Transforming PPh3 into Bidentate Phosphine Ligands at Ru-Zn Heterobimetallic Complexes

The reaction of [Ru(PPh 3) 3Cl 2] with excess ZnMe 2 led to P-C/C-H bond activation and P-C/C-C bond formation to generate a chelating diphenylphosphinobenzene ligand as well as a cyclometallated (diphenylphosphino)biphenyl group in the final product of the reaction, [Ru(dppbz)(PPh 2(biphenyl)′)(ZnMe)] (1; dppbz = 1,2-bis(diphenylphosphino)benzene); PPh 2(biphenyl)′ = cyclometallated PPh 2(biphenyl). The mechanism of reaction was studied and C-C coupling to give a bidentate 2,2′-bis(diphenylphosphino)biphenyl (BIPHEP) ligand was suggested to be one of the key steps of the process. This was confirmed by the reaction of [Ru(BIPHEP)(PPh 3)HCl] with ZnMe 2, which also gave 1. An analogous set of steps took place upon addition of ZnMe 2 to [Ru(rac-BINAP)(PPh 3)HCl] (rac-BINAP = racemic(2,2′-bis(diphenylphosphino)-1,1′-binaphthyl) to give [Ru(dppbz)(PPh 2(binaphthyl)′)ZnMe] (3). H 2 and the C-H bond of PhCCH added across the Ru-Zn bond of 1, and also reversed the phosphine cyclometallation, to give [Ru(dppbz)(Ph 2P(biphenyl))(H) 2(H)(ZnMe)] (4) and [Ru(dppbz)(Ph 2P(biphenyl))(CCPh) 2(H)(ZnMe)] (5) respectively. </p

Mesenchymal stem cell pre-treatment with cytokines and hydrogen peroxide modifies their adhesion to cardiac endothelium and murine heart sections AND A preliminary study of the molecular regulation of placental growth factor and vascular endothelial growth factor in murine preadipocytes

Project 1. I investigated whether pretreating mesechymal stem cells (MSCs) with inflammatory cytokines or hydrogen peroxide (H$_2$O$_2$) improves their adhesion to cardiovascular endothelium. Pre-treating mMSCs (passage 7-9) with H$_2$O$_2$ and IFNγ increased their static adhesion to murine cardiac endothelial cells. This effect was lost when passage 10 mMSCs were used. hMSC adhesion to human coronary artery endothelial cells increased with SDF-1α and IL-8 pre-treatment. Adhesion of mMSCs to healthy heart sections was decreased by mMSC pretreatment with H$_2$O$_2$, murine IL-8 and SDF-1α. The species of MSC origin, MSC passage number and the adhesive surface influence the manner in which MSC endothelial adhesion is altered by cytokines. Project 2. Placental growth factor (PlGF) is an attractive anti-obesity target. We aimed to explore the molecular regulation of PlGF and VEGF in murine preadipocytes. Wild-type FoxO1 transfection of murine preadipocytes induced considerable increases in PlGF and VEGF expression. H$_2$O$_2$, VEGF, cAMP and forskolin treatments promoted an increase in preadipocyte FoxO1, VEGF and PlGF expression. Enhanced PlGF expression was not accompanied by an increase PlGF secretion. These preliminary results should be interpreted cautiously and their underlying mechanisms characterised

The role of adipose and skeletal muscle derived cytokines in primary human myogenesis: implications for ageing skeletal muscle

Sarcopenia is the age-related loss of skeletal muscle mass and function; inflammation is thought to be one aetiological factor in its development. Adipose tissue accumulates with advancing age and adipose-derived cytokines (adipokines) contribute to inflammaging. Skeletal muscle myogenesis is one adaptaive mechanism by which skeletal muscle mass is sustained throughout the human lifespan. The effect of the adipose inflammatory milieu on such myogenesis is unknown, as is the relative importance of its constituent adipokines to myogenesis. This work demonstrates that conditioned medium generated from obese subcutaneuous adipose tissue has a detrimental effect on in vitro primary human myogenesis. Resistin is shown to be – in part – responsible for this phenomenon and is demonstrated to inhibit myogenesis by activating the classical NFκB pathway. Resistin is further shown to be a metabolic stressor of primary human myotubes, promoting increased oxygen consumption, fatty acid oxidation and lipid accumulation. It is important to identify more avenues for the development of pharmacological interventions in sarcopenia. To that end, this thesis also demonstrates for the first time that the myokine IL-15: 1) is pro-myogenic in primary human cultures; 2) can mitigate the detrimental effects of an inflammatory environment on myogenesis; and 3) supports myogenesis at autocrine concentrations

Report on the Traveller women's food, physical activity and health study

Research report of Community Based Participatory Research project on Traveller women's food choice and exercis

MyoCount:A software tool for the automated quantification of myotube surface area and nuclear fusion index [version 1; referees: 2 approved]

This is the author accepted manuscript. The final version is available on open access from F1000Research via the DOI in this recordData availability: The data underlying the results presented in Figure 4 and Figure 5, (.csv and .tif files) are available as ‘Myocount Validation Data’ via OSF. DOI: https://doi.org/10.17605/OSF.IO/F5DXE18. Data are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication). Software availability: Latest source code and Myocount version are available at: https://github.com/MurphyDavid/MyoCount/releases. Archived source code at time of publication: https://doi.org/10.5281/zenodo.254281111. License: MIT License. https://opensource.org/licenses/MITIt is often desirable to characterise the morphology of myogenic cultures. To achieve this, the surface area of myotubes is often quantified, along with the nuclear fusion index (NFI). Existing methods of such quantification are time-consuming and subject to error-prone human input. We have developed MyoCount, an open-source program that runs via the freely available MATLAB Runtime and quantifies myotube surface area and NFI. MyoCount allows the user to adjust its parameters to account for differences in image quality, magnification and the colour channels used in generating the image. MyoCount measures of myotube surface area and NFI were compared to the mean of measures performed by two blinded investigators using ImageJ software (surface area R2 = 0.89, NFI R2 =0.87). For NFI, the mean coefficient of variation (CV) between two investigators (17.6 ± 2.3%) was significantly higher than that between the investigator mean and MyoCount (13.5 ± 1.4%). For measurements of myotube area, the CV did not differ between both analysis methods. Given these results and the advantages of applying the same image analysis method uniformly across all images in an experiment, we suggest that MyoCount will be a useful research tool and we publish its source code and instructions for its use alongside this article.Wellcome Trus

Agreement between telehealth and in-person assessment of patients with chronic musculoskeletal conditions presenting to an advanced-practice physiotherapy screening clinic

Objective: To determine the level of agreement between a telehealth and in-person assessment of a representative sample of patients with chronic musculoskeletal conditions referred to an advanced-practice physiotherapy screening clinic. Design: Repeated-measures study design. Participants: 42 patients referred to the Neurosurgical & Orthopaedic Physiotherapy Screening Clinic (Queensland, Australia) for assessment of their chronic lumbar spine, knee or shoulder condition. Intervention: Participants underwent two consecutive assessments by different physiotherapists within a single clinic session. In-person assessments were conducted as per standard clinical practice. Telehealth assessments took place remotely via videoconferencing. Six Musculoskeletal Physiotherapists were paired together to perform both assessment types. Main outcome measures: Clinical management decisions including (i) recommended management pathways, (ii) referral to allied health professions, (iii) clinical diagnostics, and (iv) requirement for further investigations were compared using reliability and agreement statistics. Results: There was substantial agreement (83.3%; 35/42 cases) between in-person and telehealth assessments for recommended management pathways. Moderate to near perfect agreement (AC1 = 0.58–0.9) was reached for referral to individual allied health professionals. Diagnostic agreement was 83.3% between the two delivery mediums, whilst there was substantial agreement (81%; AC1 = 0.74) when requesting further investigations. Overall, participants were satisfied with the telehealth assessment. Conclusion: There is a high level of agreement between telehealth and in-person assessments with respect to clinical management decisions and diagnosis of patients with chronic musculoskeletal conditions managed in an advanced-practice physiotherapy screening clinic. Telehealth can be considered as a viable and effective medium to assess those patients who are unable to attend these services in person

Lysosomal Disorders Drive Susceptibility to Tuberculosis by Compromising Macrophage Migration.

A zebrafish genetic screen for determinants of susceptibility to Mycobacterium marinum identified a hypersusceptible mutant deficient in lysosomal cysteine cathepsins that manifests hallmarks of human lysosomal storage diseases. Under homeostatic conditions, mutant macrophages accumulate undigested lysosomal material, which disrupts endocytic recycling and impairs their migration to, and thus engulfment of, dying cells. This causes a buildup of unengulfed cell debris. During mycobacterial infection, macrophages with lysosomal storage cannot migrate toward infected macrophages undergoing apoptosis in the tuberculous granuloma. The unengulfed apoptotic macrophages undergo secondary necrosis, causing granuloma breakdown and increased mycobacterial growth. Macrophage lysosomal storage similarly impairs migration to newly infecting mycobacteria. This phenotype is recapitulated in human smokers, who are at increased risk for tuberculosis. A majority of their alveolar macrophages exhibit lysosomal accumulations of tobacco smoke particulates and do not migrate to Mycobacterium tuberculosis. The incapacitation of highly microbicidal first-responding macrophages may contribute to smokers' susceptibility to tuberculosis.This work was supported by the National Institutes of Health (R37AI054503, LR, R01NS082567, CBM, 5F30HL110455, RB, 1DP2-OD008614, DMT), the Wellcome Trust (LR), the National Institute of Health Research Cambridge Biomedical Research Centre (LR), the Health Research Board of Ireland (HRA_POR/2013/387, MO’S and CSA/2012/16, JK), and The Royal City of Dublin Hospital Trust (Grant 146, JK).This is the final version of the article. It first appeared from Cell Press via http://dx.doi.org/10.1016/j.cell.2016.02.034

Resistance exercise initiates mechanistic target of rapamycin (mTOR) translocation and protein complex co-localisation in human skeletal muscle

The mechanistic target of rapamycin (mTOR) is a central mediator of protein synthesis in skeletal muscle. We utilized immunofluorescence approaches to study mTOR cellular distribution and protein-protein co-localisation in human skeletal muscle in the basal state as well as immediately, 1 and 3 h after an acute bout of resistance exercise in a fed (FED; 20 g Protein/40 g carbohydrate/1 g fat) or energy-free control (CON) state. mTOR and the lysosomal protein LAMP2 were highly co-localised in basal samples. Resistance exercise resulted in rapid translocation of mTOR/LAMP2 towards the cell membrane. Concurrently, resistance exercise led to the dissociation of TSC2 from Rheb and increased in the co-localisation of mTOR and Rheb post exercise in both FED and CON. In addition, mTOR co-localised with Eukaryotic translation initiation factor 3 subunit F (eIF3F) at the cell membrane post-exercise in both groups, with the response significantly greater at 1 h of recovery in the FED compared to CON. Collectively our data demonstrate that cellular trafficking of mTOR occurs in human muscle in response to an anabolic stimulus, events that appear to be primarily influenced by muscle contraction. The translocation and association of mTOR with positive regulators (i.e. Rheb and eIF3F) is consistent with an enhanced mRNA translational capacity after resistance exercise

Treating Adolescents Together or Individually? Issues in Adolescent Substance Abuse Interventions

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66302/1/j.1530-0277.2002.tb02619.x.pd