Temperature dependency of whole blood viscosity and red cell properties in desert ungulates : studies on scimitar-horned oryx and dromedary camel

BACKGROUND: The dromedary camel and the oryx antelope are exposed to excessive heat and solar radiation in their desert habitat. Desertification of areas with by now little rainfall may occur eventually. Well-adapted large animal species show us what is needed to survive in scorching regions. METHODS : Four scimitar-horned oryx antelopes (Oryx dammah), 10 camels (Camelus dromedarius), nine South African Merino sheep, and 17 Nguni cows were tested for RBC aggregation, RBC elongation, and plasma viscosity. The temperature dependency of blood viscosity was tested in 10 camels and compared to human reference values. RESULTS : Unlike sheep, Nguni cow, and dromedary camel, oryx RBCs aggregate in native plasma (M0:5.2 (3.3/6.7); M1:18.1 (16.7/27.9); Myrenne MA1). Elongation indices of oryx RBCs were intermediate to low (EImax: 22.6 (19.2/25.3); SS½ 3.67 (2.52/4.95); Rheodyn SSD). Camel RBCs did not display the typical SS/EI curve by rotational ektacytometry. In-vitro blood viscosity (Physica MCR302) was lower in camels than in human blood at equal hematocrit. A decrease of temperature had only little effect on camel blood. At 10s−1, blood viscosity in camel increased from 2.18mPa*s (2.01/2.37) at 42◦C to 4.39mPa*s (4.22/4.51) at 12◦C. In human blood, viscosity ranged from 8.21mPa*s (6.95/8.25) at 37◦C to 15.52mPa*s (14.25/16.03) at 12◦C. At 1000s−1, blood viscosity in camel ranged from 2.00mPa*s (1.95/2.04) at 42◦C to 3.98mPa*s (3.88/4.08) at 12◦C. In human blood, viscosity ranged from 5.35mPa*s (4.96/5.87) at 37◦C to 11.24mPa*s (10.06/11.17) at 12◦C. CONCLUSIONS : Desert ungulates may need RBC membranes, which are fortified to withstand changes in osmolality during dehydration-rehydration cycles. This reduces RBC deformability. Dromedary camel blood does not undergo stark changes in viscosity with changes in temperature. Therefore, blood fluidity could be rather maintained during the day and night cycle. This should reduce the need of the vascularity to rhythmically adapt to changing shear forces when camels experience heterothermy.The data was presented in part at the 1st Hemorheology Days 2017 in Puchberg am Schneeberg, Austria.https://content.iospress.com/journals/clinical-hemorheology-and-microcirculationhj2018Production Animal Studie

Inhibition of profibrotic microRNA-21 affects platelets and their releasate.

Fibrosis is a major contributor to organ disease for which no specific therapy is available. MicroRNA-21 (miR-21) has been implicated in the fibrogenetic response, and inhibitors of miR-21 are currently undergoing clinical trials. Here, we explore how miR-21 inhibition may attenuate fibrosis using a proteomics approach. Transfection of miR-21 mimic or inhibitor in murine cardiac fibroblasts revealed limited effects on extracellular matrix (ECM) protein secretion. Similarly, miR-21-null mouse hearts showed an unaltered ECM composition. Thus, we searched for additional explanations as to how miR-21 might regulate fibrosis. In plasma samples from the community-based Bruneck Study, we found a marked correlation of miR-21 levels with several platelet-derived profibrotic factors, including TGF-β1. Pharmacological miR-21 inhibition with an antagomiR reduced the platelet release of TGF-β1 in mice. Mechanistically, Wiskott-Aldrich syndrome protein, a negative regulator of platelet TGF-β1 secretion, was identified as a direct target of miR-21. miR-21-null mice had lower platelet and leukocyte counts compared with littermate controls but higher megakaryocyte numbers in the bone marrow. Thus, to our knowledge this study reports a previously unrecognized effect of miR-21 inhibition on platelets. The effect of antagomiR-21 treatment on platelet TGF-β1 release, in particular, may contribute to the antifibrotic effects of miR-21 inhibitors

Measurement of Flow Properties of Mammalian Blood with Different Hematocrit Values Using Falling Needle Rheometer

The development of viscometry with high accuracy and quick operation, as well as the establishment of a data evaluation method by pathology are largely required. Especially, the flow properties of human blood are an important factor in the evaluation of blood disease on the medicine, but the method of viscometry and the data collection are not so easy. This study has been described on the viscosity measurement and their evaluations for mammalian blood (rabbit, pig and horse) including human blood. A compact-sized falling needle rheometer (FNR) and a flow analysis method using this device for blood have been developed, and the relationship between the apparent viscosity and physical properties (density, hematocrit value) of blood have also been evaluated. Measured flow properties of blood are evaluated as a flow curve showing the relationship between the shear stress and shear rate. Observed flow curves of mammalian bloods show three typical fluid regions, these are, the Non-newtonian fluid region for a low shear rate range, the transition region and the Newtonian fluid region for a high shear rate range. Flow properties of blood in the Casson fluid region and the apparent viscosity (μ) in the Newtonian fluid region are measured, and they are compared between mammals

The Rise of Three Rs Centres and Platforms in Europe

Public awareness and discussion about animal experiments and replacement methods has greatly increased in recent years. The term 'the Three Rs', which stands for the Replacement, Reduction and Refinement of animal experiments, is inseparably linked in this context. A common goal within the Three Rs scientific community is to develop predictive non-animal models and to better integrate all available data from in vitro, in silico and omics technologies into regulatory decision-making processes regarding, for example, the toxicity of chemicals, drugs or food ingredients. In addition, it is a general concern to implement (human) non-animal methods in basic research. Toward these efforts, there has been an ever-increasing number of Three Rs centres and platforms established over recent years - not only to develop novel methods, but also to disseminate knowledge and help to implement the Three Rs principles in policies and education. The adoption of Directive 2010/63/EU on the protection of animals used for scientific purposes gave a strong impetus to the creation of Three Rs initiatives, in the form of centres and platforms. As the first of a series of papers, this article gives an overview of the European Three Rs centres and platforms, and their historical development. The subsequent articles, to be published over the course of ATLA's 50th Anniversary year, will summarise the current focus and tasks as well as the future and the plans of the Three Rs centres and platforms. The Three Rs centres and platforms are very important points of contact and play an immense role in their respective countries as 'on the ground' facilitators of Directive 2010/63/EU. They are also invaluable for the widespread dissemination of information and for promoting implementation of the Three Rs in general.publishe

Markers for DNA damage are induced in the rat colon by the Alternaria toxin altertoxin-II, but not a complex extract of cultured Alternaria alternata

Mycotoxins produced by Alternaria spp. act genotoxic in cell-based studies, but data on their toxicity in vivo is scarce and urgently required for risk assessment. Thus, male Sprague-Dawley rats received single doses of a complex Alternaria toxin extract (CE; 50 mg/kg bw), altertoxin II (ATX-II; 0.21 mg/kg bw) or vehicle by gavage, one of the most genotoxic metabolites in vitro and were sacrificed after 3 or 24 h, respectively. Using SDS-PAGE/Western Blot, a significant increase of histone 2a.X phosphorylation and depletion of the native protein was observed for rats that were exposed to ATX-II for 24 h. Applying RT-PCR array technology we identified genes of interest for qRT-PCR testing, which in turn confirmed an induction of Rnf8 transcription in the colon of rats treated with ATX-II for 3 h and CE for 24 h. A decrease of Cdkn1a transcription was observed in rats exposed to ATX-II for 24 h, possibly indicating tissue repair after chemical injury. In contrast to the observed response in the colon, no markers for genotoxicity were induced in the liver of treated animals. We hereby provide the first report of ATX-II as a genotoxicant in vivo. Deviating results for similar concentrations of ATX-II in a natural Alternaria toxin mixture argue for substantial mixture effects.ISSN:2673-308

The Current Status and Work of Three Rs Centres and Platforms in Europe

The adoption of Directive 2010/63/EU on the protection of animals used for scientific purposes has given a major push to the formation of Three Rs initiatives in the form of centres and platforms. These centres and platforms are dedicated to the so-called Three Rs, which are the Replacement, Reduction and Refinement of animal use in experiments. ATLA's 50th Anniversary year has seen the publication of two articles on European Three Rs centres and platforms. The first of these was about the progressive rise in their numbers and about their founding history; this second part focuses on their current status and activities. This article takes a closer look at their financial and organisational structures, describes their Three Rs focus and core activities (dissemination, education, implementation, scientific quality/translatability, ethics), and presents their areas of responsibility and projects in detail. This overview of the work and diverse structures of the Three Rs centres and platforms is not only intended to bring them closer to the reader, but also to provide role models and show examples of how such Three Rs centres and platforms could be made sustainable. The Three Rs centres and platforms are very important focal points and play an immense role as facilitators of Directive 2010/63/EU 'on the ground' in their respective countries. They are also invaluable for the wide dissemination of information and for promoting the implementation of the Three Rs in general.publishe

The Multifaceted Roles of Mast Cells in Immune Homeostasis, Infections and Cancers

Mast cells (MCs) play important roles in normal immune responses and pathological states. The location of MCs on the boundaries between tissues and the external environment, including gut mucosal surfaces, lungs, skin, and around blood vessels, suggests a multitude of immunological functions. Thus, MCs are pivotal for host defense against different antigens, including allergens and microbial pathogens. MCs can produce and respond to physiological mediators and chemokines to modulate inflammation. As long-lived, tissue-resident cells, MCs indeed mediate acute inflammatory responses such as those evident in allergic reactions. Furthermore, MCs participate in innate and adaptive immune responses to bacteria, viruses, fungi, and parasites. The control of MC activation or stabilization is a powerful tool in regulating tissue homeostasis and pathogen clearance. Moreover, MCs contribute to maintaining the homeostatic equilibrium between host and resident microbiota, and they engage in crosstalk between the resident and recruited hematopoietic cells. In this review, we provide a comprehensive overview of the functions of MCs in health and disease. Further, we discuss how mouse models of MC deficiency have become useful tools for establishing MCs as a potential cellular target for treating inflammatory disorders

DMM Disease Models and Mechanisms / Cartilage damage and bone erosion are more prominent determinants of functional impairment in longstanding experimental arthritis than synovial inflammation

Chronic inflammation of articular joints causing bone and cartilage destruction consequently leads to functional impairment or loss of mobility in affected joints from individuals affected by rheumatoid arthritis (RA). Even successful treatment with complete resolution of synovial inflammatory processes does not lead to full reversal of joint functionality, pointing to the crucial contribution of irreversibly damaged structural components, such as bone and cartilage, to restricted joint mobility. In this context, we investigated the impact of the distinct components, including synovial inflammation, bone erosion or cartilage damage, as well as the effect of blocking tumor necrosis factor (TNF) on functional impairment in human-TNF transgenic (hTNFtg) mice, a chronic inflammatory erosive animal model of RA. We determined CatWalk-assisted gait profiles as objective quantitative measurements of functional impairment. We first determined body-weight-independent gait parameters, including maximum intensity, print length, print width and print area in wild-type mice. We observed early changes in those gait parameters in hTNFtg mice at week 5 the first clinical signs of arthritis. Moreover, we found further gait changes during chronic disease development, indicating progressive functional impairment in hTNFtg mice. By investigating the association of gait parameters with inflammation-mediated joint pathologies at different time points of the disease course, we found a relationship between gait parameters and the extent of cartilage damage and bone erosions, but not with the extent of synovitis in this chronic model. Next, we observed a significant improvement of functional impairment upon blocking TNF, even at progressed stages of disease. However, blocking TNF did not restore full functionality owing to remaining subclinical inflammation and structural microdamage. In conclusion, CatWalk gait analysis provides a useful tool for quantitative assessment of functional impairment in inflammatory destructive arthritis. Our findings indicate that cartilage damage and bone erosion, but not synovial inflammation, are the most important determinants for progressive functional impairment in this chronic erosive arthritis model.(VLID)329846

CCR6 controls autoimmune but not innate immunitydriven experimental arthritis

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease, characterized by synovial infiltration of various inflammatory cells. Chemokines are involved in controlling the recruitment of different cell types into the synovial membrane. The role of CCR6 in the development of arthritis so far remains unclear. In this study, we investigated the role of CCR6 in the pathogenesis of arthritis using three different murine arthritis models. Compared to WT animals, CCR6/ mice developed less clinical signs of arthritis in the collageninduced arthritis model but not in the K/BxN serum transfer arthritis model and in the human tumour necrosis factor transgenic arthritis model, suggesting a defect in adaptive effector functions but intact innate effector functions in the development of arthritis in CCR6/ animals. In line with this, anticollagen antibody levels were significantly reduced in CCR6/ mice compared with WT mice. Moreover, we demonstrate enhanced osteoclastogenesis in vitro in CCR6/ mice compared with WT mice. However, we did not detect differences in bone mass under steady state conditions in vivo between WT and CCR6deficient mice. These data suggest that CCR6 is crucially involved in adaptive but not in innate immunitydriven arthritis. CCR6 or its chemokine ligand CCL20 might represent a possible new target for the treatment of RA.(VLID)481838