Novel dipeptidyl peptidase IV resistant analogues of glucagon-like peptide-1(7-36)amide have preserved biological activities in vitro conferring improved glucose-lowering action in vivo

Although the incretin hormone glucagon-like peptide-1 (GLP-1) is a potent stimulator of insulin release, its rapid degradation in vivo by the enzyme dipeptidyl peptidase IV (DPP IV) greatly limits its potential for treatment of type 2 diabetes. Here, we report two novel Ala8-substituted analogues of GLP-1, (Abu8)GLP-1 and (Val8)GLP-1 which were completely resistant to inactivation by DPP IV or human plasma. (Abu8)GLP-1 and (Val8)GLP-1 exhibited moderate affinities (IC50: 4.76 and 81.1 nM, respectively) for the human GLP-1 receptor compared with native GLP-1 (IC50: 0.37 nM). (Abu8)GLP-1 and (Val8)GLP-1 dose-dependently stimulated cAMP in insulin-secreting BRIN BD11 cells with reduced potency compared with native GLP-1 (1.5- and 3.5-fold, respectively). Consistent with other mechanisms of action, the analogues showed similar, or in the case of (Val8)GLP-1 slightly impaired insulin releasing activity in BRIN BD11 cells. Using adult obese (ob/ob) mice, (Abu8 )GLP-1 had similar glucose-lowering potency to native GLP-1 whereas the action of (Val8)GLP-1 was enhanced by 37%. The in vivo insulin-releasing activities were similar. These data indicate that substitution of Ala8 in GLP-1 with Abu or Val confers resistance to DPP IV inactivation and that (Val8)GLP-1 is a particularly potent N-terminally modified GLP-1 analogue of possible use in type 2 diabetes.

A Note on Preserved Smoothness

* Supported by NSERC (Canada)

Three-dimensional tomographic reconstruction of an exceptionally well preserved ichnological assemblage from the Stainmore Foundation, Carboniferous, UK

This Ph.D. thesis addresses current issues with ichnotaxonomic practice, and characterizes an exceptionally well preserved ichnological assemblage from the Carboniferous Stainmore Formation, Northumberland, United Kingdom. Samples were collected from closely localized float representative of various units throughout the succession, which was deposited in a storm-dominated marine shoreface. Three dominant ichnotaxa were selected for three-dimensional morphological analysis due to their complicated morphology and/or unclear taxonomic status: 1) Dactyloidites jordii isp. nov.; 2) Beaconites capronus, and; 3) Neoeione moniliformis comb. nov. Using serial grinding and photography, these ichnotaxa were ground and modelled in true colour. High-resolution models of three taxa produced in this study are the basis of the first complete three-dimensional consideration of the traces, and forms the basis for refined palaeobiological and ethological analysis of these taxa. Dactyloidites jordii isp. nov. is a stellate to palmate burrow composed of numerous long, narrow rays that exhibit three orders of branching arranged into tiered galleries radiating from a central shaft. It is considered to be the feeding structure produced by a vermiform organism. Beaconites capronus is a winding trace with distinctly chevron-shaped, meniscate backfill demonstrated herein to backfill the vertical shafts associated with its burrows in a comparable fashion to the horizontal portion of the burrow. This lack of a surface connection would result in the trace making organism being exposed to low-oxygen porewater. Coping with this porewater dysoxia could be approached by burrowing organisms in a number of ways: 1) revisiting the sediment-water interface; 2) creating periodic shafts; or 3) employing anaerobic metabolism. Neoeione moniliformis was originally introduced as Eione moniliformis, however, the genus Eione Tate, 1859 is a junior homonym of Eione Rafinesque, 1814. This led to the transfer of Eione moniliformis to Parataenidium. Through careful examination and three-dimensional characterization of topotypes, the transfer to Parataenidium moniliformis is demonstrated herein to be problematic, as Parataenidium refers to primarily horizontal burrows with two distinct layers and Eione moniliformis is composed of one distinct level. As such, the new ichnogenus Neoeione is created to accommodate Neoeione moniliformis.

The Effect of Selective Heart Rate Slowing in Heart Failure with Preserved Ejection Fraction

Acknowledgments Dr Ashrafian acknowledges support from the BHF Center of Research Excellence, Oxford, UK. The research was also supported by the National Institute for Health Research Oxford Biomedical Research Center Program and by the National Institute for Health Research Rare Diseases Translational Research Collaboration (NIHR RD-TRC)

The Effect of Selective Heart Rate Slowing in Heart Failure with Preserved Ejection Fraction

Acknowledgments Dr Ashrafian acknowledges support from the BHF Center of Research Excellence, Oxford, UK. The research was also supported by the National Institute for Health Research Oxford Biomedical Research Center Program and by the National Institute for Health Research Rare Diseases Translational Research Collaboration (NIHR RD-TRC)

“Living on the edge” : the role of field margins for common vole (Microtus arvalis) populations in recently colonised Mediterranean farmland

Acknowledgments RRP was supported by a PhD-studentship from the University of Valladolid (co-funded by Banco Santander, RR 30/04/2014). Financial support was provided by ECOCYCLES (BIODIVERSA 2008, Era-net European project, EUI2008-03658 and NERC NE/G002045/1 to XL) and ECOVOLE projects (CGL2012-35348; Ministerio de Economía y Competitividad of Spain). The article also contributes to project ECOTULA (CGL2015-66962-C2-1-R). We held all the necessary licenses and permits for conducting this work (JJLL, FM and RRP held animal experimentation permits of level B for Spain, and a capture permit was provided by the Consejería de Fomento y Medio Ambiente, Junta de Castilla y León (Expte: EP/CYL/665/2014)). We thank two anonymous reviewers for providing and constructive comments to improve the manuscript.

MicroRNA signatures differentiate preserved from reduced ejection fraction heart failure

AIMS: Differentiation of heart failure with reduced (HFrEF) or preserved (HFpEF) ejection fraction independent of echocardiography is challenging in the community. Diagnostic strategies based on monitoring circulating microRNA (miRNA) levels may prove to be of clinical value in the near future. The aim of this study was to identify a novel miRNA signature that could be a useful HF diagnostic tool and provide valuable clinical information on whether a patient has HFrEF or HFpEF.

METHODS AND RESULTS: MiRNA biomarker discovery was carried out on three patient cohorts, no heart failure (no-HF), HFrEF, and HFpEF, using Taqman miRNA arrays. The top five miRNA candidates were selected based on differential expression in HFpEF and HFrEF (miR-30c, -146a, -221, -328, and -375), and their expression levels were also different between HF and no-HF. These selected miRNAs were further verified and validated in an independent cohort consisting of 225 patients. The discriminative value of BNP as a HF diagnostic could be improved by use in combination with any of the miRNA candidates alone or in a panel. Combinations of two or more miRNA candidates with BNP had the ability to improve significantly predictive models to distinguish HFpEF from HFrEF compared with using BNP alone (area under the receiver operating characteristic curve >0.82).

CONCLUSION: This study has shown for the first time that various miRNA combinations are useful biomarkers for HF, and also in the differentiation of HFpEF from HFrEF. The utility of these biomarker combinations can be altered by inclusion of natriuretic peptide. MiRNA biomarkers may support diagnostic strategies in subpopulations of patients with HF.

Exaggerated inflammation and monocytosis associate with diastolic dysfunction in heart failure with preserved ejection fraction: evidence of M2 macrophage activation in disease pathogenesis

BACKGROUND: Heart failure with preserved ejection fraction (HFPEF) is a major health problem associated with myocardial leukocyte infiltration, inflammation, and fibrosis. Monocyte and macrophage subsets play a role in HFPEF but have not been studied. We analyzed peripheral blood monocyte phenotype and plasma markers of monocyte activation in patients with HFPEF, asymptomatic LV diastolic dysfunction (aLVDD), and asymptomatic hypertension (aHTN).

METHODS AND RESULTS: Peripheral blood was collected from 23 aHTN, 30 aLVDD, and 30 HFPEF patients. Peripheral cytokines of classic/pro-inflammatory (tumor necrosis factor alpha, interleukin (IL) 12, IL-6, monocyte chemoattractant protein 1, C-X-C motif chemokine 10) and alternative/anti-inflammatory monocytes (chemokine-C-C motif ligand (CCL) 17, CCL-18, soluble CD163) were increased in aLVDD and HFPEF. Peripheral blood mononuclear cells and monocytes were purified and surface-stained for CD14, CD16, CD163, and CD206. Peripheral monocyte percentage was increased in aLVDD and HFPEF and correlated with echocardiographic LVDD indices. Classic/pro-inflammatory monocyte numbers were increased in aLVDD and HFPEF, and alternative/anti-inflammatory monocyte numbers were increased in HFPEF. CD163 M2-macrophage receptor was reduced in HFPEF. Culture of healthy donor monocytes (n = 3) with HFPEF patient-derived sera (n = 6) promoted M2 macrophage features as evidenced by altered morphology and genes (CD206, IL-10).

CONCLUSIONS: Increased peripheral inflammation, monocytosis, and monocyte differentiation to anti-inflammatory/profibrotic M2 macrophages likely associate with HFPEF and its precedent asymptomatic LVDD phase.

Murine Models of Diastolic Dysfunction and Heart Failure With Preserved Ejection Fraction

Left ventricular diastolic dysfunction leads to heart failure with preserved ejection fraction, an increasingly prevalent condition largely driven by modern day lifestyle risk factors. As heart failure with preserved ejection fraction accounts for almost one-half of all patients with heart failure, appropriate nonhuman animal models are required to improve our understanding of the pathophysiology of this syndrome and to provide a platform for preclinical investigation of potential therapies. Hypertension, obesity, and diabetes are major risk factors for diastolic dysfunction and heart failure with preserved ejection fraction. This review focuses on murine models reflecting this disease continuum driven by the aforementioned common risk factors. We describe various models of diastolic dysfunction and highlight models of heart failure with preserved ejection fraction reported in the literature. Strengths and weaknesses of the different models are discussed to provide an aid to translational scientists when selecting an appropriate model. We also bring attention to the fact that heart failure with preserved ejection fraction is difficult to diagnose in animal models and that, therefore, there is a paucity of well described animal models of this increasingly important condition.

Diel variation in movement patterns and habitat use by the near-threatened cabrera vole in mediterranean farmland

As variações circadianas no comportamento animal e o seu impacto nas populações constituem desafios importantes em ecologia e conservação. Nesta tese documentam-se as variações circadianas no uso do habitat e padrões de movimento pelo rato de Cabrera, em habitats Mediterrânicos fragmentados. O estudo baseou-se no radio-seguimento de indivíduos em habitats dominados por herbáceas e arbustos. Os resultados indicaram que a proporção de tempo despendido em deslocações, a distância percorrida, e a selecção do tipo de vegetação, estão fortemente interrelacionados, variando consideravelmente ao longo de diferentes períodos do dia. Os ratos movimentaram-se mais frequentemente e maiores distâncias nos períodos diurnos, durante os quais as áreas dominadas por herbáceas foram usadas mais intensivamente. Durante a estação seca houve alguma tendência para a diminuição dos movimentos durante as horas mais quentes. Estes resultados são discutidos no sentido de mostrar como indicadores comportamentais podem contribuir para melhorar a gestão e conservação da espécie; ABSTRACT: Understanding the circadian variations in species behaviour and its impacts on population is a challenging topic in ecology and conservation. This thesis documents the circadian variations in habitat use and movement patterns by Cabrera voles in fragmented Mediterranean farmland. The study was based on radiotracking data of individuals living in habitat patches dominated by wet grasses and shrubs. Results indicated that the proportion of time animals spent moving, the distance moved and the selection strength of vegetation were closely linked behavioural traits, which varied considerably across the 24 hour cycle. Voles moved more frequently and over larger distances during daytime, which was when wet grasses were used more intensively. During the dry season there was some tendency for a decrease in movement activity during the hottest hours of the day. These results are used to discuss how behavioural indicators may be useful to improve conservation management of the species.

Factors affecting southern water vole (Arvicola sapidus) detection and occupancy probabilities in Mediterranean farmland

Failure to detect a species at sites where it is present (i.e. imperfect detection) is known to occur frequently, but this is often disregarded in monitoring programs and metapopulation studies. Here we modelled for the first time the probability of patch occupancy by a threatened small mammal, the southern water vole (Arvicola sapidus, while accounting for the probability of detection given occupancy. Based on replicated presence sign surveys conducted in autumn (November–December 2013) and winter (February–March 2014) in a farmland landscape, we used occupancy detection modelling to test the effects of vegetation, sampling effort, observer experience, and rainfall on detection probability. We then assessed whether occupancy was related to patch size, isolation, vegetation, or presence of water, after correcting for imperfect detection. The mean detection probabilities of water vole signs in autumn (0.71) and winter (0.81) indicated that false absences may be generated in about 20–30% of occupied patches surveyed by a single observer on a single occasion. There was no statistical support for the effects of covariates on detectability. After controlling for imperfect detection, the mean probabilities of occupancy in autumn (0.31) and winter (0.29) were positively related to patch size and presence of water, and negatively so, albeit weakly, to patch isolation. Overall, our study underlined the importance of accounting for imperfect detection in sign surveys of small mammals such as water voles, pointing out the need to use occupancy detection modelling together with replicate surveys for accurately estimating occupancy and the factors affecting it.