text:
The interaction of IgE with its high-affinity Fc receptor (FcεRI) followed by an antigenic challenge is the principal pathway in IgE mediated allergic reactions. As a consequence of the high affinity binding between IgE and FcεRI, along with the continuous production of IgE by B cells, allergies usually persist throughout life, with currently no permanent cure available. Horses, especially race horses, which are commonly inbred, are a species of mammals that are very prone to the development of hypersensitivity responses, which can seriously affect their performance. Physiological responses to allergic sensitization in horses mirror that observed in humans and dogs. In this paper we describe the development of an in situ assay system for the quantitative assessment of the release of mediators of the allergic response pertaining to the equine system. To this end, the gene encoding equine FcεRIα was transfected into and expressed onto the surface of parental Rat Basophil Leukemia (RBL-2H3.1) cells. The gene product of the transfected equine α-chain formed a functional receptor complex with the endogenous rat β- and γ-chains 1. The resultant assay system facilitated an assessment of the quantity of mediator secreted from equine FcεRIα transfected RBL-2H3.1 cells following sensitization with equine IgE and antigenic challenge using β-hexosaminidase release as a readout 2, 3. Mediator release peaked at 36.68% ± 4.88% at 100 ng ml-1 of antigen. This assay was modified from previous assays used to study human and canine allergic responses 4, 5. We have also shown that this type of assay system has multiple applications for the development of diagnostic tools and the safety assessment of potential therapeutic intervention strategies in allergic disease 6, 2, 3.
24 Related JoVE Articles!
Computer-assisted Large-scale Visualization and Quantification of Pancreatic Islet Mass, Size Distribution and Architecture
Institutions: University of Chicago, National Institutes of Health, University of Chicago, University of Massachusetts.
The pancreatic islet is a unique micro-organ composed of several hormone secreting endocrine cells such as beta-cells (insulin), alpha-cells (glucagon), and delta-cells (somatostatin) that are embedded in the exocrine tissues and comprise 1-2% of the entire pancreas. There is a close correlation between body and pancreas weight. Total beta-cell mass also increases proportionately to compensate for the demand for insulin in the body. What escapes this proportionate expansion is the size distribution of islets. Large animals such as humans share similar islet size distributions with mice, suggesting that this micro-organ has a certain size limit to be functional. The inability of large animal pancreata to generate proportionately larger islets is compensated for by an increase in the number of islets and by an increase in the proportion of larger islets in their overall islet size distribution. Furthermore, islets exhibit a striking plasticity in cellular composition and architecture among different species and also within the same species under various pathophysiological conditions. In the present study, we describe novel approaches for the analysis of biological image data in order to facilitate the automation of analytic processes, which allow for the analysis of large and heterogeneous data collections in the study of such dynamic biological processes and complex structures. Such studies have been hampered due to technical difficulties of unbiased sampling and generating large-scale data sets to precisely capture the complexity of biological processes of islet biology. Here we show methods to collect unbiased "representative" data within the limited availability of samples (or to minimize the sample collection) and the standard experimental settings, and to precisely analyze the complex three-dimensional structure of the islet. Computer-assisted automation allows for the collection and analysis of large-scale data sets and also assures unbiased interpretation of the data. Furthermore, the precise quantification of islet size distribution and spatial coordinates (i.e. X, Y, Z-positions) not only leads to an accurate visualization of pancreatic islet structure and composition, but also allows us to identify patterns during development and adaptation to altering conditions through mathematical modeling. The methods developed in this study are applicable to studies of many other systems and organisms as well.
Cellular Biology, Issue 49, beta-cells, islets, large-scale analysis, pancreas
Testing Protozoacidal Activity of Ligand-lytic Peptides Against Termite Gut Protozoa in vitro (Protozoa Culture) and in vivo (Microinjection into Termite Hindgut)
Institutions: Louisiana State University Agricultural Center.
We are developing a novel approach to subterranean termite control that would lead to reduced reliance on the use of chemical pesticides. Subterranean termites are dependent on protozoa in the hindguts of workers to efficiently digest wood. Lytic peptides have been shown to kill a variety of protozoan parasites (Mutwiri et al.
2000) and also protozoa in the gut of the Formosan subterranean termite, Coptotermes formosanus
(Husseneder and Collier 2009). Lytic peptides are part of the nonspecific immune system of eukaryotes, and destroy the membranes of microorganisms (Leuschner and Hansel 2004). Most lytic peptides are not likely to harm higher eukaryotes, because they do not affect the electrically neutral cholesterol-containing cell membranes of higher eukaryotes (Javadpour et al.
1996). Lytic peptide action can be targeted to specific cell types by the addition of a ligand. For example, Hansel et al.
(2007) reported that lytic peptides conjugated with cancer cell membrane receptor ligands could be used to destroy breast cancer cells, while lytic peptides alone or conjugated with non-specific peptides were not effective. Lytic peptides also have been conjugated to human hormones that bind to receptors on tumor cells for targeted destruction of prostate and testicular cancer cells (Leuschner and Hansel 2004).
In this article we present techniques used to demonstrate the protozoacidal activity of a lytic peptide (Hecate) coupled to a heptapeptide ligand that binds to the surface membrane of protozoa from the gut of the Formosan subterranean termite. These techniques include extirpation of the gut from termite workers, anaerobic culture of gut protozoa (Pseudotrichonympha grassii
, Holomastigotoides hartmanni
), microscopic confirmation that the ligand marked with a fluorescent dye binds to the termite gut protozoa and other free-living protozoa but not to bacteria or gut tissue. We also demonstrate that the same ligand coupled to a lytic peptide efficiently kills termite gut protozoa in vitro
(protozoa culture) and in vivo
(microinjection into hindgut of workers), but is less bacteriacidal than the lytic peptide alone. The loss of protozoa leads to the death of the termites in less than two weeks.
In the future, we will genetically engineer microorganisms that can survive in the termite hindgut and spread through a termite colony as "Trojan Horses" to express ligand-lytic peptides that would kill the protozoa in the termite gut and subsequently kill the termites in the colony. Ligand-lytic peptides also could be useful for drug development against protozoan parasites.
Microbiology, Issue 46, Isoptera, Coptotermes formosanus, Formosan subterranean termite, termite control, paratransgenesis, symbionts, anaerobic, fluorescence
Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study
Institutions: Virginia Commonwealth University, Virginia Commonwealth University, University Hospital of Geneva.
Throughout native artery, collagen and elastin play an important role, providing a mechanical backbone, preventing vessel rupture, and promoting recovery under pulsatile deformations. The goal of this study was to mimic the structure of native artery by fabricating a multi-layered electrospun conduit composed of poly(caprolactone) (PCL) with the addition of elastin and collagen with blends of 45-45-10, 55-35-10, and 65-25-10 PCL-ELAS-COL to demonstrate mechanical properties indicative of native arterial tissue, while remaining conducive to tissue regeneration. Whole grafts and individual layers were analyzed using uniaxial tensile testing, dynamic compliance, suture retention, and burst strength. Compliance results revealed that changes to the middle/medial layer changed overall graft behavior with whole graft compliance values ranging from 0.8 - 2.8 % / 100 mmHg, while uniaxial results demonstrated an average modulus range of 2.0 - 11.8 MPa. Both modulus and compliance data displayed values within the range of native artery. Mathematical modeling was implemented to show how changes in layer stiffness affect the overall circumferential wall stress, and as a design aid to achieve the best mechanical combination of materials. Overall, the results indicated that a graft can be designed to mimic a tri-layered structure by altering layer properties.
Bioengineering, Issue 47, Electrospinning, Vascular Graft, Multilayer, Polycaprolactone, Elastin
Transabdominal Ultrasound for Pregnancy Diagnosis in Reeves' Muntjac Deer
Institutions: Colorado State University.
Reeves' muntjac deer (Muntiacus reevesi
) are a small cervid species native to southeast Asia, and are currently being investigated as a potential model of prion disease transmission and pathogenesis. Vertical transmission is an area of interest among researchers studying infectious diseases, including prion disease, and these investigations require efficient methods for evaluating the effects of maternal infection on reproductive performance. Ultrasonographic examination is a well-established tool for diagnosing pregnancy and assessing fetal health in many animal species1-7
, including several species of farmed cervids8-19
, however this technique has not been described in Reeves' muntjac deer. Here we describe the application of transabdominal ultrasound to detect pregnancy in muntjac does and to evaluate fetal growth and development throughout the gestational period. Using this procedure, pregnant animals were identified as early as 35 days following doe-buck pairing and this was an effective means to safely monitor the pregnancy at regular intervals. Future goals of this work will include establishing normal fetal measurement references for estimation of gestational age, determining sensitivity and specificity of the technique for diagnosing pregnancy at various stages of gestation, and identifying variations in fetal growth and development under different experimental conditions.
Medicine, Issue 83, Ultrasound, Reeves' muntjac deer, Muntiacus reevesi, fetal development, fetal growth, captive cervids
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
The Bovine Lung in Biomedical Research: Visually Guided Bronchoscopy, Intrabronchial Inoculation and In Vivo Sampling Techniques
There is an ongoing search for alternative animal models in research of respiratory medicine. Depending on the goal of the research, large animals as models of pulmonary disease often resemble the situation of the human lung much better than mice do. Working with large animals also offers the opportunity to sample the same animal repeatedly over a certain course of time, which allows long-term studies without sacrificing the animals.
The aim was to establish in vivo
sampling methods for the use in a bovine model of a respiratory Chlamydia psittaci
infection. Sampling should be performed at various time points in each animal during the study, and the samples should be suitable to study the host response, as well as the pathogen under experimental conditions.
Bronchoscopy is a valuable diagnostic tool in human and veterinary medicine. It is a safe and minimally invasive procedure. This article describes the intrabronchial inoculation of calves as well as sampling methods for the lower respiratory tract. Videoendoscopic, intrabronchial inoculation leads to very consistent clinical and pathological findings in all inoculated animals and is, therefore, well-suited for use in models of infectious lung disease. The sampling methods described are bronchoalveolar lavage, bronchial brushing and transbronchial lung biopsy. All of these are valuable diagnostic tools in human medicine and could be adapted for experimental purposes to calves aged 6-8 weeks. The samples obtained were suitable for both pathogen detection and characterization of the severity of lung inflammation in the host.
Medicine, Issue 89, translational medicine, respiratory models, bovine lung, bronchoscopy, transbronchial lung biopsy, bronchoalveolar lavage, bronchial brushing, cytology brush
Whole Cell Recording from an Organotypic Slice Preparation of Neocortex
Institutions: University of Tennessee Health Science Center.
We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex. Because of the lack of specific pharmacological agents for these channels, we have taken a genetic approach to manipulating channel expression. We use an organotypic culture preparation (16) in order to maintain cell morphology and the laminar pattern of cortex. We typically isolate acute neocortical slices at postnatal days 8-10 and maintain the slices in culture for 3-7 days. This allows us to study neurons at a similar age to those in our work with acute slices and minimizes the development of exuberant excitatory connections in the slice. We record from visually-identified pyramidal neurons in layers II/III or V using infrared illumination (IR-) and differential interference contrast microscopy (DIC) with whole cell patch clamp in current- or voltage-clamp. We use biolistic (Gene gun) transfection of wild type or mutant potassium channel DNA to manipulate expression of the channels to study their function. The transfected cells are easily identified by epifluorescence microscopy after co-transfection with cDNA for green fluorescent protein (GFP). We compare recordings of transfected cells to adjacent, untransfected neurons in the same layer from the same slice.
Neuroscience, Issue 52, Organotypic, neocortex, somatosensory, pyramidal cell, slice preparation, biolistic transfection
Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells
Institutions: Erasmus MC - University Medical Center.
Fluorescent in situ
hybridization (FISH) is a molecular technique which enables the detection of nucleic acids in cells. DNA FISH is often used in cytogenetics and cancer diagnostics, and can detect aberrations of the genome, which often has important clinical implications. RNA FISH can be used to detect RNA molecules in cells and has provided important insights in regulation of gene expression. Combining DNA and RNA FISH within the same cell is technically challenging, as conditions suitable for DNA FISH might be too harsh for fragile, single stranded RNA molecules. We here present an easily applicable protocol which enables the combined, simultaneous detection of Xist
RNA and DNA encoded by the X chromosomes. This combined DNA-RNA FISH protocol can likely be applied to other systems where both RNA and DNA need to be detected.
Biochemistry, Issue 88, Fluorescent in situ hybridization (FISH), combined DNA-RNA FISH, ES cell, cytogenetics, single cell analysis, X chromosome inactivation (XCI), Xist, Bacterial artificial chromosome (BAC), DNA-probe, Rnf12
Cell Death Associated with Abnormal Mitosis Observed by Confocal Imaging in Live Cancer Cells
Institutions: Sheba Medical Center, Tel-Aviv University, Tel-Aviv University, Tel-Aviv University, Ecole Superieure de Biotechnologie Strasbourg, Tel-Aviv University.
Phenanthrene derivatives acting as potent PARP1 inhibitors prevented the bi-focal clustering of supernumerary centrosomes in multi-centrosomal human cancer cells in mitosis. The phenanthridine PJ-34 was the most potent molecule. Declustering of extra-centrosomes causes mitotic failure and cell death in multi-centrosomal cells. Most solid human cancers have high occurrence of extra-centrosomes. The activity of PJ-34 was documented in real-time by confocal imaging of live human breast cancer MDA-MB-231 cells transfected with vectors encoding for fluorescent γ-tubulin, which is highly abundant in the centrosomes and for fluorescent histone H2b present in the chromosomes. Aberrant chromosomes arrangements and de-clustered γ-tubulin foci representing declustered centrosomes were detected in the transfected MDA-MB-231 cells after treatment with PJ-34. Un-clustered extra-centrosomes in the two spindle poles preceded their cell death. These results linked for the first time the recently detected exclusive cytotoxic activity of PJ-34 in human cancer cells with extra-centrosomes de-clustering in mitosis, and mitotic failure leading to cell death. According to previous findings observed by confocal imaging of fixed cells, PJ-34 exclusively eradicated cancer cells with multi-centrosomes without impairing normal cells undergoing mitosis with two centrosomes and bi-focal spindles. This cytotoxic activity of PJ-34 was not shared by other potent PARP1 inhibitors, and was observed in PARP1 deficient MEF harboring extracentrosomes, suggesting its independency of PARP1 inhibition. Live confocal imaging offered a useful tool for identifying new molecules eradicating cells during mitosis.
Cancer Biology, Issue 78, Medicine, Cellular Biology, Molecular Biology, Biomedical Engineering, Anatomy, Physiology, Genetics, Neoplastic Processes, Pharmacologic Actions, Live confocal imaging, Extra-centrosomes clustering/de-clustering, Mitotic Catastrophe cell death, PJ-34, myocardial infarction, microscopy, imaging
Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Institutions: Seattle University, Seattle University.
In contrast to other chromatographic methods for purifying proteins (e.g. gel filtration, affinity, and ion exchange), hydrophobic interaction chromatography (HIC) commonly requires experimental determination (referred to as screening or "scouting") in order to select the most suitable chromatographic medium for purifying a given protein 1
. The method presented here describes an automated approach to scouting for an optimal HIC media to be used in protein purification.
HIC separates proteins and other biomolecules from a crude lysate based on differences in hydrophobicity. Similar to affinity chromatography (AC) and ion exchange chromatography (IEX), HIC is capable of concentrating the protein of interest as it progresses through the chromatographic process. Proteins best suited for purification by HIC include those with hydrophobic surface regions and able to withstand exposure to salt concentrations in excess of 2 M ammonium sulfate ((NH4
). HIC is often chosen as a purification method for proteins lacking an affinity tag, and thus unsuitable for AC, and when IEX fails to provide adequate purification. Hydrophobic moieties on the protein surface temporarily bind to a nonpolar ligand coupled to an inert, immobile matrix. The interaction between protein and ligand are highly dependent on the salt concentration of the buffer flowing through the chromatography column, with high ionic concentrations strengthening the protein-ligand interaction and making the protein immobile (i.e. bound inside the column) 2
. As salt concentrations decrease, the protein-ligand interaction dissipates, the protein again becomes mobile and elutes from the column. Several HIC media are commercially available in pre-packed columns, each containing one of several hydrophobic ligands (e.g. S-butyl, butyl, octyl, and phenyl) cross-linked at varying densities to agarose beads of a specific diameter 3
. Automated column scouting allows for an efficient approach for determining which HIC media should be employed for future, more exhaustive optimization experiments and protein purification runs 4
The specific protein being purified here is recombinant green fluorescent protein (GFP); however, the approach may be adapted for purifying other proteins with one or more hydrophobic surface regions. GFP serves as a useful model protein, due to its stability, unique light absorbance peak at 397 nm, and fluorescence when exposed to UV light 5
. Bacterial lysate containing wild type GFP was prepared in a high-salt buffer, loaded into a Bio-Rad DuoFlow medium pressure liquid chromatography system, and adsorbed to HiTrap HIC columns containing different HIC media. The protein was eluted from the columns and analyzed by in-line and post-run detection methods. Buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection increased the functionality of the system and reproducibility of the experimental approach.
Biochemistry, Issue 55, hydrophobic interaction chromatography, liquid chromatography, green fluorescent protein, GFP, scouting, protein purification, Bio-Rad DuoFlow, FPLC
Kinematics and Ground Reaction Force Determination: A Demonstration Quantifying Locomotor Abilities of Young Adult, Middle-aged, and Geriatric Rats
Institutions: Riverview, NB, University of Calgary, University of Calgary, University of Calgary.
Behavior, in its broadest definition, can be defined as the motor manifestation of physiologic processes. As such, all behaviors manifest through the motor system. In the fields of neuroscience and orthopedics, locomotion is a commonly evaluated behavior for a variety of disease models. For example, locomotor recovery after traumatic injury to the nervous system is one of the most commonly evaluated behaviors 1-3
. Though locomotion can be evaluated using a variety of endpoint measurements (e.g. time taken to complete a locomotor task, etc), semiquantitative kinematic measures (e.g. ordinal rating scales (e.g. Basso Beattie and Bresnahan locomotor (BBB) rating scale, etc)) and surrogate measures of behaviour (e.g. muscle force, nerve conduction velocity, etc), only kinetics (force measurements) and kinematics (measurements of body segments in space) provide a detailed description of the strategy by which an animal is able to locomote 1
. Though not new, kinematic and kinetic measurements of locomoting rodents is now more readily accessible due to the availability of commercially available equipment designed for this purpose. Importantly, however, experimenters need to be very familiar with theory of biomechanical analyses and understand the benefits and limitations of these forms of analyses prior to embarking on what will become a relatively labor-intensive study. The present paper aims to describe a method for collecting kinematic and ground reaction force data using commercially available equipment. Details of equipment and apparatus set-up, pre-training of animals, inclusion and exclusion criteria of acceptable runs, and methods for collecting the data are described. We illustrate the utility of this behavioral analysis technique by describing the kinematics and kinetics of strain-matched young adult, middle-aged, and geriatric rats.
Neuroscience, Issue 48, Locomotion, kinetics, kinematics, aging
Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice
Institutions: Boston University, Harvard Medical School.
Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high molecular weight compounds from the bloodstream and into brain tissue. Some direct delivery success in humans has been achieved via implantation of transcranial catheters; however this method is highly invasive and associated with numerous complications. A less invasive alternative would be to dose the brain through a surgically implanted, semipermeable membrane such as the nasal mucosa that is used to repair skull base defects following endoscopic transnasal tumor removal surgery in humans. Drug transfer though this membrane would effectively bypass the BBB and diffuse directly into the brain and cerebrospinal fluid. Inspired by this approach, a surgical approach in mice was developed that uses a donor septal mucosal membrane engrafted over an extracranial surgical BBB defect. This model has been shown to effectively allow the passage of high molecular weight compounds into the brain. Since numerous drug candidates are incapable of crossing the BBB, this model is valuable for performing preclinical testing of novel therapies for neurological and psychiatric diseases.
Medicine, Issue 89, drug delivery, mucosa membrane, blood-brain barrier, neurosurgery, transnasal, mouse model
Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans
Institutions: The Children's Hospital of Philadelphia, University of Pennsylvania.
Stable isotopic profiling has long permitted sensitive investigations of the metabolic consequences of genetic mutations and/or pharmacologic therapies in cellular and mammalian models. Here, we describe detailed methods to perform stable isotopic profiling of intermediary metabolism and metabolic flux in the nematode, Caenorhabditis elegans
. Methods are described for profiling whole worm free amino acids, labeled carbon dioxide, labeled organic acids, and labeled amino acids in animals exposed to stable isotopes either from early development on nematode growth media agar plates or beginning as young adults while exposed to various pharmacologic treatments in liquid culture. Free amino acids are quantified by high performance liquid chromatography (HPLC) in whole worm aliquots extracted in 4% perchloric acid. Universally labeled 13
C-glucose or 1,6-13
-glucose is utilized as the stable isotopic precursor whose labeled carbon is traced by mass spectrometry in carbon dioxide (both atmospheric and dissolved) as well as in metabolites indicative of flux through glycolysis, pyruvate metabolism, and the tricarboxylic acid cycle. Representative results are included to demonstrate effects of isotope exposure time, various bacterial clearing protocols, and alternative worm disruption methods in wild-type nematodes, as well as the relative extent of isotopic incorporation in mitochondrial complex III mutant worms (isp-1(qm150)
) relative to wild-type worms. Application of stable isotopic profiling in living nematodes provides a novel capacity to investigate at the whole animal level real-time metabolic alterations that are caused by individual genetic disorders and/or pharmacologic therapies.
Developmental Biology, Issue 48, Stable isotope, amino acid quantitation, organic acid quantitation, nematodes, metabolism
Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro
Institutions: The University of Chicago Medical Center, The University of Chicago Medical Center.
Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro
using spreading depression (SD), as a means to develop novel therapeutic targets for episodic and chronic migraine. SD is the likely cause of migraine aura and migraine pain. It is a paroxysmal loss of neuronal function triggered by initially increased neuronal activity, which slowly propagates within susceptible brain regions. Normal brain function is exquisitely sensitive to, and relies on, coincident low-level immune signaling. Thus, neural immune signaling likely affects electrical activity of SD, and therefore migraine. Pain perception studies of SD in whole animals are fraught with difficulties, but whole animals are well suited to examine systems biology aspects of migraine since SD activates trigeminal nociceptive pathways. However, whole animal studies alone cannot be used to decipher the cellular and neural circuit mechanisms of SD. Instead, in vitro
preparations where environmental conditions can be controlled are necessary. Here, it is important to recognize limitations of acute slices and distinct advantages of hippocampal slice cultures. Acute brain slices cannot reveal subtle changes in immune signaling since preparing the slices alone triggers: pro-inflammatory changes that last days, epileptiform behavior due to high levels of oxygen tension needed to vitalize the slices, and irreversible cell injury at anoxic slice centers.
In contrast, we examine immune signaling in mature hippocampal slice cultures since the cultures closely parallel their in vivo
counterpart with mature trisynaptic function; show quiescent astrocytes, microglia, and cytokine levels; and SD is easily induced in an unanesthetized preparation. Furthermore, the slices are long-lived and SD can be induced on consecutive days without injury, making this preparation the sole means to-date capable of modeling the neuroimmune consequences of chronic SD, and thus perhaps chronic migraine. We use electrophysiological techniques and non-invasive imaging to measure
neuronal cell and circuit functions coincident with SD. Neural immune gene expression variables are measured with qPCR screening, qPCR arrays, and, importantly, use of cDNA preamplification for detection of ultra-low level targets such as interferon-gamma using whole, regional, or specific cell enhanced (via laser dissection microscopy) sampling. Cytokine cascade signaling is further assessed with multiplexed phosphoprotein related targets with gene expression and phosphoprotein changes confirmed via cell-specific immunostaining. Pharmacological and siRNA strategies are used to mimic
SD immune signaling.
Neuroscience, Issue 52, innate immunity, hormesis, microglia, T-cells, hippocampus, slice culture, gene expression, laser dissection microscopy, real-time qPCR, interferon-gamma
High-throughput Fluorometric Measurement of Potential Soil Extracellular Enzyme Activities
Institutions: Colorado State University, Oak Ridge National Laboratory, University of Colorado.
Microbes in soils and other environments produce extracellular enzymes to depolymerize and hydrolyze organic macromolecules so that they can be assimilated for energy and nutrients. Measuring soil microbial enzyme activity is crucial in understanding soil ecosystem functional dynamics. The general concept of the fluorescence enzyme assay is that synthetic C-, N-, or P-rich substrates bound with a fluorescent dye are added to soil samples. When intact, the labeled substrates do not fluoresce. Enzyme activity is measured as the increase in fluorescence as the fluorescent dyes are cleaved from their substrates, which allows them to fluoresce. Enzyme measurements can be expressed in units of molarity or activity. To perform this assay, soil slurries are prepared by combining soil with a pH buffer. The pH buffer (typically a 50 mM sodium acetate or 50 mM Tris buffer), is chosen for the buffer's particular acid dissociation constant (pKa) to best match the soil sample pH. The soil slurries are inoculated with a nonlimiting amount of fluorescently labeled (i.e.
C-, N-, or P-rich) substrate. Using soil slurries in the assay serves to minimize limitations on enzyme and substrate diffusion. Therefore, this assay controls for differences in substrate limitation, diffusion rates, and soil pH conditions; thus detecting potential enzyme activity rates as a function of the difference in enzyme concentrations (per sample).
Fluorescence enzyme assays are typically more sensitive than spectrophotometric (i.e.
colorimetric) assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light; so caution is required when handling fluorescent substrates. Likewise, this method only assesses potential enzyme activities under laboratory conditions when substrates are not limiting. Caution should be used when interpreting the data representing cross-site comparisons with differing temperatures or soil types, as in situ
soil type and temperature can influence enzyme kinetics.
Environmental Sciences, Issue 81, Ecological and Environmental Phenomena, Environment, Biochemistry, Environmental Microbiology, Soil Microbiology, Ecology, Eukaryota, Archaea, Bacteria, Soil extracellular enzyme activities (EEAs), fluorometric enzyme assays, substrate degradation, 4-methylumbelliferone (MUB), 7-amino-4-methylcoumarin (MUC), enzyme temperature kinetics, soil
Design and Construction of an Urban Runoff Research Facility
Institutions: Texas A&M University, The Scotts Miracle-Gro Company.
As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use due to increased demand for landscape irrigation. Improper irrigation practices and large rainfall events can result in runoff from urban landscapes which has potential to carry nutrients and sediments into local streams and lakes where they may contribute to eutrophication. A 1,000 m2
facility was constructed which consists of 24 individual 33.6 m2
field plots, each equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated urban landscapes. Runoff volumes from the first and second trials had coefficient of variability (CV) values of 38.2 and 28.7%, respectively. CV values for runoff pH, EC, and Na concentration for both trials were all under 10%. Concentrations of DOC, TDN, DON, PO4
, and Ca2+
had CV values less than 50% in both trials. Overall, the results of testing performed after sod installation at the facility indicated good uniformity between plots for runoff volumes and chemical constituents. The large plot size is sufficient to include much of the natural variability and therefore provides better simulation of urban landscape ecosystems.
Environmental Sciences, Issue 90, urban runoff, landscapes, home lawns, turfgrass, St. Augustinegrass, carbon, nitrogen, phosphorus, sodium
Diffusion Tensor Magnetic Resonance Imaging in the Analysis of Neurodegenerative Diseases
Institutions: University of Ulm.
Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo
. The present applications are designed to investigate differences of WM involvement patterns in different brain diseases, especially neurodegenerative disorders, by use of different DTI analyses in comparison with matched controls.
DTI data analysis is performed in a variate fashion, i.e.
voxelwise comparison of regional diffusion direction-based metrics such as fractional anisotropy (FA), together with fiber tracking (FT) accompanied by tractwise fractional anisotropy statistics (TFAS) at the group level in order to identify differences in FA along WM structures, aiming at the definition of regional patterns of WM alterations at the group level. Transformation into a stereotaxic standard space is a prerequisite for group studies and requires thorough data processing to preserve directional inter-dependencies. The present applications show optimized technical approaches for this preservation of quantitative and directional information during spatial normalization in data analyses at the group level. On this basis, FT techniques can be applied to group averaged data in order to quantify metrics information as defined by FT. Additionally, application of DTI methods, i.e.
differences in FA-maps after stereotaxic alignment, in a longitudinal analysis at an individual subject basis reveal information about the progression of neurological disorders. Further quality improvement of DTI based results can be obtained during preprocessing by application of a controlled elimination of gradient directions with high noise levels.
In summary, DTI is used to define a distinct WM pathoanatomy of different brain diseases by the combination of whole brain-based and tract-based DTI analysis.
Medicine, Issue 77, Neuroscience, Neurobiology, Molecular Biology, Biomedical Engineering, Anatomy, Physiology, Neurodegenerative Diseases, nuclear magnetic resonance, NMR, MR, MRI, diffusion tensor imaging, fiber tracking, group level comparison, neurodegenerative diseases, brain, imaging, clinical techniques
Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling
Institutions: Colorado State University, USDA-ARS, Colorado State University.
Tracing rare stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2
fluxes and soil organic matter formation to small-scale stable-isotope biomarker probing. Coupling multiple stable isotopes such as 13
C with 15
O or 2
H has the potential to reveal even more information about complex stoichiometric relationships during biogeochemical transformations. Isotope labeled plant material has been used in various studies of litter decomposition and soil organic matter formation1-4
. From these and other studies, however, it has become apparent that structural components of plant material behave differently than metabolic components (i.e
. leachable low molecular weight compounds) in terms of microbial utilization and long-term carbon storage5-7
. The ability to study structural and metabolic components separately provides a powerful new tool for advancing the forefront of ecosystem biogeochemical studies. Here we describe a method for producing 13
C and 15
N labeled plant material that is either uniformly labeled throughout the plant or differentially labeled in structural and metabolic plant components.
Here, we present the construction and operation of a continuous 13
C and 15
N labeling chamber that can be modified to meet various research needs. Uniformly labeled plant material is produced by continuous labeling from seedling to harvest, while differential labeling is achieved by removing the growing plants from the chamber weeks prior to harvest. Representative results from growing Andropogon gerardii
Kaw demonstrate the system's ability to efficiently label plant material at the targeted levels. Through this method we have produced plant material with a 4.4 atom%13
C and 6.7 atom%15
N uniform plant label, or material that is differentially labeled by up to 1.29 atom%13
C and 0.56 atom%15
N in its metabolic and structural components (hot water extractable and hot water residual components, respectively). Challenges lie in maintaining proper temperature, humidity, CO2
concentration, and light levels in an airtight 13
atmosphere for successful plant production. This chamber description represents a useful research tool to effectively produce uniformly or differentially multi-isotope labeled plant material for use in experiments on ecosystem biogeochemical cycling.
Environmental Sciences, Issue 83, 13C, 15N, plant, stable isotope labeling, Andropogon gerardii, metabolic compounds, structural compounds, hot water extraction
Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease
Institutions: Louisiana State University Health Sciences Center.
There is a major increase in the awareness of the positive impact of exercise on improving several disease states with neurobiological basis; these include improving cognitive function and physical performance. As a result, there is an increase in the number of animal studies employing exercise. It is argued that one intrinsic value of forced exercise is that the investigator has control over the factors that can influence the impact of exercise on behavioral outcomes, notably exercise frequency, duration, and intensity of the exercise regimen. However, compliance in forced exercise regimens may be an issue, particularly if potential confounds of employing foot-shock are to be avoided. It is also important to consider that since most cognitive and locomotor impairments strike in the aged individual, determining impact of exercise on these impairments should consider using aged rodents with a highest possible level of compliance to ensure minimal need for test subjects. Here, the pertinent steps and considerations necessary to achieve nearly 100% compliance to treadmill exercise in an aged rodent model will be presented and discussed. Notwithstanding the particular exercise regimen being employed by the investigator, our protocol should be of use to investigators that are particularly interested in the potential impact of forced exercise on aging-related impairments, including aging-related Parkinsonism and Parkinson’s disease.
Behavior, Issue 90, Exercise, locomotor, Parkinson’s disease, aging, treadmill, bradykinesia, Parkinsonism
Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA Receptors
Institutions: University College London.
Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA (γ-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain. It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA
Rs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons. Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials.
During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAA
Rs subtypes. This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other.
To elucidate the underlying molecular mechanisms, a novel in vitro
co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAA
R subtypes. Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification. Our results indicate that various GABAA
R subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro
model system can be used to reproduce, at least in part, the in vivo
conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses. Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAA
Rs are first described, followed by detailed instructions on how to combine these two cell types in co-culture and analyze the formation of synaptic contacts.
Neuroscience, Issue 93, Developmental neuroscience, synaptogenesis, synaptic inhibition, co-culture, stable cell lines, GABAergic, medium spiny neurons, HEK 293 cell line
In Situ Neutron Powder Diffraction Using Custom-made Lithium-ion Batteries
Institutions: University of Sydney, University of Wollongong, Australian Synchrotron, Australian Nuclear Science and Technology Organisation, University of Wollongong, University of New South Wales.
Li-ion batteries are widely used in portable electronic devices and are considered as promising candidates for higher-energy applications such as electric vehicles.1,2
However, many challenges, such as energy density and battery lifetimes, need to be overcome before this particular battery technology can be widely implemented in such applications.3
This research is challenging, and we outline a method to address these challenges using in situ
NPD to probe the crystal structure of electrodes undergoing electrochemical cycling (charge/discharge) in a battery. NPD data help determine the underlying structural mechanism responsible for a range of electrode properties, and this information can direct the development of better electrodes and batteries.
We briefly review six types of battery designs custom-made for NPD experiments and detail the method to construct the ‘roll-over’ cell that we have successfully used on the high-intensity NPD instrument, WOMBAT, at the Australian Nuclear Science and Technology Organisation (ANSTO). The design considerations and materials used for cell construction are discussed in conjunction with aspects of the actual in situ
NPD experiment and initial directions are presented on how to analyze such complex in situ
Physics, Issue 93, In operando, structure-property relationships, electrochemical cycling, electrochemical cells, crystallography, battery performance
Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation
Institutions: North Carolina State University, North Carolina State University.
Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals is often not well understood. Here we demonstrate an integrated field lysimetry and porewater sampling method for evaluating the mobility of chemicals applied to soils and established vegetation. Lysimeters, open columns made of metal or plastic, are driven into bareground or vegetated soils. Porewater samplers, which are commercially available and use vacuum to collect percolating soil water, are installed at predetermined depths within the lysimeters. At prearranged times following chemical application to experimental plots, porewater is collected, and lysimeters, containing soil and vegetation, are exhumed. By analyzing chemical concentrations in the lysimeter soil, vegetation, and porewater, downward leaching rates, soil retention capacities, and plant uptake for the chemical of interest may be quantified. Because field lysimetry and porewater sampling are conducted under natural environmental conditions and with minimal soil disturbance, derived results project real-case scenarios and provide valuable information for chemical management. As chemicals are increasingly applied to land worldwide, the described techniques may be utilized to determine whether applied chemicals pose adverse effects to human health or the environment.
Environmental Sciences, Issue 89,
Lysimetry, porewater, soil, chemical leaching, pesticides, turfgrass, waste
Isolation and Culture of Neonatal Mouse Cardiomyocytes
Institutions: King’s College London, University of California San Diego .
Cultured neonatal cardiomyocytes have long been used to study myofibrillogenesis and myofibrillar functions. Cultured cardiomyocytes allow for easy investigation and manipulation of biochemical pathways, and their effect on the biomechanical properties of spontaneously beating cardiomyocytes.
The following 2-day protocol describes the isolation and culture of neonatal mouse cardiomyocytes. We show how to easily dissect hearts from neonates, dissociate the cardiac tissue and enrich cardiomyocytes from the cardiac cell-population. We discuss the usage of different enzyme mixes for cell-dissociation, and their effects on cell-viability. The isolated cardiomyocytes can be subsequently used for a variety of morphological, electrophysiological, biochemical, cell-biological or biomechanical assays. We optimized the protocol for robustness and reproducibility, by using only commercially available solutions and enzyme mixes that show little lot-to-lot variability. We also address common problems associated with the isolation and culture of cardiomyocytes, and offer a variety of options for the optimization of isolation and culture conditions.
Cellular Biology, Issue 79, Biomedical Engineering, Bioengineering, Molecular Biology, Cell Culture Techniques, Primary Cell Culture, Cell Culture Techniques, Primary Cell Culture, Cell Culture Techniques, Primary Cell Culture, Cell Culture Techniques, Disease Models, Animal, Models, Cardiovascular, Cell Biology, neonatal mouse, cardiomyocytes, isolation, culture, primary cells, NMC, heart cells, animal model
Spatial Multiobjective Optimization of Agricultural Conservation Practices using a SWAT Model and an Evolutionary Algorithm
Institutions: University of Washington, Iowa State University, North Carolina A&T University, Iowa Geological and Water Survey.
Finding the cost-efficient (i.e.
, lowest-cost) ways of targeting conservation practice investments for the achievement of specific water quality goals across the landscape is of primary importance in watershed management. Traditional economics methods of finding the lowest-cost solution in the watershed context (e.g.
) assume that off-site impacts can be accurately described as a proportion of on-site pollution generated. Such approaches are unlikely to be representative of the actual pollution process in a watershed, where the impacts of polluting sources are often determined by complex biophysical processes. The use of modern physically-based, spatially distributed hydrologic simulation models allows for a greater degree of realism in terms of process representation but requires a development of a simulation-optimization framework where the model becomes an integral part of optimization.
Evolutionary algorithms appear to be a particularly useful optimization tool, able to deal with the combinatorial nature of a watershed simulation-optimization problem and allowing the use of the full water quality model. Evolutionary algorithms treat a particular spatial allocation of conservation practices in a watershed as a candidate solution and utilize sets (populations) of candidate solutions iteratively applying stochastic operators of selection, recombination, and mutation to find improvements with respect to the optimization objectives. The optimization objectives in this case are to minimize nonpoint-source pollution in the watershed, simultaneously minimizing the cost of conservation practices. A recent and expanding set of research is attempting to use similar methods and integrates water quality models with broadly defined evolutionary optimization methods3,4,9,10,13-15,17-19,22,23,25
. In this application, we demonstrate a program which follows Rabotyagov et al.'s approach and integrates a modern and commonly used SWAT water quality model7
with a multiobjective evolutionary algorithm SPEA226
, and user-specified set of conservation practices and their costs to search for the complete tradeoff frontiers between costs of conservation practices and user-specified water quality objectives. The frontiers quantify the tradeoffs faced by the watershed managers by presenting the full range of costs associated with various water quality improvement goals. The program allows for a selection of watershed configurations achieving specified water quality improvement goals and a production of maps of optimized placement of conservation practices.
Environmental Sciences, Issue 70, Plant Biology, Civil Engineering, Forest Sciences, Water quality, multiobjective optimization, evolutionary algorithms, cost efficiency, agriculture, development

domain: biology


text:
It’s Never Easy These Days: the Candid and Often Comical Memoir of a Hospital Manager
Similar reminiscences by hospital managers are much rarer, so this slim volume is to be welcomed in helping to complete a more realistic view of how the NHS functioned.
Moaning about managers is a common currency among clinical staff, but their unglamorous role in ensuring that buildings are fit for purpose, supplies have been ordered and despatched, and the budget is spent wisely, are critical to the effectiveness of any health system.
The author began his management career at the age of 22 when he was put in charge of the medical and surgical stores for a group of hospitals. As he had little idea what a catheter was, a steep learning curve was essential.
A long career in a wide range of posts saw him coping with the challenges posed by independent sector and NHS hospitals in the 1980s and 90s. Everything from the smooth functioning...
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domain: medical


text:
Objectives To evaluate the health-condition of military police officers and firefighters. To identify risk factors for not being medically ready for duty.
Design Cross-sectional study.
Setting Data were extracted from medical records during annual periodic health assessments of police officers and firefighters serving with the military police in Paraná, Brazil.
Participants 6621 police officers (5927 men and 694 women) and 1347 firefighters (1257 men and 90 women) who underwent health assessments between July 2018 and June 2019 were analysed. Pregnant women were excluded.
Outcome measures Data included variables such as sex, age, anthropometric measurements, lifestyle, comorbidities and laboratory tests. Multiple logistic regression was used to estimate the probability of not being medically ready for active duty.
Results Overall, police officers had worse health status than firefighters and greater prevalence of overweight and obesity, regardless of sex. Musculoskeletal diseases were the most commonly reported disease by police officers and firefighters of both sexes. Among men, hypertension was the second most prevalent disease, followed by psychiatric diseases and dyslipidaemia. Among women, psychiatric diseases were the second most prevalent. Male police officers ≥40 years old presented the highest probability of not being considered ready for duty (40.1%). The probability of male police officers between the ages of 31 and 40 not being ready was similar to that for male firefighters >40 years old. There was a higher chance of not being medically ready professionals with diseases such as diabetes mellitus (OR 2.95, 95% CI 1.97 to 5.03), dyslipidaemia (OR 2.65, 95% CI 1.96 to 3.58), hypertension (OR 2.29, 95% CI 1.85 to 4.70), high total cholesterol (OR 2.16, 95% CI 1.93 to 2.42), and heart disease (OR 2.13, 95% CI 1.32 to 3.45).
Conclusions There was a high prevalence of chronic diseases and modifiable cardiovascular risk factors among police officers and firefighters. Healthy protective measures should be offered frequently, particularly to police officers at an earlier age.
- preventive medicine
- internal medicine
Data availability statement
Data are available upon reasonable request. The data are available from the corresponding author upon reasonable request.
This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.
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Strengths and limitations of this study
Few studies have compared the health condition of military police officers and firefighters.
This study included female police officers and firefighters therefore our findings may contribute to research in this subset of the population.
The study used a cross-sectional design, wherein casual relationships could not be inferenced from the results.
Our findings may not be generalised to other professional groups.
New fields of research can be developed from the results.
The public safety sector is essential for the maintenance of social order. The military police in Paraná, South of Brazil, has been responsible for training military police officers and firefighters to fulfil this noble mission for 167 years. Although they are considered to be physically and psychologically fit when they start their careers, these workers are exposed to various work-related risk factors that can cause their physical and mental health to deteriorate.1 2
Police work involves long working days and very stressful situations, often in violent areas.1 Police officers also face unexpected situations that require strength, dexterity and physical fitness.2 Firefighters face additional risks, when they are exposed to smoke from chemical and biological hazards.1 Studies have shown that police officers and firefighters have a shorter life expectancy than the general population and high mortality and morbidity, especially due to cardiovascular disease (CVD).2
Although firefighters are subjected to physical demands, the prevalence of being overweight and obese among them is similar to the general population. The high prevalence of CVD risk factors remains a concern since firefighters are called on to exert maximal effort under stressful conditions.3 4 There is a variable prevalence of cardiovascular risk factors such as hypertension, dyslipidaemia, obesity, diabetes mellitus, smoking and metabolic syndrome among police officers.5
Several studies have investigated the health status of police officers and firefighters, focusing primarily on cardiovascular, chronic and psychiatric diseases. However, few have investigated the prevalence of their morbidities with sex and age in that population, or have estimated their risk of having unsuitable health status for their daily work activities. It is imperative that police officers and firefighters attain and maintain physical fitness for both their overall health and ability to perform job tasks safely and effectively. The present study sought to evaluate the health condition of military police officers and firefighters and to identify risk factors for not being medically ready for active duty.6
Participants and methods
Study design and evaluated population
This observational study investigated the general health status of military police officers and firefighters in Paraná between June 2018 and July 2019. Pregnant women were excluded from the study. A total of 7968 individuals were included: 6621 police officers (5927 men, 694 women) and 1347 firefighters (1257 men, 90 women), which comprised 43.4% of the military police officers and firefighters in Paraná. Participants underwent a medical assessment during the annual health inspections that comprised clinical history, physical examination and laboratory tests. An exercise stress test (ET) on a treadmill was performed by individuals aged 40 years or over. Participants were divided according to sex and age (for males: ≤30 years old, 31–40 years old and ≥41 years old; for females: ≤30 years old and ≥31 years old) to identify a subset of the study population with the greatest prevalence of chronic diseases. Women were analysed separately because of their biological and behavioural characteristics, highlighting the sex differences in cardiometabolic, obesity and physical performance parameters.
The participants were military police officers and firefighters performing regular duties who underwent an elective medical assessment during the annual health inspections. Data were collected from standardised electronic health records.
Race category was determined according to the answer given to a standard question on self-declared ethnicity, based on the method used by the Brazilian Institute of Geography and Statistics national census survey which classifies individuals as white, brown, black, yellow or indigenous.7 A semistructured anamnesis was used to investigate the presence of chronic diseases, such as diabetes mellitus, hypertension, musculoskeletal, cardiovascular, endocrine, respiratory, neurological and psychiatric diseases, among others. If the answer was yes for any disease, the treatment used was asked. Smoking status was examined in the patient interview.
The International Physical Activity Questionnaire was applied to assess physical activity. It is widely used to estimate the frequency and duration of physical activity levels and has been validated for Brazil.8 Participants were classified as inactive or insufficiently active if they did no physical activity or less than 150 min per week; as active if they did between 150 and 300 min of moderate physical activity or 75 min of intense activity per week or an equivalent combination; and as highly active if they did more than 300 min per week of moderate to intense activity.9
The Alcohol Use Disorders Identification Test–Concise questionnaire was used to assess alcohol consumption. The questionnaire assigns a score on a scale from 0 to 12. For men, a score of 0–3 indicates low risk, 4–5 indicates moderate risk, 6–7 indicates high risk and 8–12 indicates severe risk. For women, a score of 0–2 indicates low risk, 3–5 indicates moderate risk, 6–7 indicates high risk and 8–12 indicates severe risk.10
Blood pressure was measured using a portable device (Welch Allyn Tycos DS48 Pocket aneroid sphygmomanometer) attached to the left arm while the participant was in a sitting position and after at least 5 min of rest. Participants were asked to refrain from smoking or consuming caffeine or alcohol for 30 min prior to measurement. They were instructed by a doctor or trained nurse to relax as much as possible and not to talk during the measurement procedure. The arm was supported at heart level and the bladder of the cuff encircled at least 80% of the arm circumference. When the two measurements were divergent, a third measurement was performed and the mean of the measurements was used for statistical analysis.11 High blood pressure was defined by systolic blood pressure ≥140 mm Hg and/or diastolic blood pressure ≥90 mm Hg. Uncontrolled hypertension was defined as systolic blood pressure ≥140 mm Hg and/or diastolic blood pressure ≥90 mm Hg in patients taking antihypertensive medications.12
Body mass index (BMI) was calculated according to the formula (weight (kg)/height squared (m2)). Participants were classified into four categories based on WHO guidelines: low weight (<18.5 kg/m2), normal weight (18.5–24.9 kg/m2), overweight (25–29.9 kg/m2) and obese (≥30 kg/m2).13 Waist circumference (WC) was measured horizontally at the midpoint between the lower edge of the ribs and the top of the iliac crest. A WC≥94 cm for men or ≥80 cm for women was considered as a risk factor for metabolic complications.14
Participants were considered as not medically ready for duty if the results of their ET were suggestive of ischaemia, cardiac arrhythmia, or if they presented with poor health condition, such as disabling musculoskeletal diseases, uncontrolled hypertension or diabetes mellitus. Whenever necessary, they were referred for specialised medical care.
Blood samples were collected by venepuncture after 12 hours of fasting. Total cholesterol and triglycerides were determined by colorimetric assay, and glucose by the hexokinase method. Total cholesterol and triglycerides serum levels were considered ideal if they were <200 mg/dL and <150 mg/dL, respectively.15 A glucose serum level <99 mg/dL was considered normal.16 Uncontrolled diabetes was defined by fasting plasma glucose >130 mg/dL in patients taking antidiabetic medications.16
Participants aged 40 years old or older without a history of CVD were monitored by a cardiologist while they underwent an ET on a treadmill. Participants were instructed to avoid exhausting activities in the 24 hours prior to the examination, and to eat light food, and not to smoke or drink coffee in the 3 hours prior to the examination. Electrodes were placed on the chest and attached to an ECG machine. Resting ECG, heart rate and blood pressure were measured prior to starting the exercise regimen. Once no limiting factors were detected on the baseline ECG, the participant was placed on a treadmill with a protocol designed to increases in intervals during the exercise. Blood pressure and heart rate were monitored throughout, and the participant was monitored for any developing symptoms, such as chest pain, shortness of breath, dizziness or extreme fatigue.
The most common protocol used for treadmill exercise stress testing was the Bruce protocol. This protocol is divided into successive 3-min stages, which requires the patient to walk faster and at a steeper gradient. The testing protocol could be adjusted according to the participant’s tolerance, aiming for 6–12 min of exercise duration.17 The Ellestad protocol for treadmill testing has a fixed inclination of 10% and a velocity of 1.7 mph for 3 min, 3 mph for 2 min, 4 mph for 2 min and finally 5 mph for 3 min.18 Oxygen consumption was estimated by the formula (male: VO2=1.11×(60–0.55×age); female: VO2=1.11×(48–0.37×age)).
Participants were questioned about any symptoms that they experienced during exercise. All participants were monitored closely during the recovery period until their heart rate and ECG were back to baseline. Indications for terminating the test were: participant request due to severe symptoms (ie, chest pain, shortness of breath or fatigue), severe exercise-induced hypotension or hypertension, horizontal or down sloping ST depression ≥1 mm or ST-segment elevation, new bundle branch block, atrioventricular block, ventricular arrhythmia, reaching maximal heart rate or completing all stages.
Statement of patient and public involvement
Participants were not involved in the design, conduction or analysis of this study. The results will be shared with the participants and the general population through the publication of this article.
Data are shown as mean±SD, median (minimum and maximum values) or percentages. An independent samples Student’s t test was used to compare continuous variables, while Fisher’s exact test or the χ2 test was used to compare categorical variables. For the univariate and multivariate analysis, logistic regression models were adjusted for factors associated with the results of the clinical assessment (ready or not ready). Variables with p<0.05 in the univariate analysis were included in the multivariate model. Interaction was assessed for firefighters/police officers and each variable was considered for the multivariate model. If present, the interaction term was included in the model adjusted for other variables. The Wald test was used to assess statistical significance. The OR for not being medically ready for active duties and 95% CI are shown. The Kolmogorov-Smirnov normality test was used to examine whether variables were normally distributed and two-sided p<0.05 were considered statistically significant. Data were analysed using Stata V.14.1 StataCorp LP.
Among the 6621 police officers, 74.1% declared themselves as white, 20,3% as brown (mixed heritage) and 4.6% as black, while among the 1347 firefighters, 78.8% declared themselves as white, 15.9% as brown (mixed heritage) and 3.4% as black. Regarding rank, 12.4% of all individuals held the rank of officer and 87.6% were privates. Table 1 shows the general, clinical, and laboratory characteristics of the police officers and firefighters by sex. Both male and female police officers were significantly older, had longer length of service and higher BMI and WC than firefighters. Systolic and diastolic blood pressure was significantly higher in male police officers than in male firefighters. In addition, the prevalence of overweight and obesity was greater among police officers than firefighters of both sexes. A significantly higher percentage of police officers had WC above the ideal, whereas a significantly higher percentage of firefighters was within the ideal BMI range.
Musculoskeletal diseases were the most commonly self-reported diseases in all groups. In men, hypertension was the second most prevalent disease, followed by psychiatric diseases and dyslipidaemia. Among women, psychiatric diseases were the second most prevalent.
Morbidities, except for musculoskeletal diseases, were significantly more prevalent among male police officers than among male firefighters while there was no difference in the prevalence of comorbidities between female police officers and female firefighters. The most widely used medications were antihypertensive drugs and antidepressants while smoking was more prevalent among police officers than firefighters. Regarding the laboratory assessment, a significantly higher percentage of male police officers had fasting glucose, total cholesterol and triglyceride serum levels above the normal range in comparison to male firefighters.
Table 2 shows the clinical and laboratory characteristics of male police officers and male firefighters by age group, in an attempt to identify a subset of the population with the highest prevalence of risk factors for CVD chronic diseases. Increased WC was more prevalent among police officers in all age groups. Musculoskeletal diseases were the most prevalent disorder in police officers and firefighters in all age groups, without statistically significant differences between them. The prevalence of hypertension was significantly greater among police officers ≥31 years old while psychiatric diseases, dyslipidaemia and diabetes mellitus were more prevalent among police officers >40 years old with their prevalence increasing with age. Excessive alcohol consumption was more frequent among firefighters >40 years old. Police officers did significantly less physical activity than firefighters in all age groups. Cardiac arrhythmia, the abnormality most frequently detected by the ET, was significantly more prevalent among police officers than among firefighters >40 years old. Finally, a significantly greater number of police officers than firefighters were considered to not be medically ready for duty across all age groups.
All of the clinical characteristics were similar between female police officers and female firefighters ≤30 years old, save for physical activity. Female police officers >30 years old had longer length of service, higher BMI and higher WC. There was no significant difference in the prevalence of not being ready for duty between female police officers and female firefighters (table 3).
After adjusting the multivariate model for predicting unhealthy conditions and using the explanatory variables of police officer/firefighter, sex and age, the probability of these workers being considered as not ready for active duty could be estimated (table 4).
The multivariate logistic regression model included the explanatory variables shown in table 5. The best health profiles were found among female firefighters while the worst health profiles were found among male firefighters ≥40 years old and male police officers ≥30 years old. Furthermore, male police officers >40 years old presented the highest probability of not being medically ready for active duty.
The main non-modifiable risk factors associated with not being medically ready for duty were: being a police officer (OR 1.50, 95% CI 1.27 to 1.77), being male aged 31–40 years (OR 2.03, 95% CI 1.43 to 2.89), and being≥40 years old (OR 2.55 95% CI 1.79 to 3.62). Moreover, the following modifiable risk factors were identified: high WC (OR 1.63, 95% CI 1.45 to 1.83), physical inactivity (OR 1.67, 95% CI 1.44 to 1.93), smoking (OR 1.75, 95% CI 1.45 to 2.12) and excessive consumption of alcohol (OR 1.44, 95% CI 1.22 to 1.70). The medical conditions associated with not being medically ready for duty were hypertension (OR 2.29, 95% CI 1.85 to 4.70), dyslipidaemia (OR 2.65, 95% CI 1.96 to 3.58), diabetes mellitus (OR 2.95, 95% CI 1.97 to 5.03), heart disease (OR 2.13, 95% CI 1.32 to 3.45), psychiatric diseases (OR 1.33, 95% CI 1.02 to 1.74) and high serum levels of total cholesterol (OR 2.16, 95% CI 1.93 to 2.42) (figure 1).
This is the largest study of the epidemiological profile of military police officers and firefighters in Paraná. The probability of male police officers between the ages of 31 and 40 not being ready was similar to that of male firefighters >40 years old, indicating that the health condition of male police officers deteriorates earlier. Our findings indicate that the priority for regular medical assessment should be given to this subset of male police officers >30 years old and firefighters >40 years old, who in some cases already had increased WC, diabetes mellitus, CVD and dyslipidaemia, in order to facilitate access to healthcare interventions and to stimulate healthy daily habits.
Several modifiable risk factors for CVD were observed, including increased WC, obesity, physical inactivity and smoking. They were more common among participants over 30 years old, which suggests that in addition to ageing, work-related risk factors may play a central to role in the deterioration of health condition.2–5 This finding underlies the need for better planning and investment in prevention and early diagnosis in this population. Furthermore, behaviour interventions to encourage healthy habits should be implemented in order to help to modify the profile of chronic diseases.
Similar to our findings, Nogueira et al found a prevalence of 54.3% for overweight and 14.7% for obesity in a study that investigated the relationship between cardiorespiratory fitness and body composition in 4237 Brazilian firefighters in the Federal District.19 Damacena et al recently reported that the prevalence of obesity was 10.99% among male firefighters (N=892) and that there was an overall high prevalence of overweight of 48.65%. Recent online survey of 2398 female career firefighters, one may consider female firefighters to have been successful in overcoming the occupational risks, such as a challenging nutritional environment, inconsistent schedules, limited time for fitness and the metabolic impact of shift work, that expose firefighters to increased risk of obesity.20 21 Although females still represent a small percentage of the police force and firefighting service, they are an integral part of the workforce and medical attention tailored to their specific needs may lead to better health outcomes.
A study that investigated the prevalence of metabolic syndrome and its risk factors among 10 348 Chinese police officers, found a high prevalence of hypertriglyceridaemia (40.8%), hypertension (38.2%) and abdominal obesity (25.7%).22 In our study, total cholesterol, triglycerides and glucose serum levels were above the ideal range in a large number of police officers and firefighters of both sexes and all age groups, but particularly among male police officers, suggesting a greater risk for metabolic diseases in this population. The analysis by age group revealed that the prevalence of obesity and high WC increased with age. The association of weight status and increased risk for atherosclerotic CVD was greater among older firefighters highlighting the aging-related increase in CVD risk, and that additional screening may be necessary among older firefighters to ensure that they are healthy enough to continue with the strenuous and stressful work of firefighting.23 Low levels of physical activity, unhealthy eating habits, demanding jobs, sleep disorders and stress, which are common during the course of a police officer’s or firefighter’s working life, may also help to explain the high prevalence of CVD in these populations.24 25
The duties performed by police officers and firefighters were associated with musculoskeletal disorders, particularly lower back pain, suggesting the need for preventive measures and ergonomic control programmes in the workplace.26–28
A study by Han et al compared the prevalence of chronic diseases among 860 221 public-sector workers in South Korea and found that police officers had a significantly higher risk for angina pectoris, acute myocardial infarction and cerebrovascular disease, while firefighters were more susceptible to post-traumatic stress disorders.1 In the present study, psychiatric diseases were more prevalent among police officers, particularly among women over 30 years old. The close correlation between occupational stress and mental illness has been observed in previous studies.29 30 The different instruments used for work-related psychological assessments in these studies prevent comparison of these findings. Although police officers and firefighters reported that they were receiving psychiatric treatment, this may not reflect the true prevalence of this health issue, as some participants may have omitted this information for fear of stigmatisation. The overall prevalence of smoking among police officers and firefighters was lower than in the general population in the state of Paraná which is 11.3%.31 The high prevalence of mental diseases, alcoholism, smoking and substance abuse among police officers and firefighters highlights not only the need for better medical assessment to ensure timely diagnosis and treatment, but also the need for public policies to promote preventive health measures.
The present study has certain limitations that must be mentioned. First, the cross-sectional design does not allow causal relationship between risk factors and comorbidities to be established. Nevertheless, the study population was recruited after a thorough medical assessment to ensure that healthy candidates are accepted in the police and firefighting services. In addition, the study population underwent an elective medical assessment and had not requested a medical examination of their own accord. The fact that only the serum levels of total cholesterol were measured, and not high-density lipoprotein or low-density lipoprotein, prevented a more detailed assessment of CVD risk.
Furthermore, the low number of women (11.7% of police officers and 7.2% of firefighters) underpowered the statistical analysis of this subgroup of workers. Lastly, the specific nature of the study population does not allow for the generalisation of the findings to other populations.
Long-term longitudinal studies are needed to understand the effect of the different activities carried out by police officers and firefighters on their health status. The identification of the causes of the deterioration of their health condition may allow for the adoption of prompt interventions to prevent disease evolution and promote both physical and mental health.
Our study highlights the need for regular medical assessments and follow-ups of police officers and firefighters, as well as for the promotion of preventive healthcare. These measures need to be offered earlier and more frequently for police officers. The health condition of female police officers and firefighters should not be neglected as they are exposed to the same work-related risk factors as men and, consequently, may also present a high prevalence of CVD risk factors and chronic diseases. It should be stressed that the police officers and firefighters included in this study were working under normal conditions and were unaware of their medical status. This scenario may adversely affect both the workers’ own health and the health of others when they are performing their duties.
Data availability statement
Data are available upon reasonable request. The data are available from the corresponding author upon reasonable request.
Patient consent for publication
This study involves human participants and was approved by Health Sciences Department, Federal University of Paraná Reference number 85780418.0.0000.0102. Participants gave informed consent to participate in the study before taking part.
The authors would like to thank all the police officers and firefighters who took part in this study.
Contributors ARdS was responsible for data collection, planning the study, analysis and interpretation of the data and writing the manuscript. MFKI and MO assisted with the planning of the study and interpretation of the results. MO was responsible for statistical analysis. FCB took part in the interpretation of the data and supervised the writing of the manuscript. All authors contributed to the editing and reviewing of this paper. ARdS is the guarantor.
Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.
Competing interests None declared.
Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.
Provenance and peer review Not commissioned; externally peer reviewed.

domain: health


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Title: Life Begins at Forty
Author: Jessie Blackwood
Cover Artist: Paul Richmond
Publisher: Dreamspinner Press
Buy Link: Buy Link Life Begins at Forty
Genre: M/M historical (1940’s) romance
Length: Novella (112 pages)
Rating: 3.5 stars out of 5
A guest review by Jenre
For fans of Ifan and Jack from Per Ardua, this continuation of their HEA will appeal but I found it a little bland and uninspiring, despite some nicely written scenes.
After months of physiotherapy, Group Captain Jack Ratigan has regained some of the mobility lost in plane crash at the end of World War II. But six years later, he still requires the care of his cousin’s butler, Ifan—who is also Jack’s secret lover. In an era when homosexuality is an imprisonable offence, they have to maintain the utmost discretion or risk prosecution.
Insecurities, outside attacks, and misunderstandings are close to tearing Jack and Ifan apart: Jack’s impending middle age, an act of violence in their house, a letter threatening the close-knit community Jack now calls home—and the detective inspector from another jurisdiction investigating a similar unsolved case. The threat of exposure is growing, and for their love to survive, Jack and Ifan must determine who their true friends are—and if they are strongest together or apart.
This story follows on from the end of Per Ardua which I reviewed here. Jack is approaching his 40th birthday and his cousin Bronwen has plans for a big party. Jack and Ifan are horrified when they received a poison pen letter, which tells them that they will be exposed as ‘sodomites’, especially when Bronwen is injured by a stone which has been deliberately thrown through their door window. The two men know that they have been very careful in public not to show the true nature of their feelings, but when a police inspector from Cardiff arrives to investigate it puts a strain on Jack’s nerves and his relationship with Ifan.
Whilst I didn’t dislike this story, it suffers from a lack of story or plot. The above summary basically tells you what happens. I was a bit baffled really as to why I was reading a bit of a non-story, no matter how nicely written, when there was scope within the first few pages for something a bit less bland and more exciting. When the story begins with the poison pen letter and the stone through the door window, I thought that maybe the story was going to take on a bit of mystery story line with our heroes helping the police by doing a bit of sleuthing on the side, but no, all the solving of the mystery is done elsewhere and off page. Instead the letters are used as a focus for a mini-breakdown by Jack and the cause of a minor bust-up between our heroes. The argument surfaces over nothing – a storm in a teacup really – and things get blown out of proportion, Ifan totally overreacts and the next day things settle down after Jack comes to his senses. That pretty much sums up all the complications in the story, the rest of the time is made up of lots of prettily written filler scenes where Jack writes his memoirs, takes part in a birthday surprise and has a party. Even the sudden, rather unrealistic, insertion of a dramatic scene at the end of the book, or the way that the book dealt a little with the problems of being gay in the 1940’s, didn’t really lift the story out of its blandness.
The above paragraph may lead to believe that I didn’t really like the book. That wasn’t the case at all. It was nice and pleasant to revisit the characters of Jack and Ifan. The author has a fluid way of writing that makes everything flow nicely and I read the novella quickly. Jack is an easy character to like and even his minor breakdown was handled in a sensitive fashion. I liked that Ifan was starting to stand up for himself a bit and although there’s not that much change in their relationship dynamics, Jack begins to understand some of Ifan’s needs that he’d not seen before. the characters and the dialogue fitted well with the time period and the whole book was firmly grounded in the historical setting.
This really is a story for those who loved the previous book and to understand how Jack and Ifan come to love each other you need to read Per Ardua first. For me, it was a pleasant interlude but nothing that left me with a lasting impression of the story which is why I’ve gone for a rating in the middle of three stars.

domain: library


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Mental health blog Furious Seasons has just alerted me to some recent revelations about conflicts of interest in psychiatry. More than half of the new committee members in charge of the next edition of the psychiatrists’ diagnostic ‘bible’, the Diagnostic and Statistical Manual of Mental Disorders (DSM), have ties to the drug industry.
Furthermore, an article for Slate reveals that a supposedly ‘independent’ NPR national radio show on the safety of antidepressants had three guests, a host and the production company, all of whom received money from drug companies.
While the financial interests of the DSM committee must be declared, drug company links were not revealed in relation to the radio programme and the production company seem to be being evasive about discussing the situation.
Apparently, the committee for the new DSM (the DSM-V) largely parallels the situation with the previous version, where over half of the members had drug company ties.
Because of the way the US health system woks, US health insurers tend only to pay for treatments when a specific diagnosis has been made, so it is in the drug companies’ interest to influence the classification of mental illnesses to make prescribing more likely.
However, there are rumours that the insurance industry and getting rather fed up of having to pay out on potentially drug company influenced diagnoses, and are considering funding research of their own into the validity of diagnoses to counter this trend.
The two news items mentioned above seem to be the work of the Centre for Science in the Public Interest that campaigns for transparency in science education and policy.
At the end of the Slate article, the organisation note that they have a list of leading scientists who do not have links to industry. Journalists are welcome to contact them if they want a source free of potential biases.


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August 8, 2007 – Nearly a year after putting some of its lithography metrology patents up for auction, Nova Measuring Instruments says it has signed a >$1 million deal with an unnamed “top-10 semiconductor manufacturer” for a license covering use of its technologies for integrated metrology and integrated process control before and during the photolithography manufacturing step.
The deal, which the company says follows response from several semiconductor manufacturing companies interested in the patents, “is clear evidence of the strength of the IP and its importance to semiconductor manufacturing,” said Gabi Seligsohn, president/CEO of Nova, in a statement, adding that the company will “continue discussions with additional companies interested in licensing or purchasing these patents.”
In an e-mail exchange, Seligsohn clarified to WaferNEWS that Nova is still “in active discussions” regarding both licensing and outright sale of the patents, but that this first license was crafted to be “in perpetuity” to guard against possible strategic maneuverings, “such that the possible new proprietor will have no ability to enforce the patents against a licensee.”
In Sept. 2006 the company started soliciting bids from approximately 100 companies to license six of its patents relating to use of a lithography tool with integrated metrology, including bids for outright ownership of the technology. Four of the patents relate to a lithography track with integrated optical measurement capability, used with overlay registration, critical dimensions, and macrodefect inspection. Two others, from the company’s advanced process control group, are for methods for photolithographic processing involving making a spectrophotometric measurement and using it to influence the processing time, focus or exposure of a processing tool.

domain: law


text:
Producer strove to add all-new gameplay loop to Madden diehards’ favorite mode Madden NFL 22’s latest patch is a big one.
Scouting should, basically, make the NFL Draft a gameplay cycle throughout the whole season, similar to how Draft news is a yearlong story IRL.
“Draft stories” will be a part of news feed Madden NFL 22 generates about the season in progress.
The Scouting update starts by extending the staff management tasks that Franchise introduced this year, with the hiring and firing of assistant coaches.
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domain: news


KATHMANDU: As many as 2,253 new cases of coronavirus infection have been detected in the last 24 hours taking the nationwide tally to 86,823.
As such, the number of total active cases in the country as of today is 22,219.
Similarly, 4,203 people are placed in quarantine facilities in the various parts across the country.
The Health Ministry reported a good number of 1,329 recoveries in the last 24-hours. So far 64,069 people have been discharged from the health facilities following an improvement in their condition.
Seven new fatalities were reported on Saturday, taking the tally to 535.
On Saturday, Nepal’s coronavirus tally stood at 84,570 with 2,120 new infections.


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Tom Krattenmaker is part of a growing conversation centered at Yale University that acknowledges - and seeks to address - the abiding need for meaning and inspiration in postreligious America. What, they ask, gives a life meaning? What constitutes a life well led? In Confessions of a Secular Jesus Follower, Krattenmaker shares his surprising conclusion about where input and inspiration might best be found: in the figure of Jesus.
"this should be required reading for life"


Earlier this year, Waxwork Records released a 40th anniversary vinyl reissue of the score to George A. Romero’s Dawn of the Dead. With the reissue, Italian prog legends Goblin saw all 17 of their tracks for the film released on vinyl for the first time. Now, the band itself has returned for the true Dead heads, releasing a 40th anniversary release that’s distinctly their own.
(Read: 10 Essential Horror Movie Scores)
The commemorative studio album, Fearless (37513 Zombie Ave), includes “reimagined versions” of the best tunes from the iconic soundtrack, as well as new arrangements of two tracks from their also-seminal Suspiria score. It will be a merry, bloody Christmas indeed, as the album is slated to arrive this December 24th on both vinyl and CD.
Pre-orders are ongoing here.


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Woman Becomes the First Known Case of a Person Urinating Alcohol
Pittsburgh woman has become the first person to be diagnosed with a rare syndrome which causes alcohol to brew in the bladder due to the fermentation of yeast and sugar.
The anonymous patient, 61, has diabetes and liver cirrhosis and was on the donor waiting list for a liver transplant. However, she was taken off the list and referred to alcohol abuse treatment instead.
Doctors thought her problem was alcohol addiction; however, the patient denied ever drinking alcohol.
The doctors were understandably skeptical since the repeated urine tests for alcohol showed positive. As the next step, the doctors ran a blood test which surprisingly showed no alcohol in her blood.
In order to crack the mystery, Kenichi Tamama of the University of Pittsburgh Medical Center in Pennysylvania conducted some basic tests and saw that the woman’s urine contained yeast.
It is not unusual for the urine to have yeast. However, it is also known that diabetes causes a high amount of sugar in the urine. In order to see these two components in action, Tamama ran tests on her urine and discovered that the yeast was fermenting sugar in her bladder.
The doctors named the condition “urinary auto-brewery syndrome”. This yeast, identified as Candida glabrata, is normally found in the body and it is related to what breweries use.
The process that resulted in alcohol is almost exactly the same as the one used by beer makers. Needless to say, this is extraordinary since now we know that the same process can also happen in the human body.
All in all, peeing alcohol might not be that big of a deal after all. According to Tamama, it doesn't affect the patient's health. Efforts were made to eliminate the yeast, but they weren't successful due to the patient’s diabetes.
According to the paper, the patient was reconsidered for liver transplantation; however, her whereabouts are unknown at the moment.
The findings were reported in Annals of Internal Medicine.
This summer saw a surge in travelers, and their lost or mishandled luggage. Can AI save the mounting baggage crisis?


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Fifty years after English referee Kenneth Aston introduced the red and yellow cards to football, refereeing still remains one of the controversial aspects of the game.
Like the knight in shining armor, technology has come to save the day again in form of the Video Assistant Referee (VAR).
The VAR technology uses cameras and the help of an Assistant Video Assistant Review (AVAR) to review play in a Video Operation Room(VOR).
This week, the International Football Association Board(IFAB) unanimously greenlighted the technology in its 132nd annual general meeting.
The first live trial of the Video Assisted Referee (VAR) happened in a United Soccer League (USL) march in August 2016.
Since then, it has been used in the Bundesliga, Seria A, and Copa Libertadores leagues.
It was most recently trialled in an FA cup clash between Brighton and Crystal Palace.
Approval by the IFAB means the VAR will most likely be used in the 2018 FIFA World Cup. But a FIFA Council meeting in Colombia on the 16th of March will make a final decision on the matter.
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MoD announces winner of robot challenge
The Ministry of Defence has announced the winner of its Grand Challenge competition to produce a vehicle capable of identifying threats encountered by UK troops on current operations.
Team Stellar beat off six other hopefuls, whittled down from 23, in the final competition set in Copehill Down, a military training facility on Salisbury Plain.
Stellar's Saturn unmanned ground vehicle works with two other types of unmanned aerial vehicle to gather data about the battlefield from a variety of perspectives. The system is managed by a control station that combines visual, thermal and radar data.
Stellar Research has designed a system that relies on three separate, autonomous platforms: an unmanned ground vehicle, up to six medium-level unmanned aerial vehicles (UAVs) and one high-level UAV. The intention is to build a system that can be used to analyse the battlefield from a macro and micro level.
For more photos, see the full story on sister site silicon.com