Quantifying Target Occupancy of Small Molecules Within Living Cells.

The binding affinity and kinetics of target engagement are fundamental to establishing structure-activity relationships (SARs) for prospective therapeutic agents. Enhancing these binding parameters for operative targets, while minimizing binding to off-target sites, can translate to improved drug efficacy and a widened therapeutic window. Compound activity is typically assessed through modulation of an observed phenotype in cultured cells. Quantifying the corresponding binding properties under common cellular conditions can provide more meaningful interpretation of the cellular SAR analysis. Consequently, methods for assessing drug binding in living cells have advanced and are now integral to medicinal chemistry workflows. In this review, we survey key technological advancements that support quantitative assessments of target occupancy in cultured cells, emphasizing generalizable methodologies able to deliver analytical precision that heretofore required reductionist biochemical approaches.

Autopolyploidy, Allopolyploidy, and Phylogenetic Networks with Horizontal Arcs.

Polyploidization is an evolutionary process by which a species acquires multiple copies of its complete set of chromosomes. The reticulate nature of the signal left behind by it means that phylogenetic networks offer themselves as a framework to reconstruct the evolutionary past of species affected by it. The main strategy for doing this is to first construct a so-called multiple-labelled tree and to then somehow derive such a network from it. The following question therefore arises: How much can be said about that past if such a tree is not readily available? By viewing a polyploid dataset as a certain vector which we call a ploidy (level) profile, we show that among other results, there always exists a phylogenetic network in the form of a beaded phylogenetic tree with additional arcs that realizes a given ploidy profile. Intriguingly, the two end vertices of almost all of these additional arcs can be interpreted as having co-existed in time thereby adding biological realism to our network, a feature that is, in general, not enjoyed by phylogenetic networks. In addition, we show that our network may be viewed as a generator of ploidy profile space, a novel concept similar to phylogenetic tree space that we introduce to be able to compare phylogenetic networks that realize one and the same ploidy profile. We illustrate our findings in terms of a publicly available Viola dataset.

Blood and liver telomere length, mitochondrial DNA copy number, and hepatic gene expression of mitochondrial dynamics in mid-lactation cows supplemented with l-carnitine under systemic inflammation.

The current study was conducted to examine the effect of l-carnitine (LC) supplementation on telomere length and mitochondrial DNA copy number (mtDNAcn) per cell in mid-lactation cows challenged by lipopolysaccharide (LPS) in blood and liver. The mRNA abundance of 31 genes related to inflammation, oxidative stress, and the corresponding stress response mechanisms, the mitochondrial quality control and the protein import system, as well as the phosphatidylinositol 3-kinase/protein kinase B pathway, were assessed using microfluidics integrated fluidic circuit chips (96.96 dynamic arrays). In addition to comparing the responses in cows with or without LC, our objectives were to characterize the oxidative and inflammatory status by assessing the circulating concentration of lactoferrin (Lf), haptoglobin (Hp), fibrinogen, derivates of reactive oxygen metabolites (dROM), and arylesterase activity (AEA), and to extend the measurement of Lf and Hp to milk. Pluriparous Holstein cows were assigned to either a control group (CON, n = 26) or an LC-supplemented group (CAR; 25 g LC/cow per day; d 42 ante partum to d 126 postpartum (PP), n = 27). On d 111 PP, each cow was injected intravenously with LPS (Escherichia coli O111:B4, 0.5 µg/kg). The mRNA abundance was examined in liver biopsies of d -11 and +1 relative to LPS administration. Plasma and milk samples were frequently collected before and after the challenge. After LPS administration, circulating plasma fibrinogen and serum dROM concentrations increased, whereas AEA decreased. Moreover, serum P4 initially increased by 3 h after LPS administration and declined thereafter irrespective of grouping. The Lf concentrations increased in both groups after LPS administration, with the CAR group showing greater concentrations in serum and milk than the CON group. After LPS administration, telomere length in blood increased, whereas mtDNAcn per cell decreased; however, both remained unaffected in liver. For mitochondrial protein import genes, the hepatic mRNA abundance of the translocase of the mitochondrial inner membrane (TIM)-17B was increased in CAR cows. Moreover, TIM23 increased in both groups after LPS administration. Regarding the mRNA abundance of genes related to stress response mechanisms, 7 out of 14 genes showed group × time interactions, indicating a (local) protective effect due to the dietary LC supplementation against oxidative stress in mid-lactating dairy cows. For mtDNAcn and telomere length, the effects of the LPS-induced inflammation were more pronounced than the dietary supplementation of LC. Dietary LC supplementation affected the response to LPS primarily by altering mitochondrial dynamics. Regarding mRNA abundance of genes related to the mitochondrial protein import system, the inner mitochondrial membrane translocase (TIM complex) seemed to be more sensitive to dietary LC than the outer mitochondrial membrane translocase (TOM complex).

Forest-Based Networks.

In evolutionary studies, it is common to use phylogenetic trees to represent the evolutionary history of a set of species. However, in case the transfer of genes or other genetic information between the species or their ancestors has occurred in the past, a tree may not provide a complete picture of their history. In such cases, tree-based phylogenetic networks can provide a useful, more refined representation of the species' evolution. Such a network is essentially a phylogenetic tree with some arcs added between the tree's edges so as to represent reticulate events such as gene transfer, hybridization and recombination. Even so, this model does not permit the direct representation of evolutionary scenarios where reticulate events have taken place between different subfamilies or lineages of species. To represent such scenarios, in this paper we introduce the notion of a forest-based network, that is, a collection of leaf-disjoint phylogenetic trees on a set of species with arcs added between the edges of distinct trees within the collection. Forest-based networks include the recently introduced class of overlaid species forests which can be used to model introgression. As we shall see, even though the definition of forest-based networks is closely related to that of tree-based networks, they lead to new mathematical theory which complements that of tree-based networks. As well as studying the relationship of forest-based networks with other classes of phylogenetic networks, such as tree-child networks and universal tree-based networks, we present some characterizations of some special classes of forest-based networks. We expect that our results will be useful for developing new models and algorithms to understand reticulate evolution, such as introgression and gene transfer between species.

The hybrid number of a ploidy profile.

Polyploidization, whereby an organism inherits multiple copies of the genome of their parents, is an important evolutionary event that has been observed in plants and animals. One way to study such events is in terms of the ploidy number of the species that make up a dataset of interest. It is therefore natural to ask: How much information about the evolutionary past of the set of species that form a dataset can be gleaned from the ploidy numbers of the species? To help answer this question, we introduce and study the novel concept of a ploidy profile which allows us to formalize it in terms of a multiplicity vector indexed by the species the dataset is comprised of. Using the framework of a phylogenetic network, we present a closed formula for computing the hybrid number (i.e. the minimal number of polyploidization events required to explain a ploidy profile) of a large class of ploidy profiles. This formula relies on the construction of a certain phylogenetic network from the simplification sequence of a ploidy profile and the hybrid number of the ploidy profile with which this construction is initialized. Both of them can be computed easily in case the ploidy numbers that make up the ploidy profile are not too large. To help illustrate the applicability of our approach, we apply it to a simplified version of a publicly available Viola dataset.

Investigation of a potential electrogenic transport-system for myo-inositol in the small intestine of laying hens.

1. Myo-inositol (MI) is an essential metabolite for cell function in animals and humans. The aim of this study was to characterise the transport mechanism of MI in the small intestine of laying hens as there is a lack of knowledge about the MI uptake mechanisms. The hypothesised secondary active, cation coupled transport of MI was assessed by electrophysiological measurements with Ussing chambers, and was compared to the electrophysiology of glucose transport.2. Twenty-six laying hens were used. The potential ion-dependent transport was tested in tissue of the small intestine. Barrier function of the tissue was shown by determining the transepithelial resistance. During the experiments, mucosal and serosal buffers were sampled to measure time-dependent changes in MI concentrations. Samples from eight hens were further used for Western blot analyses of the jejunal apical membranes.3. Active MI transport, indicated by changes in the short circuit current after MI addition, could not be demonstrated in the Ussing chambers experiments. MI was further not detectable in the serosal buffer, nor in the lysates of mucosal tissue cytoplasm nor lipids. Thus, there was no evidence for a MI transport or absorption. However, Western blot analyses of the jejunal apical membrane revealed signals indicated the expression of the MI transport proteins SMIT-1 and SMIT-2.4. In conclusion, the MI transport process in the chicken intestine is more complex than it was presumed and is probably influenced by still unknown regulations or metabolic processes.

Effects of dietary l-carnitine supplementation on the response to an inflammatory challenge in mid-lactating dairy cows: Hepatic mRNA abundance of genes involved in fatty acid metabolism.

This study aimed at characterizing the effects of dietary l-carnitine supplementation on hepatic fatty acid (FA) metabolism during inflammation in mid-lactating cows. Fifty-three pluriparous Holstein dairy cows were randomly assigned to either a control (CON, n = 26) or an l-carnitine supplemented (CAR; n = 27) group. The CAR cows received 125 g of a rumen-protected l-carnitine product per cow per day (corresponding to 25 g of l-carnitine/cow per day) from d 42 antepartum (AP) until the end of the trial on d 126 postpartum (PP). Aside from the supplementation, the same basal diets were fed in the dry period and during lactation to all cows. In mid lactation, each cow was immune-challenged by a single intravenous injection of 0.5 µg of LPS/kg of BW at d 111 PP. Blood samples were collected before and after LPS administration. The mRNA abundance of in total 39 genes related to FA metabolism was assessed in liver biopsies taken at d -11, 1, and 14 relative to LPS (d 111 PP) and also on d 42 AP as an individual covariate using microfluidics integrated fluidic circuit chips (96.96 dynamic arrays). In addition to the concentrations of 3 selected proteins related to FA metabolism, acetyl-CoA carboxylase α (ACACA), 5' AMP-activated protein kinase (AMPK), and solute carrier family 25 member 20 (SLC25A20) were assessed by a capillary Western blot method in liver biopsies from d -11 and 1 relative to LPS from 11 cows each of CAR and CON. On d -11 relative to LPS, differences between the mRNA abundance in CON and CAR were limited to acyl-CoA dehydrogenase (ACAD) very-long-chain (ACADVL) with greater mRNA abundance in the CAR than in the CON group. The liver fat content decreased from d -11 to d 1 relative to the LPS injection and remained at the lower level until d 14 in both groups. One day after the LPS challenge, lower mRNA abundance of carnitine palmitoyltransferase 1 (CPT1), CPT2, ACADVL, ACAD short-chain (ACADS), and solute carrier family 22 member 5 (SLC22A5) were observed in the CAR group as compared with the CON group. However, the mRNA abundance of protein kinase AMP-activated noncatalytic subunit gamma 1 (PRKAG1), ACAD medium-chain (ACADM), ACACA, and FA binding protein 1 (FABP1) were greater in the CAR group than in the CON group on d 1 relative to LPS. Two weeks after the LPS challenge, differences between the groups were no longer detectable. The altered mRNA abundance before and 1 d after LPS pointed to increased transport of FA into hepatic mitochondria during systemic inflammation in both groups. The protein abundance of AMPK was lower in CAR than in CON before the LPS administration. The protein abundance of SLC25A20 was neither changing with time nor treatment and the ACACA protein abundance was only affected by time. In conclusion, l-carnitine supplementation temporally altered the hepatic mRNA abundance of some genes related to mitochondrial biogenesis and very-low-density lipoprotein export in response to an inflammatory challenge, but with largely lacking effects before and 2 wk after LPS.

Covid-19 shelter-at-home and work, lifestyle and well-being in desk workers.

BACKGROUND: Emerging cross-sectional reports find that the COVID-19 pandemic and related social restrictions negatively affect lifestyle behaviours and mental health in general populations. AIMS: To study the longitudinal impact of COVID-19 on work practices, lifestyle and well-being among desk workers during shelter-at-home restrictions. METHODS: We added follow-up after completion of a clinical trial among desk workers to longitudinally measure sedentary behaviour, physical activity, sleep, diet, mood, quality of life and work-related health using validated questionnaires and surveys. We compared outcomes assessed before and during COVID-19 shelter-at-home restrictions. We assessed whether changes in outcomes differed by remote working status (always, changed to or never remote) using analysis of covariance (ANCOVA). RESULTS: Participants (N = 112; 69% female; mean (SD) age = 45.4 (12.3) years; follow-up = 13.5 (6.8) months) had substantial changes to work practices, including 72% changing to remote work. Deleterious changes from before to during shelter-at-home included: 1.3 (3.5)-h increase in non-workday sedentary behaviour; 0.7 (2.8)-point worsening of sleep quality; 8.5 (21.2)-point increase in mood disturbance; reductions in five of eight quality of life subscales; 0.5 (1.1)-point decrease in work-related health (P < 0.05). Other outcomes, including diet, physical activity and workday sedentary behaviour, remained stable (P ≥ 0.05). Workers who were remote before and during the pandemic had greater increases in non-workday sedentary behaviour and stress, with greater declines in physical functioning. Wake time was delayed overall by 41 (61) min, and more so in workers who changed to remote. CONCLUSIONS: Employers should consider supporting healthy lifestyle and well-being among desk workers during pandemic-related social restrictions, regardless of remote working status.

Correction to: Tree-Based Unrooted Phylogenetic Networks.

The level-5 example of a network presented in Fig. 4 of Francis et al. (2018) is tree-based even though it states in the caption and in the text that this is not the case.

Insulin signaling and insulin response in subcutaneous and retroperitoneal adipose tissue in Holstein cows during the periparturient period.

Adipose tissue response to endocrine stimuli, such as insulin, is crucial for metabolic adaptation at the onset of lactation in dairy cows. However, the exact molecular mechanisms behind this response are not well understood. Thus, the aim of this study was to determine the dynamics in protein expression and phosphorylation of key components in insulin signaling in subcutaneous (SCAT) and retroperitoneal (RPAT) adipose tissues of Holstein dairy cows. Furthermore, by ex vivo examinations, response to insulin was assessed in SCAT and RPAT at different time points during the periparturient period. Biopsy samples were taken 42 d prepartum, and 1, 21, and 100 d postpartum. Insulin and glucose concentrations were measured in blood serum in consecutive serum samples from d -42 until d +100. After parturition, the majority of the key components were downregulated in both adipose tissues but recovered by d +100. The extent of hormone-sensitive lipase phosphorylation increased postpartum and remained high throughout the experimental period. Strong differences in molecular response were observed between the 2 depots. The RPAT expressed a remarkably greater extent of AMP-activated kinase phosphorylation compared with SCAT, indicating that AMP-activated kinase as an energy sensor is highly active particularly in RPAT in times of energy scarcity. Consequently, this depot expressed a greater extent of hormone-sensitive lipase phosphorylation over the whole experimental period. Insulin response after parturition appeared to be greater in RPAT too, due to the significantly greater expression of the insulin receptor at d +21 and +100. Although insulin concentrations in plasma were low postpartum, the depot-specific changes in molecular modulation of insulin signaling and insulin response suggested that both adipose tissue depots studied were contributing to the periparturient homeorhetic adaptation, although most likely to a different extent.

Tree-Based Unrooted Phylogenetic Networks.

Phylogenetic networks are a generalization of phylogenetic trees that are used to represent non-tree-like evolutionary histories that arise in organisms such as plants and bacteria, or uncertainty in evolutionary histories. An unrooted phylogenetic network on a non-empty, finite set X of taxa, or network, is a connected, simple graph in which every vertex has degree 1 or 3 and whose leaf set is X. It is called a phylogenetic tree if the underlying graph is a tree. In this paper we consider properties of tree-based networks, that is, networks that can be constructed by adding edges into a phylogenetic tree. We show that although they have some properties in common with their rooted analogues which have recently drawn much attention in the literature, they have some striking differences in terms of both their structural and computational properties. We expect that our results could eventually have applications to, for example, detecting horizontal gene transfer or hybridization which are important factors in the evolution of many organisms.

Different milk feeding intensities during the first 4 weeks of rearing dairy calves: Part 3: Plasma metabolomics analysis reveals long-term metabolic imprinting in Holstein heifers.

Adequate nutrition of calves is a fundamental requirement for efficient production in later life. Suboptimal nutrition before weaning could have detrimental long-term effects on metabolic health and could thereby decrease production efficiency. In this study, the metabolomic profiles of German Holstein calves reared on whole milk ad libitum (n = 10), milk replacer ad libitum (n = 9), and milk replacer in restricted amounts (n = 9) were compared. Furthermore, this profiling approach was extended to the first lactation in the same animals for characterizing the long-term effect of quantitative and qualitative dietary manipulations affecting calves during development in a period that is sensitive to metabolic imprinting. Blood plasma samples were collected on d 3, 22, and 52 of life as well as during wk 4 before and wk 3 and 8 after the first calving. Samples were subjected to a targeted metabolomics analysis using the AbsoluteIDQ p180 kit of Biocrates Life Science AG (Innsbruck, Austria). Profiling of metabolomics data was performed by principal component analysis and heatmap visualization of the metabolome, as well as by comparing fold changes and t-test statistics of metabolites. A quantitative identification of 180 plasma metabolites was possible, belonging to the metabolite classes of acyl-carnitines, AA, biogenic amines, phosphatidylcholines, lysophosphatidylcholines, sphingomyelins, and hexoses. Comparing metabolite concentrations between ad libitum-reared and restrictively reared animals revealed significant differences both during calfhood as well as during first lactation. Most dominantly, acylcarnitines of both short- and long-chain length were more abundant in ad libitum reared animals in the long-term, suggesting alterations in mitochondrial function, most likely indicating adaptive mechanisms of energy expenditure. Furthermore, plasma sphingomyelin concentrations were affected by ad libitum versus restricted milk replacer feeding, which can imply long-term modulatory mechanisms affecting insulin sensitivity. The functional characterization of the identified metabolic patterns, particularly the alterations of single lipid species, is required for further improving our understanding of the links between early nutrition shaping metabolic development and a healthy productive life of Holstein dairy cows.

Effects of energy supply and nicotinic acid supplementation on serum anti-oxidative capacity and on expression of oxidative stress-related genes in blood leucocytes of periparturient primi- and pluriparous dairy cows.

The periparturient period is accompanied by metabolic and oxidative stress. Niacin is known to decrease lipolysis but is also reported to have anti-oxidative effects. Therefore, we examined the effects of energy supply and a nicotinic acid (NA) supplementation on anti-oxidative serum parameters and on the expression of oxidative stress-related genes in blood leucocytes of periparturient dairy cows, differing in parity. Twenty-nine pluriparous and 18 primiparous cows were allocated to four different feeding groups 42 days before expected parturition until 100 days postpartum and fed a ration with either a low concentrate proportion of 30% (LC) or a high concentrate proportion of 60% (HC). After parturition, all animals received 30% concentrate which was increased to 50% either within 16 (LC group) or 24 days (HC group). Half of the animals per group were supplemented with 24 g NA per day from 42 days prepartum until 24 days postpartum. All investigated parameters varied significantly over time compared to parturition (p < .05). Ferric reducing ability (FRA) exhibited a nadir before parturition, and the antioxidant enzymes glutathione peroxidase (GPX) and superoxide dismutase (SOD) showed peak activities around parturition. Expression levels of GPX1, SOD2, xanthine dehydrogenase (XDH) and nuclear factor (erythroid-derived 2)-like 2 (NRF2) peaked before calving. The concentrate level influenced GPX activity and mRNA abundance of SOD2, XDH and poly (ADP-ribose) polymerase 1 (PARP1). Pluriparous animals exhibited higher serum GPX activities, a more distinct nadir for FRA and higher expression levels for GPX1, SOD2 and XDH. Primiparous cows displayed higher serum SOD activities. NA supplementation increased serum SOD activity antepartum in LC animals. Parturition was characterised by an increased need for antioxidants and an increased expression of oxidative stress-related genes that clearly differed with parity and was influenced by energy supply while NA exerted only minor effects on the investigated parameters.

Age intrinsic loss of telomere protection via TRF1 reduction in endothelial cells.

Aging is a major factor predisposing for multiple diseases. Telomeres at the ends of chromosomes protect the integrity of chromosomal DNA. A specialized six-protein complex termed shelterin protects the telomere from unwanted interaction with DNA damage pathways. The aim of our study was to evaluate the integrity of telomeres and the stability of telomere protection during aging in endothelial cells (EC). We describe that aging EC can be characterized by an increased cell size (40%, p=0.02) and increased expression of PAI 1 (4 fold, p=0.02), MCP1 (10 fold, p=0.001) and GMCSF (15 fold, p=0.004). Telomeric state in aging cells is defined by an increased telomere oxidation (27%, p=0.01), reduced telomere length (62%, p=0.02), and increased DNA damage foci formation (5% in young EC versus 16% in aged EC, p=0.003). This telomeric dysfunction is accompanied by a reduction in the shelterin component TRF1 (33% mRNA, p=0.001; 24% protein, p=0.007). Overexpression of TRF1 in aging EC reduced telomere-associated DNA damage foci to 5% (p=0.02) and reduced expression levels of MCP1 (18% reduction, p=0.008). Aged EC have increased telomere damage and an intrinsic loss of telomere protection. Reestablishing telomere integrity could therefore be a target for rejuvenating endothelial cell function.

Silica Polymerization from Supersaturated Dilute Aqueous Solutions in the Presence of Alkaline Earth Salts.

The early stages of silica polymerization in aqueous solution proceed according to a mechanism based on three steps: nucleation, particle growth, and agglomeration of the particles. Application of time-resolved static and dynamic light scattering as a powerful in situ technique in combination with spectrophotometric analysis of the monomer consumption based on the molybdenum blue method was carried out to further investigate this 3-step process. Experiments were carried out at four different initial silicic acid contents covering a range between 350 and 750 ppm in the presence of either 10 mM NaCl or 5 mM of a mixture of CaCl2 and MgCl2. The process in all cases was initiated with a drop of pH to 7. Addition of the salts made possible an analysis of the impact of an electrolyte on the process. Independent of the presence or absence of salt, particle growth in step two proceeded as a monomer-addition process without being interfered significantly by Ostwald-ripening. The growing particles were compact with a homogeneous density. The size of the particles approached final values between 5 and 20 nm with the actual value increasing with decreasing initial silicic acid content. Above a certain concentration of initial silica content, which depends on the level of added salt, particle-particle interactions caused agglomeration. The presence of electrolyte shifted this level from ∼2000 ppm to a range between 500 and 750 ppm. The resulting agglomerates had a fractal dimension of 2. Independent of the conditions, particle growth could be described with a simple nucleation and growth model.

Uprooted Phylogenetic Networks.

The need for structures capable of accommodating complex evolutionary signals such as those found in, for example, wheat has fueled research into phylogenetic networks. Such structures generalize the standard model of a phylogenetic tree by also allowing for cycles and have been introduced in rooted and unrooted form. In contrast to phylogenetic trees or their unrooted versions, rooted phylogenetic networks are notoriously difficult to understand. To help alleviate this, recent work on them has also centered on their "uprooted" versions. By focusing on such graphs and the combinatorial concept of a split system which underpins an unrooted phylogenetic network, we show that not only can a so-called (uprooted) 1-nested network N be obtained from the Buneman graph (sometimes also called a median network) associated with the split system [Formula: see text] induced on the set of leaves of N but also that that graph is, in a well-defined sense, optimal. Along the way, we establish the 1-nested analogue of the fundamental "splits equivalence theorem" for phylogenetic trees and characterize maximal circular split systems.

Minimum triplet covers of binary phylogenetic X-trees.

Trees with labelled leaves and with all other vertices of degree three play an important role in systematic biology and other areas of classification. A classical combinatorial result ensures that such trees can be uniquely reconstructed from the distances between the leaves (when the edges are given any strictly positive lengths). Moreover, a linear number of these pairwise distance values suffices to determine both the tree and its edge lengths. A natural set of pairs of leaves is provided by any 'triplet cover' of the tree (based on the fact that each non-leaf vertex is the median vertex of three leaves). In this paper we describe a number of new results concerning triplet covers of minimum size. In particular, we characterize such covers in terms of an associated graph being a 2-tree. Also, we show that minimum triplet covers are 'shellable' and thereby provide a set of pairs for which the inter-leaf distance values will uniquely determine the underlying tree and its associated branch lengths.

On the challenge of reconstructing level-1 phylogenetic networks from triplets and clusters.

Phylogenetic networks have gained prominence over the years due to their ability to represent complex non-treelike evolutionary events such as recombination or hybridization. Popular combinatorial objects used to construct them are triplet systems and cluster systems, the motivation being that any network N induces a triplet system [Formula: see text] and a softwired cluster system [Formula: see text]. Since in real-world studies it cannot be guaranteed that all triplets/softwired clusters induced by a network are available, it is of particular interest to understand whether subsets of [Formula: see text] or [Formula: see text] allow one to uniquely reconstruct the underlying network N. Here we show that even within the highly restricted yet biologically interesting space of level-1 phylogenetic networks it is not always possible to uniquely reconstruct a level-1 network N, even when all triplets in [Formula: see text] or all clusters in [Formula: see text] are available. On the positive side, we introduce a reasonably large subclass of level-1 networks the members of which are uniquely determined by their induced triplet/softwired cluster systems. Along the way, we also establish various enumerative results, both positive and negative, including results which show that certain special subclasses of level-1 networks N can be uniquely reconstructed from proper subsets of [Formula: see text] and [Formula: see text]. We anticipate these results to be of use in the design of algorithms for phylogenetic network inference.

[Cardioprotection via the arm? : How a blood pressure cuff decreases infarct sizes]. / Kardioprotektion über den Arm? : Wenn eine Blutdruckmanschette den Herzinfarkt verkleinert.

Cardiovascular diseases and especially myocardial infarctions are responsible for a high morbidity and mortality throughout Europe. An essential aspect of myocardial infarction is ischemia/reperfusion injury which represents the necrosis of myocytes following reperfusion. One possible option to counteract ischemia/reperfusion injury is the much researched process of remote ischemic conditioning (RIC), whereby a certain tissue (e.g. skeletal muscle) is subjected to several cycles of short periods (e.g. 5 min) of ischemia and reperfusion and leads to the protection of another organ (e.g. the heart). Despite substantial efforts to elucidate the underlying mechanisms during the last decades, this phenomenon is not yet completely understood. Clinical studies mainly concentrated on laboratory and radiological parameters, which led to better understanding of RIC; however, large clinical studies evaluating the possible influence on mortality are still lacking. This review article provides an introduction to RIC and summarizes the current understanding of known pathomechanisms and the results of important clinical studies.

Reduced Ang2 expression in aging endothelial cells.

Aging endothelial cells are characterized by increased cell size, reduced telomere length and increased expression of proinflammatory cytokines. In addition, we describe here that aging reduces the migratory distance of endothelial cells. Furthermore, we observe an increase of the quiescence protein Ang1 and a decrease of the endothelial activation protein Ang2 upon aging. Supplementing Ang2 to aged endothelial cells restored their migratory capacity. We conclude that aging shifts the balance of the Ang1/Ang2 network favouring a quiescent state. Activation of endothelial cells in aging might be necessary to enhance wound healing capacities.