NCR World headquarters in Midtown, Atlanta is LEED Platinum certified and uses a newly installed unique water system designed and supplied by Ecovie.
Ecovie worked with engineering firm Integral Group to develop the overall design. Georgia Water Tanks, the local Georgia representative for Ecovie implemented the design and worked with mechanical contractor McKenney’s through installation and start up. Ecovie and Georgia Water Tanks supplied equipment for rainwater and nuisance groundwater collection working closely with Integral Group on conceptual design and layout while providing and integrating equipment from our key suppliers Intewa, Aqua and QuantumFlo. This is a two-phase project with the Phase 1 tower complete and the Phase 2 tower coming on line soon.
From the 48,000 square foot Phase 1 tower, rainwater is directed through Intewa PURAIN Filters for storage in the 48,000 gallon concrete cistern in the building lower utility area. As water is needed for cooling tower make-up, irrigation, and toilet flushing, water is transferred through sediment filtration and UV disinfection as per Georgia State plumbing code to a 3,000 gallon buffer tank. The system also captured condensate from the air conditioners which can add up to thousands of gallons or clean water every day in the summer. Soon Phase 2 will be adding another 30,000 square feet of catchment.
One of the more unique features of this water system is that foundation water is collected for use on the property. This in itself is not unique, but environmental tests on-site showed that the water was contaminated with several VOC’s (volatile organic compounds) and would require treatment before discharge. Typically, in such a case, treated water would be discharged and not used. In this case, we are treating up to 20,000 gallons a day for non-potable use on site depending on how much groundwater is present. The purification method used is an advanced oxidation process (AOP) with high-intensity UV and carbon filtration to bring the water into environmental compliance.
Building Water Use:
With cooling tower make up, toilet flushing, and irrigation projected water demand is over 10,000 gallons per day in winter and over 20,000 gallons per day in the summer. Collected water can supply almost all of the 5+ million gallons needed for these uses. The triplex, 330 GPM, QuantumFlo booster system assures all demands are met. Treatment flow of 12 GPM to the 3,000 gallon buffer tank is plenty big enough to keep up with this demand.
With Phase 2, collection area from the second tower is being added as well as water supply for non-potable use. An additional treated groundwater cistern has been added to expand on-site water use.


Today the European Parliament has finally stood on the side of women and showed governments that are undermining women’s rights that we no longer accept this.
Despite enormous mobilisation from anti-choice and anti-gender movements and last-minute manoeuvres by the EPP, the plenary has adopted the progressive resolution recognising sexual and reproductive health and rights (SRHR) as fundamental human rights. Drafted by S&D MEP Fred Matić, the text calls for guaranteed access to SRHRs in the EU without discrimination, including to reliable sex education, fertility treatment, modern contraceptives and safe and legal abortion. It also urges the European Commission to appoint an EU special envoy for SRHRs, to protect them at the EU level.
Fred Matić, S&D MEP and the European Parliament’s negotiator on the sexual and reproductive health and rights, said:
“A historic day! I am proud that the majority of our house recognises that in the 21st century we must no longer treat women as second-class citizens, but respect their feelings and freedoms, and make sure all women across Europe get the access to healthcare they deserve.
“Contrary to what the far right has said, I am not advocating abortion. What my report is saying is that every woman has the right to choose. The number of abortions performed is similar in countries where abortion is legal and in countries where it is not. But abortions performed clandestinely are dangerous for women’s health, often leading to fatal consequences. That is why we are calling for abortion to be decriminalised in all member states.
“But there is much more than abortion. We call for appropriate sex education for all children in primary and secondary schools, and for access to fertility treatment without discrimination, especially for single women and female same-sex couples.
“Anti-choice and anti-gender movements have spared no effort to prevent the adoption of my report. They spread disinformation about it, launched a petition against it and even went so far as to send little plastic foetuses to many MEPs. But they did not intimidate us! I hope that one day they also will walk off the edge of their flat earth.”
S&D spokesperson on women’s rights and gender equality Maria Noichl MEP added:
“I am glad that despite numerous attempts to block this report in the last minute, not only by the far right but also the EPP, it was adopted. We have been waiting for almost a decade to send this clear message from the European Parliament that we are putting women and their rights centre stage.
“We are making it clear: sexual and reproductive rights, meaning access to abortion, but also sexual education, contraception, fertility treatment, health services during pregnancy and birth, and the treatment of related diseases, must remain rights in the entire EU. They cannot be withdrawn by member states. We trust women themselves to make the right decisions and we want to facilitate this through information, education and support.
“During the negotiations and shortly before the vote, it became obvious why this report is so urgently needed. The opposition against women has increased. Fake news and flimsy excuses are used to try to undo years of progress and to send women back to the home. To all governments and organisations in the EU that are trying to undermine women’s rights, we are saying today: we will not allow this.”
Source S&D: https://www.socialistsanddemocrats.eu/newsroom/european-parliament-says-womens-rights-and-srhr-are-human-rights-and-calls-decriminalise


text:
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The bodice is black and features silver appliques with diamonds and clear stones. There is a very deep nude insert in the bodice and black lace sleeves.
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domain: design


text:
Electronic muscle stimulation is a technique that is widely used by elite athletes all over the world. It has been shown to be effective and successful toward toning and building muscle mass. This is why devices have been developed to accomplish this type of workout.
The way it works is fairly straight forward. The electrical impulses control the muscles via the motor nerves. Programming these impulses allows users to target their workouts for power, endurance, or resistance. Here, powerful contractions can be received without the pain of a traditional workout.
A microprocessor controls the work of the muscles, while adhering to the principles of traditional training physiology. Not only does electrical muscle stimulation help build muscles, but it also encompasses the toning and shaping that improves overall physical fitness. While it is not meant to take the place of a traditional workout, it can help supplement it and add to the benefits it can offer. It can also be a viable means of exercise for those who are unable to participate in the more traditional methods of exercise due to health problems or joint and muscle restrictions. It is advisable to consult a doctor before using an electronic muscle stimulation machine if this is the case.
Electronic muscle stimulation is effective for fitness training, as well as for muscle rehabilitation. It helps build endurance so that the muscles may be used for a longer period of time. The resistance it also provides, will assist in expanding the range of muscle movement and help to promote strength. The recovery process is also shortened by this method, as the patient is able to exercise in a non-restricted environment without too much strain or tension being place on the body.
This type of equipment has been successfully used in the physical therapy and sports medicine fields for many years, and has nothing to do with the various other pieces of exercise equipment that are commonly found on the market. As with any other workout, there are stages that must be accomplished in order to reap the full benefits of this type of program. Many machines also come with DVD-based training material that is designed to walk the user through the various steps involved in the process. Here, users will also be able to create an individualized training program that will best fit their individual needs. Before trying electronic muscle stimulation, it is advisable to check with a physician to make sure this is right for you.

domain: sports


text:
Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.
25 Related JoVE Articles!
The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects
Institutions: University and Bern University Hospital (Inselspital), Kantonsspital Aarau, Boston Children's Hospital, Boston Children's Hospital, University and Bern University Hospital (Inselspital), University Hospital Cologne, Länggasse Bern.
Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.
Medicine, Issue 92,
Subarachnoid hemorrhage, animal models, rabbit, extracorporeal blood shunt, early brain injury, delayed cerebral vasospasm, microsurgery.
Videomorphometric Analysis of Hypoxic Pulmonary Vasoconstriction of Intra-pulmonary Arteries Using Murine Precision Cut Lung Slices
Acute alveolar hypoxia causes pulmonary vasoconstriction (HPV) - also known as von Euler-Liljestrand mechanism - which serves to match lung perfusion to ventilation. Up to now, the underlying mechanisms are not fully understood. The major vascular segment contributing to HPV is the intra-acinar artery. This vessel section is responsible for the blood supply of an individual acinus, which is defined as the portion of lung distal to a terminal bronchiole. Intra-acinar arteries are mostly located in that part of the lung that cannot be selectively reached by a number of commonly used techniques such as measurement of the pulmonary artery pressure in isolated perfused lungs or force recordings from dissected proximal pulmonary artery segments1,2
. The analysis of subpleural vessels by real-time confocal laser scanning luminescence microscopy is limited to vessels with up to 50 µm in diameter3
We provide a technique to study HPV of murine intra-pulmonary arteries in the range of 20-100 µm inner diameters. It is based on the videomorphometric analysis of cross-sectioned arteries in precision cut lung slices (PCLS). This method allows the quantitative measurement of vasoreactivity of small intra-acinar
arteries with inner diameter between 20-40 µm which are located at gussets of alveolar septa next to alveolar ducts and of larger pre-acinar
arteries with inner diameters between 40-100 µm which run adjacent to bronchi and bronchioles. In contrast to real-time imaging of subpleural vessels in anesthetized and ventilated mice, videomorphometric analysis of PCLS occurs under conditions free of shear stress. In our experimental model both arterial segments exhibit a monophasic HPV when exposed to medium gassed with 1% O2
and the response fades after 30-40 min at hypoxia.
Medicine, Issue 83, Hypoxic pulmonary vasoconstriction, murine lungs, precision cut lung slices, intra-pulmonary, pre- and intra-acinar arteries, videomorphometry
The Helsinki Rat Microsurgical Sidewall Aneurysm Model
Institutions: University of Helsinki, Helsinki, Finland.
Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced into clinical practice. Furthermore, experimental models are needed to elucidate the complex aneurysm biology leading to rupture of saccular aneurysms.
Several different kinds of experimental models for saccular aneurysms have been established in different species. Many of them, however, require special skills, expensive equipment, or special environments, which limits their widespread use. A simple, robust, and inexpensive experimental model is needed as a standardized tool that can be used in a standardized manner in various institutions.
The microsurgical rat abdominal aortic sidewall aneurysm model combines the possibility to study both novel endovascular treatment strategies and the molecular basis of aneurysm biology in a standardized and inexpensive manner. Standardized grafts by means of shape, size, and geometry are harvested from a donor rat's descending thoracic aorta and then transplanted to a syngenic recipient rat. The aneurysms are sutured end-to-side with continuous or interrupted 9-0 nylon sutures to the infrarenal abdominal aorta.
We present step-by-step procedural instructions, information on necessary equipment, and discuss important anatomical and surgical details for successful microsurgical creation of an abdominal aortic sidewall aneurysm in the rat.
Medicine, Issue 92, Animal models, Rat, Sidewall saccular aneurysms, Microsurgery, aneurysm wall
Assessing Cerebral Autoregulation via Oscillatory Lower Body Negative Pressure and Projection Pursuit Regression
Institutions: Harvard Medical School, Spaulding Hospital Cambridge.
The process by which cerebral perfusion is maintained constant over a wide range of systemic pressures is known as “cerebral autoregulation.” Effective dampening of flow against pressure changes occurs over periods as short as ~15 sec and becomes progressively greater over longer time periods. Thus, slower changes in blood pressure are effectively blunted and faster changes or fluctuations pass through to cerebral blood flow relatively unaffected. The primary difficulty in characterizing the frequency dependence of cerebral autoregulation is the lack of prominent spontaneous fluctuations in arterial pressure around the frequencies of interest (less than ~0.07 Hz or ~15 sec). Oscillatory lower body negative pressure (OLBNP) can be employed to generate oscillations in central venous return that result in arterial pressure fluctuations at the frequency of OLBNP. Moreover, Projection Pursuit Regression (PPR) provides a nonparametric method to characterize nonlinear relations inherent in the system without a priori
assumptions and reveals the characteristic non-linearity of cerebral autoregulation. OLBNP generates larger fluctuations in arterial pressure as the frequency of negative pressure oscillations become slower; however, fluctuations in cerebral blood flow become progressively lesser. Hence, the PPR shows an increasingly more prominent autoregulatory region at OLBNP frequencies of 0.05 Hz and below (20 sec cycles). The goal of this approach it to allow laboratory-based determination of the characteristic nonlinear relationship between pressure and cerebral flow and could provide unique insight to integrated cerebrovascular control as well as to physiological alterations underlying impaired cerebral autoregulation (e.g.
, after traumatic brain injury, stroke, etc.
Medicine, Issue 94, cerebral blood flow, lower body negative pressure, autoregulation, sympathetic nervous system
Myocardial Infarction and Functional Outcome Assessment in Pigs
Institutions: University Medical Center Utrecht, Interuniversity Cardiology Institute of the Netherlands.
Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e.
directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.
Medicine, Issue 86, myocardial infarction (MI), AMI, large animal model, pig, translational medicine, ischemic heart disease
Setting-up an In Vitro Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated Transport
Institutions: VECT-HORUS SAS, CNRS, NICN UMR 7259.
The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue. Our aim was to develop and characterize a highly reproducible rat syngeneic in vitro
model of the BBB using co-cultures of primary rat brain endothelial cells (RBEC) and astrocytes to study receptors involved in transcytosis across the endothelial cell monolayer. Astrocytes were isolated by mechanical dissection following trypsin digestion and were frozen for later co-culture. RBEC were isolated from 5-week-old rat cortices. The brains were cleaned of meninges and white matter, and mechanically dissociated following enzymatic digestion. Thereafter, the tissue homogenate was centrifuged in bovine serum albumin to separate vessel fragments from nervous tissue. The vessel fragments underwent a second enzymatic digestion to free endothelial cells from their extracellular matrix. The remaining contaminating cells such as pericytes were further eliminated by plating the microvessel fragments in puromycin-containing medium. They were then passaged onto filters for co-culture with astrocytes grown on the bottom of the wells. RBEC expressed high levels of tight junction (TJ) proteins such as occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. The transendothelial electrical resistance (TEER) of brain endothelial monolayers, indicating the tightness of TJs reached 300 ohm·cm2
on average. The endothelial permeability coefficients (Pe) for lucifer yellow (LY) was highly reproducible with an average of 0.26 ± 0.11 x 10-3
cm/min. Brain endothelial cells organized in monolayers expressed the efflux transporter P-glycoprotein (P-gp), showed a polarized transport of rhodamine 123, a ligand for P-gp, and showed specific transport of transferrin-Cy3 and DiILDL across the endothelial cell monolayer. In conclusion, we provide a protocol for setting up an in vitro
BBB model that is highly reproducible due to the quality assurance methods, and that is suitable for research on BBB transporters and receptors.
Medicine, Issue 88, rat brain endothelial cells (RBEC), mouse, spinal cord, tight junction (TJ), receptor-mediated transport (RMT), low density lipoprotein (LDL), LDLR, transferrin, TfR, P-glycoprotein (P-gp), transendothelial electrical resistance (TEER),
Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery
Institutions: University Hospital Munich, Munich Cluster for Systems Neurology (SyNergy), University Heidelberg, Charing Cross Hospital.
Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called “coagulation model”. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the “coagulation model” and report histological and functional analysis tools.
Medicine, Issue 89, stroke, brain ischemia, animal model, middle cerebral artery, electrocoagulation
Analysis of Tubular Membrane Networks in Cardiac Myocytes from Atria and Ventricles
Institutions: Heart Research Center Goettingen, University Medical Center Goettingen, German Center for Cardiovascular Research (DZHK) partner site Goettingen, University of Maryland School of Medicine.
In cardiac myocytes a complex network of membrane tubules - the transverse-axial tubule system (TATS) - controls deep intracellular signaling functions. While the outer surface membrane and associated TATS membrane components appear to be continuous, there are substantial differences in lipid and protein content. In ventricular myocytes (VMs), certain TATS components are highly abundant contributing to rectilinear tubule networks and regular branching 3D architectures. It is thought that peripheral TATS components propagate action potentials from the cell surface to thousands of remote intracellular sarcoendoplasmic reticulum (SER) membrane contact domains, thereby activating intracellular Ca2+
release units (CRUs). In contrast to VMs, the organization and functional role of TATS membranes in atrial myocytes (AMs) is significantly different and much less understood. Taken together, quantitative structural characterization of TATS membrane networks in healthy and diseased myocytes is an essential prerequisite towards better understanding of functional plasticity and pathophysiological reorganization. Here, we present a strategic combination of protocols for direct quantitative analysis of TATS membrane networks in living VMs and AMs. For this, we accompany primary cell isolations of mouse VMs and/or AMs with critical quality control steps and direct membrane staining protocols for fluorescence imaging of TATS membranes. Using an optimized workflow for confocal or superresolution TATS image processing, binarized and skeletonized data are generated for quantitative analysis of the TATS network and its components. Unlike previously published indirect regional aggregate image analysis strategies, our protocols enable direct characterization of specific components and derive complex physiological properties of TATS membrane networks in living myocytes with high throughput and open access software tools. In summary, the combined protocol strategy can be readily applied for quantitative TATS network studies during physiological myocyte adaptation or disease changes, comparison of different cardiac or skeletal muscle cell types, phenotyping of transgenic models, and pharmacological or therapeutic interventions.
Bioengineering, Issue 92, cardiac myocyte, atria, ventricle, heart, primary cell isolation, fluorescence microscopy, membrane tubule, transverse-axial tubule system, image analysis, image processing, T-tubule, collagenase
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
Ultrasound Assessment of Endothelial-Dependent Flow-Mediated Vasodilation of the Brachial Artery in Clinical Research
Institutions: University of California, San Francisco, Veterans Affairs Medical Center, San Francisco, Veterans Affairs Medical Center, San Francisco.
The vascular endothelium is a monolayer of cells that cover the interior of blood vessels and provide both structural and functional roles. The endothelium acts as a barrier, preventing leukocyte adhesion and aggregation, as well as controlling permeability to plasma components. Functionally, the endothelium affects vessel tone.
Endothelial dysfunction is an imbalance between the chemical species which regulate vessel tone, thombroresistance, cellular proliferation and mitosis. It is the first step in atherosclerosis and is associated with coronary artery disease, peripheral artery disease, heart failure, hypertension, and hyperlipidemia.
The first demonstration of endothelial dysfunction involved direct infusion of acetylcholine and quantitative coronary angiography. Acetylcholine binds to muscarinic receptors on the endothelial cell surface, leading to an increase of intracellular calcium and increased nitric oxide (NO) production. In subjects with an intact endothelium, vasodilation was observed while subjects with endothelial damage experienced paradoxical vasoconstriction.
There exists a non-invasive, in vivo
method for measuring endothelial function in peripheral arteries using high-resolution B-mode ultrasound. The endothelial function of peripheral arteries is closely related to coronary artery function. This technique measures the percent diameter change in the brachial artery during a period of reactive hyperemia following limb ischemia.
This technique, known as endothelium-dependent, flow-mediated vasodilation (FMD) has value in clinical research settings. However, a number of physiological and technical issues can affect the accuracy of the results and appropriate guidelines for the technique have been published. Despite the guidelines, FMD remains heavily operator dependent and presents a steep learning curve. This article presents a standardized method for measuring FMD in the brachial artery on the upper arm and offers suggestions to reduce intra-operator variability.
Medicine, Issue 92, endothelial function, endothelial dysfunction, brachial artery, peripheral artery disease, ultrasound, vascular, endothelium, cardiovascular disease.
Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model
Institutions: University Medical Center Utrecht, Vascular Connect b.v., University Medical Center Utrecht, University Medical Center Utrecht.
To simplify and facilitate beating heart (i.e.,
off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the Trinity Clip, is developed based on the excimer laser-assisted nonocclusive anastomosis technique. The Trinity Clip connector enables simplified, sutureless, and nonocclusive connection of the graft to the coronary artery, and an excimer laser catheter laser-punches the opening of the anastomosis. Consequently, owing to the complete nonocclusive anastomosis construction, coronary conditioning (i.e.,
occluding or shunting) is not necessary, in contrast to the conventional anastomotic technique, hence simplifying the off-pump bypass procedure. Prior to clinical application in coronary artery bypass grafting, the safety and quality of this novel connector will be evaluated in a long-term experimental porcine off-pump coronary artery bypass (OPCAB) study. In this paper, we describe how to evaluate the coronary anastomosis in the porcine OPCAB model using various techniques to assess its quality. Representative results are summarized and visually demonstrated.
Medicine, Issue 93, Anastomosis, coronary, anastomotic connector, anastomotic coupler, excimer laser-assisted nonocclusive anastomosis (ELANA), coronary artery bypass graft (CABG), off-pump coronary artery bypass (OPCAB), beating heart surgery, excimer laser, porcine model, experimental, medical device
Isolation of Microvascular Endothelial Tubes from Mouse Resistance Arteries
Institutions: University of Missouri, Dalton Cardiovascular Research Center.
The control of blood flow by the resistance vasculature regulates the supply of oxygen and nutrients concomitant with the removal of metabolic by-products, as exemplified by exercising skeletal muscle. Endothelial cells (ECs) line the intima of all resistance vessels and serve a key role in controlling diameter (e.g.
endothelium-dependent vasodilation) and, thereby, the magnitude and distribution of tissue blood flow. The regulation of vascular resistance by ECs is effected by intracellular Ca2+
signaling, which leads to production of diffusible autacoids (e.g.
nitric oxide and arachidonic acid metabolites)1-3
that elicit smooth muscle cell relaxation. Thus understanding the dynamics of endothelial Ca2+
signaling is a key step towards understanding mechanisms governing blood flow control. Isolating endothelial tubes eliminates confounding variables associated with blood in the vessel lumen and with surrounding smooth muscle cells and perivascular nerves, which otherwise influence EC structure and function. Here we present the isolation of endothelial tubes from the superior epigastric artery (SEA) using a protocol optimized for this vessel.
To isolate endothelial tubes from an anesthetized mouse, the SEA is ligated in situ
to maintain blood within the vessel lumen (to facilitate visualizing it during dissection), and the entire sheet of abdominal muscle is excised. The SEA is dissected free from surrounding skeletal muscle fibers and connective tissue, blood is flushed from the lumen, and mild enzymatic digestion is performed to enable removal of adventitia, nerves and smooth muscle cells using gentle trituration. These freshly-isolated preparations of intact endothelium retain their native morphology, with individual ECs remaining functionally coupled to one another, able to transfer chemical and electrical signals intercellularly through gap junctions6,7
. In addition to providing new insight into calcium signaling and membrane biophysics, these preparations enable molecular studies of gene expression and protein localization within native microvascular endothelium.
Basic Protocol, Issue 81, endothelial tubes, microcirculation, calcium signaling, resistance vasculature, Confocal microscopy
Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling
Institutions: University of Virginia, California Polytechnic State University, University of Virginia, Johns Hopkins University.
The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3
. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 μm). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.
Biomedical Engineering, Issue 73, Medicine, Anatomy, Physiology, Surgery, Immunology, Hematology, Microvessels, Capillaries, Arterioles, Venules, Vascular Diseases, Ischemia, spinotrapezius, peripheral vascular disease, functional vasodilation, arteriolar ligation, vessels, circulation, confocal microscopy, animal model
Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion
Institutions: University of Milano Bicocca.
The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.
Medicine, Issue 72, Neuroscience, Neurobiology, Biomedical Engineering, Anatomy, Physiology, Surgery, Brain Ischemia, Stroke, Hemodynamics, middle cerebral artery occlusion, cerebral hemodynamics, perfusion monitoring, Laser Doppler, intracranial collaterals, ischemic penumbra, rat, animal model
Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography
Institutions: North Carolina Central University, Durham, North Carolina Central University, Durham, Wake Forest University School of Medicine.
Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance. These small vessels of between 100-400 μm in diameter function primarily in directing blood flow to various organs according to the overall requirements of the body. The rat mesenteric artery has a diameter greater than 100 μm. The myography technique, first described by Mulvay and Halpern1
, was based on the method proposed by Bevan and Osher2
. The technique provides information about small vessels under isometric conditions, where substantial shortening of the muscle preparation is prevented. Since force production and sensitivity of vessels to different agonists is dependent on the extent of stretch, according to active tension-length relation, it is essential to conduct contraction studies under isometric conditions to prevent compliance of the mounting wires. Stainless steel wires are preferred to tungsten wires because of oxidation of the latter, which affects recorded responses3
.The technique allows for the comparison of agonist-induced contractions of mounted vessels to obtain evidence for normal function of vascular smooth muscle cell receptors.
We have shown in several studies that isolated mesenteric arteries that are contracted with phenylyephrine relax upon addition of cumulative concentrations of extracellular calcium (Ca2+e
). The findings led us to conclude that perivascular sensory nerves, which express the G protein-coupled Ca2+
-sensing receptor (CaR), mediate this vasorelaxation response. Using an automated wire myography method, we show here that mesenteric arteries from Wistar, Dahl salt-sensitive(DS) and Dahl salt-resistant (DR) rats respond differently to Ca2+e
. Tissues from Wistar rats showed higher Ca2+
-sensitivity compared to those from DR and DS. Reduced CaR expression in mesenteric arteries from DS rats correlates with reduced Ca2+e
-induced relaxation of isolated, pre-contracted arteries. The data suggest that the CaR is required for relaxation of mesenteric arteries under increased adrenergic tone, as occurs in hypertension, and indicate an inherent defect in the CaR signaling pathway in Dahl animals, which is much more severe in DS.
The method is useful in determining vascular reactivity ex vivo
in mesenteric resistance arteries and similar small blood vessels and comparisons between different agonists and/or antagonists can be easily and consistently assessed side-by-side6,7,8
Medicine, Issue 55, cardiovascular, resistant arteries, contraction, relaxation, myography
Imaging of Estrogen Receptor-α in Rat Pial Arterioles using a Digital Immunofluorescent Microscope
Institutions: Uniformed Services University of the Health Sciences.
Many of estrogen's effects on vascular reactivity are mediated through interaction with estrogen receptors 1, 2, 3
. Although two sub-types exist (estrogen receptor -α and β),estrogen receptor-α has been identified in both the smooth muscle and in endothelial cells of pial arterial segments using fluorescent staining combined with confocal laser scanning microscopy 4
. Furthermore, ER-α is located in the nuclei and in the cytoplasm of rat basilar arteries 5
. The receptors are abundant and fluoresce brightly, but clear visualization of discrete groups of receptors is difficult likely due to the numbers located in many cell layers of pial vessel segments. Additionally, many reports using immunohistochemical techniques paired with confocal microscopy poorly detail the requirements critical for reproduction of experiments 6
. Our purpose for this article is to describe a simple technique to optimize the staining and visualization of ER-α using cross-sectional slices of pial arterioles obtain from female rat brains. We first perfuse rats with Evans blue dye to easily identify surface pial arteries which we isolate under a dissecting microscope. Use of a cryostat to slice 8 μm cross sections of the arteries allows us to obtain thin vessel sections so that different vessel planes are more clearly visualized. Cutting across the vessel rather than use of a small vessel segment has the advantage of easier viewing of the endothelial and smooth muscle layers. In addition, use of a digital immunofluorescent microscope with extended depth software produces clear images of ten to twelve different vessel planes and is less costly than use of a confocal laser scanning microscope.
Molecular Biology, Issue 57, digital immunofluorescent microscopy, brain, estrogen receptor-α, cerebral microvasculature, rat, immunohistochemistry
The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity
Institutions: West Virginia University .
The isolated microvessel preparation is an ex vivo
preparation that allows for examination of the different contributions of factors that control vessel diameter, and thus, perfusion resistance1-5
. This is a classic experimental preparation that was, in large measure, initially described by Uchida et al.15
several decades ago. This initial description provided the basis for the techniques that was extensively modified and enhanced, primarily in the laboratory of Dr. Brian Duling at the University of Virginia6-8
, and we present a current approach in the following pages. This preparation will specifically refer to the gracilis arteriole in a rat as the microvessel of choice, but the basic preparation can readily be applied to vessels isolated from nearly any other tissue or organ across species9-13
. Mechanical (i.e., dimensional) changes in the isolated microvessels can easily be evaluated in response to a broad array of physiological (e.g., hypoxia, intravascular pressure, or shear) or pharmacological challenges, and can provide insight into mechanistic elements comprising integrated responses in an intact, although ex vivo
, tissue. The significance of this method is that it allows for facile manipulation of the influences on the integrated regulation of microvessel diameter, while also allowing for the control of many of the contributions from other sources, including intravascular pressure (myogenic), autonomic innervation, hemodynamic (e.g., shear stress), endothelial dependent or independent stimuli, hormonal, and parenchymal influences, to provide a partial list. Under appropriate experimental conditions and with appropriate goals, this can serve as an advantage over in vivo
or in situ
tissue/organ preparations, which do not readily allow for the facile control of broader systemic variables.
The major limitation of this preparation is essentially the consequence of its strengths. By definition, the behavior of these vessels is being studied under conditions where many of the most significant contributors to the regulation of vascular resistance have been removed, including neural, humoral, metabolic, etc. As such, the investigator is cautioned to avoid over-interpretation and extrapolation of the data that are collected utilizing this preparation. The other significant area of concern with regard to this preparation is that it can be very easy to damage cellular components such as the endothelial lining or the vascular smooth muscle, such that variable source of error can be introduced. It is strongly recommended that the individual investigator utilize appropriate measurements to ensure the quality of the preparation, both at the initiation of the experiment and periodically throughout the course of a protocol.
Physiology, Issue 62, isolated microvessel preparation, skeletal muscle arterioles, resistance arteriole, microcirculation, arteriolar wall mechanics
Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining
Institutions: Clinical Research Institute of Montreal, Laval University.
Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al.
in 1986 1
. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3
Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4
. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via
a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.
Medicine, Issue 69, Neuroscience, Biochemistry, Anatomy, Physiology, transient ischemic stroke, middle cerebral artery occlusion, intraluminal model, neuroscore, cresyl violet staining, mice, imaging
A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage
Institutions: SUNY Upstate Medical University, SUNY Upstate Medical University.
Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 μm ± 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 ± 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8
. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.
Medicine, Issue 71, Anatomy, Physiology, Neurobiology, Neuroscience, Immunology, Surgery, Aneurysm, cerebral, hemorrhage, model, mortality, rat, rodent, subarachnoid, vasospasm, animal model
Tilt Testing with Combined Lower Body Negative Pressure: a "Gold Standard" for Measuring Orthostatic Tolerance
Institutions: Simon Fraser University .
Orthostatic tolerance (OT) refers to the ability to maintain cardiovascular stability when upright, against the hydrostatic effects of gravity, and hence to maintain cerebral perfusion and prevent syncope (fainting). Various techniques are available to assess OT and the effects of gravitational stress upon the circulation, typically by reproducing a presyncopal event (near-fainting episode) in a controlled laboratory environment. The time and/or degree of stress required to provoke this response provides the measure of OT. Any technique used to determine OT should: enable distinction between patients with orthostatic intolerance (of various causes) and asymptomatic control subjects; be highly reproducible, enabling evaluation of therapeutic interventions; avoid invasive procedures, which are known to impair OT1
In the late 1980s head-upright tilt testing was first utilized for diagnosing syncope2
. Since then it has been used to assess OT in patients with syncope of unknown cause, as well as in healthy subjects to study postural cardiovascular reflexes2-6
. Tilting protocols comprise three categories: passive tilt; passive tilt accompanied by pharmacological provocation; and passive tilt with combined lower body negative pressure (LBNP). However, the effects of tilt testing (and other orthostatic stress testing modalities) are often poorly reproducible, with low sensitivity and specificity to diagnose orthostatic intolerance7
Typically, a passive tilt includes 20-60 min of orthostatic stress continued until the onset of presyncope in patients2-6
. However, the main drawback of this procedure is its inability to invoke presyncope in all individuals undergoing the test, and corresponding low sensitivity8,9
. Thus, different methods were explored to increase the orthostatic stress and improve sensitivity.
Pharmacological provocation has been used to increase the orthostatic challenge, for example using isoprenaline4,7,10,11
or sublingual nitrate12,13
. However, the main drawback of these approaches are increases in sensitivity at the cost of unacceptable decreases in specificity10,14
, with a high positive response rate immediately after administration15
. Furthermore, invasive procedures associated with some pharmacological provocations greatly increase the false positive rate1
Another approach is to combine passive tilt testing with LBNP, providing a stronger orthostatic stress without invasive procedures or drug side-effects, using the technique pioneered by Professor Roger Hainsworth in the 1990s16-18
. This approach provokes presyncope in almost all subjects (allowing for symptom recognition in patients with syncope), while discriminating between patients with syncope and healthy controls, with a specificity of 92%, sensitivity of 85%, and repeatability of 1.1±0.6 min16,17
. This allows not only diagnosis and pathophysiological assessment19-22
, but also the evaluation of treatments for orthostatic intolerance due to its high repeatability23-30
. For these reasons, we argue this should be the "gold standard" for orthostatic stress testing, and accordingly this will be the method described in this paper.
Medicine, Issue 73, Anatomy, Physiology, Biomedical Engineering, Neurobiology, Kinesiology, Cardiology, tilt test, lower body negative pressure, orthostatic stress, syncope, orthostatic tolerance, fainting, gravitational stress, head upright, stroke, clinical techniques
Intravascular Perfusion of Carbon Black Ink Allows Reliable Visualization of Cerebral Vessels
Institutions: University of Duisburg-Essen Medical School.
The anatomical structure of cerebral vessels is a key determinant for brain hemodynamics as well as the severity of injury following ischemic insults. The cerebral vasculature dynamically responds to various pathophysiological states and it exhibits considerable differences between strains and under conditions of genetic manipulations. Essentially, a reliable technique for intracranial vessel staining is essential in order to study the pathogenesis of ischemic stroke. Until recently, a set of different techniques has been employed to visualize the cerebral vasculature including injection of low viscosity resin, araldite F, gelatin mixed with various dyes1
carmine red, India ink) or latex with2
carbon black. Perfusion of white latex compound through the ascending aorta has been first reported by Coyle and Jokelainen3
. Maeda et al.2
have modified the protocol by adding carbon black ink to the latex compound for improved contrast visualization of the vessels after saline perfusion of the brain. However, inefficient perfusion and inadequate filling of the vessels are frequently experienced due to high viscosity of the latex compound4
. Therefore, we have described a simple and cost-effective technique using a mixture of two commercially available carbon black inks (CB1 and CB2) to visualize the cerebral vasculature in a reproducible manner5
. We have shown that perfusion with CB1+CB2 in mice results in staining of significantly smaller cerebral vessels at a higher density in comparison to latex perfusion5
. Here, we describe our protocol to identify the anastomotic points between the anterior (ACA) and middle cerebral arteries (MCA) to study vessel variations in mice with different genetic backgrounds. Finally, we demonstrate the feasibility of our technique in a transient focal cerebral ischemia model in mice by combining CB1+CB2-mediated vessel staining with TTC staining in various degrees of ischemic injuries.
Neuroscience, Issue 71, Neurobiology, Medicine, Anatomy, Physiology, Cellular Biology, Immunology, Neurology, Cerebral vascular anatomy, colored latex, carbon black, ink, stroke, vascular territories, brain, vessels, imaging, animal model
Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion
Institutions: University of Florida , University of Florida , Shiraz University of Medical Sciences, Shiraz, Iran .
Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3
. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5
. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1
). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.
Medicine, Issue 72, Neurobiology, Neuroscience, Biomedical Engineering, Anatomy, Physiology, Genetics, Genomics, Surgery, Ischemic stroke, Cerebrovascular, Brain, Cranial window, Rat, Endothelin-1, temporal craniectomy, animal model
2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia
Institutions: Wayne State University School of Medicine, Wayne State University School of Medicine, Wayne State University School of Medicine.
Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ischemia produces damage to specific brain regions shown to be highly sensitive to ischemia 1
. Hippocampal neurons have higher sensitivity to ischemic insults compared to other cell populations, and specifically, the CA1 region of the hippocampus is particularly vulnerable to ischemia/reperfusion 2
The design of therapeutic interventions, or study of mechanisms involved in cerebral damage, requires a model that produces damage similar to the clinical condition and in a reproducible manner. Bilateral carotid vessel occlusion with hypotension (2VOH) is a model that produces reversible forebrain ischemia, emulating the cerebral events that can occur during cardiac arrest and resuscitation. We describe a model modified from Smith et al
. (1984) 2
, as first presented in its current form in Sanderson, et al.
, which produces reproducible injury to selectively vulnerable brain regions 3-6
. The reliability of this model is dictated by precise control of systemic blood pressure during applied hypotension, the duration of ischemia, close temperature control, a specific anesthesia regimen, and diligent post-operative care. An 8-minute ischemic insult produces cell death of CA1 hippocampal neurons that progresses over the course of 6 to 24 hr of reperfusion, while less vulnerable brain regions are spared. This progressive cell death is easily quantified after 7-14 days of reperfusion, as a near complete loss of CA1 neurons is evident at this time.
In addition to this brain injury model, we present a method for CA1 damage quantification using a simple, yet thorough, methodology. Importantly, quantification can be accomplished using a simple camera-mounted microscope, and a free ImageJ (NIH) software plugin, obviating the need for cost-prohibitive stereology software programs and a motorized microscopic stage for damage assessment.
Medicine, Issue 76, Biomedical Engineering, Neurobiology, Neuroscience, Immunology, Anatomy, Physiology, Cardiology, Brain Ischemia, ischemia, reperfusion, cardiac arrest, resuscitation, 2VOH, brain injury model, CA1 hippocampal neurons, brain, neuron, blood vessel, occlusion, hypotension, animal model
Mouse Model of Middle Cerebral Artery Occlusion
Institutions: Ernest Gallo Clinic and Research Center, University of California, San Francisco, Kent State University.
Stroke is the most common fatal neurological disease in the United States 1
. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2
. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3
, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4
. The technique can be used to model permanent or transient occlusion 5
. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6
. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8
. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9
. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.
Medicine, Issue 48, Neurology, Stroke, mice, ischemia
Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment
Institutions: Havard Medical School, Massachusetts General Hospital.
Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel. Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion. Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction. Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm. This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.
Medicine, Issue 31, Aneurysm, intraoperative, brain, surgery, surgical clipping, blood flow, aneurysmal segment, ultrasonic flow probe

domain: medical


Your chance to remix one of their biggest tracks
Plump DJs 'fess up to a 'Dirty Weekend'
London-based independent bass label notches up half a century
Plus latest DJ mix
Massive Mason Competition
The details announced
Yeah we know it's 2007, but voting for Breakspoll, the international breakbeat awards, has begun...


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Paintbrush Cookies Recipe
Instructions:1. Preheat oven to 375F.
2. Roll out dough using floured pastry cloth to 1/4" thickness.
3. Cut into different shapes using cookie cutters.
4. Set on greased baking sheet.
5. Paint designs with egg yolk paints. (Blend egg yolk and water.
6. Divide among several custard cups.
7. Add different food colouring to each cup.
8. Paint designs using small paintbrushes.
9. If egg yolk, thickens while standing, add few drops water.
10. Bake 8-10 minutes at 375F.
11. For clear colours, do not let cookies brown.

domain: food


(MENAFN) According to a recent report by Bloomberg, the United States is strategizing to enhance its artillery shell production by increasing its procurement of military-grade explosives from Turkey. This move comes in response to the depletion of United States stockpiles due to the conflict in Ukraine, which has intensified the demand for munitions worldwide.
With Western nations facing challenges in supplying sufficient munitions to support Ukraine's defense efforts against Russia, the United States is seeking to address the shortfall by leveraging Turkish resources. Bloomberg highlights that the surge in global demand for weapons components has hindered efforts by the United States and its allies to scale up artillery shell production for Ukraine.
Anonymous officials cited by Bloomberg reveal that the United States plans to rely on Turkey for the supply of propellants such as trinitrotoluene and nitroguanidine, crucial ingredients for domestic defense manufacturing. This strategic shift contrasts with the approach taken by the European Union, where resistance from countries like France, Greece, and Cyprus has hindered purchases from Turkey, despite member states falling behind on their commitments to provide munitions to Ukraine.
The report underscores the significance of the United States-Turkey partnership in addressing critical defense needs, despite past tensions between the two countries. While disputes over Ankara's acquisition of Russian air defense systems strained relations, recent developments suggest a thaw in bilateral ties. This includes Turkey's authorization of Sweden to join NATO, which paved the way for the United States to approve the long-awaited sale of F-16 fighter jets and upgrade kits to Turkey in January.
Despite opposition from some quarters, such as Senator Rand Paul's attempt to block the deal citing concerns over Turkish conduct, the United States Congress ultimately approved the USD23 billion agreement. Bloomberg suggests that this collaboration over artillery shells represents a continuation of the positive momentum in United States-Turkey relations.
As the global arms race intensifies and geopolitical tensions persist, the strategic partnership between the United States and Turkey in the realm of defense procurement underscores the evolving dynamics of international alliances and the imperative of addressing critical security needs through cooperation and strategic partnerships.
MENAFN provides the information “as is” without warranty of any kind. We do not accept any responsibility or liability for the accuracy, content, images, videos, licenses, completeness, legality, or reliability of the information contained in this article. If you have any complaints or copyright issues related to this article, kindly contact the provider above.


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Directions to DoubleTree suites from MSP Airport:
MSP Airport is located 12 miles S.E. of this downtown Minneapolis hotel. Take I-494 west to I-35W north. Take I-35W north to Downtown exits. Take 11th Street exit 6 blocks to hotel. Hotel is located on the corner of 11th Street and LaSalle Avenue.
The hotel does not provide ground transportation to/from The Minneapolis-Saint Paul International Airport. Taxis await guests at either terminal after all arriving flights as well as light-rail transit to downtown Minneapolis. Rail users should take the BLUE line to Minneapolis.

domain: geography


text:
Explore 18 Collections - D Matching
This collection contains issues of the bulletin published by the Dallas/Fort Worth Section of the American Chemical Society. Included are The Activator (1944-1948) and the subsequent title, the Southwest Retort. The bulletins include division information, regional meetings, news about members, editorials, and other items of interest to members.Added: July 2012.
Selected photographs of quilt panels for the AIDS Memorial Quilt.Added: January 2018.
Materials from small collections associated with The Dallas Way archive include clippings, publications, papers, photographs, photo albums, buttons, and other artifacts from a variety of individuals and organizations.Added: October 2019.
Correspondence, newsletters, and grant reports related to art education and the North Texas Institute for Educators on the Visual Arts (NTIEVA), headquartered at the UNT. Materials were scanned by the donor, D. Jack Davis, and provided in a digital format to UNT Special Collections. Additional materials in an analog format can be viewed in the UNT Special Collections Reading Room.Added: January 2018.
The Defense Base Closure and Realignment Commissions (also known as the BRAC Commissions) were established by Congress to review and analyze Department of Defense recommendations for reorganizing U.S. military base structure "to more efficiently and effectively support our forces, increase operational readiness and facilitate new ways of doing business." This collection includes reports, documents, data, correspondence, and public comments from both the 1995 and 2005 Commissions.Added: October 2010.
Video from the 2017 Dene/Athabaskan Language Conference and Workshop.Added: March 2019.
News and viewpoints related to the Fracking Referendum passed in the city of Denton, Texas on November 4, 2014. Materials include a compilation of web pages crawled in late 2014 and a collection of oral histories conducted in Fall 2015 by UNT students.Added: August 2017.
This collection includes materials from yearly meetings since 2017 to build an infrastructure and workflow for developing CoRSAL, such as slides and recordings of presentations.Added: July 2020.
Documents related to the planning of the DFW Archives Bazaar, a recurring community outreach and education event focused on resources available at cultural heritage institutions in the Dallas-Fort Worth area. Photographs and videos of the event are also included.Added: January 2019.
Serving as virtual proceedings for the Digital Frontiers Conference, this collection contains abstracts, presentations, video, workshops, student responses, supporting materials, flyers, and other items from the conference and related activities.Added: October 2012.
These images document the disappearance of food and beverage items by photographing trays before and after schoolchildren ate lunch. The images were taken in North Texas during lunchtime in urban middle schools that participated in the National School Lunch and School Breakfast Programs.Added: May 2011.
Documenting The History of the Civil Rights Movement in Dallas County, Texas Oral History Collection
Transcripts of oral histories with civil rights figures from Dallas, Texas. Each interview focuses on the interviewee's family and educational history, organizational involvement, and career or personal milestones.Added: June 2020.

domain: history


text:
Provided by Brandpoint
Do you love the snap that chili peppers give to your food? Are you a fan of the heat they impart to a variety of dishes? Even if you’re not, you may want to take another look at the humble chili pepper. From migraine relief to weight control, researchers are verifying what some cultures have known for centuries: the chili pepper and its key constituents, like capsaicin, deliver a host of health benefits. Here are just some of the ways chili peppers are helping to fight some of the country’s most common health conditions:
Obesity is also a worldwide issue, making effective weight-control remedies a priority for health advocates around the world. Science Daily reports that University of Wyoming researchers are investigating capsaicin as a weight-control supplement. Researchers believe the chili pepper constituent may help boost metabolism by inhibiting cells that store fat and encouraging the type of cells that turn fat into energy.
Multiple studies aim at exploring the effect of capsaicin on cancer cells. One notable investigation from Marshall University found evidence that the compound inhibited the growth of lung cancer tumors through an animal-based study, according to the American Institute for Cancer Research. In another study, researchers at Nottingham University in the United Kingdom also found that capsaicin killed cancer cells without harming healthy cells in cultures of human lung and pancreatic cancer cells.
About 30 million Americans live with diabetes, and the disease is the seventh-leading cause of death in the U.S., according to the American Diabetes Association. While multiple medicines are available to treat diabetes, many people still find it difficult to control the disease. A study published in the American Journal of Clinical Nutrition found that eating chili peppers as part of a regular, healthy diet could help control diabetes.
Chili peppers also deliver health benefits beyond medicinal ones. The peppers are highly nutritious, delivering a punch of nutrients that are also associated with better health, including vitamins A, C and B-6, iron, copper and potassium.


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This lovely oval shaped dresser box or trinket box is the epitome of elegance. The portrait miniature is so well done and the box itself is patterned beautifully all around the portrait, as well as the outside of the bottom.
Size: 3 1/4 by 2 3/4 by 1 1/4 inches.
It is in very good condition with less than the expected wear. I was assured by the certified experts at Kodner Gallery that this is an antique.
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Unique Collectibles, Antiques and Fine Arts from Around the World
Never the ordinary...unique items chosen over the last 50 years of travel around the world.


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Cades Cove History: Wallet of Long Time Resident Elijah Oliver Donated to the Collections Preservation Center
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The Collections Preservation Center at Great Smoky Mountains National Park recently received an amazing donation! The great-great-granddaughter of long time Cades Cove resident Elijah Oliver had in her possession Oliver’s wallet. Elijah Oliver had used this wallet for many years! What made it even more special was that important papers had been tucked inside. Do you want to learn more about the recent donation filled with Cades Cove history? Here is more information on the wallet of long time Cades Cove resident Elijah Oliver:
Who was Elijah Oliver?
Born in the 1820s, Elijah Oliver was the son of the first European settler John Oliver. Elijah grew up in Cades Cove and had a big impact on Cades Cove history. He went on to build his cabin (Elijah Oliver Cabin) in 1865 which still stands today. The cabin is in a remote part of Cades Cove and includes a unique guest room called “The Stranger Room.” Oliver was also the deacon and clerk of the Primitive Baptist Church in Cades Cove and served in the positions for many years. Like most of the residents of the area, Elijah supported the Union.
What Does the Wallet Look Like?
Elijah Oliver’s wallet was actually quite large for the wallets of the time period. It is made of leather and appeared to be more of a pocketbook than a traditional wallet. For its age, the wallet is in great condition and even has the leather strap intact. The wallet itself and the documents that were found inside provide more information on Cades Cove history. The wallet was photographed, documented, and then added to the collection at the climate controlled Great Smoky Mountains National Park Collections Preservation Center.
What Documents Were Inside the Wallet?
Elijah Oliver’s wallet was crammed with folded-up papers! The papers which hold Cades Cove history had to be carefully unfolded by a professional archivist at the Collections Preservation Center. Special brushes were used to clean the papers that dated from 1840 to 1910. One of the documents inside the wallet dates to the 1860s and was issued by the State of Tennessee. It verified that Oliver was not involved with the Confederacy. At that time, there was a Tennessee law for men over 21 to carry official documentation like the document found in Elijah’s wallet in order to vote. Another document found in his wallet was an official pass dating to 1863. This pass was issued by the Union Military Authority and proved the person was not a Confederate spy or deserter. The document had to be shown when traveling on the road. It is great that the wallet and the documents inside will now be preserved for future generations to learn about Cades Cove history!
The donation of Elijah Oliver’s wallet is fantastic for the further understanding of Cades Cove history. Are you interested in learning more about Cades Cove history and the people who once lived there? Check out our Cades Cove ultimate recommended reading guide to learn about first hand accounts of Cades Cove residents.

domain: library


text: Colorado Economics. Our new prison chief spent a day in solitary confinement and wrote about it.http://www.nytimes.com/2014/02/21/opinion/my-night-in-solitary.html?_r=0Total spending on prisons is 654 million -8% of the CO. budget. 25 million more than what is spent on higher education. There are 20,000 prisoners in Colorado, 250,000 students in higher education.On a more personal note, there was a big table in the entrance to Whole Foods this week featuring "Sumo Tangerines". They have samples that are delicious. Looked like a deal - I bought six. Checking out I thought, that looks like alot for lunch and some tangerines. Looked at the receipt. The tangerines are $3.50 each. Taking out a loan for the next half dozen.

domain: economics


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From Othering to Belonging
How can we, as a society, move from “othering” to belonging. What and whom does othering actually benefit? How can we expand the circle of human concern and concern for nature? How can we live into our innate interconnection to create true inclusivity and wholeness? How do we build the structures, institutions, policies, cultures and stories that will support that inclusivity? Join Angela Glover Blackwell, Founder-in Residence at PolicyLink, which works to improve access and opportunity for all low-income people and communities of color, and john a. powell, renowned law professor, activist, and founder of the Othering and Belonging Institute at UC Berkeley, for a deep one-on-one conversation about these critically important, existential questions.
Note: This remarkable dialogue will be preceded by a special private screening of “From Othering to Belonging: The Circle of Human Concern,” a cathartic episode from the new docu-series “Changing of the Gods” directed by Kenny Ausubel and Louie Schwartzberg. The episode is a transformative passage through the valley of the shadow of Othering that illuminates the complex dynamics of the human psyche, ruthless political manipulations by elites using “divide-and-conquer” strategies to maintain their wealth and power, and the rising social movements for truth, reconciliation and healing.
May 13th | 2:45 pm to 4:15 pm
Keep Your Finger on the Pulse
Our bi-weekly newsletter provides insights into the people, projects, and organizations creating lasting change in the world.

domain: sociology


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The other day, Dinesh Karthik was forthright. But stand-in skipper Ajinkya Rahane chose to be more polite on Wednesday. Asked about the Afghanistan captain Asghar Stanikzai’s claim that the visitors having better spinners, the comeback man had pointed towards the experience-overload in favour of Ravi Ashwin and company.
But Rahane offered a more considerate view: “Every member of a team will always believe that his team is good, batting is good and all that. We all know stats but we don’t want to focus on stats. In a Test match, each and every session matters a lot. We should give our best all the time. Ashwin, (Ravindra) Jadeja, Kuldeep (Yadav) – they all are experienced and quality spinners. They back each other and enjoy each other’s company,” Rahane said.
A cut-throat comparison between the two teams, making a detour on the historical significance of India hosting Afghanistan for their first-ever Test, will highlight the gulf in class. India have a three-way competition between Murali Vijay, Shikhar Dhawan and KL Rahul for the two opening slots. Vijay has an experience of 56 Tests, with 3802 runs and 11 centuries. The South Africa tour earlier this year had witnessed his form slump, but the challenge was stiffer, and he is likelier to retain his spot. Dhawan had lost his place in the playing XI on the heels of jus-t two low scores in South Africa. But the left-hander’s average is 42.62 over 29 Tests. The excess of riches makes KL Rahul’s inclusion uncertain, notwithstanding the fact that three of his four Test centuries so far had been scored overseas. Besides, Rahul is fresh from a purple patch-hitting IPL for Kings XI Punjab. Maybe, he could be slotted into the middle-order, though it’s not as straightforward.
Cheteshwar Pujara is one of the best number threes in the long-form, along with Joe Root and Kane Williamson.
Steve Smith’s fall from grace has temporarily ruled him out of contention. Pujara has scored 1,240 runs in 14 Tests in the last one-odd year. In Virat Kohli’s absence, Rahane is likely to follow Pujara in the batting order. Karun Nair, brought back to the fold at the expense of Rohit Sharma, remains the only batsman in the history of Test cricket to be dropped straight after hitting a triple ton. In fact, the Karnataka boy faded into domestic cricket after his 303 not out against England at Chennai in December 2016.
A healthy rivalry between Rahul and Nair, if at all the former is considered for a middle-order slot, is good for the Indian cricket. Against Afghanistan though, the playing XI is incidental.
Whoever plays, the hosts will boast of a far stronger batting than their rivals. Also, it’s sort of inconsequential if the team management picks three spinners and includes Kuldeep, or goes with three fast bowlers. Jadeja and Ashwin are ranked No. 4 and No. 5 in the world. The duo has a collective experience of 92 Tests, accounting for 476 scalps. Kuldeep had bowled a match-winning spell on his debut against Australia last year. The chinaman bowler is keeping his senior colleagues on their toes. Even after a few rounds of mowing, the Chinnaswamy pitch retains a tinge of green. Playing three seamers appears a likely option. And weather permitting, India will go into the game as the outright favourites.
No disrespect to Afghanistan though. The best part is that they are showing enough positivity to prevent nerves. “What nerves! I’m hearing this for the first time,” Stanikzai said at the pre-match press conference. An excellent spin attack led by Rashid Khan is a reason for Afghanistan’s confidence. “Right now Rashid is the most difficult spinner to play around. He hasn’t played Test cricket so we need to look and see what happens. But I’m sure his professionalism will help him adjust and he will come out well,” coach Phil Simmons observed.
Rubbing shoulders with the big guns and attaining the IPL stardom has made Rashid arguably the most talked about spinner at the moment. The leggie, like his team mates, however, is a novice in red-ball international cricket. In fact, the 19-year-old has turned up in only four first-class matches. Stanikzai, Mohammad Nabi and Mohammad Shahzad are the veterans in the squads, first-class experience wise, with 23, 32 and 20 matches respectively in their bags. But the Afghans are game, making the Test build-up fascinating.
ICC to fete Afghanistan
Ahead of the historic Test, the ICC chief executive David Richardson will hand over the Afghanistan cricket team a souvenir/memento at Chinnaswamy tomorrow, it is learnt. Afghanistan’s rise to Test status is a serious shot in the arm for the world body, which has globalisation high on its agenda. Live from 9.30 am on Star Sports


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Timber a major Branch County crop
Timber is a large cash crop in Branch County, one that does not get noticed as much as corn and soybeans.
Especially during the winter, you can see large piles of cut timber worth thousands of dollars piled along fields ready for transport.
The best quality hardwoods usually go to Coldwater Veneer, world recognized for producing the thin layer of wood used in furniture and cabinet manufacturing.
Those not selected there often head to hardwood saw mills like Frick Lumber in Wawaka, Ind., according to Kurt France, whose France Timber company cut piles of hardwood each year in Branch County.
“Right now, we have stopped logging because most of the mills are full and are not buying,” France said. “We are just clearing land, fence rows and grinding.”
Jason Reichhart, another major logger agrees.
“I have informed my timber lot owners to let it be and push to cut until earliest this fall or winter," he said.
Like the softwood used in construction, some prices were up in the last year but now have leveled off as mills are filled with more wood than they can process.
Walnut trees can bring in between $500 to $5,000 depending the size and quality.
“Those 60-foot tall walnut, you just don’t see those trees anymore,” France said. He saw hardwood prices increase about 15% since the pandemic began.
There is a market for even the roughest timber. The bottom grades usually head to smaller mills to be turned into pallet wood.
Matt Delagrange, of S and M Pallet, buys wood year round to keep his Amish run saw mill on Quincy-Grange Road running year round. His prices rose 5 cents a board foot to 38 cents over the last five months.
“The pallet companies can’t get pine now. The price has skyrocketed,” Delagrange said.
This also has happened to building lumber. You would not think the hard woods like hard maple, oak, and walnut would be used for pallets. but they are, he said. These are the short pieces, those with knots, cracks and wormy are not suitable for veneer or good lumber.
“I cut the board and runners and they are used in pallets. I’ve got all hardwoods right now," he said.
His company supplies Union Pallet in Coldwater, as well as a Toledo company.
“I got more companies that want wood, but I can’t keep up,” Delagrange said.
What does he get paid? That’s his trade secret. S and M has been in business for 15 years supplying pallet makers.
Some farmers report that in bad years with low crop prices, the timber wood can make a difference in turning a profit.

domain: news


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It's one thing to cook good food and another feel good in the apron you're in while doing it. The Banker apron is Shokunin chef/owner Darren Maclean's everyday go-to in the kitchen. Designed by Medium Rare Chef Apparel, this apron has always been a favourite for chefs. Starting up in Calgary in 2008, the company's first customers were local chefs and that still rings true in 2019. Today you'll find their apparel line on industry professional and home cooks alike all around the world.
This black cotton canvas apron pictured boasts leather accents, cross-back straps, antique brass grommets, 2 chest pockets, 3 lap pockets, and 1 hidden cell phone pocket because food photos are important, right?
To win a Medium Rare Banker apron of your very own, let us know below what your signature dish to cook at home is.


Excellent service. Received ordered in just a few days. Quicker than I expected. Trolling motor works great so far. Easy to mount, although I was replacing my current motor and wiring was intact. I really like the rubber mounting inserts, although, I wasn't crazy about having to drill the larger holes (1/2") in my fiberglass. I have been wanting to swab to a motorguide for a long time.
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Direct and indirect effects of subclinical mastitis treatment in dairy herds
Barlow J., Nightingale C., Zadoks R., White L., Schukken Y.
The objective of this research is to evaluate the impact of treatment of subclinical mastitis on duration of infection and mastitis transmission in dairy herds. We describe the use of a deterministic state-transition model of intramammary infections (IMI) to predict the impact of subclinical mastitis treatment programs in populations of lactating dairy cattle. Results of model simulations indicate that the parameters duration of infection and the transmission coefficients influence the steady state values for the proportion of cows infected. The theoretical model predicts that a reduction in the duration of subclinical infections caused by contagious pathogens leads to a decrease in the prevalence and incidence of new IMI among susceptible individuals. Thus the model demonstrates potential indirect effects of mastitis treatment interventions resulting from changes in the intensity of pathogen transmission within a herd. © Wageningen Academic Publishers The Netherlands, 2005.


The MTV VMAs is one the biggest nights in music, and this year was no different. Justin Bieber, Billie Eilish and Olivia Rodrigo all won top accolades at the 2021 MTV Video Music Awards.
It was held at the Barclays Centre in New York City with the ceremony hosted by Doja Cat. Madonna and Jennifer Lopez opened the show paying tribute to MTV’s 40th birthday.
Olivia Rodrigo took home multiple VMAs this year, but how many did the teen win and which ones?
Read on to find out!
How many VMAs did Olivia Rodrigo win?
Teen sensation, Olivia Roderigo took home three awards for Best New Artist, Song of the Year for her hit ‘Drivers License’ in the fan-voted awards, and Push Performance of the Year.
She also performed her track ‘Good 4 U’ in which she lit up the stage. Olivia is actually a Disney Channel actress, and her album ‘Sour’ topped the charts in the US and UK and has been one of the biggest selling albums of the year.
Accepting the Song of the Year VMA she said: “This has been the most magical year of my life and it’s all because of you.
“I want to dedicate this award to all the other girls who write songs on their bedroom floor.
“There are a lot of people who will try to dim your light but speaking your mind and sharing your heart are the most beautiful things in the world and here’s to that.”
Fans react to Olivia Rodrigo’s VMA wins
While another fan tweeted: “No because my girl killed it last night and SHE DESERVES ALL THE LOVE SHE’S GETTING. I love her so freakin’ much.”
One user cannot wait to see what the future holds for the promising young singer: “18 year old Olivia Rodrigo just won Best New Artist of The Year at the VMAS omg we gon see her getting nominated for Grammys and Brits and Amas just now omg I can’t f—–g wait. I love her”.
Which other artists won VMAs?
Justin Bieber, BTS and Lil Nas X all had an extremely successful night at the 2021 VMAs.
Bieber came in with a leading seven nominations and he bagged the awards for Artist of the Year and Best Pop Video for ‘Peaches’ along with Daniel Caesar and Giveon.
BTS won the award for ‘Best K-Pop’ and Lil Nas X secured three VMAs, including Video of the Year which was the subject of controversy due to its explicit nature. In his acceptance speech, Nas said: “First, I want to say thank you to the gay agenda! Let’s go, gay agenda!” As well as Video of the Year, he also won trophies for Best Direction and Best Visual Effects for the music video of his hit single ‘Montero’.


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Coinbase (COIN) begins trading today on the NASQAD. COIN hits a high of $429…. Valued at over 100 billion dollars USD on first day of trading. Should you buy Coinbase today or tomorrow or should you wait to buy Coinbase? S&P500 (SPY) starts daily consolidation after losing the low of yesterday breaking the 13 day streak of a higher low everyday. News & events for today… Daily market recap & MJ Sector Review. Charting and technical analysis for Wednesday April 14th, 2021.
For a 7 day FREE trial to the POW Group community visit: https://pursuitofwealthgroup.com
POW Algo Alerts has launched! Visit https://pursuitofwealthgroup.com to learn more and find out about a free trial.
– Bybit: https://www.bybit.com/en-US?affiliate_id=17927&group_id=0&group_type=1
#Stocks, #Crypto, #Coinbase
00:44 News & Events
04:25 Crypto Market Review
08:20 Broader Market Review
13:00 Individual Stock Reviews
14:42 CAD MJ Sector Review
15:50 US MJ Sector Review


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CATCH YOUR CAT !
Dear friends, you can CATCH YOUR CAT-ARTWORK with little help from MOSCOW CAT MUSEUM and put it in your cozy interior at home or at office space.
Our CAT-ARTWORKS is very interesting, very nice, very funny, very collectable and made by best Russian artists.
CATCH YOUR CAT is so simple.
You can choose some works from THE TODAY'S ART EXHIBITION and send us a fax (e-mail) with the names of the works you want to get and your own info (fax, tel., e-mail).
We will send you a bill and after you pay we will send you work(works) by express mail (DHL, EMS GARANTPOST, TNT, UPS, FED EX).
You will get "YOUR CAT" as soon as possible.
Every CAT-ARTWORK have the museum's certifiCATe and the museum's special stamp. Also we usually send information about artist or artist's CATalogue.
PLEASE DON'T BE LATE - THE FIRST PERSON TAKES IT ALL.
Monthly we will show you 20-30 new and interesting CAT-ARTWORKS.
WE WISH YOU SUCCESS AND - CATCH YOUR CAT!
Each of pictures of the galleries is a reference to more detailed scene with description

domain: artistic


Thorlos Experia Multi-Activity Micro-Mini (2 Pairs) Product Description
Designed for feet that don't hurt. High performance runners, cyclists, hikers and fitness walkers who prefer a minimal amount of foot protection. These socks are lghtly padded that provides the feel of near "barefoot" and not recommended for people with any distracting foot issues or long-time wearers of Thorlos dependent or relies on thick or moderate cushioned products.
Fiber Contents | 66% THOR?WICK® COOL | 20% Nylon | 1% Elastic | 13% Polyester
2019 New Thorlos Experia Multi-Activity Micro-Mini 2 Pairs Black On Black FVXVIJN