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Marine plankton is an important component of the global carbon cycle. Whereas the production and seafloor export of organic carbon produced by the plankton, the biological pump, has received much attention, the long-term variability in plankton calcification, controlling the carbonate counter pump, remains less well understood. However, it has been shown that changes in pelagic calcification (biological compensation) could affect the ocean's buffering capacity and thus regulate global carbon budget on geological timescales. Here we use Neogene pelagic sediments deposited on the Ceara Rise in the tropical Atlantic to characterize the variability in pelagic carbonate production with a focus on warm climates. A re-evaluation of published records of carbonate accumulation at the Ceara Rise reveals a systematic increase in sedimentation rates since the late Miocene, but the carbonate accumulation rate does not show a clear trend. Instead, we observe substantial orbital timescale variability in carbonate accumulation, combined with a trend towards less carbonate on average at sites located below 4 km, likely due to the effect of carbonate dissolution. To evaluate long-term changes against possible orbital-scale variability, we generated new high-resolution records of carbonate accumulation rate at Ocean Drilling Program (ODP) Site 927 across two Quaternary interglacials (MIS 5 and MIS 9), the Pliocene warm period (MIS KM5) and the Miocene Climatic Optimum (MCO). We observe that the highest carbonate accumulation rates occurred during the Pliocene but that each of the studied intervals was characterized by large-magnitude orbital variability. Prominent variations in carbonate accumulation prior to the Quaternary preservation cycles appear to follow Earth obliquity and eccentricity. These results imply that pelagic carbonate accumulation in the tropical ocean, buffered from large temperature changes, varied on orbital timescales. The magnitude of the orbital-scale variability was similar or even higher than the long-term mean differences among the studied intervals. Since preservation can be excluded as a driver of these changes prior to the Quaternary, the observed variations must reflect changes in the export flux of pelagic biogenic carbonate. We conclude that the overall carbonate production by pelagic calcifiers responded to local changes in light, temperature, and nutrients delivered by upwelling, which followed long orbital cycles, as well as to long-term shifts in climate and/or ocean chemistry. The inferred changes on both timescales were sufficiently large such that when extrapolated on a global scale, they could have played a role in the regulation of the carbon cycle and global climate evolution during the transition from the Miocene warm climates into the Quaternary icehouse.

Abstract Thanks to their outstanding mechanical properties, Bulk Metallic Glasses (BMGs) are new alternatives to traditional crystalline metals for mechanical and micromechanical applications including power transmission. However, the tribological properties of BMGs are still poorly understood, mostly because their amorphous nature induces counter intuitive responses to friction and wear. In the present study, four different BMGs (Cu47Zr46Al7, Zr46Cu45Al7Nb2, Zr60Cu28Al12, and Zr61Cu25Al12Ti2) underwent ball-on-disc friction tests against 100Cr6 steel balls (American Iron and Steel Institute (AISI) 52100) at different relative humidities (RHs) ranging from 20% to 80%. Controlling humidity enabled to observe a high repeatability of the friction and wear responses of the BMG. Interestingly, the friction coefficient decreased by a factor of 2 when the humidity was increased, and the wear rate of BMGs was particularly low thanks to a 3rd-body tribolayer that forms on the BMG surface, composed of oxidized wear particles originating from the ball. The morphology of this tribolayer is highly correlated to humidity. The study also identifies how the tribolayer is built up from the initial contact until the steady state is achieved.

Background Although the critical role of pericytes in maintaining vascular integrity has been extensively demonstrated in the brain and the retina, little is known about their role in the heart. We aim to investigate structural and functional consequences of partial pericyte depletion (≈60%) in the heart of adult mice. Methods and Results To deplete pericytes in adult mice, we used platelet‐derived growth factor receptor β–Cre/ERT2; RosaDTA mice and compared their phenotype with that of control mice (RosaDTA) chosen among their littermates. Cardiac function was assessed via echocardiography and left ventricular catheterization 1 month after the first tamoxifen injection. We found mice depleted with pericytes had a reduced left ventricular ejection fraction and an increased end‐diastolic pressure, demonstrating both systolic and diastolic dysfunction. Consistently, mice depleted with pericytes presented a decreased left ventricular contractility and an increased left ventricular relaxation time (dP/dtmin). At the tissue level, mice depleted of pericytes displayed increased coronary endothelium leakage and activation, which was associated with increased CD45+ cell infiltration. Consistent with systolic dysfunction, pericyte depletion was associated with an increased expression of myosin heavy chain 7 and decreased expression of ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 2 and connexin 43. More important, coculture assays demonstrated, for the first time, that the decreased expression of connexin 43 is likely attributable to a direct effect of pericytes on cardiomyocytes. Besides, this study reveals that cardiac pericytes may undergo strong remodeling on injury. Conclusions Cardiac pericyte depletion induces both systolic and diastolic dysfunction, suggesting that pericyte dysfunction may contribute to the occurrence of cardiac diseases.

Adherent-invasive Escherichia coli (AIEC) strains are frequently recovered from stools of patients with dysbiotic microbiota. They have remarkable properties of adherence to the intestinal epithelium, and survive better than other E. coli in macrophages. The best studied of these AIEC is probably strain LF82, which was isolated from a Crohn's disease patient. This strain contains five complete prophages, which have not been studied until now. We undertook their analysis, both in vitro and inside macrophages, and show that all of them form virions. The Gally prophage is by far the most active, generating spontaneously over 108 viral particles per mL of culture supernatants in vitro, more than 100-fold higher than the other phages. Gally is also over-induced after a genotoxic stress generated by ciprofloxacin and trimethoprim. However, upon macrophage infection, a genotoxic environment, this over-induction is not observed. Analysis of the transcriptome and key steps of its lytic cycle in macrophages suggests that the excision of the Gally prophage continues to be repressed in macrophages. We conclude that strain LF82 has evolved an efficient way to block the lytic cycle of its most active prophage upon macrophage infection, which may participate to its good survival in macrophages.

Heart failure with preserved ejection fraction (HFpEF) has been recognized as the greatest single unmet need in cardiovascular medicine. Indeed, the morbi-mortality of HFpEF is high and as the population ages and the comorbidities increase, so considerably does the prevalence of HFpEF. However, HFpEF pathophysiology is still poorly understood and therapeutic targets are missing. An unifying, but untested, theory of the pathophysiology of HFpEF, proposed in 2013, suggests that cardiovascular risk factors lead to a systemic inflammation, which triggers endothelial cells (EC) and coronary microvascular dysfunction. This cardiac small vessel disease is proposed to be responsible for cardiac wall stiffening and diastolic dysfunction. This paradigm is based on the fact that microvascular dysfunction is highly prevalent in HFpEF patients. More specifically, HFpEF patients have been shown to have decreased cardiac microvascular density, systemic endothelial dysfunction and a lower mean coronary flow reserve. Importantly, impaired coronary microvascular function has been associated with the severity of HF. This review discusses evidence supporting the causal role of endothelial dysfunction in the pathophysiology of HFpEF in human and experimental models.

Sliding electrical contacts are commonly used with a slip ring to collect the current in moving system generators, alternators, or electrical motors. These contacts are also found in electrical transports without batteries, which are mostly supplied by means of a pantograph–catenary system. These systems are fraught with numerous issues. Among them, it is worth highlighting wear and heating, which lead to failures and pre-worn materials. Moreover, with the increase in speed and improvements in technologies and materials, new problems emerge. This is the case with the substitution of the classic copper strip with graphite or copper-impregnated graphite. Multiple works that studied sliding electrical contacts have been achieved recently, some by trying to create a model of the system based on experimental results, and others only based on experimental works and measurements. This paper aims to review articles from this last category by making a synthesis of different test benches used and then by opening a discussion based on different results highlighted by scholars. This discussion is divided into five points that constitute the system inputs. These are the environment, material, normal load, sliding speed, and current. Based on this discussion, a conclusion attempts to evaluate topics where results and trends are commonly established by authors and topics where there is a lack of work or some conflicts in the results or trends between different articles. For this last point, some perspectives are given for further experimental works.

Inflammation of the central nervous system (CNS) induces endothelial blood-brain barrier (BBB) opening as well as the formation of a tight junction barrier between reactive astrocytes at the Glia Limitans. We hypothesized that the CNS parenchyma may acquire protection from the reactive astrocytic Glia Limitans not only during neuroinflammation but also when BBB integrity is compromised in the resting state. Previous studies found that astrocyte-derived Sonic hedgehog (SHH) stabilizes the BBB during CNS inflammatory disease, while endothelial-derived desert hedgehog (DHH) is expressed at the BBB under resting conditions. Here, we investigated the effects of endothelial Dhh on the integrity of the BBB and Glia Limitans. We first characterized DHH expression within endothelial cells at the BBB, then demonstrated that DHH is down-regulated during experimental autoimmune encephalomyelitis (EAE). Using a mouse model in which endothelial Dhh is inducibly deleted, we found that endothelial Dhh both opens the BBB via the modulation of forkhead box O1 (FoxO1) transcriptional activity and induces a tight junctional barrier at the Glia Limitans. We confirmed the relevance of this glial barrier system in human multiple sclerosis active lesions. These results provide evidence for the novel concept of "chronic neuroinflammatory tolerance" in which BBB opening in the resting state is sufficient to stimulate a protective barrier at the Glia Limitans that limits the severity of subsequent neuroinflammatory disease. In summary, genetic disruption of the BBB generates endothelial signals that drive the formation under resting conditions of a secondary barrier at the Glia Limitans with protective effects against subsequent CNS inflammation. The concept of a reciprocally regulated CNS double barrier system has implications for treatment strategies in both the acute and chronic phases of multiple sclerosis pathophysiology.

Continental atmospheric relative humidity (RH) is a key climate parameter. Combined with atmospheric temperature, it allows us to estimate the concentration of atmospheric water vapor, which is one of the main components of the global water cycle and the most important gas contributing to the natural greenhouse effect. However, there is a lack of proxies suitable for reconstructing, in a quantitative way, past changes of continental atmospheric humidity. This reduces the possibility of making model–data comparisons necessary for the implementation of climate models. Over the past 10 years, analytical developments have enabled a few laboratories to reach sufficient precision for measuring the triple oxygen isotopes, expressed by the 17O-excess (17O-excess = ln (δ17O + 1) – 0.528 × ln (δ18O + 1)), in water, water vapor and minerals. The 17O-excess represents an alternative to deuterium-excess for investigating relative humidity conditions that prevail during water evaporation. Phytoliths are micrometric amorphous silica particles that form continuously in living plants. Phytolith morphological assemblages from soils and sediments are commonly used as past vegetation and hydrous stress indicators. In the present study, we examine whether changes in atmospheric RH imprint the 17O-excess of phytoliths in a measurable way and whether this imprint offers a potential for reconstructing past RH. For that purpose, we first monitored the 17O-excess evolution of soil water, grass leaf water and grass phytoliths in response to changes in RH (from 40 to 100 %) in a growth chamber experiment where transpiration reached a steady state. Decreasing RH from 80 to 40 % decreases the 17O-excess of phytoliths by 4.1 per meg/% as a result of kinetic fractionation of the leaf water subject to evaporation. In order to model with accuracy the triple oxygen isotope fractionation in play in plant water and in phytoliths we recommend direct and continuous measurements of the triple isotope composition of water vapor. Then, we measured the 17O-excess of 57 phytolith assemblages collected from top soils along a RH and vegetation transect in inter-tropical West and Central Africa. Although scattered, the 17O-excess of phytoliths decreases with RH by 3.4 per meg/%. The similarity of the trends observed in the growth chamber and nature supports that RH is an important control of 17O-excess of phytoliths in the natural environment. However, other parameters such as changes in the triple isotope composition of the soil water or phytolith origin in the plant may come into play. Assessment of these parameters through additional growth chambers experiments and field campaigns will bring us closer to an accurate proxy of changes in relative humidity.

Abstract Background Viral metagenomic studies have suggested a role for bacteriophages in intestinal dysbiosis associated with several human diseases. However, interpretation of viral metagenomic studies is limited by the lack of knowledge of phages infecting major human gut commensal bacteria, such as Faecalibacterium prausnitzii, a bacterial symbiont repeatedly found depleted in inflammatory bowel disease (IBD) patients. In particular, no complete genomes of phages infecting F. prausnitzii are present in viral databases. Methods We identified 18 prophages in 15 genomes of F. prausnitzii, used comparative genomics to define eight phage clades, and annotated the genome of the type phage of each clade. For two of the phages, we studied prophage induction in vitro and in vivo in mice. Finally, we aligned reads from already published viral metagenomic data onto the newly identified phages. Results We show that each phage clade represents a novel viral genus and that a surprisingly large fraction of them (10 of the 18 phages) codes for a diversity-generating retroelement, which could contribute to their adaptation to the digestive tract environment. We obtained either experimental or in silico evidence of activity for at least one member of each genus. In addition, four of these phages are either significantly more prevalent or more abundant in stools of IBD patients than in those of healthy controls. Conclusion Since IBD patients generally have less F. prausnitzii in their microbiota than healthy controls, the higher prevalence or abundance of some of its phages may indicate that they are activated during disease. This in turn suggests that phages could trigger or aggravate F. prausnitzii depletion in patients. Our results show that prophage detection in sequenced strains of the microbiota can usefully complement viral metagenomic studies.

The data included in this article are related to the research article entitled “Disentangling pectic homogalacturonan and rhamnogalacturonan-I polysaccharides: evidence for sub-populations in fruit parenchyma systems” (Cornuault et al., 2018) [1]. Cell wall properties are an important contributor to fruit texture. These datasets compile textural and immunochemical analysis of polysaccharides of four economically important fruit crops: tomato, strawberry, aubergine and apple with contrasting textures and related taxonomical origins. Cell wall components and their extractability were assessed using characterized monoclonal antibodies. In addition, textural data obtained for the four parenchyma systems show variations in the mechanical properties. The two datasets are a basis to relate cell wall composition and organization to the mechanical properties of the fruit parenchyma tissues. Keywords: Pectic polysaccharides, Fruits, Cell wall, Texture, Monoclonal antibodies, Solanaceae, Rosaceae, Tomato, Aubergine, Apple, Strawberry

Introduction: Tactile sensitivity is impaired in older adults, which contributes to the loss of manual dexterity and mobility function. The reliability of classical psychophysical tests, such as two-point gap discrimination, has been questioned. Here we tested a new method to determine tactile acuity during dynamic touch, which is more functional than static touch. The aim was to validate a method providing a high level of discrimination of tactile acuity in the elderly.Methods: We tested the ability of subjects to evaluate the distance between bands printed on poly-methyl-methacrylate (PMMA) sheets. Pairs of sheets were compared in two groups of participants aged from 60 to 74 years; the test group was required to apply a cosmetic foam with an active ingredient on both their hands twice a day for 1 month, the control group had an identical task but used the same cosmetic foam without any active ingredient. The tests were run in a double-blind, placebo-controlled study.Results: The tactile discrimination threshold decreased by 83 μm after 1 month of cosmetic application in the group using the active ingredient, while it was unchanged in the control group.Discussion: The test presented here provided highly accurate results and should be useful to determine tactile performance. It allows the monitoring of tactile rehabilitation and/or skin treatments used to restore tactile acuity in the elderly.

True oceanic islands typically host reduced species diversity together with high levels of endemism, which make these environmental set-ups ideal for the exploration of species diversification drivers. In the present study, we used black fly species (Diptera: Simuliidae) from Reunion Island as a model to highlight the main drivers of insect species diversification in this young and remote volcanic island located in the Southwestern Indian Ocean. Using local and regional (Comoros and Seychelles archipelagos) samples as well as specimens from continental Africa, we tested the likelihood of two distinct scenarios, i.e. multiple colonizations vs. in-situ diversification. For this, posterior odds were used to test whether species from Reunion did form a monophyletic group and we estimated divergence times between species. Three out of the four previously described Reunion black fly species could be sampled, namely Simulium ruficorne, Simulium borbonense and Simulium triplex. The phylogenies based on nuclear and mitochondrial markers showed that S. ruficorne and S. borbonense are the most closely related species. Interestingly, we report a probable mitochondrial introgression between these two species although they diverged almost six million years ago. Finally, we showed that the three Reunion species did not form a monophyletic group, and, combined with the molecular datation, the results indicated that Reunion black fly diversity resulted from multiple colonization events. Thus, multiple colonizations, rather than in-situ diversification, are likely responsible for an important part of black fly diversity found on this young Darwinian island.

Holocene eastern Mediterranean Sea sediments contain an organic-rich sapropel S1 layer that was formed in oxygen-depleted waters. The spatial distribution of this layer revealed that during S1 deposition, deep waters were anoxic below a depth of 1800 m. However, whether this boundary permanently existed from the early to the mid-Holocene has not been examined yet. To answer this question, a multi-proxy approach was applied to a core retrieved close to the 1800 m boundary (at 1780 m). We measured the bulk sediment elemental composition, the stable isotopic composition of the planktonic foraminifer Globigerinoides ruber and the abundance of benthic foraminifera since the last deglaciation. The result indicates that authigenic U and Mo accumulation began around 13–12 cal ka BP, in concert with surface water freshening estimated from the G. ruber δ18O record. The onset of bottom and pore water oxygen depletion occurred prior to S1 deposition inferred from barium enrichment. In the middle of the S1 deposition period, reduced authigenic V, Fe and As contents and the Br/Cl ratio indicated short-term bottom-water re-oxygenation. A sharp Mn peak and maximal abundance for benthic foraminifera marked a total recovery for circulation at approximately 7 cal ka BP. Based on our results and existing data, we suggest that S1 formation within the upper 1780 m of the eastern Mediterranean Sea was preconditioned by reduced ventilation, resulting from excess freshwater inputs due to insolation changes under deglacial conditions that initiated between 15 and 12 cal ka BP within the upper 1780 m. Short-term re-oxygenation in the Levantine Basin is estimated to have affected bottom water at least as deep as 1780 m in response to cooling and/or the reduction of freshwater inputs. We tentatively propose that complete ventilation recovery at the S1 termination was depth-dependent, with earlier oxygenation within the upper 1780 m. Our results provide new constraints on vertical water column structure in the eastern Mediterranean Sea since the last deglaciation.

Understanding the mechanisms responsible for phenotypic diversification within and among species ultimately rests with linking naturally occurring mutations to functionally and ecologically significant traits. Colour polymorphisms are of great interest in this context because discrete colour patterns within a population are often controlled by just a few genes in a common environment. We investigated how and why phenotypic diversity arose and persists in the Zosterops borbonicus white-eye of Reunion (Mascarene archipelago), a colour polymorphic songbird in which all highland populations contain individuals belonging to either a brown or a grey plumage morph. Using extensive phenotypic and genomic data, we demonstrate that this melanin-based colour polymorphism is controlled by a single locus on chromosome 1 with two large-effect alleles, which was not previously described as affecting hair or feather colour. Differences between colour morphs appear to rely upon complex cis-regulatory variation that either prevents the synthesis of pheomelanin in grey feathers, or increases its production in brown ones. We used coalescent analyses to show that, from a ‘brown’ ancestral population, the dominant ‘grey’ allele spread quickly once it arose from a new mutation. Since colour morphs are always found in mixture, this implies that the selected allele does not go to fixation, but instead reaches an intermediate frequency, as would be expected under balancing selection.

Sandwich ELISA is a highly sensitive method that can be used to determine if two epitopes are part of the same macromolecule or supramolecular complex. In the case of plant cell wall glycans, it can reveal the existence of inter-polymers linkages, leading to better understanding of overall cell wall architectures. This development of a conventional sandwich ELISA protocol uses a carbohydrate-binding module (CBM), a small protein domain found in some carbohydrate catalysing or activating enzymes, and rat monoclonal antibodies (mAbs) which can be combined in the same ELISA plate without risk of cross reaction; the secondary anti-rat HRP antibody being only able to bind to the rat mAb and not the CBM. This protocol was developed and modified in the Prof. J. Paul Knox lab at the University of Leeds.

A protected population of the white-clawed crayfish, Austropotamobius pallipes pallipes (Lereboullet), has been studied between 1995 and 1996. 1,511 crayfish were collected over the period of study including trap and hand catches. 356 animals exceeded the minimum legal size -90 mm TL (Total Length)-, representing 23.6% of the total catches. Catch Per Unit Effort (C.P.U.E.) ranged from 1.2 in February to 8.6 in October. The sex ratio of total catch was biased in favour of females (1:l.g). Males are only dominant during the cold period where water temperature is less than 10°C. Thelohania seems to be cyclic in this population with high levels of infected crayfish during both summers. No infected crayfish were found between January and March. Estimated densities ranged from 4.4 to 2.2 adults.m2 of brook area. Mating activity started in mid to end November as day-length shortened and water temperature fell below 10°C. Berried females ranged from 58.5 to 100.4 mm in TL. Mean pleopodal egg number declined over the duration of the incubation period (mean = 55 eggs per female in December to 30 eggs per female in May). The maximum number of eggs was 85 for a female (85 mm TL) captured in November. In the beginning of May, 40% of females sampled had no eggs. All the results seem to show an over-density of crayfish in this population and recommendations to regulate it are discussed.

Pericytes are perivascular cells imbedded within the basement membrane of microvascular endothelial cells. Importantly, pericytes are critical for microvascular integrity especially endothelial barrier integrity and immune quiescence and may participate in organ disease notably vascular dementia or heart failure. However, little is known about pericyte homeostasis in adult tissues. Our aim is to investigate the behavior of pericytes after they have been reduced by 50% in the brain and heart of adult mice. To deplete pericytes in mice, we used diphtheria toxin (DTA)-mediated conditional cell ablation. More specifically, we used Pdgfrb-Cre/ERT2; Rosa-DTA mice, in which DTA expression is induced specifically in pericytes by Pdgfrb promoter-driven expression of Cre/ERT2 and may be temporally controlled by tamoxifen injections. To induce a 50% pericyte depletion, adult mice were injected with 1 mg tamoxifen for 2 consecutive days and sacrificed 4, 7, 14, 21 and 28 days after the first injection to follow their fate over time. We found pericytes are renewed both in the brain and in the heart. Their renewal follows the same kinetic in both organs, i.e. their number starts to increase between 7 and 14 days after the first tamoxifen injection and pericytes are fully renewed 21 days after they were depleted. The origin of the new pericytes is still under investigation, however, what we found is that pericytes being renewed, transiently express smooth muscle actin alpha 2 (ACTA2), at least in the heart. This study indicates for the first time that pericytes can undergo extensive remodeling after tissue injury in adults. [ABSTRACT FROM AUTHOR]

Many species distribution models (SDMs) are built with precise but geographically restricted presence–absence data sets (e.g., a country) where only a subset of the environmental conditions experienced by a species across its range is considered (i.e., spatial niche truncation). This type of truncation is worrisome because it can lead to incorrect predictions e.g., when projecting to future climatic conditions belonging to the species niche but unavailable in the calibration area. Data from citizen‐science programs, species range maps or atlases covering the full species range can be used to capture those parts of the species' niche that are missing regionally. However, these data usually are too coarse or too biased to support regional management. Here, we aim to (1) demonstrate how varying degrees of spatial niche truncation affect SDMs projections when calibrated with climatically truncated regional data sets and (2) test the performance of different methods to harness information from larger‐scale data sets presenting different spatial resolutions to solve the spatial niche truncation problem. We used simulations to compare the performance of the different methods, and applied them to a real data set to predict the future distribution of a plant species (Potentilla aurea) in Switzerland. SDMs calibrated with geographically restricted data sets expectedly provided biased predictions when projected outside the calibration area or time period. Approaches integrating information from larger‐scale data sets using hierarchical data integration methods usually reduced this bias. However, their performance varied depending on the level of spatial niche truncation and how data were combined. Interestingly, while some methods (e.g., data pooling, downscaling) performed well on both simulated and real data, others (e.g., those based on a Poisson point process) performed better on real data, indicating a dependency of model performance on the simulation process (e.g., shape of simulated response curves). Based on our results, we recommend to use different data integration methods and, whenever possible, to make a choice depending on model performance. In any case, an ensemble modeling approach can be used to account for uncertainty in how niche truncation is accounted for and identify areas where similarities/dissimilarities exist across methods. [ABSTRACT FROM AUTHOR]

Heart failure with preserved ejection fraction (HFpEF) is proposed to be caused by endothelial dysfunction in cardiac small vessels. We previously identified Hhipl1, as a gene upregulated in the coronary vasculature of Leptin receptor deficient mice (Leprdb/db) a well-established mouse model of HFpEF. Importantly, Hhipl1 encodes for a decoy receptor of Desert Hedgehog (DHH) which is known to be critical for endothelial integrity. Our objective is to investigate the functional consequences of impaired Hedgehog (HH) signaling in the adult heart in order to identify novel mechanisms underlying the development of diastolic dysfunction. To do so, Cdh5-Cre/ERT2, DhhFlox/Flox (DhhECKO) mice and their control littermates were administered with tamoxifen at 8 weeks of age to induce Dhh KO. Their cardiac function, exercise tolerance, and the phenotype of their coronary vasculature were assessed one month later. DhhECKO mice presented significantly reduced exercise tolerance, increased end-diastolic pressure (EDP) and Tau, with no change in their ejection fraction consistent with diastolic dysfunction. At molecular and cellular level, impaired cardiac relaxation in DhhECKO mice was associated with a significantly decreased phospholamban phosphorylation on Thr17 and an alteration of sarcomeric shortening in ex-vivo. Besides, as expected, DhhECKO mice exhibited phenotypic changes in their coronary vasculature including a prominent pro-thrombotic phenotype (63 ± 6.2 vs. 25 ± 5.2 thrombi/mm2; P < 0.001) leading to an impaired capillary perfusion and local hypoxia. Notably, antiaggregant therapies (aspirin and clopidogrel) prevented the occurrence of both diastolic dysfunction and exercise intolerance in DhhECKO mice demonstrating for the first time that thrombosis may promote diastolic dysfunction. Importantly, we confirmed the critical role of thrombosis in Leprdb/db mice which also displayed increased cardiac small vessel thrombosis in comparison to control mice. Alike DhhECKO mice, we found that antiaggregants decreased EDP (6.3 ± 0.4 mmHg in aspirin-treated vs. 11.3 ± 0.79 in control mice; P = 0.001) and improved exercise tolerance in Leprdb/db mice (34 ± 2.52 min in aspirin-treated vs. 24 ± 3.55 in control mice; P = 0.004). Altogether, these results demonstrate that small vessel thrombosis may participate in the pathophysiology of heart failure with preserved ejection fraction. [ABSTRACT FROM AUTHOR]

Chronic limb threatening ischemia (CLTI), which is one of the major complications of diabetes, is associated with poor limb and cardiovascular prognoses, along with a dramatic decrease in life expectancy. However, the pathophysiology of this disease is poorly understood and therapeutic targets are missing. Our objective is to explore the contribution of endothelial dysfunction in the development of CLTI in diabetic mice. Hind limb ischemia (HLI) was induced by ligation and resection of the femoral artery in C57BL6/J male mice, in which insulin resistance was induced by a high fat diet, and hyperglycaemia was induced one month later by low dose streptozotocin administrations. According to our previous investigations, ischemic foot re-perfusion, assessed via laser doppler perfusion imaging, was significantly reduced in diabetic animal 28 days after HLI surgery was performed (ratio blood flow in the ischemic leg vs non ischemic leg = 0.27 ± 0.09 vs 0.44 ± 0.21 in control mice; P = 0.03), even though angiogenesis was identical in both groups. On the contrary, we found that impaired ischemic foot re-perfusion was associated with an increased endothelial cell activation attested by an increased ICAM1 expression (P < 0.0001). We then hypothesized that ICAM1, by interacting with white blood cells (WBC), may compromise the perfusion of capillaries with a diameter smaller than a WBC. Accordingly, we found that WBC circulation velocity in the microvasculature was significantly diminished in diabetic mice (533 ± 169 μm/s vs 960 ± 188 in control mice; P = 0.004) and associated with a decreased percentage of capillaries perfusion by BS-1 lectin (78.5 ± 10.5% vs 96.1 ± 5.7 in control mice; P = 0.03). With the aim to test whether ICAM1 overexpression may be responsible for impaired ischemic foot re-perfusion, diabetic mice were administered with anti-ICAM1 antibodies or isotype control for 14 days, starting 14 days after HLI surgery was performed. We found that anti-ICAM1 therapy significantly increased WBC circulation velocity within the microvasculature (779 ± 113 μm/s vs 447 ± 107 in mice treated with isotype control; P < 0.0001) and the percentage of perfused capillary (P = 0.009). Altogether, our results demonstrate that ICAM1 overexpression may compromise hind limb ischemia recovery in diabetic mice by decreasing WBC circulation velocity and impairing capillary perfusion. [ABSTRACT FROM AUTHOR]

Coronary microvascular disease has been proposed to be responsible for heart failure with preserved ejection fraction (HFpEF) about 10 years ago. However, to date the role and phenotype of the coronary microvasculature have still been poorly considered and investigated in animal models of HFpEF. To characterize the phenotype of the coronary microvasculature in male and female mice with diastolic dysfunction. We assessed cardiac function and characterized the coronary microvasculature in two mouse models of HFpEF: mice fed with a high fat diet (HFD) + L-NAME regimen and Leptin receptor deficient (Leprdb/db) mice. Notably, our study was done in males, females and ovariectomized (OVX) female mice in order to search for possible sexual dysmorphism. Upon a HFD + L-NAME regimen, both males and OVX females but not non OVX females develop diastolic dysfunction attested by an increased end diastolic pressure. In Leprdb/db mice, both male and non OVX female mice develop diastolic dysfunction. Notably, female mice have reduced estradiol and progesterone level mice mimicking ovariectomy. We found that both OVX and non OVX females but not males display increased endothelial activation attested by increased ICAM1 expression, endothelium leakage attested by increased Fibrinogen and IgG extravasation and decreased arteriole diameter suggesting vasoconstriction. The same results were found in Leprdb/db mice. Diastolic dysfunction is not always associated with cardiac small vessel disease since Leprdb/db males and C57BL/6 J males fed with a high fat diet (HFD) + L-NAME regimen develop diastolic dysfunction in the absence of endothelial dysfunction. Also endothelial dysfunction may not be sufficient to induce diastolic dysfunction since non OVX female mice fed with a high fat diet (HFD) + L-NAME regimen display endothelial dysfunction while they do not develop diastolic dysfunction. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases which are the leading cause of death worldwide are promoted by well-known risk factors that include high blood pressure, age, sex, diabetes and/or obesity. Pericytes are mural cells wrapped around microvessels in many organs. They are mostly described as essential for microvascular integrity in the brain and retina. While the detrimental effects of cardiovascular risk factors on endothelial cells have been extensively studied, little is known about their impact on the biology of pericytes, especially in the heart. To determine how cardiac pericytes are modified by some cardiovascular risk factors. To do so, we used mice that were exposed to a high-fat diet (HFD) only to recapitulate obesity, to a HFD together with low dose-streptozotocin injections to mimic type 2 diabetes or to a HFD and L-NAME to recapitulate obesity with a mild hypertension. Mice were sacrificed 3 or 6 months after establishing these risk factors for histological and gene expression analyses. In mice exposed to the HFD + L-NAME regimen for 3 months, the percentage of capillaries covered by pericytes was not modified. However their phenotype appeared to be modified. Indeed, we isolated pericytes (CD31–, CD146+ cells) in the heart of both HFD L-NAME fed mice and control mice using magnetic beads and performed a bulk RNA-sequencing analysis on these cells which identified 3 genes of which the expression was significantly different in the 2 groups. In particular, Angiopoietin-like 4 mRNA expression was increased in the pericytes of HFD + L-NAME fed mice. This protein plays an important role in the regulation of lipid metabolism which makes it attractive to explore. In mice exposed to either HFD alone of HFD + streptozotocin for 6 months, the percentage of capillaries covered by pericytes was significantly diminished suggesting pericyte death. Cardiovascular risk factors, such as obesity and diabetes, may change the phenotype and/or the number of pericytes in the heart suggesting that these cells, known to be critical for microvascular integrity, could contribute to the onset of cardiac diseases. [ABSTRACT FROM AUTHOR]

Lower-limb peripheral artery disease (PAD) is a prevalent and challenging complication of diabetes, requiring innovating therapies. Praliciguat is an orally available stimulator of soluble guanylate cyclase (sGC) reported to have favorable effects on metabolic and hemodynamic endpoints in preclinical and clinical studies, suggesting the potential for benefit in PAD. We evaluated the effect of praliciguat on hindlimb ischemia recovery in a mouse model of diabetes and investigated the molecular mechanism of sGC stimulation. Hindlimb ischemia was induced in db/db mice by ligation and excision of the left femoral artery. Praliciguat 10 mg/kg/day (n = 10 mice) or vehicle (n = 10) were administered in the diet for 31 days starting 3 days before surgery. Foot perfusion was assessed with a Laser Doppler Imager and reported as a ratio in the ischemic versus non-ischemic limb. Ischemic leg function was assessed with a 4-point scale: 0, plantar/toe flexion in response to tail traction; 1, plantar but not toe flexion; 2, no flexion; 3, foot dragging. Ischemic foot perfusion and function were better 28 days after surgery in praliciguat than in vehicle treated mice (mean ± SD: perfusion 1.05 ± 0.23 vs. 0.38 ± 0.16; P < 0.0001; function 1.14 ± 0.36 vs. 1.73 ± 0.80; P = 0.03). Praliciguat did not impact angiogenesis. Arteriolar diameter was higher (9.88 ± 0.96 vs. 8.57 ± 0.64 μm; P = 0.004) and ICAM-1 expression lower (186 ± 41 vs. 403 ± 177% of healthy muscle; P = 0.005) in the ischemic limb of mice treated with praliciguat than with vehicle. Praliciguat attenuated accumulation of oxidative pro-angiogenic and pro-inflammatory muscle fibers. In cultured myoblasts praliciguat significantly downregulated Myh2 and Cxcl12 mRNA expression. Conditioned medium from praliciguat-treated myoblast decreased ICAM-1 mRNA expression in HUVECs. These results suggest that praliciguat restored perfusion and function in the ischemic muscle of db/db mice by increasing arteriolar diameter and decreasing ICAM-1 expression in endothelial cells via downregulation of Cxcl12 in myocytes. [ABSTRACT FROM AUTHOR]

Heart failure with preserved ejection fraction is proposed to be caused by endothelial dysfunction in cardiac small vessels. Our goal is to demonstrate that endothelial dysfunction is sufficient to induce diastolic dysfunction using mice with genetically induced endothelial dysfunction. We used Desert Hedgehog (Dhh) endothelial deficient mice (DhhECKO) as such a model. Indeed, we recently demonstrated that Dhh is critical for endothelial integrity. Practically, DhhECKO mice were administered with tamoxifen at 8 weeks of age to induce Dhh KO and endothelial dysfunction. Mice were sacrificed one month later. As expected, DhhECKO mice exhibited phenotypic changes in their cardiac small vessels characterized by a pro-inflammatory phenotype, a pro-thrombotic phenotype and abnormal endothelium permeability. Notably, these changes are sufficient to induce diastolic dysfunction since DhhECKO mice presented significantly increased end-diastolic pressures, while their left ventricular ejection fraction was comparable to control mice. Moreover, DhhECKO have a reduced exercise tolerance, suggesting a heart failure. Importantly, Aspirin therapy prevented the occurrence of both diastolic dysfunction and exercise intolerance in these mice. Altogether, these results demonstrate that small vessel thrombosis may participate in the pathophysiology of diastolic dysfunction. [ABSTRACT FROM AUTHOR]