Highly cytotoxic trithiophenolatodiruthenium complexes of the type [(η6- p -MeC6H4Pr i )2Ru2(SC6H4- p -X)3]+: synthesis, molecular structure, electrochemistry, cytotoxicity, and glutathione oxidation potential

A series of cationic dinuclear p-cymene ruthenium trithiophenolato complexes of the type [(η6-p-MeC6H4Pr i )2Ru2(SC6H4-p-X)3]+ (1 Xis H, 2 Xis Me, 3 Xis Ph, 4 Xis Br, 5 Xis OH, 6 Xis NO2, 7 Xis OMe, 8 Xis CF3, 9 Xis F, 10 Xis Pr i , 11 Xis Bu t ) have been synthesized from the reaction of [(η6-p-MeC6H4Pr i )RuCl2]2 with the corresponding thiol, isolated as the chloride salts, and further studied for their electrochemical properties, cytotoxicity towards human ovarian cancer cells, and catalytic activity for glutathione (GSH) oxidation. Complex 1 was also compared with the benzene and hexamethylbenzene analogues [(η6-C6H6)2Ru2(SC6H5)3]+ (12) and [(η6-C6Me6)2Ru2(SC6H5)3]+ (13). The most active compound [11]Cl was structurally studied by single-crystal X-ray diffraction analysis. The concentrations corresponding to 50% inhibition of cancer cell growth (IC50 values) in the A2780 and A2780cisR cell lines of these complexes except for 6 were in the submicromolar range, complex 11 showing an IC50 value of 0.03µM in both cell lines. The high in vitro anticancer activity of these complexes may be at least partially due to their catalytic potential for the oxidation of GSH, although there is no clear correlation between the IC50 values and the turnover frequencies at about 50% conversion. However, the cytotoxicity is tentatively correlated to the physicochemical properties of the compounds determined by the electronic influence of the substituents X (Hammett constants σ p) and the lipophilicity of the thiols p-XC6H4SH (calculated logP parameters

Fate of systemically and locally administered adipose-derived mesenchymal stromal cells and their effect on wound healing

There is increasing interest in the use of adipose-derived mesenchymal stromal cells (ASCs) for wound repair. As the fate of administered cells is still poorly defined, we aimed to establish the location, survival, and effect of ASCs when administered either systemically or locally during wound repair under physiological conditions. To determine the behavior of ASCs, a rat model with wounds on the dorsal aspect of the hind paws was used and two treatment modes were assessed: ASCs administered systemically into the tail vein or locally around the wound. ASCs were transduced to express both firefly luciferase (Fluc) and green fluorescent protein to enable tracking by bioluminescence imaging and immunohistological analysis. Systemically administered ASCs were detected in the lungs 3 hours after injection with a decrease in luminescent signal at 48 hours and signal disappearance from 72 hours. No ASCs were detected in the wound. Locally administered ASCs remained strongly detectable for 7 days at the injection site and became distributed within the wound bed as early as 24 hours post injection with a significant increase observed at 72 hours. Systemically administered ASCs were filtered out in the lungs, whereas ASCs administered locally remained and survived not only at the injection site but were also detected within the wound bed. Both treatments led to enhanced wound closure. It appears that systemically administered ASCs have the potential to enhance wound repair distally from their site of entrapment in the lungs whereas locally administered ASCs enhanced wound repair as they became redistributed within the wound bed.The Swiss National Science Foundation Project (#310030_170132), Medical Faculty of University of Geneva, the South African Medical Research Council University Flagship Program (SAMRC-RFA-UFSP-01-2013/STEM CELLS), the SAMRC Extramural Unit for Stem Cell Research and Therapy, and the Institute for Cellular and MolecularMedicine (ICMM) of the University of Pretoria. The Fonds National Suisse de la Recherche Scientifique (FNSNF) and the Faculty of Medicine of the University of Geneva for contributing to the funding in Geneva and the South African Medical Research Council (Flagship and Extramural Unit awards), the National Research Foundation (Scarce Skills Doctoral Scholarship, Grant UID: 88799), Ernst & Ethel Eriksen Trust (Doctoral Scholarship), University of Pretoria Bursary Office (Postgraduate Bursary), and the Institute for Cellular and Molecular Medicine (ICMM) of the University of Pretoria for contributing to the funding in South Africa.https://stemcellsjournals.onlinelibrary.wiley.com/journal/21576580am2020Immunolog

From the Lena Goldfields Case to the Yukos Shareholders Cases: State Control over Strategic Subsoil Vs Protection of Foreign Investors' Interests

International investment law and arbitration have become attractive to solve expropriation cases between a state and foreign investors. Such evolution is mainly based on key cases. The 1930 Lena Goldfields, Ltd vs USSR case directly contributed to the recognition of arbitration and the definition of several international legal principles that have gradually been acknowledged as the minimum standards of international investment law. Moreover, the Lena Goldfields case has clearly showed that legal considerations and enforcement of the arbitration award are influenced by geopolitical matters, especially when strategic subsoil is involved. If societies with different (ideological) references are in latent conflict, arbitration in international investment law turns out to be a weapon. Such approach continues to be valid when capitalism prevails and private entities gain economical strength in a weakened state. The 2000's Yukos Shareholders vs Russian Federation cases further illustrate it, as the geopolitical force of a country is notably defined by its standing on the energy market. To fully grasp the way in which the above-mentioned cases have contributed to the international jurisprudence, the two parts of the current manuscript will follow the same structure. First, will be presented the global historical context and the local history. Then, will be described the chronological phases of the cases and briefly analyzed the key legal aspects of the cases

Role of LRH-1/NR5A2 in islet physiology and pathophysiology

The nuclear receptor Liver Receptor Homolog-1 (LRH-1) plays a key role in several biological processes. The correlative association between LRH-1 function induced by estrogens and the protective role of the hormone on pancreatic insulin-secreting ß-cells led us to investigate the potential implication of LRH-1 in islet ß-cell physiology. We demonstrate that LRH-1 is expressed in ß-cells and that transcript levels are modulated by 17ß-estradiol through the ERa signalling pathway. Adenoviral-mediated overexpression of LRH-1 in human islets did not alter proliferation but conferred protection against cytokines and streptozotocin-induced apoptosis. More importantly, incubation of human islets with an agonist of LRH-1 (BL001) also protected islets against cell death caused by cytokines and streptozotocin. BL001 also increased overall ß-cell performance in islets isolated from Type 2 diabetic donors. Our results indicate that LRH-1 could be an attractive target for stimulating islet survival and performance in diabetic patients

Involvement of Fumarase C and NADH Oxidase in Metabolic Adaptation of Pseudomonas fluorescens Cells Evoked by Aluminum and Gallium Toxicity▿

Iron (Fe) is a critical element in all aerobic organisms as it participates in a variety of metabolic networks. In this study, aluminum (Al) and gallium (Ga), two Fe mimetics, severely impeded the ability of the soil microbe Pseudomonas fluorescens to perform oxidative phosphorylation. This was achieved by disrupting the activity and expression of complexes I, II, and IV. These toxic metals also inactivated aconitase (ACN) and fumarase A (FUM A), two tricarboxylic acid cycle enzymes dependent on Fe for their catalytic activity, while FUM C, an Fe-independent enzyme, displayed an increase in activity and expression under these stressed situations. Furthermore, in the Al- and Ga-exposed cells, the activity and expression of an H2O-forming NADH oxidase were markedly increased. The incubation of the Al- and Ga-challenged cells in an Fe-containing medium led to the recovery of the affected enzymatic activities. Taken together, these data provide novel insights into how environmental pollutants such as Al and Ga interfere with cellular Fe metabolism and also illustrate the ability of Pseudomonas fluorescens to modulate metabolic networks to combat this situation

Fate of systemically and locally administered adipose-derived mesenchymal stromal cells and their effect on wound healing

There is increasing interest in the use of adipose-derived mesenchymal stromal cells (ASCs) for wound repair. As the fate of administered cells is still poorly defined, we aimed to establish the location, survival, and effect of ASCs when administered either systemically or locally during wound repair under physiological conditions. To determine the behavior of ASCs, a rat model with wounds on the dorsal aspect of the hind paws was used and two treatment modes were assessed: ASCs administered systemically into the tail vein or locally around the wound. ASCs were transduced to express both firefly luciferase (Fluc) and green fluorescent protein to enable tracking by bioluminescence imaging and immunohistological analysis. Systemically administered ASCs were detected in the lungs 3 h after injection with a decrease in luminescent signal at 48 h and signal disappearance from 72 h. No ASCs were detected in the wound. Locally administered ASCs remained strongly detectable for 7 days at the injection site and became distributed within the wound bed as early as 24 h post injection with a significant increase observed at 72 h. Systemically administered ASCs were filtered out in the lungs, whereas ASCs administered locally remained and survived not only at the injection site but were also detected within the wound bed. Both treatments led to enhanced wound closure. It appears that systemically administered ASCs have the potential to enhance wound repair distally from their site of entrapment in the lungs whereas locally administered ASCs enhanced wound repair as they became redistributed within the wound bed

Hepatic nuclear factor 1alpha (HNF1alpha) dysfunction down-regulates X-box-binding protein 1 (XBP1) and sensitizes beta-cells to endoplasmic reticulum stress

Correct endoplasmic reticulum (ER) function is critical for the health of secretory cells, such as the pancreatic β-cell, and ER stress is often a contributory factor to β-cell death in type 2 diabetes. We have used an insulin-secreting cell line with inducible expression of dominant negative (DN) HNF1α, a transcription factor vital for correct β-cell development and function, to show that HNF1α is required for Xbp1 transcription and maintenance of the normal ER stress response. DN HNF1α expression sensitizes the β-cell to ER stress by directly down-regulating Xbp1 transcription, whereas Atf6 is unaffected. Furthermore, DN HNF1α alters calcium homeostasis, resulting in elevated cytoplasmic calcium and increased store-operated calcium entry, whereas mitochondrial calcium uptake is normal. Loss of function of XBP1 is toxic to the β-cell and decreases production of the ER chaperone BiP, even in the absence of ER stress. DN HNF1α-induced sensitivity to cyclopiazonic acid can be partially rescued with the chemical chaperone tauroursodeoxycholate. Rat insulin 2 promoter-DN HNF1α mouse islets express lower levels of BiP mRNA, synthesize less insulin, and are sensitized to ER stress relative to matched control mouse islets, suggesting that this mechanism is also operating in vivo

Highly cytotoxic trithiophenolatodiruthenium complexes of the type [(η6-p-MeC6H4Pri)2Ru2(SC6H4-p-X)3]+: synthesis, molecular structure, electrochemistry, cytotoxicity, and glutathione oxidation potential

A series of cationic dinuclear p-cymene ruthenium trithiophenolato complexes of the type [(eta(6)-p-MeC6H4Pr (i) )(2)Ru-2(SC6H4-p-X)(3)](+) (1 X is H, 2 X is Me, 3 X is Ph, 4 X is Br, 5 X is OH, 6 X is NO2, 7 X is OMe, 8 X is CF3, 9 X is F, 10 X is Pr (i) , 11 X is Bu (t) ) have been synthesized from the reaction of [(eta(6)-p-MeC6H4Pr (i) )RuCl2](2) with the corresponding thiol, isolated as the chloride salts, and further studied for their electrochemical properties, cytotoxicity towards human ovarian cancer cells, and catalytic activity for glutathione (GSH) oxidation. Complex 1 was also compared with the benzene and hexamethylbenzene analogues [(eta(6)-C6H6)(2)Ru-2(SC6H5)(3)](+) (12) and [(eta(6)-C6Me6)(2)Ru-2(SC6H5)(3)](+) (13). The most active compound [11]Cl was structurally studied by single-crystal X-ray diffraction analysis. The concentrations corresponding to 50 % inhibition of cancer cell growth (IC50 values) in the A2780 and A2780cisR cell lines of these complexes except for 6 were in the submicromolar range, complex 11 showing an IC50 value of 0.03 A mu M in both cell lines. The high in vitro anticancer activity of these complexes may be at least partially due to their catalytic potential for the oxidation of GSH, although there is no clear correlation between the IC50 values and the turnover frequencies at about 50 % conversion. However, the cytotoxicity is tentatively correlated to the physicochemical properties of the compounds determined by the electronic influence of the substituents X (Hammett constants sigma (p)) and the lipophilicity of the thiols p-XC6H4SH (calculated log P parameters)

Embryonic stem cell-based screen for small molecules: cluster analysis reveals four response patterns in developing neural cells

Neural differentiation of embryonic stem cells (ESC) is considered a promising model to perform in vitro testing for neuroactive and neurotoxic compounds. We studied the potential of a dual reporter murine ESC line to identify bioactive and/or toxic compounds. This line expressed firefly luciferase under the control of the neural cell-specific tubulin alpha promoter (TUBA1A), and renilla luciferase under the control of the ubiquitous translation elongation factor 1-alpha-1 (EEF1A1) promoter. During neural differentiation, TUBA1A activity increased, while EEF1A1 activity decreased. We first validated our test system using the known neurotoxin methyl mercury. This compound altered expression of both reporter genes, with ESC-derived neural precursors being affected at markedly lower concentrations than undifferentiated ESCs. Analysis of a library of 1040 bioactive compounds picked up 127 compounds with altered EEF1A1 and/or TUBA1A promoter activity, which were classified in 4 clusters. Cluster 1 (low EEF1A1 and TUBA1A) was the largest cluster, containing many cytostatic drugs, as well as known neurodevelopmental toxicants, psychotropic drugs and endocrine disruptors. Cluster 2 (high EEF1A1, stable TUBA1A) was limited to three sulfonamides. Cluster 3 (high EEF1A1 and TUBA1A) was small, but markedly enriched in neuroactive and neurotoxic compounds. Cluster 4 (stable EEF1A1, high TUBA1A) was heterogeneous, containing endocrine disruptors, neurotoxic and cytostatic drugs. The dual reporter gene assay described here might be a useful addition to in vitro drug testing panels. Our two-dimensional testing strategy provides information on complex response patterns, which could not be achieved by a single marker approach