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Gill Hermelin started teaching at Rock School in 2018. He teaches bass, guitar, drums, piano, voice, and band. A previous student of the school, his favorite instruments to play are guitar, piano, and bass. Gill has received private lessons in audio engineering and has extensive knowledge of music composition and songwriting techniques. Gill recorded his first album of original songs titled “Dreamer” in 2017. He is now in the process of recording his second album.
Check out Gill’s music on Spotify:


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The seven-piece Grateful Dead tribute band, Dark Star Orchestra has revealed that they will be hitting the road in the spring months of the upcoming new year. The historic commemorative Grateful Dead band will be selling out amphitheaters across the nation but this tour will be the first time ever that the collective will be performing in an underground cave in Pelham, Tennessee during their upcoming 2020 excursion.
The Dark Star Orchestra 2020 North American Spring Tour will kick off in the month of March and make its way to cities like St. Petersburg (Tickets), St. Augustine (Tickets), Pompano Beach (Tickets), Orlando (Tickets), and more. Tickets will be available for purchase to the general public on Friday, Dec. 13 (Tickets).
Since coming together in 1997, Dark Star Orchestra has been performing improvisational original sets of the legendary rock group Grateful Dead while continuously touring for the band's amazing cult-like following that have unwaveringly celebrated the band's artistry since the 1960s. With the support of the Deadheads, Dark Star Orchestra has been on multiple tour stints this year that will eventually bleed into the new year and continue for many more.
Dark Star Orchestra 2020 North American Spring Tour
March 27, 2020 – Raleigh, NC – The Ritz
March 28-29 – Wilmington, NC – Greenfield Lake Amphitheater
March 31 – Columbia, SC – The Senate
April 2 – St. Petersburg, FL – Jannus Live (Tickets)
April 3 – St. Augustine, FL – St. Augustine Amphitheater (Tickets)
April 4 – Pompano Beach, FL – Pompano Beach Amphitheater (Tickets)
April 5 – Orlando, FL – Plaza Live (Tickets)
April 7 – Savannah, GA – Victory North
April 9 – Charleston, SC – Charleston Music Hall
April 10 – Atlanta, GA – The Tabernacle
April 11 – Pelham, TN – The Caverns
April 12 – Birmingham, AL – Avondale Brewing Co.
Dates, venues, and prices are subject to change. Please check individual event pages for more details.

domain: artistic


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Synesthesia is a rare condition in which a stimulus from one modality automatically and consistently triggers unusual sensations in the same and/or other modalities. A relatively common and well-studied type is grapheme-color synesthesia, defined as the consistent experience of color when viewing, hearing and thinking about letters, words and numbers. We describe our method for investigating to what extent synesthetic associations between letters and colors can be learned by reading in color in nonsynesthetes. Reading in color is a special method for training associations in the sense that the associations are learned implicitly while the reader reads text as he or she normally would and it does not require explicit computer-directed training methods. In this protocol, participants are given specially prepared books to read in which four high-frequency letters are paired with four high-frequency colors. Participants receive unique sets of letter-color pairs based on their pre-existing preferences for colored letters. A modified Stroop task is administered before and after reading in order to test for learned letter-color associations and changes in brain activation. In addition to objective testing, a reading experience questionnaire is administered that is designed to probe for differences in subjective experience. A subset of questions may predict how well an individual learned the associations from reading in color. Importantly, we are not claiming that this method will cause each individual to develop grapheme-color synesthesia, only that it is possible for certain individuals to form letter-color associations by reading in color and these associations are similar in some aspects to those seen in developmental grapheme-color synesthetes. The method is quite flexible and can be used to investigate different aspects and outcomes of training synesthetic associations, including learning-induced changes in brain function and structure.
24 Related JoVE Articles!
Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure
Institutions: Massachusetts General Hospital, Harvard Medical School, Massachusetts General Hospital, Harvard Medical School, Massachusetts General Hospital, Harvard Medical School, Massachusetts General Hospital, Harvard Medical School, The Methodist Hospital Research Institute, The Methodist Hospital, Weill Cornell Medical College, Bejing University of Chinese Medicine, The Hong Kong Polytechnic University, Brigham and Women's Hospital, Harvard Medical School.
Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase
transgenic mice reported in this manuscript provides a rapid in vivo
assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs.
Medicine, Issue 30, Gua Sha, blood extravasation, bruises, heme oxygenase-1, gene expression, systems biology, small animal molecular imaging, optical and bioluminescence imaging, HO-1-luciferase transgenic mice, Chinese folk therapy
Developing Custom Chinese Hamster Ovary-host Cell Protein Assays using Acoustic Membrane Microparticle Technology
Institutions: BioScale, Inc., BioScale, Inc..
Custom assays for unique proteins are often limited to time consuming manual detection and quantitation techniques such as ELISA or Western blots due to the complexity of development on alternate platforms. BioScale's proprietary Acoustic Membrane MicroParticle (AMMP) technology allows sandwich immunoassays to be easily developed for use on the ViBE platform, providing better sensitivity, reproducibility, and automated operation. Provided as an example, this protocol outlines the procedure for developing a custom Chinese Hamster Ovary- Host Cell Protein (CHO-HCP) assay. The general principles outlined here can be followed for the development of a wide variety of immunoassays.
An AMMP assay measures antigen concentration by measuring changes in oscillation frequency caused by the binding of microparticles to the sensor surface to calculate. It consists of four major components: (1) a cartridge that contains a functionalized eight sensor chip (2) antibody labeled magnetic microparticles, (3) hapten tagged antibody that binds to the surface of the functionalized chip (4) samples containing the antigen of interest. BioScale's biosensor is a resonant device that contains eight individual membranes with separate fluidic paths. The membranes change oscillation frequency in response to mass accumulating on the surface and this frequency change is used to quantitate the amount of added mass.
To facilitate use in a wide variety of immunoassays the sensor is functionalized with an anti-hapten antibody. Assay specific antibodies are modified through the covalent conjugation of a hapten tag to one antibody and biotin to the other. The biotin label is used to bind the antibody to streptavidin coupled magnetic beads which, in combination with the hapten-tagged antibody, are used to capture the analyte in a sandwich. The complex binds to the chip through the anti-hapten/hapten interaction. At the end of each assay run the sensors are cleaned with a dilute acid enabling the sequential analysis of columns from a 96-well plate.
Here, we present the method for developing a custom CHO-HCP AMMP assay for bioprocess development. Developing AMMP assays or modifying existing assays into AMMP assays can provide better performance (reproducibility, sensitivity) in complex samples and reduced operator time. The protocol shows the steps for development and the discussion section reviews representative results. For a more in-depth explanation of assay optimization and customization parameters contact BioScale. This kit offers generic bioprocess development assays such as Residual Protein A, Product titer, and CHO-HCP.
Bioengineering, Issue 48, Immunoassays, Chinese Hamster Ovary Host Cell Protein, Residual Protein A assay, Assay development, Biomarker detection and quantitation, Phospho-AKT, Gadd34, tissue sample, tumor sample, bioreactor sample
Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay
Institutions: University of California, Berkeley .
Fluorescence in situ
hybridization (FISH) is a technique that allows specific DNA sequences to be detected on metaphase or interphase chromosomes in cell nuclei1
. The technique uses DNA probes with unique sequences that hybridize to whole chromosomes or specific chromosomal regions, and serves as a powerful adjunct to classic cytogenetics. For instance, many earlier studies reported the frequent detection of increased chromosome aberrations in leukemia patients related with benzene exposure, benzene-poisoning patients, and healthy workers exposed to benzene, using classic cytogenetic analysis2
. Using FISH, leukemia-specific chromosomal alterations have been observed to be elevated in apparently healthy workers exposed to benzene3-6
, indicating the critical roles of cytogentic changes in benzene-induced leukemogenesis.
Generally, a single FISH assay examines only one or a few whole chromosomes or specific loci per slide, so multiple hybridizations need to be conducted on multiple slides to cover all of the human chromosomes. Spectral karyotyping (SKY) allows visualization of the whole genome simultaneously, but the requirement for special software and equipment limits its application7
. Here, we describe a novel FISH assay, OctoChrome-FISH, which can be applied for Chromosomics
, which we define here as the simultaneous analysis of all 24 human chromosomes on one slide in human studies, such as chromosome-wide aneuploidy study (CWAS)8
. The basis of the method, marketed by Cytocell as the Chromoprobe Multiprobe System, is an OctoChrome device that is divided into 8 squares, each of which carries three different whole chromosome painting probes (Figure 1). Each of the three probes is directly labeled with a different colored fluorophore, green (FITC), red (Texas Red), and blue (Coumarin). The arrangement of chromosome combinations on the OctoChrome device has been designed to facilitate the identification of the non-random structural chromosome alterations (translocations) found in the most common leukemias and lymphomas, for instance t(9;22), t(15;17), t(8;21), t(14;18)9
. Moreover, numerical changes (aneuploidy) in chromosomes can be detected concurrently. The corresponding template slide is also divided into 8 squares onto which metaphase spreads are bound (Figure 2), and is positioned over the OctoChrome device. The probes and target DNA are denatured at high-temperature and hybridized in a humid chamber, and then all 24 human chromosomes can be visualized simultaneously.
OctoChrome FISH is a promising technique for the clinical diagnosis of leukemia and lymphoma and for detection of aneuploidies in all chromosomes. We have applied this new Chromosomic
approach in a CWAS study of benzene-exposed Chinese workers8,10
Genetics, Issue 60, Chromosomics, OctoChrome-FISH, fluorescence in situ hybridization (FISH), Chromosome-wide aneuploidy study (CWAS), aneuploidy, chromosomal translocations, leukemia, lymphoma
Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung
Institutions: University of Liverpool , University of Liverpool .
There is growing concern about the relevance of in vitro
antimicrobial susceptibility tests when applied to isolates of P. aeruginosa
from cystic fibrosis (CF) patients. Existing methods rely on single or a few isolates grown aerobically and planktonically. Predetermined cut-offs are used to define whether the bacteria are sensitive or resistant to any given antibiotic1
. However, during chronic lung infections in CF, P. aeruginosa
populations exist in biofilms and there is evidence that the environment is largely microaerophilic2
. The stark difference in conditions between bacteria in the lung and those during diagnostic testing has called into question the reliability and even relevance of these tests3
Artificial sputum medium (ASM) is a culture medium containing the components of CF patient sputum, including amino acids, mucin and free DNA. P. aeruginosa
growth in ASM mimics growth during CF infections, with the formation of self-aggregating biofilm structures and population divergence4,5,6
. The aim of this study was to develop a microtitre-plate assay to study antimicrobial susceptibility of P. aeruginosa
based on growth in ASM, which is applicable to both microaerophilic and aerobic conditions.
An ASM assay was developed in a microtitre plate format. P. aeruginosa
biofilms were allowed to develop for 3 days prior to incubation with antimicrobial agents at different concentrations for 24 hours. After biofilm disruption, cell viability was measured by staining with resazurin. This assay was used to ascertain the sessile cell minimum inhibitory concentration (SMIC) of tobramycin for 15 different P. aeruginosa
isolates under aerobic and microaerophilic conditions and SMIC values were compared to those obtained with standard broth growth. Whilst there was some evidence for increased MIC values for isolates grown in ASM when compared to their planktonic counterparts, the biggest differences were found with bacteria tested in microaerophilic conditions, which showed a much increased resistance up to a >128 fold, towards tobramycin in the ASM system when compared to assays carried out in aerobic conditions.
The lack of association between current susceptibility testing methods and clinical outcome has questioned the validity of current methods3
. Several in vitro
models have been used previously to study P. aeruginosa
. However, these methods rely on surface attached biofilms, whereas the ASM biofilms resemble those observed in the CF lung9
. In addition, reduced oxygen concentration in the mucus has been shown to alter the behavior of P. aeruginosa2
and affect antibiotic susceptibility10
. Therefore using ASM under microaerophilic conditions may provide a more realistic environment in which to study antimicrobial susceptibility.
Immunology, Issue 64, Microbiology, Pseudomonas aeruginosa, antimicrobial susceptibility, artificial sputum media, lung infection, cystic fibrosis, diagnostics, plankton
Stress-induced Antibiotic Susceptibility Testing on a Chip
Institutions: Fraunhofer USA Center for Manufacturing Innovation, Harvard Medical School, Boston University, Boston University.
We have developed a rapid microfluidic method for antibiotic susceptibility testing in a stress-based environment. Fluid is passed at high speeds over bacteria immobilized on the bottom of a microfluidic channel. In the presence of stress and antibiotic, susceptible strains of bacteria die rapidly. However, resistant bacteria survive these stressful conditions. The hypothesis behind this method is new: stress activation of biochemical pathways, which are targets of antibiotics, can accelerate antibiotic susceptibility testing. As compared to standard antibiotic susceptibility testing methods, the rate-limiting step - bacterial growth - is omitted during antibiotic application. The technical implementation of the method is in a combination of standard techniques and innovative approaches. The standard parts of the method include bacterial culture protocols, defining microfluidic channels in polydimethylsiloxane (PDMS), cell viability monitoring with fluorescence, and batch image processing for bacteria counting. Innovative parts of the method are in the use of culture media flow for mechanical stress application, use of enzymes to damage but not kill the bacteria, and use of microarray substrates for bacterial attachment. The developed platform can be used in antibiotic and nonantibiotic related drug development and testing. As compared to the standard bacterial suspension experiments, the effect of the drug can be turned on and off repeatedly over controlled time periods. Repetitive observation of the same bacterial population is possible over the course of the same experiment.
Bioengineering, Issue 83, antibiotic, susceptibility, resistance, microfluidics, microscopy, rapid, testing, stress, bacteria, fluorescence
gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair
Institutions: Centre Georges-François Leclerc.
The widespread use of Next Generation Sequencing has opened up new avenues for cancer research and diagnosis. NGS will bring huge amounts of new data on cancer, and especially cancer genetics. Current knowledge and future discoveries will make it necessary to study a huge number of genes that could be involved in a genetic predisposition to cancer. In this regard, we developed a Nextera design to study 11 complete genes involved in DNA damage repair. This protocol was developed to safely study 11 genes (ATM
, and TP53
) from promoter to 3'-UTR in 24 patients simultaneously. This protocol, based on transposase technology and gDNA enrichment, gives a great advantage in terms of time for the genetic diagnosis thanks to sample multiplexing. This protocol can be safely used with blood gDNA.
Genetics, Issue 92, gDNA enrichment, Nextera, NGS, DNA damage, BRCA1, BRCA2
Analysis of Nephron Composition and Function in the Adult Zebrafish Kidney
Institutions: University of Notre Dame.
The zebrafish model has emerged as a relevant system to study kidney development, regeneration and disease. Both the embryonic and adult zebrafish kidneys are composed of functional units known as nephrons, which are highly conserved with other vertebrates, including mammals. Research in zebrafish has recently demonstrated that two distinctive phenomena transpire after adult nephrons incur damage: first, there is robust regeneration within existing nephrons that replaces the destroyed tubule epithelial cells; second, entirely new nephrons are produced from renal progenitors in a process known as neonephrogenesis. In contrast, humans and other mammals seem to have only a limited ability for nephron epithelial regeneration. To date, the mechanisms responsible for these kidney regeneration phenomena remain poorly understood. Since adult zebrafish kidneys undergo both nephron epithelial regeneration and neonephrogenesis, they provide an outstanding experimental paradigm to study these events. Further, there is a wide range of genetic and pharmacological tools available in the zebrafish model that can be used to delineate the cellular and molecular mechanisms that regulate renal regeneration. One essential aspect of such research is the evaluation of nephron structure and function. This protocol describes a set of labeling techniques that can be used to gauge renal composition and test nephron functionality in the adult zebrafish kidney. Thus, these methods are widely applicable to the future phenotypic characterization of adult zebrafish kidney injury paradigms, which include but are not limited to, nephrotoxicant exposure regimes or genetic methods of targeted cell death such as the nitroreductase mediated cell ablation technique. Further, these methods could be used to study genetic perturbations in adult kidney formation and could also be applied to assess renal status during chronic disease modeling.
Cellular Biology, Issue 90,
zebrafish; kidney; nephron; nephrology; renal; regeneration; proximal tubule; distal tubule; segment; mesonephros; physiology; acute kidney injury (AKI)
Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes
Institutions: Charité Medical Faculty and Max-Delbrück Center for Molecular Medicine (MDC), Charité - Universitätsmedizin Berlin, Charité - Universitätsmedizin Berlin.
KCNE genes encode for a small family of Kv channel ancillary subunits that form heteromeric complexes with Kv channel alpha subunits to modify their functional properties. Mutations in KCNE genes have been found in patients with cardiac arrhythmias such as the long QT syndrome and/or atrial fibrillation. However, the precise molecular pathophysiology that leads to these diseases remains elusive. In previous studies the electrophysiological properties of the disease causing mutations in these genes have mostly been studied in heterologous expression systems and we cannot be sure if the reported effects can directly be translated into native cardiomyocytes. In our laboratory we therefore use a different approach. We directly study the effects of KCNE gene deletion in isolated cardiomyocytes from knockout mice by cellular electrophysiology - a unique technique that we describe in this issue of the Journal of Visualized Experiments
. The hearts from genetically engineered KCNE mice are rapidly excised and mounted onto a Langendorff apparatus by aortic cannulation. Free Ca2+
in the myocardium is bound by EGTA, and dissociation of cardiac myocytes is then achieved by retrograde perfusion of the coronary arteries with a specialized low Ca2+
buffer containing collagenase. Atria, free right ventricular wall and the left ventricle can then be separated by microsurgical techniques. Calcium is then slowly added back to isolated cardiomyocytes in a multiple step comprising washing procedure. Atrial and ventricular cardiomyocytes of healthy appearance with no spontaneous contractions are then immediately subjected to electrophysiological analyses by patch clamp technique or other biochemical analyses within the first 6 hours following isolation.
Physiology, Issue 73, Medicine, Cellular Biology, Molecular Biology, Genetics, Biomedical Engineering, Anatomy, Cardiology, Cardiac Output, Low, Cardiomyopathies, Heart Failure, Arrhythmias, Cardiac, Ventricular Dysfunction, Cardiomyocytes, Kv channel, cardiac arrythmia, electrophysiology, patch clamp, mouse, animal model
Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples
Institutions: University of Manitoba, University of Manitoba.
Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo
flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina.
Medicine, Issue 89, mucosal, immunology, FGT, lavage, cervical, CMC
Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies
Institutions: The University of Western Ontario, Instituto de Ciencias de la Vid y el Vino, Ghent University, University of Amsterdam.
The two-spotted spider mite, Tetranychus urticae
, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with more than 150 of economic value. It is a major pest of greenhouse crops, especially in Solanaceae
, tomatoes, eggplants, peppers, cucumbers, zucchini) and greenhouse ornamentals (e.g.
, roses, chrysanthemum, carnations), annual field crops (such as maize, cotton, soybean, and sugar beet), and in perennial cultures (alfalfa, strawberries, grapes, citruses, and plums)1,2
. In addition to the extreme polyphagy that makes it an important agricultural pest, T. urticae
has a tendency to develop resistance to a wide array of insecticides and acaricides that are used for its control3-7
is an excellent experimental organism, as it has a rapid life cycle (7 days at 27 °C) and can be easily maintained at high density in the laboratory. Methods to assay gene expression (including in situ
hybridization and antibody staining) and to inactivate expression of spider mite endogenous genes using RNA interference have been developed8-10
. Recently, the whole genome sequence of T. urticae
has been reported, creating an opportunity to develop this pest herbivore as a model organism with equivalent genomic resources that already exist in some of its host plants (Arabidopsis thaliana
and the tomato Solanum lycopersicum
. Together, these model organisms could provide insights into molecular bases of plant-pest interactions.
Here, an efficient method for quick and easy collection of a large number of adult female mites, their application on an experimental plant host, and the assessment of the plant damage due to spider mite feeding are described. The presented protocol enables fast and efficient collection of hundreds of individuals at any developmental stage (eggs, larvae, nymphs, adult males, and females) that can be used for subsequent experimental application.
Environmental Sciences, Issue 89, two-spotted spider mite, plant-herbivore interaction, Tetranychus urticae, Arabidopsis thaliana, plant damage analysis, herbivory, plant pests
Making Sense of Listening: The IMAP Test Battery
Institutions: MRC Institute of Hearing Research, National Biomedical Research Unit in Hearing.
The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.
Neuroscience, Issue 44, Listening skills, auditory processing, auditory psychophysics, clinical assessment, child-friendly testing
Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays
Institutions: University of Toronto, University of Toronto, University of Regina.
Phenotypes are determined by a complex series of physical (e.g.
protein-protein) and functional (e.g.
gene-gene or genetic) interactions (GI)1
. While physical interactions can indicate which bacterial proteins are associated as complexes, they do not necessarily reveal pathway-level functional relationships1. GI screens, in which the growth of double mutants bearing two deleted or inactivated genes is measured and compared to the corresponding single mutants, can illuminate epistatic dependencies between loci and hence provide a means to query and discover novel functional relationships2
. Large-scale GI maps have been reported for eukaryotic organisms like yeast3-7
, but GI information remains sparse for prokaryotes8
, which hinders the functional annotation of bacterial genomes. To this end, we and others have developed high-throughput quantitative bacterial GI screening methods9, 10
Here, we present the key steps required to perform quantitative E. coli
Synthetic Genetic Array (eSGA) screening procedure on a genome-scale9
, using natural bacterial conjugation and homologous recombination to systemically generate and measure the fitness of large numbers of double mutants in a colony array format.
Briefly, a robot is used to transfer, through conjugation, chloramphenicol (Cm) - marked mutant alleles from engineered Hfr (High frequency of recombination) 'donor strains' into an ordered array of kanamycin (Kan) - marked F- recipient strains. Typically, we use loss-of-function single mutants bearing non-essential gene deletions (e.g.
the 'Keio' collection11
) and essential gene hypomorphic mutations (i.e.
alleles conferring reduced protein expression, stability, or activity9, 12, 13
) to query the functional associations of non-essential and essential genes, respectively. After conjugation and ensuing genetic exchange mediated by homologous recombination, the resulting double mutants are selected on solid medium containing both antibiotics. After outgrowth, the plates are digitally imaged and colony sizes are quantitatively scored using an in-house automated image processing system14
. GIs are revealed when the growth rate of a double mutant is either significantly better or worse than expected9
. Aggravating (or negative) GIs often result between loss-of-function mutations in pairs of genes from compensatory pathways that impinge on the same essential process2
. Here, the loss of a single gene is buffered, such that either single mutant is viable. However, the loss of both pathways is deleterious and results in synthetic lethality or sickness (i.e.
slow growth). Conversely, alleviating (or positive) interactions can occur between genes in the same pathway or protein complex2
as the deletion of either gene alone is often sufficient to perturb the normal function of the pathway or complex such that additional perturbations do not reduce activity, and hence growth, further. Overall, systematically identifying and analyzing GI networks can provide unbiased, global maps of the functional relationships between large numbers of genes, from which pathway-level information missed by other approaches can be inferred9
Genetics, Issue 69, Molecular Biology, Medicine, Biochemistry, Microbiology, Aggravating, alleviating, conjugation, double mutant, Escherichia coli, genetic interaction, Gram-negative bacteria, homologous recombination, network, synthetic lethality or sickness, suppression
Using Eye Movements to Evaluate the Cognitive Processes Involved in Text Comprehension
Institutions: University of Illinois at Chicago.
The present article describes how to use eye tracking methodologies to study the cognitive processes involved in text comprehension. Measuring eye movements during reading is one of the most precise methods for measuring moment-by-moment (online) processing demands during text comprehension. Cognitive processing demands are reflected by several aspects of eye movement behavior, such as fixation duration, number of fixations, and number of regressions (returning to prior parts of a text). Important properties of eye tracking equipment that researchers need to consider are described, including how frequently the eye position is measured (sampling rate), accuracy of determining eye position, how much head movement is allowed, and ease of use. Also described are properties of stimuli that influence eye movements that need to be controlled in studies of text comprehension, such as the position, frequency, and length of target words. Procedural recommendations related to preparing the participant, setting up and calibrating the equipment, and running a study are given. Representative results are presented to illustrate how data can be evaluated. Although the methodology is described in terms of reading comprehension, much of the information presented can be applied to any study in which participants read verbal stimuli.
Behavior, Issue 83, Eye movements, Eye tracking, Text comprehension, Reading, Cognition
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Institutions: Emory University, Emory University.
The protective effect of many HLA class I alleles on HIV-1 pathogenesis and disease progression is, in part, attributed to their ability to target conserved portions of the HIV-1 genome that escape with difficulty. Sequence changes attributed to cellular immune pressure arise across the genome during infection, and if found within conserved regions of the genome such as Gag, can affect the ability of the virus to replicate in vitro
. Transmission of HLA-linked polymorphisms in Gag to HLA-mismatched recipients has been associated with reduced set point viral loads. We hypothesized this may be due to a reduced replication capacity of the virus. Here we present a novel method for assessing the in vitro
replication of HIV-1 as influenced by the gag
gene isolated from acute time points from subtype C infected Zambians. This method uses restriction enzyme based cloning to insert the gag
gene into a common subtype C HIV-1 proviral backbone, MJ4. This makes it more appropriate to the study of subtype C sequences than previous recombination based methods that have assessed the in vitro
replication of chronically derived gag-pro
sequences. Nevertheless, the protocol could be readily modified for studies of viruses from other subtypes. Moreover, this protocol details a robust and reproducible method for assessing the replication capacity of the Gag-MJ4 chimeric viruses on a CEM-based T cell line. This method was utilized for the study of Gag-MJ4 chimeric viruses derived from 149 subtype C acutely infected Zambians, and has allowed for the identification of residues in Gag that affect replication. More importantly, the implementation of this technique has facilitated a deeper understanding of how viral replication defines parameters of early HIV-1 pathogenesis such as set point viral load and longitudinal CD4+ T cell decline.
Infectious Diseases, Issue 90, HIV-1, Gag, viral replication, replication capacity, viral fitness, MJ4, CEM, GXR25
An Allele-specific Gene Expression Assay to Test the Functional Basis of Genetic Associations
Institutions: University of Oxford.
The number of significant genetic associations with common complex traits is constantly increasing. However, most of these associations have not been understood at molecular level. One of the mechanisms mediating the effect of DNA variants on phenotypes is gene expression, which has been shown to be particularly relevant for complex traits1
This method tests in a cellular context the effect of specific DNA sequences on gene expression. The principle is to measure the relative abundance of transcripts arising from the two alleles of a gene, analysing cells which carry one copy of the DNA sequences associated with disease (the risk variants)2,3
. Therefore, the cells used for this method should meet two fundamental genotypic requirements: they have to be heterozygous both for DNA risk variants and for DNA markers, typically coding polymorphisms, which can distinguish transcripts based on their chromosomal origin (Figure 1). DNA risk variants and DNA markers do not need to have the same allele frequency but the phase (haplotypic) relationship of the genetic markers needs to be understood. It is also important to choose cell types which express the gene of interest. This protocol refers specifically to the procedure adopted to extract nucleic acids from fibroblasts but the method is equally applicable to other cells types including primary cells.
DNA and RNA are extracted from the selected cell lines and cDNA is generated. DNA and cDNA are analysed with a primer extension assay, designed to target the coding DNA markers4
. The primer extension assay is carried out using the MassARRAY (Sequenom)5
platform according to the manufacturer's specifications. Primer extension products are then analysed by matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF/MS). Because the selected markers are heterozygous they will generate two peaks on the MS profiles. The area of each peak is proportional to the transcript abundance and can be measured with a function of the MassARRAY Typer software to generate an allelic ratio (allele 1: allele 2) calculation. The allelic ratio obtained for cDNA is normalized using that measured from genomic DNA, where the allelic ratio is expected to be 1:1 to correct for technical artifacts. Markers with a normalised allelic ratio significantly different to 1 indicate that the amount of transcript generated from the two chromosomes in the same cell is different, suggesting that the DNA variants associated with the phenotype have an effect on gene expression. Experimental controls should be used to confirm the results.
Cellular Biology, Issue 45, Gene expression, regulatory variant, haplotype, association study, primer extension, MALDI-TOF mass spectrometry, single nucleotide polymorphism, allele-specific
Perceptual and Category Processing of the Uncanny Valley Hypothesis' Dimension of Human Likeness: Some Methodological Issues
Institutions: University of Zurich.
Mori's Uncanny Valley Hypothesis1,2
proposes that the perception of humanlike characters such as robots and, by extension, avatars (computer-generated characters) can evoke negative or positive affect (valence) depending on the object's degree of visual and behavioral realism along a dimension of human likeness
) (Figure 1
). But studies of affective valence of subjective responses to variously realistic non-human characters have produced inconsistent findings 3, 4, 5, 6
. One of a number of reasons for this is that human likeness is not perceived as the hypothesis assumes. While the DHL can be defined following Mori's description as a smooth linear change in the degree of physical humanlike similarity, subjective perception of objects along the DHL can be understood in terms of the psychological effects of categorical perception (CP) 7
. Further behavioral and neuroimaging investigations of category processing and CP along the DHL and of the potential influence of the dimension's underlying category structure on affective experience are needed. This protocol therefore focuses on the DHL and allows examination of CP. Based on the protocol presented in the video as an example, issues surrounding the methodology in the protocol and the use in "uncanny" research of stimuli drawn from morph continua to represent the DHL are discussed in the article that accompanies the video. The use of neuroimaging and morph stimuli to represent the DHL in order to disentangle brain regions neurally responsive to physical human-like similarity from those responsive to category change and category processing is briefly illustrated.
Behavior, Issue 76, Neuroscience, Neurobiology, Molecular Biology, Psychology, Neuropsychology, uncanny valley, functional magnetic resonance imaging, fMRI, categorical perception, virtual reality, avatar, human likeness, Mori, uncanny valley hypothesis, perception, magnetic resonance imaging, MRI, imaging, clinical techniques
Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study
Institutions: RWTH Aachen University, Fraunhofer Gesellschaft.
Plants provide multiple benefits for the production of biopharmaceuticals including low costs, scalability, and safety. Transient expression offers the additional advantage of short development and production times, but expression levels can vary significantly between batches thus giving rise to regulatory concerns in the context of good manufacturing practice. We used a design of experiments (DoE) approach to determine the impact of major factors such as regulatory elements in the expression construct, plant growth and development parameters, and the incubation conditions during expression, on the variability of expression between batches. We tested plants expressing a model anti-HIV monoclonal antibody (2G12) and a fluorescent marker protein (DsRed). We discuss the rationale for selecting certain properties of the model and identify its potential limitations. The general approach can easily be transferred to other problems because the principles of the model are broadly applicable: knowledge-based parameter selection, complexity reduction by splitting the initial problem into smaller modules, software-guided setup of optimal experiment combinations and step-wise design augmentation. Therefore, the methodology is not only useful for characterizing protein expression in plants but also for the investigation of other complex systems lacking a mechanistic description. The predictive equations describing the interconnectivity between parameters can be used to establish mechanistic models for other complex systems.
Bioengineering, Issue 83, design of experiments (DoE), transient protein expression, plant-derived biopharmaceuticals, promoter, 5'UTR, fluorescent reporter protein, model building, incubation conditions, monoclonal antibody
Cortical Source Analysis of High-Density EEG Recordings in Children
Institutions: UCL Institute of Child Health, University College London.
EEG is traditionally described as a neuroimaging technique with high temporal and low spatial resolution. Recent advances in biophysical modelling and signal processing make it possible to exploit information from other imaging modalities like structural MRI that provide high spatial resolution to overcome this constraint1
. This is especially useful for investigations that require high resolution in the temporal as well as spatial domain. In addition, due to the easy application and low cost of EEG recordings, EEG is often the method of choice when working with populations, such as young children, that do not tolerate functional MRI scans well. However, in order to investigate which neural substrates are involved, anatomical information from structural MRI is still needed. Most EEG analysis packages work with standard head models that are based on adult anatomy. The accuracy of these models when used for children is limited2
, because the composition and spatial configuration of head tissues changes dramatically over development3
In the present paper, we provide an overview of our recent work in utilizing head models based on individual structural MRI scans or age specific head models to reconstruct the cortical generators of high density EEG. This article describes how EEG recordings are acquired, processed, and analyzed with pediatric populations at the London Baby Lab, including laboratory setup, task design, EEG preprocessing, MRI processing, and EEG channel level and source analysis.
Behavior, Issue 88, EEG, electroencephalogram, development, source analysis, pediatric, minimum-norm estimation, cognitive neuroscience, event-related potentials
Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies
Institutions: Flemish Institute for Technological Research (VITO), Hasselt University, Hasselt University, Leuven University.
The microcirculation consists of blood vessels with diameters less than 150 µm. It makes up a large part of the circulatory system and plays an important role in maintaining cardiovascular health. The retina is a tissue that lines the interior of the eye and it is the only tissue that allows for a non-invasive analysis of the microvasculature. Nowadays, high-quality fundus images can be acquired using digital cameras. Retinal images can be collected in 5 min or less, even without dilatation of the pupils. This unobtrusive and fast procedure for visualizing the microcirculation is attractive to apply in epidemiological studies and to monitor cardiovascular health from early age up to old age.
Systemic diseases that affect the circulation can result in progressive morphological changes in the retinal vasculature. For example, changes in the vessel calibers of retinal arteries and veins have been associated with hypertension, atherosclerosis, and increased risk of stroke and myocardial infarction. The vessel widths are derived using image analysis software and the width of the six largest arteries and veins are summarized in the Central Retinal Arteriolar Equivalent (CRAE) and the Central Retinal Venular Equivalent (CRVE). The latter features have been shown useful to study the impact of modifiable lifestyle and environmental cardiovascular disease risk factors.
The procedures to acquire fundus images and the analysis steps to obtain CRAE and CRVE are described. Coefficients of variation of repeated measures of CRAE and CRVE are less than 2% and within-rater reliability is very high. Using a panel study, the rapid response of the retinal vessel calibers to short-term changes in particulate air pollution, a known risk factor for cardiovascular mortality and morbidity, is reported. In conclusion, retinal imaging is proposed as a convenient and instrumental tool for epidemiological studies to study microvascular responses to cardiovascular disease risk factors.
Medicine, Issue 92, retina, microvasculature, image analysis, Central Retinal Arteriolar Equivalent, Central Retinal Venular Equivalent, air pollution, particulate matter, black carbon
Oscillation and Reaction Board Techniques for Estimating Inertial Properties of a Below-knee Prosthesis
Institutions: University of Northern Colorado, Arizona State University, Iowa State University.
The purpose of this study was two-fold: 1) demonstrate a technique that can be used to directly estimate the inertial properties of a below-knee prosthesis, and 2) contrast the effects of the proposed technique and that of using intact limb inertial properties on joint kinetic estimates during walking in unilateral, transtibial amputees. An oscillation and reaction board system was validated and shown to be reliable when measuring inertial properties of known geometrical solids. When direct measurements of inertial properties of the prosthesis were used in inverse dynamics modeling of the lower extremity compared with inertial estimates based on an intact shank and foot, joint kinetics at the hip and knee were significantly lower during the swing phase of walking. Differences in joint kinetics during stance, however, were smaller than those observed during swing. Therefore, researchers focusing on the swing phase of walking should consider the impact of prosthesis inertia property estimates on study outcomes. For stance, either one of the two inertial models investigated in our study would likely lead to similar outcomes with an inverse dynamics assessment.
Bioengineering, Issue 87, prosthesis inertia, amputee locomotion, below-knee prosthesis, transtibial amputee
Methods to Explore the Influence of Top-down Visual Processes on Motor Behavior
Institutions: Rutgers University, Rutgers University, Rutgers University, Rutgers University, Rutgers University.
Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are deliberately planned, while others spontaneously occur below conscious awareness. The deliberate component of this dichotomy has been studied extensively in several contexts, while the spontaneous component remains largely under-explored. Moreover, how perceptual processes modulate these movement classes is still unclear. In particular, a currently debated issue is whether the visuomotor system is governed by the spatial percept produced by a visual illusion or whether it is not affected by the illusion and is governed instead by the veridical percept. Bistable percepts such as 3D depth inversion illusions (DIIs) provide an excellent context to study such interactions and balance, particularly when used in combination with reach-to-grasp movements. In this study, a methodology is developed that uses a DII to clarify the role of top-down processes on motor action, particularly exploring how reaches toward a target on a DII are affected in both deliberate and spontaneous movement domains.
Behavior, Issue 86, vision for action, vision for perception, motor control, reach, grasp, visuomotor, ventral stream, dorsal stream, illusion, space perception, depth inversion
Cross-Modal Multivariate Pattern Analysis
Institutions: University of Southern California.
Multivariate pattern analysis (MVPA) is an increasingly popular method of analyzing functional magnetic resonance imaging (fMRI) data1-4
. Typically, the method is used to identify a subject's perceptual experience from neural activity in certain regions of the brain. For instance, it has been employed to predict the orientation of visual gratings a subject perceives from activity in early visual cortices5
or, analogously, the content of speech from activity in early auditory cortices6
Here, we present an extension of the classical MVPA paradigm, according to which perceptual stimuli are not predicted within, but across sensory systems. Specifically, the method we describe addresses the question of whether stimuli that evoke memory associations in modalities other than the one through which they are presented induce content-specific activity patterns in the sensory cortices of those other modalities. For instance, seeing a muted video clip of a glass vase shattering on the ground automatically triggers in most observers an auditory image of the associated sound; is the experience of this image in the "mind's ear" correlated with a specific neural activity pattern in early auditory cortices? Furthermore, is this activity pattern distinct from the pattern that could be observed if the subject were, instead, watching a video clip of a howling dog?
In two previous studies7,8
, we were able to predict sound- and touch-implying video clips based on neural activity in early auditory and somatosensory cortices, respectively. Our results are in line with a neuroarchitectural framework proposed by Damasio9,10
, according to which the experience of mental images that are based on memories - such as hearing the shattering sound of a vase in the "mind's ear" upon seeing the corresponding video clip - is supported by the re-construction of content-specific neural activity patterns in early sensory cortices.
Neuroscience, Issue 57, perception, sensory, cross-modal, top-down, mental imagery, fMRI, MRI, neuroimaging, multivariate pattern analysis, MVPA
Freezing, Thawing, and Packaging Cells for Transport
Institutions: Molecular Pathology Laboratory Network, Inc.
Cultured mammalian cells are used extensively in cell biology studies. It requires a number of special skills in order to be able to preserve the structure, function, behavior, and biology of the cells in culture. This video describes the basic skills required to freeze and store cells and how to recover frozen stocks.
Basic Protocols, Issue 17, Current Protocols Wiley, Freezing Cells, Cell Culture, Thawing Cells, Storage of Cells, Suspension Cells, Adherent Cells
Pyrosequencing: A Simple Method for Accurate Genotyping
Institutions: Washington University in St. Louis.
Pharmacogenetic research benefits first-hand from the abundance of information provided by the completion of the Human Genome Project. With such a tremendous amount of data available comes an explosion of genotyping methods. Pyrosequencing(R) is one of the most thorough yet simple methods to date used to analyze polymorphisms. It also has the ability to identify tri-allelic, indels, short-repeat polymorphisms, along with determining allele percentages for methylation or pooled sample assessment. In addition, there is a standardized control sequence that provides internal quality control. This method has led to rapid and efficient single-nucleotide polymorphism evaluation including many clinically relevant polymorphisms. The technique and methodology of Pyrosequencing is explained.
Cellular Biology, Issue 11, Springer Protocols, Pyrosequencing, genotype, polymorphism, SNP, pharmacogenetics, pharmacogenomics, PCR

domain: biology


text:
There was Histrionic Laughter at the Clown's Cadaver by N. Alexsander Sidirov
Oscillating between lyrical poetry, dramatic, confessional, and abstract the only thing that seems consistent is the amount of surprise experienced every page. Calling this book a modern poetry collection feels somewhat of a misnomer considering the novel and avant-garde imagery, its ominous and often omnipresent eye, and ultimately the transcendental climax that is difficult to compare to other works in the genre. Each poem in this book has a specific place along an emotional odyssey that blisters with novelty and an almost lynchian flare for the surreal and absurd. Whether you ultimately enjoy his words you will decide, but one certainly can never forget them. This is the matter that dreams are made of...
Worldbuilding: Piqued curiosity
Plot: Mostly clear
Characters: Roles are clear
Immersion: Satisfying, fulfilling experience
Emotional Response: Engaging
Thought Provoking: New ideas came up
Cover: Matches the story well
This is a unique and imaginative collection of poetry and prose in a variety of original styles.
Submitted by rebeccacrunden on

domain: literature


- Studebaker Champion '48
- Studebaker Erskin Six '27
- Jaguar XJS Coupé 5.3 V12 HE White '85
- Vespa PX125E '83
- Ford Mustang Convertible V8 '73
- Harley Davidson Servi-Car '46
- MG A 1600 Roadster Red '60
- MG B Roadster '66
- Chevrolet Silverado Stepside '92
- Citroën Traction Avant Cabriolet (réplique) '34
- Abarth 695 Tributo Ferrari 2010
- Citroën 2CV 6 Special '87
- Porsche 356 BT5 Super 90 '61
- Mini 1000 Cooper '69
- Mercedes-Benz 220SE Ponton Cabriolet '60
- Porsche 911 3.2 Carrera Coupé '86
- Austin Healey 100/4 BN1 '53
- Jaguar E-Type OTS 4.2 1st series '66
- Mini Monte Carlo '94
- DKW F93 '57
- Oldsmobile Super 88 Green & White '55
- Range Rover Convertible Sand '85
- Willys Jeep MB43 '43
- Fiat 850 Spider '71
- Ferrari 348 Spider US Cabriolet Red '94
- Porsche 911 3.2 WTL Cabriolet '85
- Chevrolet Camaro SS Look '69
- Mercedes-Benz 220A Cabriolet '53
- Alvis TA21 Drop Head Coupé '53
- Cadillac 61 Series Touring Sedan '49
- Porsche 911 3.2 Carrera Grand Prix White '86
- Citroën SM '72
- Ford Mustang Hardtop Coupé 289ci V8 '66
- Lincoln Continental Mark VI '80
- Porsche 911 3.2 G50 '87
- Ferrari 365 GT4 2+2 Blue '75
- Nash Metropolitan '54
- Jaguar XJ6 3.4 Series 2 '77
- BMW R 1200 GS Adventure 2012
- Sima Violet 500 '24
- Corvette C2 Stingray '66
- Jaguar D-Type Replica by RAM '69
- Peugeot 203 '52
- Triumph GT6 MKII '68
- Porsche 928 S4 White '91
- Austin Healey 3000 MK1 BT7 '61
- Fiat 500 White '69
- Rolls Royce Corniche I Convertible '73
- DeLorean DMC 12 '81
- Mercedes-Benz 280SL Pagode '70
Ford V8 5 Window coupé
V8 5 Window coupé
In late 1929, Henry Ford, considering the production of his own V8 engine, instructed engineer Fred Thoms to acquire as many available eight-cylinder engines as possible. At that time, most of the eight-cylinder engines were found in high-end automobiles such as LaSalle, Cadillac and, acquired by Ford in 1922, Lincoln. Ford intended to study the design of the engines and use his research to develop a cost effective V8 that could be mass-produced.
From first impression, the 1932 Ford may have seemed like a warmed-over Model A, but upon closer inspection it became apparent that this was a new design from the grille to the rear bumper. With a lower stance and tasteful sleek lines, the new V8 Model 18 was undoubtedly the best looking car Ford had produced to date. With the promise of performance, combined with styling cues reminiscent of the luxurious Lincoln, and all at a cost that reflected a car more common, the 1932 Ford was a hit with the public.
It wasn’t long after the 1932 Ford’s debut that extensive problems with the Flathead V8 became apparent. Overheating and block cracking were common. The new Ford Flathead V8 had a reputation for consuming oil, and nearly every component of the engine experienced some failure or issue. Of the first 2,000 engines produced, almost all of them needed to have their camshafts and valve components replaced. Swamped with Flathead V8 failure issues, Ford began installing the readily available, refined 4-cylinder Model B engine into many of the early 1932 cars. Perhaps the result of bringing the engine into production without adequate testing and development, the issues with the Flathead V8 would unfortunately have to be solved by way of customer complaint.
Despite Ford’s initial growing pains with the new engine, the production of an affordable single cast Flathead V8 engine remains one his most significant achievements, and would create ripples throughout the auto industry in the years that followed. In time, the Ford Flathead V8 would become a favorite with hot rodders and to this day, the 1932 Ford, or “Deuce,” in all its forms, from full fendered five window coupe to unhaltered highboy, is considered by many to be the definitive hot rod.
Wheelbase : 269 cm (106 in). front-mounted engine : V8 3622 cc (221 ci), 65 bhp at 3400 rpm.
<<<<<< For more pictures scroll down! >>>>>>
Oldtimerfarm is going to renovate!
Due to renovations we are currently publishing less pictures per car on our website.
For more pictures or more information please contact Xavier via phone: 0472/401338
or via e-mail: firstname.lastname@example.org
Oldtimerfarm specializes in consignment sales of vintage and collection cars and we are proud to present you this car. Oldtimerfarm is located in Belgium, 9880 Aalter, Steenweg op Deinze 51C, where this car is in our showroom. We are open from Tuesday to Saturday 10-17h (also without appointment). We are closed on Sundays and Mondays. Mondays on appointment only. Make sure you scroll down to look at the extensive photo report (100 pictures). Of course, a more detailed description can be obtained by telephone.
Oldtimerfarm from Aalter would also like to sell your classic car or car collection. For more information, please contact Xavier.
Exhibitions where you can find us:
|26-29/10/2017||Auto Moto Depoca Padova - Italy|
|17-19/11/2017||InterClassics Brussel - Belgium|
|16-17/12/2017||Dreamcar International Kortrijk - Belgium|
|11-14/01/2018||InterClassics Maastricht - Holland|
Pictures of the car:


text:
My New York City Bucket List
I visited NYC for the first time almost 15 years ago. During that first visit I instantly fell in love with the city. The energy, the busyness, the buildings, the character, I love it all. Over the years I’ve got to experience and see so much of NYC. We used to plan trips to New York when we lived in other States far away. Then 3.5 years ago we moved to New Jersey and now we go to New York City all the time. So I decided to start a “New York City bucket list”. A list of all the places I want to see, the things I want to experience, and the places I want to eat in NYC. Some of the things I’ve already done so I can cross them off, but I still have quite a long list of things to do! This will give me a lot to blog about, both past experiences in addition to future ones! Some of the things I can cross of my list are things like, watching the ball drop in Times Square on New Years Eve. We did that years ago, but I’ve never written a post about it, or shared any photos. So I will have a lot of experiences to share with you! I’m looking forward to crossing more things off my NYC bucket list, in addition to sharing future experiences with you.
I’ve added a “Bucket List” tab to my home page, so you can keep checking back! I’m sure the list will keep growing!
Here are just a couple photos of me in NYC over the years.

domain: hobby


text:
Helen Thomas, Mearsheimer, Walt To Address ‘Move Over Aipac’ Conference
Helen Thomas, Stephen Walt and John Mearsheimer are scheduled to speak at a pro-Palestinian conference aiming to counter AIPAC’s annual gathering.
A coalition of about 100 leftist and pro-Palestinian groups, led by Code Pink and the U.S. Campaign to End the Israeli Occupation, will run “Move Over AIPAC” in Washington from May 21 to 24, coinciding with the American Israel Public Affairs Committee’s annual conference.
The thrust of the “Move Over AIPAC” event will be “to learn about the extraordinary influence AIPAC has on U.S. policy and how to strengthen an alternative that respects the rights of all people in the region,” according to its statement.
The conference will include protests outside the Washington Convention Center, the site of the AIPAC forum.
Thomas, a veteran Washington journalist, lost her column with the Hearst Corp. last year after she said Israelis “should get the hell out of Palestine” and “go home” to Poland, Germany and the United States. She has since said she accepts the “inevitability” of Israel,” but also told Playboy in an interview that “Israeli lobbies” have “total control” of Washington and the financial markets.
Walt and Mearsheimer, scholars who wrote the 2006 book “The Israel Lobby,” will deliver the keynote address.

domain: news


Arch Enemy, The Eagle Flies Alone - QOTSA fragrance - new Hollywood Undead - new...
6 years ago
Arch Enemy, release 2nd single The Eagle Flies Alone - Queen of The Stone Age fragrance satire - new Hollywood Undead song - new Belphegor video - new ...
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No time for breakfast? Hungry after a surf, swim or run? We created the superfood power pack with you in mind. Our premium low fat organic yoghurt is blended with antioxidant rich acai, fibre and protein from linseed, and flavoured with organic fruits such as raspberry or blueberry, or organic honey. Packaged in a durable squeezy pack designed to fit comfortably in your hand and withstand being thrown around in your sports bag, each pack contains a good amount (10g) of protein making it the perfect on the go meal or after sports snack
Organic low fat drinking yogurt with banana, acai and linseed in a Power Pak. An enterprising combination of low fat organic yogurt, acai and linseed for people on the move. Smash a Power Pak as you create your day.
Organic yogurt [organic low fat milk, organic raw sugar, milk solids], banana mix [water, organic banana puree (3.0%), organic sugar, acai puree (0.5%), linseed (0.5%), corn starch, vegetable gum (xanthan), food acid (malic acid)], live cultures (includes probiotics: Acidophilus and Bifidus).
Per Serving size: 200g, Fat: 1.9g, Protein: 5g, Total Carbohydrates: 12.7g, Energy(kj): 356, Protein(g/mg): 5g, Calcium(g/mg): 107mg, Fat(g/mg): 1.9g, servingtype: 100g, Saturated Fat(g/mg): 1.1g, Sodium(g/mg): 58mg, Total Carbohydrate(g/mg): 12.7g, sugar: 12.1g
About the company
Founded by surfer, long time yogi and successful entrepreneur David Prior, five:am is proudly Australian owned and operated. David has realised his vision to take organic yoghurt from a health food niche into the mainstream making it more affordable and accessible. All the milk used in five:am’s tasty yoghurt range is sourced from The Organic Dairy Farmer’s Cooperative in Gippsland, Victoria, a group of farms dedicated to upholding the highest standards of sustainable and organic farming, and then just up the road to the bustling home of five:am in Carrum Downs, southeast of Melbourne, where the team has refined its yoghurteering technique in order to create the best-tasting yoghurt around. And that’s just what you’ll get – all organic, no additives and always delicious.
Company: Five Am Life
Brand: Five Am
Slogan: five:am wake up, be amazing
Category: Drinking Yogurt/Fermented Beverages
Packaging: 200g Plastic Pouch
Claims: Australian certified organic. Low fat. With protein.
Variants: Blueberry, Raspberry, Honey, Banana
Where to Buy: Coming Soon

domain: health


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You’ve possibly heard that Multivitamins are essential for your health. But are they honestly important? How about Folate, Calcium citrate, or Iron? Let’s find out. This article will solve your questions and get you to the right music. A precise multivitamin can supply your body with the vitamins it wishes. It ought to also incorporate a wide form of other nutrients and minerals, consisting of calcium and iron.
While many multivitamins include important nutrients and minerals, you could not want them as multi-functional pills. Some multivitamins include excessive quantities of positive nutrients. Minerals, together with iron, manganese, copper, selenium, and molybdenum, also are risky whilst eaten up in extra. Talk to your medical doctor before beginning a multivitamin application. If you are not positive about whether or now not you must take a multivitamin, ask your health practitioner or nutritionist for a recommendation.
In addition to stopping birth defects, multivitamins can assist women in pregnancy and lactation meets their nutrient wishes. While multivitamins may additionally look like a complicated supplement, a few people are more likely to take them throughout meals for preserving men’s health, and for that, you can soak up Vidalista 40 Or Vidalista 80 mg. To make multivitamins simpler to swallow; split the dose into distinctive instances of the day. You can also take two tablets at once, instead of one.
Although multivitamins can assist prevent iron deficiency, they’re no substitute for a balanced weight loss plan that includes important nutrients and minerals. Iron is a mineral that is vital for the body’s boom and improvement, and a deficiency can lead to fatigue, lethargy, and a standard feeling of malaise. Many multivitamins no longer contain enough iron to provide an each-day endorsed allowance.
The maximum commonplace group of people who are iron-poor is ladies without menopause, ladies who are pregnant, and kids. Men are least possible to be iron-poor, although the CDC estimates that nearly 1-0.33 of guys have an extra of this mineral.
Men seldom require supplemental iron, and maximum multivitamins for older guys no longer include a whole lot of iron at all. Iron-unfastened is a popular multivitamin, but, can also comprise iron. An examination through Consumer Lab observed that less than 20% of ladies’ multivitamins did not include any iron.
Among the essential nutrients, folate is important to the wholesome development of the worried system. This diet is typically located in multivitamins, prenatal vitamins, and B-complicated dietary supplements. Folate is also available as methyl folate (five-methyl-THF) in nutritional dietary supplements. Those with the MTHFR gene mutation may additionally gain from methyl folate dietary supplements as opposed to folic acid.
Folate is a certainly occurring nutrition B9 that enables the frame to create new cells. It is observed certainly in lots of foods, mainly oranges, and darkish green leafy veggies. Folate is likewise determined in red meat liver, nuts, kidney beans, and black-eyed peas.
Folate is crucial for the improvement of pink blood cells, and it’s miles especially crucial at some stages of rapid growth. Folate is found in sufficient quantities in meals inclusive of eggs, meat, and dairy products.
There are numerous motives to take a calcium complement. People with digestive problems can also need to don’t forget a calcium citrate product. In addition to being more difficult to absorb, calcium citrate doesn’t require belly acid to dissolve.
Taking calcium citrate is considered safe for most people when taken in quantities under 2,500 milligrams day by day. Calcium citrate may be purchased over the counter and is typically advocated in small doses of 500 milligrams a day. It is to be had as a nutritional supplement and may be taken on an empty stomach or a full one.
Although calcium citrate is taken into consideration as secure for healthful human beings, it can interact with a few medications. Some medicinal drugs, including digoxin and fluoroquinolones, lower the absorption of calcium.
Calcium may additionally intrude with the absorption of a few nutrients and minerals. Those who are taking calcium citrate must contact their healthcare provider before changing their dose. Taking calcium citrate with positive drugs can increase the risk of hyperkalemia and different negative outcomes.
Folate optimizes bioavailability
Folate is an important nutrient for ultimate health, and a first-rate multivitamin containing sufficient folate can help to maintain the most effective ranges of this nutrition. However, the bioavailability of folate inside the human frame depends on numerous factors, along with genetic polymorphisms. Genetic polymorphisms may affect one-of-a-kind parameters in exclusive individuals, and thus, the recommended dietary allowance (RDA) for folate is a good deal higher for pregnant and lactating girls.
Gummy vitamins are extremely good for folks who locate it difficult to swallow tablets or drugs. They are also a fun opportunity to take pills. Children and adults alike can revel in gummy nutrients. Gummies may help human beings take into account to take their vitamins after they’d otherwise forget. They’re also excellent for kids, as they can be without difficulty absorbed and fed on. But the trouble is that gummy vitamins may be too sweet and contain too little of the essential vitamins.
A gummy diet is less nutritious than different multivitamins. It includes fewer nutrients than drugs, but it’s miles less difficult for the stomach to absorb. It’s critical to study the label to make certain that the product incorporates the vitamins you need.
Depending on the producer, gummy vitamins may additionally comprise much less of an expansion of nutrients than their tablet counterparts. There are a few blessings and disadvantages to gummy vitamins, so make sure to study the labels carefully.
Whole Source Multivitamin is a premium, fairly bioavailable multivitamin complement. It combines 14 fruits with 23 vital vitamins and minerals to support the whole, foremost health, and for more, you could additionally have Arrowmeds Treatment. It is sourced from herbal resources and is unfastened from artificial colors, flavors, and preservatives.
Read More: Mangoes are a healthy and nutritious fruit.

domain: medical


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“TRUCKS ZOOMING BY”
Author Pamela Jane has written more than 30 children’s books, two memoirs and is the coauthor of “Pride and Prejudice and Kitties: a Cat Lover’a romp through Jane Austen’s Classic”.Her essays have appeared in The Wall Street Journal, Writer’s Digest, and The Huffington Post. She lives in PA.
About "TRUCKS ZOOMING BY"
A young girl dreams of being a truck driver one day―just like her mom and dad. From sunup to sundown, they drive over city streets, busy highways, and country roads. On one special day, she joins them and gets to see all different kinds of trucks zooming by!


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Collision coverage has a deductible, which is the amount you pay before your coverage helps pay for your claim. You can typically choose the amount of your deductible when you buy coverage. So, if you choose a $1,000 deductible and your car is later damaged in a covered accident, you'd have to pay $1,000 toward repair costs. Your collision coverage would help pay the rest, up to your coverage limit.
The key difference in collision vs. comprehensive coverage is that, to a certain extent, the element of the car driver's control. As we have stated before, collision insurance will typically cover events within a motorist's control, or when another vehicle collides with your car. Comprehensive coverage generally falls under "acts of God or nature," that are typically out of your control when driving. These can include such events as a spooked deer, a heavy hailstorm, or a carjacking.
This coverage reimburses you for the cost of your rental car if your insured vehicle is in the shop or is unavailable due to an accident. You need to have comprehensive and collision on your policy in order to add rental car coverage. In certain states, Esurance offers CarMatch Rental Coverage®, which covers the rental cost of a vehicle comparable in size and body type to your regular ride.
The main difference between collision and comprehensive coverage comes down to the question of what the driver controls. Collision insurance will cover events within a motorist's control or when another vehicle collides with your car. Comprehensive coverage generally falls under "acts of God or nature," or things that are typically out of your control when driving. These can include events such as a spooked deer, a heavy hailstorm, or a carjacking.
Drive Other Car insurance is similar to an Individual Named Insured Endorsement. If you need to borrow, test drive, rent, or lease a vehicle, Drive Other Car insurance will extend the coverages you’ve purchased for your commercial auto insurance policy - like Liability insurance, Physical Damage insurance, Medical Payments, and Uninsured Motorist Insurance, to a non-owned car.
Insurers don't determine your actual cash value (ACV) settlement based on what you owe, but rather on what the car is worth just prior to the accident. Let's say you owe $20,000 on your new car, but it's only worth about $16,000. If your car is totaled, you might get a settlement check of $16,000 but still owe an additional $4,000 on your loan or lease.
Collision coverage is probably the most important coverage you need to have in order to protect your vehicle against physical damage. It is not difficult to accidentally hit something when driving. Somebody is always at fault, and that someone could be you. Some of the most significant damage to your vehicle can come from a collision with another vehicle, tree, pole or guardrail. In order to purchase collision coverage, you’ll need to purchase basic coverage as well. The higher your deductible (the amount you pay if you do get into a collision), the lower your monthly payments will often be — and this can be the best way to get the coverage you need and the savings you deserve at the same time.
We know you are busy and the last thing you want to do is spend your precious time getting a car insurance quote. That is why we have made getting an affordable car insurance quote quick and simple. In as little as 3 minutes, you can get a car insurance quote from SafeAuto with coverages that are right for you - for the right price. We offer Bodily Injury and Property Damage for liability only insurance shoppers, as well as optional coverages such as Comprehensive, Collision, Medical Payments and much more. We offer 24/7 customer and sales support for all our potential and current customers so we are there when you need us. Another perk of SafeAuto insurance – our convenient payment options can make your monthly price more affordable and your car insurance can be even cheaper with SafeAuto discounts such as Multi-car and Senior Driver Improvement Course discounts.

domain: finance


The European Space Agency (ESA) commemorated the 20th anniversary of the Mars Express spacecraft on Friday by displaying images sent by the spacecraft approximately every 50 seconds.
Given the relative positions of Earth and Mars yesterday, it took approximately 17 minutes for signals from the spacecraft to reach our planet.
This meant that the live broadcast that occurred on Friday stretched the definition of the term “live” somewhat. Nonetheless, this is the first time that near-instant images from Mars have been shown to the public.
The European Space Agency broadcast the first Mars “live” transmission.
“Oh, here it is! This is the first image from Mars, and it’s the closest you’ll ever get to the red planet unless you voyage there yourself. “This is Mars 16 minutes ago,” a commentator on ESA’s live stream said as the first images appeared shortly after the live stream began on June 2 at 9.15 PM IST.
Mars Express collects the majority of its observations when it is not in direct communication with Earth. This typically occurs for one of two reasons: either the spacecraft is on the opposite side of the Sun or Mars, or its antennas are pointed away from Earth when it is conducting observations. The recorded version of the live stream is available below.
Every couple of days, the Visual Monitoring Camera on the Mars Express takes images of the planet and transmits them to Earth, where they are processed by ESA and made public.
This system is ideally suited for scientific missions, as astronomers can examine the captured data days later. However, transmitting live images back to the planet is a difficult and technically remarkable endeavor.
This is why there have only been a few instances of obtaining live images from outer space throughout history. The breathtaking live videos transmitted by Apollo missions are excellent examples. And more recently, the DART mission where NASA crashed a spacecraft into the asteroid Dimorphos is also an excellent example.


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♦ 11) PD Universal LCD Screen Protectors
♦ 12) ImageRecall Digital Image Recovery Software
Capture your circle of relatives’s precious moments with the Canon PowerShot SX420 IS Wi-Fi Digital Camera. With a powerful 42x Optical Zoom (24-1008mm) and 24mm wide-angle lens, the SX420 IS is primed to capture the shots you want, and can easily share them with the addition of built-in Wi-Fi and NFC connectivity.
Take more high-resolution pictures faster (as much as 95MB/sec.) with this Transcend 64GB SDXC UHS-I Class 10 U3 V30 Memory Card.
Increase your shooting capacity with this powerful NB-11L/NB-11LH Lithium-ion rechargeable battery.
This Rapid Multi-voltage Charger plugs immediately in for your wall outlet or your car’s 12V outlet, and charges your battery in approximately 60 to 90 minutes.
This padded carrying case features a zippered main compartment with padded adjustable divider, plus a mesh pocket to safely hold your equipment and accessories.
The Precision Design SD & MicroSD SDHC / SDXC Memory Card Reader transfers photos and videos out of your SD, SDHC, SDXC, microSD, microSDHC, microSDXC, Memory Stick or Memory Stick Micro (M2) memory card for your computer quickly and easily.
This 5-piece cleaning kit includes Lens Cleaning Tissues, Lens Cloth, Lens Cleaning Liquid, Blower Brush and Cotton Swabs.
Make certain this fits by entering your model number.
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 Canon PowerShot SX420 IS Wi-Fi Digital Camera (Black) +  Go beyond 64GB SDXC U3 Card +  Spare NB-11L/H Battery +
 Charger for Canon NB-11L/H +  PD-C10 Camera / Camcorder Case +  PD-T14 Flexible Camera Tripod +
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Mr & Mrs Herd, Greater Manchester
After two years of other agents failing to sell our home, we had at last found the right team. You certainly put all other agents in the shade.
It is important to note that this mortgage calculator is for guidance purposes only. Mortgage lenders have their own affordability criteria therefore it is important you speak to one of our Mortgage Advisers for further information and a more accurate indication of what products may be available to you and what your monthly mortgage payments are likely to be.
This fantastic 3 bedroom property is immaculately presented throughout and been maintained to a high standard including a recently fitted kitchen, situated at the end of a very quiet cul de sac with open views to the front, it's ideal for a growing family, comprising of living room, dining room, cloak room, modern kitchen and conservatory to the ground floor, 2 double bedrooms and single, en suite to the master bedroom and modern 3 piece shower room, externally the property boasts driveway which is gated halfway, detached garage with a well presented rear garden including decking with a fitted canopy, viewing is highly recommended
Contact branch for relevant Energy Performance Certificate


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Some of the nations most celebrated writers, actors and comedians have created monologues for a range of statues across London.
Streets on the development in are meant to be named after British butterflies.
Abseiling technicians are scrubbing each of the four faces of Parliament's Great Clock, checking them for damage.
8,000 people are getting together to meet their online video idols.
The UK's biggest YouTube festival, Summer in the City, is being held at Alexandra Palace.
The event was set up by vloggers and allows those YouTube stars - many of whom are hugely popular - the opportunity to perform in a live setting, often in front of the biggest audience of their lives. The fans can watch their favourite musicians, comedians, video bloggers and filmmakers on the main stage, but also meet them face to face to get a selfie or an autograph.
– Tom Burns, Summer in the City founder
Summer in the City is an annual event that brings YouTube stars from across the world together with content creators, fans and industry personnel for the UK's largest celebration of all things YouTube. The weekend features live performances, talks and many opportunities for fans and content creators to connect with each other and meet their favourite YouTubers.


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I am super excited to say that my hubby and I just purchased a house. It’s a 1958 home on a quiet street that has never ever been updated. I’m pretty sure the shag carpet is original along with the wood panelling and pink bath tub! We wanted a project and that is exactly what we got and I am really looking forward to renovating this house. It definitely has some design challenges and I look forward to sharing my solutions with you.
I only have the one picture of the front of the house right now. It could definitely use some curb appeal but that big tree in the front is a Magnolia. 🙂
The house is not officially ours until the beginning of January but we have already started the ball rolling on the renovation. We are on a tight timeline as we will live in our current home until the renovation is done. We have submitted a form through our realtor that allows us to have access to the house if we need it so our builder and engineer can get in and we can have professional floor plans done up to submit for permits. The permit process in Victoria varies depending on the extensiveness of the work. Our permits should take about 3 weeks (fingers crossed) so the sooner we get them in the better if we want to get the ball rolling come the beginning of January.
Our plan is to make the living room the master bedroom and remove the existing fire place, which will give us more room in the basement to build a one bedroom suite. It will also give us room for a walk in closet and ensuite in the master. Switch the front entry closet to the bedroom and and use the front bedroom closet as a linen closet and then add a front entryway closet on the right. There was an addition built that we have to tear down and rebuild as it was not built properly and without a permit. This will turn into the kitchen and dining room and will flow into a great room. We will extend the patio off the dining room to the full length of the house. The main bathroom is really small and will need to be updated with a new tub/shower, toilet and I am going to do a pedestal sink and move it over slightly to give more room across from the tub/shower.
I think one of my favourite parts of my job is taking an existing floor plan an making it more functional weather it’s for my own renovation or a clients. Then of course making everything beautiful! 🙂
One thing I can’t stress more when starting a renovation is planning. Things happen quickly and it’s always good to be prepared. Use pinterest or Houzz and start your renovation boards and decide what you are going to do in each room. Check out my Pinterest renovation board HERE or Houzz board HERE
When you put together a budget and be prepared to to spend at least 10 – 15 % more. (unless you are super lucky and things all go as planned) Research research. research!!! Do your homework and compare prices and purchase items if they go on sale like appliances and flooring. The big ones! Planning is the key to a successful stress free renovation. Be prepared for some surprises and making some changes along the way as these things happen.
I could go on and on and on for days about starting a renovation but these are just a few steps you need to take. Hire a good contractor, have all the testing, engineering and drawings done up and make a plan! Right now we are in the planning stages, and I have ideas for each room and will share those as we go along.
Hope you have a great weekend!
If you would like some help making your home a place you love, contact me to book an In-Home or Online Consultation.
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domain: design


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Find out how to create simple CSS animations such as rollover buttons, 3D transforms, animated graphics and logos, load screens, and more.
- Intermediate (2)
Learn how to create typography that's beautiful, functional, and easily scalable across tablets, phones, and computers of all types and sizes.
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