The boron-oxygen core of borinate esters is responsible for the store-operated calcium entry potentiation ability

International audienceBACKGROUND: Store-Operated Calcium Entry (SOCE) is the major Ca2+ ion entry pathway in lymphocytes and is responsible of a severe combined immunodeficiency (SCID) when deficient. It has recently been observed or highlighted in other cell types such as myoblasts and neurons, suggesting a wider physiological role of this pathway. Whereas Orai1 protein is considered to be the channel allowing the SOCE in T cells, it is hypothesized that other proteins like TRPC could associate with Orai1 to form SOCE with different pharmacology and kinetics in other cell types. Unraveling SOCE cell functions requires specific effectors to be identified, just as dihydropyridines were crucial for the study of Ca2+ voltage-gated channels, or spider/snake toxins for other ion channel classes. To identify novel SOCE effectors, we analyzed the effects of 2-aminoethyl diphenylborinate (2-APB) and its analogues. 2-APB is a molecule known to both potentiate and inhibit T cell SOCE, but it is also an effector of TRP channels and endoplasmic reticulum Ca2+-ATPase. RESULTS: A structure-function analysis allowed to discover that the boron-oxygen core present in 2-APB and in the borinate ester analogues is absolutely required for the dual effects on SOCE. Indeed, a 2-APB analogue where the boron-oxygen core is replaced by a carbon-phosphorus core is devoid of potentiating capacity (while retaining inhibition capacity), highlighting the key role of the boron-oxygen core present in borinate esters for the potentiation function. However, dimesityl borinate ester, a 2-APB analogue with a terminal B-OH group showed an efficient inhibitory ability, without any potentiating capacity. The removal or addition of phenyl groups respectively decrease or increase the efficiency of the borinate esters to potentiate and inhibit the SOCE. mRNA expression revealed that Jurkat T cells mainly expressed Orai1, and were the more sensitive to 2-APB modulation of SOCE. CONCLUSIONS: This study allows the discovery of new boron-oxygen core containing compounds with the same ability as 2-APB to both potentiate and inhibit the SOCE of different leukocyte cell lines. These compounds could represent new tools to characterize the different types of SOCE and the first step in the development of new immunomodulators

A combined analytical-experimental investigation of friction in cylinder liner inserts under mixed and boundary regimes of lubrication

It is necessary to develop an analytical solution in order to combine predictions with measured tribological parameters and fundamentally understand the mechanism of lubrication in a typical region of engine cycle, using tribometric studies. This paper deals with the development of such a representative approach. An analytical, rather than a numerical approach is expounded, as it is shown to suffice for the purpose of precise time-efficient predictions, which conform well to the measurements. The effect of surface topography, material and operating conditions are ascertained for the representative case of top compression ring—cylinder liner contact at the top dead centre reversal in transition from the compression to power stroke. Stainless steel uncoated surface used as press fit cylinder liners for niche original equipment manufacturer applications are compared with those furnished with a Nickel-Silicon Carbide wear-resistant coating of choice in high performance motorsport

Double-Stranded RNA Attenuates the Barrier Function of Human Pulmonary Artery Endothelial Cells

Circulating RNA may result from excessive cell damage or acute viral infection and can interact with vascular endothelial cells. Despite the obvious clinical implications associated with the presence of circulating RNA, its pathological effects on endothelial cells and the governing molecular mechanisms are still not fully elucidated. We analyzed the effects of double stranded RNA on primary human pulmonary artery endothelial cells (hPAECs). The effect of natural and synthetic double-stranded RNA (dsRNA) on hPAECs was investigated using trans-endothelial electric resistance, molecule trafficking, calcium (Ca2+) homeostasis, gene expression and proliferation studies. Furthermore, the morphology and mechanical changes of the cells caused by synthetic dsRNA was followed by in-situ atomic force microscopy, by vascular-endothelial cadherin and F-actin staining. Our results indicated that exposure of hPAECs to synthetic dsRNA led to functional deficits. This was reflected by morphological and mechanical changes and an increase in the permeability of the endothelial monolayer. hPAECs treated with synthetic dsRNA accumulated in the G1 phase of the cell cycle. Additionally, the proliferation rate of the cells in the presence of synthetic dsRNA was significantly decreased. Furthermore, we found that natural and synthetic dsRNA modulated Ca2+ signaling in hPAECs by inhibiting the sarco-endoplasmic Ca2+-ATPase (SERCA) which is involved in the regulation of the intracellular Ca2+ homeostasis and thus cell growth. Even upon synthetic dsRNA stimulation silencing of SERCA3 preserved the endothelial monolayer integrity. Our data identify novel mechanisms by which dsRNA can disrupt endothelial barrier function and these may be relevant in inflammatory processes

Vaccinia Virus G8R Protein: A Structural Ortholog of Proliferating Cell Nuclear Antigen (PCNA)

BACKGROUND: Eukaryotic DNA replication involves the synthesis of both a DNA leading and lagging strand, the latter requiring several additional proteins including flap endonuclease (FEN-1) and proliferating cell nuclear antigen (PCNA) in order to remove RNA primers used in the synthesis of Okazaki fragments. Poxviruses are complex viruses (dsDNA genomes) that infect eukaryotes, but surprisingly little is known about the process of DNA replication. Given our previous results that the vaccinia virus (VACV) G5R protein may be structurally similar to a FEN-1-like protein and a recent finding that poxviruses encode a primase function, we undertook a series of in silico analyses to identify whether VACV also encodes a PCNA-like protein. RESULTS: An InterProScan of all VACV proteins using the JIPS software package was used to identify any PCNA-like proteins. The VACV G8R protein was identified as the only vaccinia protein that contained a PCNA-like sliding clamp motif. The VACV G8R protein plays a role in poxvirus late transcription and is known to interact with several other poxvirus proteins including itself. The secondary and tertiary structure of the VACV G8R protein was predicted and compared to the secondary and tertiary structure of both human and yeast PCNA proteins, and a high degree of similarity between all three proteins was noted. CONCLUSIONS: The structure of the VACV G8R protein is predicted to closely resemble the eukaryotic PCNA protein; it possesses several other features including a conserved ubiquitylation and SUMOylation site that suggest that, like its counterpart in T4 bacteriophage (gp45), it may function as a sliding clamp ushering transcription factors to RNA polymerase during late transcription

Comparison of Fibers for Creep Strengthening of Zinc-aluminum Foundry Alloys

A comparative evaluation is made of a variety of possible fibrous reinforcements for strengthening zinc-aluminium foundry alloys. The composites are processed by squeeze casting, using preforms of alumina, carbon, stainless steel or low carbon steel fibres. A drastic improvement of the creep strength is achieved with the use of alumina or steel fibres. However, an acceptable level of fracture toughness is maintained only in the composites reinforced with steel fibres. This property results from the low interface adhesion which allows bridging of the crack. by the fibres. Low carbon steel fibres do not exhibit more interface reaction than stainless steel fibres. It is concluded that low carbon steel fibres provide a better compromise when taking into account the creep strength, the fracture toughness and the cost of the composite

Zn-al Matrix Composites - Investigation of the Thermal-expansion, Creep Resistance and Fracture-toughness

Composites of Zn-A1 alloys (ZA8 and ZA27) reinforced by fibres of alumina, carbon or steel have been prepared by squeeze casting. Microstructural observations reveal the segregation of the zinc-rich phase around the fibres. The composites exhibit a greatly reduced creep rate in comparison with the matrix alloys. The thermal expansion coefficient is also significantly decreased. The impact toughness has been studied by Charpy tests

Processing and Properties of Metal-matrix Composites Reinforced With Continuous Fibers for the Control of Thermal-expansion, Creep Resistance and Fracture-toughness

Al-, Zn-, and Cu-based matrix composites reinforced with continuous fibres of carbon, SiC, Al2O3, or steel have been processed by squeeze casting or powder metallurgy. Interface reactions can be controlled by alloying additions in the matrix. Interface adhesion has been characterized from the distributions of fibre pull-out lengths on fracture surfaces. Thermal expansion curves reveal the magnitude of the stress transfer at interfaces. In the case of low melting point matrices, ductile steel fibres offer the best combination of fracture toughness and creep resistance