really impassioned by left-wing politics, but she always found politics really tedious till now. To quote her: "Ugh, why do I have to be so political? It's really boring."
So, yeah, I'm really glad she's got a cause she cares about now. I read The Uninhabitable Earth recently, and I had to put it down because it's so bleak. Honestly, if I think about climate change too much, it makes it hard to get out of bed in the morning because there's just no way back.
People tend to find it depressing if I tell them over a glass of wine that capitalism needs to be totally remodelled for us to survive, that we're all gonna die and should at least stop having children. The generation below us – I don't wanna call them naïve but I think they're unaware of this huge, established system that they're up against. I haven't had the heart to tell my sister. I don't want to crush her budding idealism.
Iona, 17, in Surrey finishing A‑levels
My brother Cam is so dramatic. I honestly don't see the point in thinking: the world's going to explode, now I'm going to ruin my life worrying about it. People really need to look at the science. We need to keep thinking and researching. Who knows, maybe we could pull something out of the bag, like one of those giant CO2 hoover things.
The difference between my siblings and me is that I've always been more scientific than them. I'm the only one that's done science at A‑level. I doubt most of the people who are getting so depressed about the climate have actually read the scientific reports and I doubt they'd understand them. Including my brother.
We were on holiday this summer and Cam got drunk and was, like, "the world's gonna burn – we can never have kids!" I told him to stop preaching and walked off. But I thought about it after and realised he has a point.
Since then, I've been way more engaged with climate stuff, in particular in a marine biology sense – coral bleaching is something I really care about. But it's not in the same doom and gloom way as my brother. I do believe it's reversible. I'm applying for biology at uni and I hope I'll be able to use what I learn there to help the cause. And yes, I absolutely do plan on having kids one day.


-- (c) Virginia Mayo: ' If I... I'd be a brazen hussy ' in Wonder Man (1945).
-- (d) In Who Framed Roger Rabbit , Jessica Rabbit is a: ' painted hussy '.
ETYMOLOGY: From the legitimate Middle English huswif meaning housewife or mistress of a household, a frugal, strict woman who ran all the domestic aspects of family life . Rosalie Maggio, The Dictionary of Bias-Free Usage (1991): Derived from housewife, this word has taken the low-road while its innocuous, affectionate mate , hubby , has taken the high road .' When used adjectively Rosalie Maggio recommends the following alternatives: bold , brazen , seductive, shameless, immodest, immoral , indecent , bold-faced, etc. See playgirl for synonyms.
2. In gay parlance, an unrestrained homosexual .