Characterization of the Roco Protein Family in Dictyostelium discoideum

The Roco family consists of multidomain Ras-GTPases that include LRRK2, a protein mutated in familial Parkinson's disease. The genome of the cellular slime mold Dictyostelium discoideum encodes 11 Roco proteins. To study the functions of these proteins, we systematically knocked out the roco genes. Previously described functions for GbpC, Pats1, and QkgA (Roco1 to Roco3) were confirmed, while novel developmental defects were identified in roco4- and roco11-null cells. Cells lacking Roco11 form larger fruiting bodies than wild-type cells, while roco4-null cells show strong developmental defects during the transition from mound to fruiting body; prestalk cells produce reduced levels of cellulose, leading to unstable stalks that are unable to properly lift the spore head. Detailed phylogenetic analysis of four slime mold species reveals that QkgA and Roco11 evolved relatively late by duplication of an ancestor roco4 gene (later than ∼300 million years ago), contrary to the situation with other roco genes, which were already present before the split of the common ancestor of D. discoideum and Polysphondylium pallidum (before ∼600 million years ago). Together, our data show that the Dictyostelium Roco proteins serve a surprisingly diverse set of functions and highlight Roco4 as a key protein for proper stalk cell formation

The Roco protein family:a functional perspective

In this review, we discuss the evolutionary, biochemical, and functional data available for members of the Roco protein family. They are characterized by having a conserved supradomain that contains a Ras-like GTPase domain, called Roc, and a characteristic COR (C-terminal of Roc) domain. A kinase domain and diverse regulatory and protein protein interaction domains are also often found in Roco proteins. First detected in the slime mold Dictyostelium discoideum, they have a broad phylogenetic range, being present in both prokaryotes and eukaryotes. The functions of these proteins are diverse. The best understood are Dictyostelium Rocos, which are involved in cell division, chemotaxis, and development. However, this family has received extensive attention because mutations in one of the human Roco genes (LRRK2) cause familial Parkinson disease. Other human Rocos are involved in epilepsy and cancer. Biochemical data suggest that Roc domains are capable of activating kinase domains intramolecularly. Interestingly, some of the dominant, disease-causing mutations in both the GTPase and kinase domains of LRRK2 increase kinase activity. Thus, Roco proteins may act as stand-alone transduction units, performing roles that were thought so far to require multiple proteins, as occur in the Ras transduction pathway

Advancing interoperable soil data exchange for global soil data information systems

In order to be able to address local, regional and global issues such as sustainable land management, food security, climate change mitigation and soil-related indicators of the UN Agenda for Sustainable Development the need for reliable, relevant and accurate soil information and data is increasing. Currently, ..

Intra-arterial ultrasonic imaging for recanalization by spark erosion

Presently several new methods are being developed to recanalize obstructed arteries during catheterization. Intra-arterial high frequency ultrasonic imaging may be used as a guidance for these new techniques. Spark erosion is a new obstruction removal technology. Experiments have shown that this method can be applied in a selective way. An ultrasonic intra-arterial imaging system allows for the proper indication of the spark erosion catheter relative to the obstruction. The first in vitro results of this study illustrate that integration of catheter tip imaging and spark erosion is possible

Targeting endocannabinoid signaling: FAAH and MAG lipase inhibitors

Inspired by the medicinal properties of the plant Cannabis sativa and its principal component (−)-trans-Δ9-tetrahydrocannabinol (THC), researchers have developed a variety of compounds to modulate the endocannabinoid system in the human brain. Inhibitors of fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), which are the enzymes responsible for the inactivation of the endogenous cannabinoids anandamide and 2-arachidonoylglycerol, respectively, may exert therapeutic effects without inducing the adverse side effects associated with direct cannabinoid CB1 receptor stimulation by THC. Here we review the FAAH and MAGL inhibitors that have reached clinical trials, discuss potential caveats, and provide an outlook on where the field is headed.NWOMolecular Physiolog

Clickable Vitamins as a New Tool to Track Vitamin A and Retinoic Acid in Immune Cells.

The vitamin A derivative, retinoid acid (RA) is key player in guiding adaptive mucosal immune responses. However, data on the uptake and metabolism of vitamin A within human immune cells has remained largely elusive because retinoids are small, lipophilic molecules which are difficult to detect. To overcome this problem and to be able to study the effect of vitamin A metabolism in human immune cell subsets, we have synthesized novel bio-orthogonal retinoid-based probes (clickable probes), which are structurally and functionally indistinguishable from vitamin A. The probes contain a functional group (an alkyne) to conjugate to a fluorogenic dye to monitor retinoid molecules in real-time in immune cells. We demonstrate, by using flow cytometry and microscopy, that multiple immune cells have the capacity to internalize retinoids to varying degrees, including human monocyte-derived dendritic cells (DCs) and naïve B lymphocytes. We observed that naïve B cells lack the enzymatic machinery to produce RA, but use exogenous retinoic acid to enhance CD38 expression. Furthermore, we showed that human DCs metabolize retinal into retinoic acid, which in co-culture with naïve B cells led to of the induction of CD38 expression. These data demonstrate that in humans, DCs can serve as an exogenous source of RA for naïve B cells. Taken together, through the use of clickable vitamins our data provide valuable insight in the mechanism of vitamin A metabolism and its importance for human adaptive immunity.Molecular Physiolog

Detection of vascular morphology by high frequency intravascular ultrasonic imaging

This study was designed to validate the potential clinical utility of intravascular ultrasonic imaging in vitro and in vivo. In vitro studies were performed to assess the accuracy of dimensional and morphological information. In vitro images of human vessels (n = 75) demonstrated that lesion thickness determined echographically closely related with histological samples (r = 0.83). Morphologically, muscular and elastic arteries could be distinguished echographically based on the echogenicity of the arterial media. Close relation was also found in the morphological subtypes of atherosclerosis. Subsequently, intravascular ultrasound was used percutaneously in vivo in 20 patients to obtain images of the iliac and superficial femoral artery. High quality real-time images were obtained. Normal vessels were seen showing pulsatile circular images with a hypoechoic muscular media resulting in a typical three-layered appearance. Diseased arteries revealed non-obstructive and obstructive lumen. At the site of obstruction thinning of the muscular media was evident. Pulsation was not always present. Following dilatation of the obstructive lesion using balloon angioplasty the ultrasonic cross-sections changed drastically revealing plaque rupture, dissection, plaque-free wall rupture, rest stenosis and oedema. We conclude that intravascular ultrasonic imaging is a promising technique to document accurate dimensional and morphological characteristics of human vascular disease for guidance of therapeutic interventions