Activation of Rac-1 and RhoA contributes to podocyte injury in chronic kidney disease

Rho-family GTPases like RhoA and Rac-1 are potent regulators of cellular signaling that control gene expression, migration and inflammation. Activation of Rho-GTPases has been linked to podocyte dysfunction, a feature of chronic kidney diseases (CKD). We investigated the effect of Rac-1 and Rho kinase (ROCK) inhibition on progressive renal failure in mice and studied the underlying mechanisms in podocytes. SV129 mice were subjected to 5/6-nephrectomy which resulted in arterial hypertension and albuminuria. Subgroups of animals were treated with the Rac-1 inhibitor EHT1846, the ROCK inhibitor SAR407899 and the ACE inhibitor Ramipril. Only Ramipril reduced hypertension. In contrast, all inhibitors markedly attenuated albumin excretion as well as glomerular and tubulo-interstitial damage. The combination of SAR407899 and Ramipril was more effective in preventing albuminuria than Ramipril alone. To study the involved mechanisms, podocytes were cultured from SV129 mice and exposed to static stretch in the Flexcell device. This activated RhoA and Rac-1 and led via TGFβ to apoptosis and a switch of the cells into a more mesenchymal phenotype, as evident from loss of WT-1 and nephrin and induction of α-SMA and fibronectin expression. Rac-1 and ROCK inhibition as well as blockade of TGFβ dramatically attenuated all these responses. This suggests that Rac-1 and RhoA are mediators of podocyte dysfunction in CKD. Inhibition of Rho-GTPases may be a novel approach for the treatment of CKD

Reciprocal regulation of PKA and rac signaling

Activated G protein-coupled receptors (GPCRs) and receptor tyrosine kinases relay extracellular signals through spatial and temporal controlled kinase and GTPase entities. These enzymes are coordinated by multifunctional scaffolding proteins for precise intracellular signal processing. The cAMP-dependent protein kinase A (PKA) is the prime example for compartmentalized signal transmission downstream of distinct GPCRs. A-kinase anchoring proteins tether PKA to specific intracellular sites to ensure precision and directionality of PKA phosphorylation events. Here, we show that the Rho-GTPase Rac contains A-kinase anchoring protein properties and forms a dynamic cellular protein complex with PKA. The formation of this transient core complex depends on binary interactions with PKA subunits, cAMP levels and cellular GTP-loading accounting for bidirectional consequences on PKA and Rac downstream signaling. We show that GTP-Rac stabilizes the inactive PKA holoenzyme. However, β-adrenergic receptor-mediated activation of GTP-Rac–bound PKA routes signals to the Raf-Mek-Erk cascade, which is critically implicated in cell proliferation. We describe a further mechanism of how cAMP enhances nuclear Erk1/2 signaling: It emanates from transphosphorylation of p21-activated kinases in their evolutionary conserved kinase-activation loop through GTP-Rac compartmentalized PKA activities. Sole transphosphorylation of p21-activated kinases is not sufficient to activate Erk1/2. It requires complex formation of both kinases with GTP-Rac1 to unleash cAMP-PKA–boosted activation of Raf-Mek-Erk. Consequently GTP-Rac functions as a dual kinase-tuning scaffold that favors the PKA holoenzyme and contributes to potentiate Erk1/2 signaling. Our findings offer additional mechanistic insights how β-adrenergic receptor-controlled PKA activities enhance GTP-Rac–mediated activation of nuclear Erk1/2 signaling

Notes on large angle crossing graphs

A graph G is an a-angle crossing (aAC) graph if every pair of crossing edges in G intersect at an angle of at least a. The concept of right angle crossing (RAC) graphs (a=Pi/2) was recently introduced by Didimo et. al. It was shown that any RAC graph with n vertices has at most 4n-10 edges and that there are infinitely many values of n for which there exists a RAC graph with n vertices and 4n-10 edges. In this paper, we give upper and lower bounds for the number of edges in aAC graphs for all 0 < a < Pi/2

Rapid identification and differentiation of the vaccine strain Rac H from EHV 1 field isolates using a non-radioactive DNA probe

A method for rapid differentiation between the EHV 1 live vaccine strain Rac H and field isolates is described. Total DNA was isolated from virus-infected small scale cell cultures. DNA fragments digested with restriction endonuclease BamHI were separated, transfered and immobilized on filter membranes. A Digoxigenin-labeled probe derived from EHV 1 was used for hybridization. This probe hybridized specifically to sequences of the inverted terminal repeat region which in case of Rac H include a deletion of 0.8 kb. By comparing the different migration patterns after blot hybridization it could be shown that in 65 isolates from cases of abortion the live vaccine strain Rac H was not involve

Dobutamine stress MRI in pulmonary hypertension: relationships between stress pulmonary artery relative area change, RV performance, and 10-year survival

In pulmonary hypertension (PH), right ventricular (RV) performance determines survival. Pulmonary artery (PA) stiffening is an important biomechanical event in PH and also predicts survival based on the PA relative area change (RAC) measured at rest using magnetic resonance imaging (MRI). In this exploratory study, we sought to generate novel hypotheses regarding the influence of stress RAC on PH prognosis and the interaction between PA stiffening, RV performance and survival. Fifteen PH patients underwent dobutamine stress-MRI (ds-MRI) and right heart catheterization. RACREST, RACSTRESS, and ΔRAC (RAC STRESS – RAC REST) were correlated against resting invasive hemodynamics and ds-MRI data regarding RV performance and RV-PA coupling efficiency (n’vv [RV stroke volume/RV end-systolic volume]). The impact of RAC, RV data, and n’vv on ten-year survival were determined using Kaplan–Meier analysis. PH patients with a low ΔRAC (&lt;−2.6%) had a worse long-term survival (log-rank P = 0.045, HR for death = 4.46 [95% CI = 1.08–24.5]) than those with ΔRAC ≥ −2.6%. Given the small sample, these data should be interpreted with caution; however, low ΔRAC was associated with an increase in stress diastolic PA area indicating proximal PA stiffening. Associations of borderline significance were observed between low RACSTRESS and low n’vvSTRESS, Δη’VV, and ΔRVEF. Further studies are required to validate the potential prognostic impact of ΔRAC and the biomechanics potentially connecting low ΔRAC to shorter survival. Such studies may facilitate development of novel PH therapies targeted to the proximal PA

Compact Drawings of 1-Planar Graphs with Right-Angle Crossings and Few Bends

We study the following classes of beyond-planar graphs: 1-planar, IC-planar, and NIC-planar graphs. These are the graphs that admit a 1-planar, IC-planar, and NIC-planar drawing, respectively. A drawing of a graph is 1-planar if every edge is crossed at most once. A 1-planar drawing is IC-planar if no two pairs of crossing edges share a vertex. A 1-planar drawing is NIC-planar if no two pairs of crossing edges share two vertices. We study the relations of these beyond-planar graph classes (beyond-planar graphs is a collective term for the primary attempts to generalize the planar graphs) to right-angle crossing (RAC) graphs that admit compact drawings on the grid with few bends. We present four drawing algorithms that preserve the given embeddings. First, we show that every $n$-vertex NIC-planar graph admits a NIC-planar RAC drawing with at most one bend per edge on a grid of size $O(n) \times O(n)$. Then, we show that every $n$-vertex 1-planar graph admits a 1-planar RAC drawing with at most two bends per edge on a grid of size $O(n^3) \times O(n^3)$. Finally, we make two known algorithms embedding-preserving; for drawing 1-planar RAC graphs with at most one bend per edge and for drawing IC-planar RAC graphs straight-line

Permeability of Concrete with Recycled Concrete Aggregate and Pozzolanic Materials under Stress.

The research reported herein studied the permeability of concrete containing recycled-concrete aggregate (RA), superfine phosphorous slag (PHS), and ground granulated blast-furnace slag (GGBS) with and without stress. Test results showed that the chloride diffusion coefficient of RA concrete (RAC) without external loads decreased with time, and the permeability of RAC is much lower than that of the reference concrete due to the on-going hydration and the pozzolanic reaction provided by the PHS and GGBS additives in the RAC mixture. The permeability of chloride under flexural load is much more sensitive than that under compressive load due to the differences in porosity and cracking pattern. At low compressive stress levels, the permeability of chloride decreased by the closing of pores and microcracks within RAC specimens. However, in a relatively short time the chloride diffusion coefficient and the chloride content increased rapidly with the increase of compressive stress when it exceeded a threshold stress level of approximate 35% of the ultimate compressive strength. Under flexural stress, the chloride transport capability increased with the increase of stress level and time. At high compressive and flexural stress levels, creep had a significant effect on the permeability of chloride in the RAC specimens due to the damage from the nucleation and propagation of microcracks over time. It is apparent that mortar cracking has more of a significant effect on the chloride transport in concrete than cracking in the interfacial transition zone (ITZ)

α-Tocopherol in plasma and milk from organically managed dairy cows fed natural or synthetic vitamin E or seaweed

The objective was to compare the effects of supplementing lactating dairy cows with synthetic (All-rac), natural (RRR) α-tocopheryl acetate or seaweed with a control on the concentration of α-tocopherol in blood and milk. Twenty four dairy cows in mid lactation, fed an organic feed ration, were randomly allocated to the four treatments in a replicated Latin square design. Plasma and milk α-tocopherol concentrations were higher in RRR and All-rac than in the other treatments and higher in RRR than in All-rac. RRR-α-tocopherol was the predominant steroisomer (> 86%), in both plasma and milk, whereas the remaining part was largely made up by the three synthetic 2R isomers. In cows fed the control, seaweed and RRR, the proportion of RRR-α-tocopherol in plasma and milk constituted more than 97% of the total α-tocopherol. The study demonstrated that dairy cows in mid and late lactation have preferential uptake of RRR-α-tocopherol compared to other stereoisomer

Integrins engage mitochondrial function for signal transduction by a mechanism dependent on Rho GTPases.

We show here the transient activation of the small GTPase Rac, followed by a rise in reactive oxygen species (ROS), as necessary early steps in a signal transduction cascade that lead to NFkappaB activation and collagenase-1 (CL-1)/matrix metalloproteinase-1 production after integrin-mediated cell shape changes. We show evidence indicating that this constitutes a new mechanism for ROS production mediated by small GTPases. Activated RhoA also induced ROS production and up-regulated CL-1 expression. A Rac mutant (L37) that prevents reorganization of the actin cytoskeleton prevented integrin-induced CL-1 expression, whereas mutations that abrogate Rac binding to the neutrophil NADPH membrane oxidase in vitro (H26 and N130) did not. Instead, ROS were produced by integrin-induced changes in mitochondrial function, which were inhibited by Bcl-2 and involved transient membrane potential loss. The cells showing this transient decrease in mitochondrial membrane potential were already committed to CL-1 expression. These results unveil a new molecular mechanism of signal transduction triggered by integrin engagement where a global mitochondrial metabolic response leads to gene expression rather than apoptosis

Optimising nutrition in residential aged care: A narrative review

In developed countries the prevalence of protein-energy malnutrition increases with age and multi-morbidities increase nutritional risk in aged care residents in particular. This paper presents a narrative review of the current literature on the identification, prevalence, associated risk factors, consequences, and management of malnutrition in the <i>residential aged care (RAC)</i> setting. We performed searches of English-language publications on <i>Medline, PubMed, Ovid and the Cochrane Library</i> from January 1 1990 to November 25 2015. We found that, on average, half of all residents in aged care are malnourished as a result of factors affecting appetite, dietary intake and nutrient absorption. Malnutrition is associated with a multitude of adverse outcomes, including increased risk of infections, falls, pressure ulcers and hospital admissions, all of which can lead to increased health care costs and poorer quality of life. A number of food and nutrition strategies have demonstrated positive nutritional and clinical outcomes in the <i>RAC</i> setting. These strategies extend beyond simply enhancing the nutritional value of foods and hence necessitate the involvement of a range of committed stakeholders. Implementing a nutritional protocol in <i>RAC</i> facilities that comprises routine nutrition screening, assessment, appropriate nutrition intervention, including attention to food service systems, and monitoring by a multidisciplinary team can help prevent decline in residents’ nutritional status. Food and nutritional issues should be identified early and managed on admission and regularly in the <i>RAC</i> setting