Mimicking exercise in three-dimensional bioengineered skeletal muscle to investigate cellular and molecular mechanisms of physiological adaptation

Bioengineering of skeletal muscle in vitro in order to produce highly aligned myofibres in relevant three dimensional (3D) matrices have allowed scientists to model the in vivo skeletal muscle niche. This review discusses essential experimental considerations for developing bioengineered muscle in order to investigate exercise mimicking stimuli. We identify current knowledge for the use of electrical stimulation and co-culture with motor neurons to enhance skeletal muscle maturation and contractile function in bioengineered systems in vitro. Importantly, we provide a current opinion on the use of acute and chronic exercise mimicking stimuli (electrical stimulation and mechanical overload) and the subsequent mechanisms underlying physiological adaptation in 3D bioengineered muscle. We also identify that future studies using the latest bioreactor technology, providing simultaneous electrical and mechanical loading and flow perfusion in vitro, may provide the basis for advancing knowledge in the future. We also envisage, that more studies using genetic, pharmacological, and hormonal modifications applied in human 3D bioengineered skeletal muscle may allow for an enhanced discovery of the in-depth mechanisms underlying the response to exercise in relevant human testing systems. Finally, 3D bioengineered skeletal muscle may provide an opportunity to be used as a pre-clinical in vitro test-bed to investigate the mechanisms underlying catabolic disease, while modelling disease itself via the use of cells derived from human patients without exposing animals or humans (in phase I trials) to the side effects of potential therapies

A deep X-ray view of the bare AGN Ark 120. I. Revealing the Soft X-ray Line Emission

The Seyfert 1 galaxy, Ark 120, is a prototype example of the so-called class of bare nucleus AGN, whereby there is no known evidence for the presence of ionized gas along the direct line of sight. Here deep ($>400$ ks exposure), high resolution X-ray spectroscopy of Ark 120 is presented, from XMM-Newton observations which were carried out in March 2014, together with simultaneous Chandra/HETG exposures. The high resolution spectra confirmed the lack of intrinsic absorbing gas associated with Ark 120, with the only X-ray absorption present originating from the ISM of our own Galaxy, with a possible slight enhancement of the Oxygen abundance required with respect to the expected ISM values in the Solar neighbourhood. However, the presence of several soft X-ray emission lines are revealed for the first time in the XMM-Newton RGS spectrum, associated to the AGN and arising from the He and H-like ions of N, O, Ne and Mg. The He-like line profiles of N, O and Ne appear velocity broadened, with typical FWHM widths of $\sim5000$ km s$^{-1}$, whereas the H-like profiles are unresolved. From the clean measurement of the He-like triplets, we deduce that the broad lines arise from gas of density $n_{\rm e}\sim10^{11}$ cm$^{-3}$, while the photoionization calculations infer that the emitting gas covers at least 10 percent of $4\pi$ steradian. Thus the broad soft X-ray profiles appear coincident with an X-ray component of the optical-UV Broad Line Region on sub-pc scales, whereas the narrow profiles originate on larger pc scales, perhaps coincident with the AGN Narrow Line Region. The observations show that Ark 120 is not intrinsically bare and substantial X-ray emitting gas exists out of our direct line of sight towards this AGN

Review of health economic models exploring and evaluating treatment and management of hospital-acquired pneumonia and ventilator-associated pneumonia

BACKGROUND: Hospital-acquired pneumonia (HAP) is pneumonia that occurs ≥48 h after hospital admission; it is the most common hospital-acquired infection contributing to death. Ventilator-associated pneumonia (VAP) arises ≥48-72 h after intubation. Opinions differ on whether VAP is a subset of HAP; the same pathogens predominate in both. Compared with VAP-free controls, patients developing VAP are twice as likely to die and have significantly longer stays in intensive care units. Guidelines recommend that microbiological cultures should guide antibiotic treatment, but these lack sensitivity and take 48-72 h to process, meaning that initial therapy must be empiric, generally with broad-spectrum agents. Given increasing pressure to improve both antibiotic stewardship and patient outcomes, the National Institute for Health and Care Excellence and the Infectious Diseases Society of America recommend research into rapid molecular diagnostic tests to identify causative organisms and their antibiotic resistances. Ideally, these would supersede culture, being quicker and more sensitive. In the UK, the INHALE research programme, funded by the National Institute for Health Research, is exploring rapid molecular diagnostics to inform treatment of HAP/VAP and, given resource implications, incorporates a health economic component. AIM: To identify previous economic modelling of HAP/VAP costs to inform this component. METHODS: Literature review of HAP/VAP studies with economic modelling identified from three databases. FINDINGS: Twenty studies were identified. Only one study specifically evaluated strategies to improve diagnosis; the remaining 19 studies omitted this important aspect. CONCLUSION: HAP/VAP modelling would be improved by better awareness of long-term outcomes and treatment complexity. To the authors' knowledge, no similar literature reviews of economic modelling for HAP/VAP have been published

The mRNA expression of SETD2 in human breast cancer: Correlation with clinico-athological parameters

BACKGROUND: SET domain containing protein 2 (SETD2) is a histone methyltransferase that is involved in transcriptional elongation. There is evidence that SETD2 interacts with p53 and selectively regulates its downstream genes. Therefore, it could be implicated in the process of carcinogenesis. Furthermore, this gene is located on the short arm of chromosome 3p and we previously demonstrated that the 3p21.31 region of chromosome 3 was associated with permanent growth arrest of breast cancer cells. This region includes closely related genes namely: MYL3, CCDC12, KIF9, KLHL18 and SETD2. Based on the biological function of these genes, SETD2 is the most likely gene to play a tumour suppressor role and explain our previous findings. Our objective was to determine, using quantitative PCR, whether the mRNA expression levels of SETD2 were consistent with a tumour suppressive function in breast cancer. This is the first study in the literature to examine the direct relationship between SETD2 and breast cancer. METHODS: A total of 153 samples were analysed. The levels of transcription of SETD2 were determined using quantitative PCR and normalized against (CK19). Transcript levels within breast cancer specimens were compared to normal background tissues and analyzed against conventional pathological parameters and clinical outcome over a 10 year follow-up period. RESULTS: The levels of SETD2 mRNA were significantly lower in malignant samples (p = 0.0345) and decreased with increasing tumour stage. SETD2 expression levels were significantly lower in samples from patients who developed metastasis, local recurrence, or died of breast cancer when compared to those who were disease free for > 10 years (p = 0.041). CONCLUSION: This study demonstrates a compelling trend for SETD2 transcription levels to be lower in cancerous tissues and in patients who developed progressive disease. These findings are consistent with a possible tumour suppressor function of this gene in breast cancer

Comparative Transcriptome and Methylome Analysis in Human Skeletal Muscle Anabolism, Hypertrophy and Epigenetic Memory

Transcriptome wide changes in human skeletal muscle after acute (anabolic) and chronic resistance exercise (RE) induced hypertrophy have been extensively determined in the literature. We have also recently undertaken DNA methylome analysis (850,000 + CpG sites) in human skeletal muscle after acute and chronic RE, detraining and retraining, where we identified an association between DNA methylation and epigenetic memory of exercise induced skeletal muscle hypertrophy. However, it is currently unknown as to whether all the genes identified in the transcriptome studies to date are also epigenetically regulated at the DNA level after acute, chronic or repeated RE exposure. We therefore aimed to undertake large scale bioinformatical analysis by pooling the publicly available transcriptome data after acute (110 samples) and chronic RE (181 samples) and comparing these large data sets with our genome-wide DNA methylation analysis in human skeletal muscle after acute and chronic RE, detraining and retraining. Indeed, after acute RE we identified 866 up- and 936 down-regulated genes at the expression level, with 270 (out of the 866 upregulated) identified as being hypomethylated, and 216 (out of 936 downregulated) as hypermethylated. After chronic RE we identified 2,018 up- and 430 down-regulated genes with 592 (out of 2,018 upregulated)identified as being hypomethylated and 98 (out of 430 genes downregulated) as hypermethylated. After KEGG pathway analysis, genes associated with ‘cancer’ pathways were significantly enriched in both bioinformatic analysis of the pooled transcriptome and methylome datasets after both acute and chronic RE. This resulted in 23 (out of 69) and 28 (out of 49) upregulated and hypomethylated and 12 (out of 37) and 2 (out of 4) downregulated and hypermethylated ‘cancer’ genes following acute and chronic RE respectively. Within skeletal muscle tissue, these ‘cancer’ genes predominant functions were associated with matrix/actin structure and remodelling, mechano-transduction (e.g. PTK2/Focal Adhesion Kinase and Phospholipase D- following chronic RE), TGF-beta signalling and protein synthesis (e.g. GSK3B after acute RE). Interestingly, 51 genes were also identified to be up/downregulated in both the acute and chronic RE pooled transcriptome analysis as well as significantly hypo/hypermethylated after acute RE, chronic RE, detraining and retraining. Five genes; FLNB, MYH9, SRGAP1, SRGN, ZMIZ1 demonstrated increased gene expression in the acute and chronic RE transcriptome and also demonstrated hypomethylation in these conditions. Importantly, these 5 genes demonstrated retained hypomethylation even during detraining (following training induced hypertrophy) when exercise was ceased and lean mass returned to baseline (pre-training) levels, identifying them as genes associated with epigenetic memory in skeletal muscle. Importantly, for the first time across the transcriptome and epigenome combined, this study identifies novel differentially methylated genes associated with human skeletal muscle anabolism, hypertrophy and epigenetic memory

Investigating the prevalence, predictors, and prognosis of suboptimal statin use early after a non-ST elevation acute coronary syndrome

BACKGROUND:High-potency statin therapy is recommended in the secondary prevention of car-diovascular disease but discontinuation, dose reduction, statin switching, and/or nonadherence occurin practice.OBJECTIVES:To determine the prevalence and predictors of deviation from high-potency statin useearly after a non-ST elevation acute coronary syndrome (NSTE-ACS) and its association with subse-quent major adverse cardiovascular events (MACE) and all-cause mortality (ACM).METHODS:A total of 1005 patients from a UK-based prospective NSTE-ACS cohort study dis-charged on high-potency statin therapy (atorvastatin 80 mg, rosuvastatin 20 mg, or 40 mg daily)were included. At 1 month, patients were divided into constant high-potency statin users, and subop-timal users incorporating statin discontinuation, dose reduction, switching statin to a lower equivalentpotency, and/or statin nonadherence. Follow-up was a median of 16 months.RESULTS:There were 156 suboptimal (w15.5%) and 849 constant statin users. Factors associatedin multivariable analysis with suboptimal statin occurrence included female sex (odds ratio 1.75, 95%confidence interval [CI] 1.14–2.68) and muscular symptoms (odds ratio 4.28, 95% CI 1.30–14.08).Suboptimal statin use was associated with increased adjusted risks of time to MACE (hazard ratio2.10, 95% CI 1.25–3.53,P5.005) and ACM (hazard ratio 2.46, 95% CI 1.38–4.39,P5.003). Sub-group analysis confirmed that the increased MACE/ACM risks were principally attributable to statindiscontinuation or nonadherence.CONCLUSIONS:Conversion to suboptimal statin use is common early after NSTE-ACS and ispartly related to muscular symptoms. Statin discontinuation or non-adherence carries an adverse prog-nosis. Interventions that preserve and enhance statin utilization could improve post NSTE-ACSoutcomes

High Resolution X-ray Spectroscopy of the Seyfert 1, Mrk 1040. Revealing the Failed Nuclear Wind with Chandra

High resolution X-ray spectroscopy of the warm absorber in the nearby X-ray bright Seyfert 1 galaxy, Mrk 1040 is presented. The observations were carried out in the 2013-2014 timeframe using the Chandra High Energy Transmission Grating with a total exposure of 200 ks. A multitude of absorption lines from Ne, Mg and Si are detected from a wide variety of ionization states. In particular, the detection of inner K-shell absorption lines from Ne, Mg and Si, from charge states ranging from F-like to Li-like ions, suggests the presence of a substantial amount of low ionization absorbing gas, illuminated by a steep soft X-ray continuum. The observations reveal at least 3 warm absorbing components ranging in ionization parameter from $\log\xi = 0-2$ and with column densities of $N_{\rm H} =1.5-4.0 \times 10^{21}$cm$^{-2}$. The velocity profiles imply that the outflow velocities of the absorbing gas are low and within $\pm100$ km s$^{-1}$ of the systemic velocity of Mrk 1040, which suggests any outflowing gas may have stalled in this AGN on large enough scales. The warm absorber is likely located far from the black hole, within 300 pc of the nucleus and is spatially coincident with emission from an extended Narrow Line Region as seen in the HST images. The iron K band spectrum reveals only narrow emission lines, with Fe K$\alpha$ at 6.4 keV consistent with originating from reflection off Compton thick pc-scale reprocessing gas

The effects of developmental trauma on theory of mind and its relationship to psychotic experiences: A behavioural study

BACKGROUND: Developmental psychological trauma induces vulnerability to psychosis. However, the mechanisms underlying this association are poorly understood. Impairments in Theory of Mind (ToM) have been observed in adult survivors of developmental trauma and individuals with psychosis. ToM is therefore a candidate mechanism underlying the association between developmental trauma and psychosis. METHODS: We used a computerised version of the Director task - where a participant is instructed by a confederate to move an object around a 4 × 4 grid, whilst taking account of whether these objects are visible to a confederate who instructs the participant - to investigate impairments in ToM in 209 participants (age: M = 37.8, SD=13.6; 56% female). Participants were divided into a) developmental trauma-positive (DT+) and control groups (DT-) based on their history of developmental trauma and b) then further into subclinical (S) and healthy groups (H) as based on psychotic experiences indexed by the CAPE-P15. After exclusion, the numbers in each group were: DT+H (47), DT+S (84), DT-H (54), DT-S (12). (Total: 197). RESULTS: Developmental trauma exposure was associated with psychotic experiences (OR: 7.89, p < .001), which remained significant after controlling for demographic and clinical confounds (adjusted R2 = 0.452, R2 change = 0.0184, p = .009). Participants with developmental trauma (F1, 194) = 5.46, p = .020, ηp2 = 0.027) and participants more prone to psychotic experiences (F1, 194) = 4.71, p = .031, ηp2 = 0.024) demonstrated significantly lower accuracy on the Director task relative to their respective control, after controlling for the effects of age. CONCLUSIONS: ToM deficits are associated with self-reported developmental trauma and psychotic experiences. Further work is needed to explore these relationships further and whether they represent generalised or specific effect effects on developmental trauma and psychopathological domains