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Topic:Animal Models

Animal models in equine research refer to the use of horses or other species to study biological and disease processes relevant to equine health. These models facilitate the investigation of various physiological and pathological conditions, allowing researchers to explore disease mechanisms, test therapeutic interventions, and understand genetic influences on health outcomes. Horses themselves are often studied as models for human diseases due to certain physiological similarities. Additionally, other species may be used to model equine-specific conditions when direct study in horses is impractical. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and findings of animal models in equine research.
Ferret Models for Henipavirus Infection.
Methods in molecular biology (Clifton, N.J.)    August 23, 2023   Volume 2682 205-217 doi: 10.1007/978-1-0716-3283-3_15
Rockx B, Mire CE.Henipaviruses are emerging zoonotic viruses that can cause outbreaks of severe respiratory and neurological disease in humans and animals such as horses. The mechanism by which these viruses can cause disease remain largely unknown and to date there are no therapeutics or vaccines approved for use in humans. Nipah virus is listed on the World Health Organization R & D Blueprint list of epidemic threats. In order to advance the availability of effective therapeutics and vaccines and medicines that can be used to save lives and avert large scale crises, animal models are required which recap...
The influence of different horseshoes and ground substrates on mid-stance hoof orientation at the walk.
Equine veterinary journal    August 17, 2023   Volume 56, Issue 3 598-606 doi: 10.1111/evj.13990
Reilly PT, van Eps A, Stefanovski D, Pfau T.Horseshoes with modified contact surfaces combined with deformable ground substrates are used to change hoof orientation during mid-stance, for example, for therapeutic reasons. Objective: To measure the effect of horseshoes and ground substrates on sagittal and transverse plane hoof orientation at mid-stance using a dorsal hoof wall mounted triaxial accelerometer. Methods: In vivo experiment, randomised crossover design. Methods: Differences in sagittal and transverse plane angles between standing and mid-stance of the left front hoof of six horses walking with regular horseshoes, egg bar, to...
The Triple-E Model: Advancing Equestrian Research with Perspectives from One Health.
Animals : an open access journal from MDPI    August 16, 2023   Volume 13, Issue 16 2642 doi: 10.3390/ani13162642
Keener MM, Tumlin KI.Equestrian sport has various welfare issues and educational needs. To address these complex interactions, we propose an integrated approach called the Triple-E Model, which focuses on the equine, equestrian, and environmental triad. A literature review of existing models suggests that complexities of these interactions are overlooked, despite the significant impact of equine industries on economics, healthcare, and animal welfare. This paper discusses current models and theories used to evaluate equine-equestrian-environmental interactions and introduces the Triple-E Model to foster multidisci...
Equine induced pluripotent stem cells are responsive to inflammatory cytokines before and after differentiation into musculoskeletal cell types.
In vitro cellular & developmental biology. Animal    August 15, 2023   doi: 10.1007/s11626-023-00800-3
Palomino Lago E, Jelbert ER, Baird A, Lam PY, Guest DJ.Persistent inflammation is associated with the poor regeneration of musculoskeletal tissues. Embryonic stem cells (ESCs) have an attenuated response to inflammatory cytokines, but there are mixed reports on the response of induced pluripotent stem cells (iPSCs) to inflammation. Horses provide a relevant large animal model for studying musculoskeletal tissue diseases and the testing of novel therapies. The aim of this study was to determine if equine iPSCs are responsive to the inflammatory cytokines IL-1β, TNFα and IFN-γ in their undifferentiated state, or following differentiation into ten...
Animal model for tendinopathy.
Journal of orthopaedic translation    August 14, 2023   Volume 42 43-56 doi: 10.1016/j.jot.2023.06.005
Luo J, Wang Z, Tang C, Yin Z, Huang J, Ruan D, Fei Y, Wang C, Mo X, Li J, Zhang J, Fang C, Li J, Chen X, Shen W.Tendinopathy is a common motor system disease that leads to pain and reduced function. Despite its prevalence, our mechanistic understanding is incomplete, leading to limited efficacy of treatment options. Animal models contribute significantly to our understanding of tendinopathy and some therapeutic options. However, the inadequacies of animal models are also evident, largely due to differences in anatomical structure and the complexity of human tendinopathy. Different animal models reproduce different aspects of human tendinopathy and are therefore suitable for different scenarios. This rev...
Regenerative Strategies in Treatment of Peripheral Nerve Injuries in Different Animal Models.
Tissue engineering and regenerative medicine    August 12, 2023   Volume 20, Issue 6 839-877 doi: 10.1007/s13770-023-00559-4
Khaled MM, Ibrahium AM, Abdelgalil AI, El-Saied MA, El-Bably SH.Peripheral nerve damage mainly resulted from traumatic or infectious causes; the main signs of a damaged nerve are the loss of sensory and/or motor functions. The injured nerve has limited regenerative capacity and is recovered by the body itself, the recovery process depends on the severity of damage to the nerve, nowadays the use of stem cells is one of the new and advanced methods for treatment of these problems. Following our review, data are collected from different databases "Google scholar, Springer, Elsevier, Egyptian Knowledge Bank, and PubMed" using different keywords such as Periphe...
Equine Gunshot Euthanasia: Creation of a 3D-Printed Model with Integrated Sensors for Training.
Animals : an open access journal from MDPI    August 9, 2023   Volume 13, Issue 16 2566 doi: 10.3390/ani13162566
Dybdal N, Horgan M, Costa L, Davis E, Lucero S, Nieves S, Quiroz V, Weberg K, Madigan JE.Challenges and issues related to the use of pentobarbital euthanasia and disposal of animal remains within the US have recently been reviewed. Environmental and public health challenges increasingly necessitate consideration of alternative methods such as gunshots, an American Veterinary Medical Association (AVMA) "acceptable with conditions" method, for the humane euthanasia of horses. A recent study reported a correctly aimed gunshot provides a humane option for euthanizing horses. However, although aiming guidelines exist, studies examining bullet trajectories in animals euthanized by gunsh...
Genetic structure and genome-wide association study of the traditional Kazakh horses.
Animal : an international journal of animal bioscience    August 2, 2023   Volume 17, Issue 9 100926 doi: 10.1016/j.animal.2023.100926
Pozharskiy A, Abdrakhmanova A, Beishova I, Shamshidin A, Nametov A, Ulyanova T, Bekova G, Kikebayev N, Kovalchuk A, Ulyanov V, Turabayev A....Horses are traditionally used in Kazakhstan as a source of food and as working and saddle animals as well. Here, for the first time, microarray-based medium-density single nucleotide polymorphism (SNP) genotyping of six traditionally defined types and breeds of indigenous Kazakh horses was conducted to reveal their genetic structure and find markers associated with animal size and weight. The results showed that the predefined separation between breeds and sampled populations was not supported by the molecular data. The lack of genetic variation between breeds and populations was revealed by t...
First Insight into the Seroepidemiology of Hepatitis E Virus (HEV) in Dogs, Cats, Horses, Cattle, Sheep, and Goats from Bulgaria.
Viruses    July 21, 2023   Volume 15, Issue 7 doi: 10.3390/v15071594
Tsachev I, Gospodinova K, Pepovich R, Takova K, Kundurzhiev T, Zahmanova G, Kaneva K, Baymakova M.In recent years, hepatitis E virus (HEV) infection has been found to be widespread among different animal species worldwide. In Bulgaria, high HEV seropositivity was found among pigs (60.3%), wild boars (40.8%), and East Balkan swine (82.5%). The aim of the present study was to establish the seroprevalence of HEV among dogs, cats, horses, cattle, sheep, and goats in Bulgaria. In total, 720 serum samples from six animal species were randomly collected: dogs-90 samples; cats-90; horses-180; cattle-180; sheep-90; and goats-90. The serum samples were collected from seven districts of the country: ...
Pharmacokinetics and plasma protein binding of a single dose of clodronate disodium are similar for juvenile sheep and horses.
American journal of veterinary research    July 21, 2023   Volume 84, Issue 8 doi: 10.2460/ajvr.23.03.0051
Vergara-Hernandez FB, Nielsen BD, Kottwitz JJ, Panek CL, Robison CI, Paris BL, Welsh TH, Bradbery AN, Leatherwood JL, Colbath AC.To determine the single-dose pharmacokinetics of clodronate disodium (CLO) in juvenile sheep and the plasma protein binding (PPB) of CLO in juvenile sheep and horses. Methods: 11 juvenile crossbred sheep (252 ± 6 days) for the pharmacokinetic study. Three juvenile crossbred sheep (281 ± 4 days) and 3 juvenile Quarter Horses (599 ± 25 days) for PPB analysis. Methods: CLO concentrations were determined using liquid chromatography-mass spectrometry. Pharmacokinetic parameters were calculated by noncompartmental analysis from plasma samples obtained at 0, 0.5, 1, 3, 6, 12, 24, 48, and 72 hours ...
Impact of Social Buffering and Restraint on Welfare Indicators during UK Commercial Horse Slaughter.
Animals : an open access journal from MDPI    July 12, 2023   Volume 13, Issue 14 doi: 10.3390/ani13142276
Fletcher KA, Limon G, Padalino B, Hall GK, Chancellor N, Grist A, Gibson TJ.Current legislation in the United Kingdom stipulates that horses should not be slaughtered within sight of one another. However, abattoir personnel anecdotally report that, for semi-feral horses unused to restraint, co-slaughtering alongside a conspecific could reduce distress through social buffering and improve safety, but there is a lack of evidence to support this. CCTV footage from an English abattoir was assessed retrospectively with welfare indicators from when horses entered the kill pen until they were killed. Of 256 horses analysed, 12% (32/256) were co-slaughtered (alongside a consp...
High-Resolution Genotyping of Expressed Equine MHC Reveals a Highly Complex MHC Structure.
Genes    July 10, 2023   Volume 14, Issue 7 doi: 10.3390/genes14071422
Vasoya D, Tzelos T, Benedictus L, Karagianni AE, Pirie S, Marr C, Oddsdóttir C, Fintl C, Connelley T.The Major Histocompatibility Complex (MHC) genes play a key role in a number of biological processes, most notably in immunological responses. The MHCI and MHCII genes incorporate a complex set of highly polymorphic and polygenic series of genes, which, due to the technical limitations of previously available technologies, have only been partially characterized in non-model but economically important species such as the horse. The advent of high-throughput sequencing platforms has provided new opportunities to develop methods to generate high-resolution sequencing data on a large scale and app...
Host cell restriction factors of equine infectious anemia virus.
Virologica Sinica    July 5, 2023   Volume 38, Issue 4 485-496 doi: 10.1016/j.virs.2023.07.001
Wang XF, Zhang X, Ma W, Li J, Wang X.Equine infectious anemia virus (EIAV) is a member of the lentivirus genus in the Retroviridae family and is considered an animal model for HIV/AIDS research. An attenuated EIAV vaccine, which was successfully developed in the 1970s by classical serial passage techniques, is the first and only lentivirus vaccine that has been widely used to date. Restriction factors are cellular proteins that provide an early line of defense against viral replication and spread by interfering with various critical steps in the viral replication cycle. However, viruses have evolved specific mechanisms to overcom...
Correction: Górski et al. Application of Two-Dimensional Entropy Measures to Detect the Radiographic Signs of Tooth Resorption and Hypercementosis in an Equine Model. Biomedicines 2022, 10, 2914.
Biomedicines    June 29, 2023   Volume 11, Issue 7 doi: 10.3390/biomedicines11071865
Górski K, Borowska M, Stefanik E, Polkowska I, Turek B, Bereznowski A, Domino M.Incorrect Reference: In the original article [1], the reference [43] was incorrectly written as Padhye, N.; Rios, D.; Fay, V.; Hanneman, S.K. Pressure Injury Link to Entropy of Abdominal Temperature; Cizik School of Nursing, The University of Texas Health Science Center at Houston: Houston, TX, USA, 2022; Preprint. It should be Flood, M.W.; Grimm, B. EntropyHub: An open-source toolkit for entropic time series analysis. PLoS ONE 2021, 16, e0259448. With this correction, the order of references has not been changed. The authors state that the scientific conclusions are unaffected. This correcti...
Fusion of visible and thermal images improves automated detection and classification of animals for drone surveys.
Scientific reports    June 27, 2023   Volume 13, Issue 1 10385 doi: 10.1038/s41598-023-37295-7
Krishnan BS, Jones LR, Elmore JA, Samiappan S, Evans KO, Pfeiffer MB, Blackwell BF, Iglay RB.Visible and thermal images acquired from drones (unoccupied aircraft systems) have substantially improved animal monitoring. Combining complementary information from both image types provides a powerful approach for automating detection and classification of multiple animal species to augment drone surveys. We compared eight image fusion methods using thermal and visible drone images combined with two supervised deep learning models, to evaluate the detection and classification of white-tailed deer (Odocoileus virginianus), domestic cow (Bos taurus), and domestic horse (Equus caballus). We cla...
Equine Crofton Weed (Ageratina spp.) Pneumotoxicity: What Do We Know and What Do We Need to Know?
Animals : an open access journal from MDPI    June 23, 2023   Volume 13, Issue 13 doi: 10.3390/ani13132082
Shapter FM, Granados-Soler JL, Stewart AJ, Bertin FR, Allavena R.Crofton weed (Ageratina adenophora) is a global and highly invasive weed, with ingestion causing severe respiratory disease in horses, leading to irreversible and untreatable pulmonary fibrosis and oedema. While reports of equine pneumotoxicity remain common in Australia and New Zealand, equine pneumotoxicity may be underdiagnosed in other countries where Crofton weed is endemic but poorly differentiated. The pathogenesis of Crofton weed toxicity following ingestion has been well described in a number of different animal models, including rodents, rabbits, and goats. However, induced toxicity ...
Machine Learning-Based Sensor Data Fusion for Animal Monitoring: Scoping Review.
Sensors (Basel, Switzerland)    June 20, 2023   Volume 23, Issue 12 5732 doi: 10.3390/s23125732
Aguilar-Lazcano CA, Espinosa-Curiel IE, Ríos-Martínez JA, Madera-Ramírez FA, Pérez-Espinosa H.The development of technology, such as the Internet of Things and artificial intelligence, has significantly advanced many fields of study. Animal research is no exception, as these technologies have enabled data collection through various sensing devices. Advanced computer systems equipped with artificial intelligence capabilities can process these data, allowing researchers to identify significant behaviors related to the detection of illnesses, discerning the emotional state of the animals, and even recognizing individual animal identities. This review includes articles in the English langu...
Towards an objective measurement of sleep quality in non-human animals; using the horse as a model species for the creation of sleep quality indices.
Biology open    June 20, 2023   Volume 12, Issue 7 bio059964 doi: 10.1242/bio.059964
Greening L, Allen S, McBride S.Sleep disturbance is observed across species, resulting in neurocognitive dysfunction and poor impulse control/regulation of negative emotion. Understanding animal sleep disturbance is thus important to understand how environmental factors influence animal sleep and day-to-day welfare. Self-reporting tools for sleep disturbance are commonly used in human research to determine sleep quality, that cannot be transferred to non-verbal animal species research. Human research has, however, successfully used frequency of awakenings to create an objective measurement of sleep quality. The aim of this ...
Horses (Equus caballus) facial micro-expressions: insight into discreet social information.
Scientific reports    May 27, 2023   Volume 13, Issue 1 8625 doi: 10.1038/s41598-023-35807-z
Tomberg C, Petagna M, de Selliers de Moranville LA.Facial micro-expressions are facial expressions expressed briefly (less than 500 ms) and involuntarily. Described only in humans, we investigated whether micro-expressions could also be expressed by non-human animal species. Using the Equine Facial action coding system (EquiFACS), an objective tool based on facial muscles actions, we demonstrated that a non-human species, Equus caballus, is expressing facial micro-expressions in a social context. The AU17, AD38 and AD1 were selectively modulated as micro-expression-but not as standard facial expression (all durations included)-in presence of ...
Impact of inbreeding and genetic parameter estimates for seminal traits in Lusitano horses.
Theriogenology    May 27, 2023   Volume 208 43-51 doi: 10.1016/j.theriogenology.2023.05.011
Gonçalves AR, Telo da Gama L, Antunes L, Guimarães H, Bliebernicht M, Duarte JC, Cosinha C, Duarte Rego B, Ferro da Costa P, Guimarães T, Rocha A....The objectives of this study were to establish baseline information for seminal traits in Lusitano stallions, to assess the impact of inbreeding, interval between collections and age on semen quality during the breeding and non-breeding seasons, and to estimate the corresponding genetic parameters. A total of 2129 ejaculates by 146 Lusitano stallions used for artificial insemination, obtained from four equine reproduction centers distributed throughout Portugal, over a period of 14 years (2008-2021), were included in the study. The seminal traits analyzed, and the corresponding means and stand...
3D T1 relaxation time measurements in an equine model of subtle post-traumatic osteoarthritis using MB-SWIFT. Pala S, Hänninen NE, Mohammadi A, Ebrahimi M, Te Moller NCR, Brommer H, René van Weeren P, Mäkelä JTA, Korhonen RK, Afara IO, Töyräs J....The aim of this study is to assess whether articular cartilage changes in an equine model of post-traumatic osteoarthritis (PTOA), induced by surgical creation of standard (blunt) grooves, and very subtle sharp grooves, could be detected with ex vivo T relaxation time mapping utilizing three-dimensional (3D) readout sequence with zero echo time. Grooves were made on the articular surfaces of the middle carpal and radiocarpal joints of nine mature Shetland ponies and osteochondral samples were harvested at 39 weeks after being euthanized under respective ethical permissions. T relaxation times ...
IgE-binding monocytes upregulate the coagulation cascade in allergic horses.
Genes and immunity    May 16, 2023   Volume 24, Issue 3 130-138 doi: 10.1038/s41435-023-00207-w
Simonin EM, Wagner B.IgE-binding monocytes are a rare peripheral immune cell type involved in the allergic response through binding of IgE on their surface. IgE-binding monocytes are present in both healthy and allergic individuals. We performed RNA sequencing to ask how the function of IgE-binding monocytes differs in the context of allergy. Using a large animal model of allergy, equine Culicoides hypersensitivity, we compared the transcriptome of IgE-binding monocytes in allergic and non-allergic horses at two seasonal timepoints: (i) when allergic animals were clinical healthy, in the winter "Remission Phase", ...
The oxygen transport cascade and exercise: Lessons from comparative physiology.
Comparative biochemistry and physiology. Part A, Molecular & integrative physiology    May 13, 2023   Volume 282 111442 doi: 10.1016/j.cbpa.2023.111442
Ramsook AH, Dominelli PB, Angus SA, Senefeld JW, Wiggins CC, Joyner MJ.Studies of animal physiology not only provide valuable knowledge for the species in question, but also offer insights into human physiology. This thought is best highlighted by the 'Krogh Principle', which states "for many problems there is an animal on which it can be most conveniently studied". This graphical review focuses on three distinct stages of the oxygen transport cascade in which human exercise physiology knowledge has been enhanced by studies carried out in animal models. We begin by exploring ventilation, and the detrimental effects of cold, dry air on the airways in two sets of e...
A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice.
Toxins    April 30, 2023   Volume 15, Issue 5 316 doi: 10.3390/toxins15050316
Fan Y, Lou J, Tam CC, Wen W, Conrad F, Leal da Silva Alves P, Cheng LW, Garcia-Rodriguez C, Farr-Jones S, Marks JD.Equine-derived antitoxin (BAT) is the only treatment for botulism from botulinum neurotoxin serotype G (BoNT/G). BAT is a foreign protein with potentially severe adverse effects and is not renewable. To develop a safe, more potent, and renewable antitoxin, humanized monoclonal antibodies (mAbs) were generated. Yeast displayed single chain Fv (scFv) libraries were prepared from mice immunized with BoNT/G and BoNT/G domains and screened with BoNT/G using fluorescence-activated cell sorting (FACS). Fourteen scFv-binding BoNT/G were isolated with K values ranging from 3.86 nM to 103 nM (median K 2...
Investigating the Market Value of Brumbies (Equus caballus) in the Australian Riding Horse Market.
Animals : an open access journal from MDPI    April 27, 2023   Volume 13, Issue 9 doi: 10.3390/ani13091481
Condon V, Wilson B, Fleming PJS, Kennedy BPA, Keeley T, Barwick J, McGreevy P.Feral horses, also known as brumbies, are widely distributed across Australia with some populations being managed largely by human intervention. Rehoming of suitable feral horses following passive trapping has wide community acceptance as a management tool. However, there is little information about the number and relative economic value of feral horses compared with cohorts in the riding horse market. We examined 15,404 advertisements of horses for sale in 53 editions of Horse Deals, published from February 2017 to July 2022. Despite the considerable media attention and public scrutiny surrou...
Extracellular vesicles in the treatment and prevention of osteoarthritis: can horses help us translate this therapy to humans?
Extracellular vesicles and circulating nucleic acids    April 17, 2023   Volume 4, Issue 2 151-169 doi: 10.20517/evcna.2023.11
O'Brien TJ, Hollinshead F, Goodrich LR.Osteoarthritis (OA) is a common joint disease affecting humans and horses, resulting in significant morbidity, financial expense, and loss of athletic use. While the pathogenesis is incompletely understood, inflammation is considered crucial in the development and progression of the disease. Mesenchymal stromal cells (MSCs) have received increasing scientific attention for their anti-inflammatory, immunomodulatory, and pro-regenerative effects. However, there are concerns about their ability to become a commercially available therapeutic. Extracellular vesicles (EVs) are now recognized to play...
Tracking the Source of Human Q Fever from a Southern French Village: Sentinel Animals and Environmental Reservoir.
Microorganisms    April 13, 2023   Volume 11, Issue 4 1016 doi: 10.3390/microorganisms11041016
Laidoudi Y, Rousset E, Dessimoulie AS, Prigent M, Raptopoulo A, Huteau Q, Chabbert E, Navarro C, Fournier PE, Davoust B., also known as the causal agent of Q fever, is a zoonotic pathogen infecting humans and several animal species. Here, we investigated the epidemiological context of from an area in the Hérault department in southern France, using the One Health paradigm. In total, 13 human cases of Q fever were diagnosed over the last three years in an area comprising four villages. Serological and molecular investigations conducted on the representative animal population, as well as wind data, indicated that some of the recent cases are likely to have originated from a sheepfold, which revealed bacterial c...
Efficacy of a brain-penetrant antiviral in lethal Venezuelan and eastern equine encephalitis mouse models.
Science translational medicine    April 12, 2023   Volume 15, Issue 691 eabl9344 doi: 10.1126/scitranslmed.abl9344
Cao X, Yang D, Parvathareddy J, Chu YK, Kim EJ, Fitz-Henley JN, Li X, Lukka PB, Parmar KR, Temrikar ZH, Dhole P, Adcock RS, Gabbard J, Bansal S....Venezuelan and eastern equine encephalitis viruses (VEEV and EEEV, respectively) are mosquito-borne, neuroinvasive human pathogens for which no FDA-approved therapeutic exists. Besides the biothreat posed by these viruses when aerosolized, arthropod transmission presents serious health risks to humans, as demonstrated by the 2019 outbreak of EEE disease in the United States that resulted in 38 confirmed cases, 19 deaths, and neurological effects in survivors. Here, we describe the discovery of a 2-pyrrolidinoquinazolinone scaffold, efficiently synthesized in two to five steps, whose structural...
Comparison of Trap and Equine Attraction to Mosquitoes.
Insects    April 11, 2023   Volume 14, Issue 4 374 doi: 10.3390/insects14040374
Dilling SC, TenBroeck SH, Hogsette JA, Kline DL.Mosquitoes are pests of horses, but mosquito trap efficacy data, especially the ability of traps to protect horses, are lacking. Studies were conducted to investigate the comparative attraction between traps and horses, increase trap attraction by adding horse odors to the airstream of a trap, determine the spatial distribution of adult mosquitoes, estimate the numbers of mosquitoes feeding on horses, determine the relative attraction of horses to mosquitoes, and estimate the range of mosquitoes' attraction between two horses. When a horse and a mosquito trap were placed 3.5 m apart, there was...
The Effect of Insect Bite Hypersensitivity on Movement Activity and Behaviour of the Horse.
Animals : an open access journal from MDPI    April 8, 2023   Volume 13, Issue 8 1283 doi: 10.3390/ani13081283
Söderroos D, Ignell R, Haubro Andersen P, Bergvall K, Riihimäki M.Insect bite hypersensitivity (IBH) associated with biting midges is a common allergic skin disease in horses, reducing the welfare of affected horses. This study investigated the effect of IBH on animal welfare and behaviour and assessed a new prophylactic insect repellent. In total, 30 horses were recruited for a prospective cross-over and case-control study. Clinical signs of IBH, inflammatory markers in skin biopsies and behavioural data (direct observations, motion index) were scored longitudinally during two consecutive summers. No differences were observed in the total number of itching...
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