Abstract Epithelial-mesenchymal transition (EMT) is closely implicated in the pathogenesis of idiopathic pulmonary fibrosis. Associated with this phenotypic transition is the acquisition of an ...elongated cell morphology and establishment of stress fibers. The extent to which these EMT-associated changes influence cellular mechanics is unclear. We assessed the biomechanical properties of alveolar epithelial cells (A549) following exposure to TGF-β1. Using atomic force microscopy, changes in cell stiffness and surface membrane features were determined. Stimulation with TGF-β1 gave rise to a significant increase in stiffness, which was augmented by a collagen I matrix. Additionally, TGF-β1-treated cells exhibited a rougher surface profile with notable protrusions. Simultaneous quantitative examination of the morphological attributes of stimulated cells using an image-based high-content analysis system revealed dramatic alterations in cell shape, F-actin content and distribution. Together, these investigations point to a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing EMT. From the Clinical Editor Epithelial-mesenchymal transition is implicated in the pathogenesis of pulmonary fibrosis. Using atomic force microscopy, the authors demonstrate a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing mesenchymal transition.
Methylenetetrahydrofolate dehydrogenase (MTHFD1) deficiency has recently been reported to cause a folate-responsive syndrome displaying a phenotype that includes megaloblastic anemia and severe ...combined immunodeficiency.
To describe our investigative approach to the molecular diagnosis and evaluation of immune dysfunction in a family with MTHFD1 deficiency.
The methods used were exome sequencing and analysis of variants in genes involved in the folate metabolic pathway in a family with 2 affected siblings. Routine laboratory and research data were analyzed to gain an in-depth understanding of innate, humoral, and cell-mediated immune function before and after folinic acid supplementation.
Interrogation of exome data for concordant variants between the siblings in the genes involved in folate metabolic pathway identified a heterozygous mutation in exon 3 of the MTHFD1 gene that was shared with their mother. In view of highly suggestive phenotype, we extended our bioinformatics interrogation for structural variants in the MTHFD1 gene by manual evaluation of the exome data for sequence depth coverage of all the exons. A deletion involving exon 13 that was shared with their father was identified. Routine laboratory data showed lymphopenia involving all subsets and poor response to vaccines. In vitro analysis of dendritic cell and lymphocyte function was comparable to that in healthy volunteers. Treatment with folinic acid led to immune reconstitution, enabling discontinuation of all prophylactic therapies.
Exome sequencing demonstrated MTHFD1 deficiency as a novel cause of a combined immunodeficiency. Folinic acid was established as precision therapy to reverse the clinical and laboratory phenotype of this primary immunodeficiency.
Background Cysteinyl leukotrienes (cys-LTs) are potent asthma-related mediators that function through their G protein–coupled receptors, cys-LT receptor type 1 (CysLT1R) and cys-LT receptor type 2 ...(CysLT2R). Objective Because many G protein–coupled receptors transactivate the epidermal growth factor receptor (EGFR) through metalloprotease-mediated ligand shedding, we investigated the effects of cys-LTs on signal transduction and proliferation of bronchial fibroblasts. Methods Human bronchial fibroblasts were grown from biopsy specimens of healthy subjects. Mitogenesis was assessed on the basis of tritiated methylthymidine incorporation. Results Leukotriene (LT) D4 alone did not increase mitogenesis but dose-dependently increased thymidine incorporation and cell proliferation in the presence of epidermal growth factor (EGF). The enhancement was not prevented by CysLT1R antagonists (MK-571 and montelukast) or by a dual antagonist (BAY u9773), which is consistent with the lack of detectable mRNA for CysLT1R and CysLT2R in bronchial fibroblasts. LTD4 did not cause EGFR transphosphorylation nor was the synergism blocked by the metalloprotease inhibitor GM6001. The EGFR-selective kinase inhibitor AG1478 suppressed the synergy between LTD4 and EGF but had no effect on synergistic interactions of LTD4 with other receptor tyrosine kinase growth factors. The effect of LTD4 involved a pertussis toxin–sensitive and protein kinase C–mediated intracellular pathway, leading to sustained growth factor–dependent phosphorylation of extracellular signal–regulated kinase 1/2 and protein kinase B (PKB/Akt). Conclusion Cys-LTs do not transactivate EGFR but have a broader capability to synergize with receptor tyrosine kinase pathways. Clinical implications This study implies a critical role of cys-LTs in airway fibrosis in asthma and other chronic airway diseases, which might not be blocked by therapy with current LT receptor antagonists.
The Physical Activity 4 Everyone Cluster Randomized Trial Sutherland, Rachel L., MPH; Campbell, Elizabeth M., PhD; Lubans, David R., PhD ...
American journal of preventive medicine,
August 2016, Letnik:
51, Številka:
2
Journal Article
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Introduction Few interventions have been successful in reducing the physical activity decline typically observed among adolescents. The aim of this paper is to report the 24-month effectiveness of a ...multicomponent school-based intervention (Physical Activity 4 Everyone) in reducing the decline in moderate to vigorous physical activity (MVPA) among secondary school students in disadvantaged areas of New South Wales, Australia. Study design A cluster RCT was conducted in five intervention and five control schools with follow-up measures taken at 24 months post-randomization. Setting/participants The trial was undertaken within secondary schools located in disadvantaged communities in New South Wales, Australia. Intervention A multicomponent school-based intervention based on the Health Promoting Schools Framework was implemented. The intervention consisted of seven physical activity promotion strategies that targeted the curriculum (teaching strategies to increase physical activity in physical education lessons, student physical activity plans, and modification of school sport program); school environment (recess/lunchtime activities, school physical activity policy); parents (parent newsletters); and community (community physical activity provider promotion). Six additional strategies supported school implementation of the physical activity intervention strategies. Main outcome measure Minutes per day spent in MVPA, objectively measured by accelerometer. Results Participants (N=1,150, 49% male) were a cohort of students aged 12 years (Grade 7) at baseline (March–June 2012) and 14 years (Grade 9) at follow-up (March–July 2014). At 24-month follow-up, there were significant effects in favor of the intervention group for daily minutes of MVPA. The adjusted mean difference in change in daily MVPA between groups was 7.0 minutes (95% CI=2.7, 11.4, p <0.002) (analysis conducted December 2014–February 2015). Sensitivity analyses based on multiple imputation were consistent with the main analysis (6.0 minutes, 95% CI=0.6, 11.3, p <0.031). Conclusions The intervention was effective in increasing adolescents’ minutes of MVPA, suggesting that implementation of the intervention by disadvantaged schools has the potential to slow the decline in physical activity. Trial registration Australian New Zealand Clinical Trials Registry ACTRN12612000382875.
Nonsyndromic Thoracic Aortic Aneurysm and Dissection Sherrah, Andrew G., BSc, MBBS; Andvik, Sarah, BMed, MD; van der Linde, Denise, MD, PhD ...
Journal of the American College of Cardiology,
02/2016, Letnik:
67, Številka:
6
Journal Article
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Abstract Background Genetic aortopathy (GA) underlies thoracic aortic aneurysms (TAA) in younger adults. Comparative survival and predictors of outcomes in nonsyndromic TAA (NS-TAA) are incompletely ...defined compared to Marfan syndrome (MFS) and bicuspid aortic valve (BAV). Objectives The study sought to compare survival and clinical outcomes for individuals with NS-TAA, MFS, and BAV. Methods From 1988 to 2014, all patients presenting with GA 16 to 60 years of age were enrolled in a prospective study of clinical outcomes. Risk factors for death and aortic dissection were identified by Cox proportional hazards modeling and a mortality risk score developed. Results Diagnosis of GA was made for 760 patients (age 36.9 ± 13.6 years, 26.8% female; NS-TAA, n = 311; MFS, n = 221; BAV, n = 228). MFS patients were younger than NS-TAA and BAV. Presentation with aortic dissection was more common for NS-TAA than MFS or BAV. The 687 patients surviving >30 days after presentation were followed for a median of 7 years. Calculated 10-year mortality was 7.8% for NS-TAA, 8.7% for MFS, and 3.5% for BAV (NS-TAA and MFS vs. BAV p <0.05). Factors associated with all-cause mortality were MFS (p = 0.04), age at presentation, and family history of dissection. Conclusions Clinical outcomes for MFS and NS-TAA are similar but worse than BAV. Independent predictors of mortality, including family history of aortic dissection and age, can be included in an Aortopathy Mortality Risk Score to predict survival. Management of NS-TAA, including surgical intervention, should be similar to that of MFS.
Abstract Background The incidence of endometrial cancer is increasing worldwide. Despite improved overall survival rates, its aetiopathogenesis remains complicated, although oestrogens are known ...causal factors. Among other factors thought to be implicated are the endocannabinoids. Anandamide (AEA), the prototypical endocannabinoid, has emerged as a possible factor in the pathogenesis of endometrial cancer because oestrogen regulates AEA concentrations and the expression of the cannabinoid receptors in other tissues. Furthermore, the endocannabinoid system is protective against the growth and spread of different malignancies. We aimed to investigate and quantify anandamide and its receptors in endometrial cancer. Methods In this single-centre study conducted at a UK teaching hospital, 31 patients with endometrial carcinomas (27 type 1, four type 2) and seven healthy age-matched controls with atrophic endometria were recruited. Biopsy samples were taken and divided into two—one for this study and the other for histological confirmation of diagnosis (graded according to the FIGO system). The biopsy sample for this study was further divided into two—one for AEA measurement and the other for quantitative RT-PCR. CB1 and CB2 transcript levels were measured in triplicate and normalised against the housekeeping genes PPIA , MRPL19 , and IPO8 . 15 of 31 cancer patients (12 type 1, three type 2) and all seven of the control group, volunteered blood for plasma collection. At hysterectomy, 31 endometrial cancer samples (27 type 1, four type 2) and seven age-matched atrophic control endometrial biopsy samples were obtained for tissue AEA measurement. Extracts from the plasma and endometrial homogenates were assessed for the presence of AEA with UHPLC-tandem mass spectrometry. Findings In the endometrial cancer tissues, both CB1 and CB2 transcript levels were significantly decreased by 90% (p<0·0004) and 80% (p< 0·0001), respectively, compared with atrophic tissues. Plasma AEA concentrations were significantly higher in patients with endometrial cancer than in the control group (mean 1·95 nM SE 0·59 vs 1·06 0·08, p=0·037). Plasma AEA concentrations in patients with type 1 disease were significantly higher than in control patients (2·17 0·72 vs 1·06 0·08, p<0·05), but in patients with type 2 disease there was no difference (1·08 0·05). Similarly, tissue AEA in patients with endometrial cancer was significantly higher than in the control group (5·45 nmol/g SE 0·91 vs 1·77 0·66, p=0·046). Tissue AEA was significantly higher in type 1 endometrial cancer than in atrophic samples (6·12 0·98 vs 1·77 0·66, p<0·05), but type 2 samples did not differ (0·90 0·16). Furthermore, in samples available for comparison (15 endometrial cancer, seven atrophic) there was a statistically significant correlation between plasma and tissue AEA concentrations ( r =0·55, p=0·008). Interpretation The results from this study suggest that downregulation of CB1 and CB2 receptor transcript levels and raised tissue and plasma AEA concentrations are linked in some way. We speculate that the endocannabinoid system is implicated in the aetiopathogenesis of endometrial cancer, but further work is warranted. The correlation between plasma and tissue AEA concentrations suggests that plasma AEA could be used as a biomarker for some forms of endometrial malignant disease, especially type 1 endometrioid adenocarcinoma. The endocannabinoid system thus presents an attractive novel target for pharmacological intervention in some endometrial malignancies. Funding University Hospitals of Leicester NHS Trust.
Summary Background Antipsychotic drugs are usually the first line of treatment for schizophrenia; however, many patients refuse or discontinue their pharmacological treatment. We aimed to establish ...whether cognitive therapy was effective in reducing psychiatric symptoms in people with schizophrenia spectrum disorders who had chosen not to take antipsychotic drugs. Methods We did a single-blind randomised controlled trial at two UK centres between Feb 15, 2010, and May 30, 2013. Participants aged 16–65 years with schizophrenia spectrum disorders, who had chosen not to take antipsychotic drugs for psychosis, were randomly assigned (1:1), by a computerised system with permuted block sizes of four or six, to receive cognitive therapy plus treatment as usual, or treatment as usual alone. Randomisation was stratified by study site. Outcome assessors were masked to group allocation. Our primary outcome was total score on the positive and negative syndrome scale (PANSS), which we assessed at baseline, and at months 3, 6, 9, 12, 15, and 18. Analysis was by intention to treat, with an ANCOVA model adjusted for site, age, sex, and baseline symptoms. This study is registered as an International Standard Randomised Controlled Trial, number 29607432. Findings 74 individuals were randomly assigned to receive either cognitive therapy plus treatment as usual (n=37), or treatment as usual alone (n=37). Mean PANSS total scores were consistently lower in the cognitive therapy group than in the treatment as usual group, with an estimated between-group effect size of −6·52 (95% CI −10·79 to −2·25; p=0·003). We recorded eight serious adverse events: two in patients in the cognitive therapy group (one attempted overdose and one patient presenting risk to others, both after therapy), and six in those in the treatment as usual group (two deaths, both of which were deemed unrelated to trial participation or mental health; three compulsory admissions to hospital for treatment under the mental health act; and one attempted overdose). Interpretation Cognitive therapy significantly reduced psychiatric symptoms and seems to be a safe and acceptable alternative for people with schizophrenia spectrum disorders who have chosen not to take antipsychotic drugs. Evidence-based treatments should be available to these individuals. A larger, definitive trial is needed. Funding National Institute for Health Research.
A multidisciplinary collaboration investigated the world's largest, most catastrophic epidemic thunderstorm asthma event that took place in Melbourne, Australia, on Nov 21, 2016, to inform mechanisms ...and preventive strategies.
Meteorological and airborne pollen data, satellite-derived vegetation index, ambulance callouts, emergency department presentations, and data on hospital admissions for Nov 21, 2016, as well as leading up to and following the event were collected between Nov 21, 2016, and March 31, 2017, and analysed. We contacted patients who presented during the epidemic thunderstorm asthma event at eight metropolitan health services (each including up to three hospitals) via telephone questionnaire to determine patient characteristics, and investigated outcomes of intensive care unit (ICU) admissions.
Grass pollen concentrations on Nov 21, 2016, were extremely high (>100 grains/m3). At 1800 AEDT, a gust front crossed Melbourne, plunging temperatures 10°C, raising humidity above 70%, and concentrating particulate matter. Within 30 h, there were 3365 (672%) excess respiratory-related presentations to emergency departments, and 476 (992%) excess asthma-related admissions to hospital, especially individuals of Indian or Sri Lankan birth (10% vs 1%, p<0·0001) and south-east Asian birth (8% vs 1%, p<0·0001) compared with previous 3 years. Questionnaire data from 1435 (64%) of 2248 emergency department presentations showed a mean age of 32·0 years (SD 18·6), 56% of whom were male. Only 28% had current doctor-diagnosed asthma. 39% of the presentations were of Asian or Indian ethnicity (25% of the Melbourne population were of this ethnicity according to the 2016 census, relative risk RR 1·93, 95% CI 1·74–2·15, p <0·0001). Of ten individuals who died, six were Asian or Indian (RR 4·54, 95% CI 1·28–16·09; p=0·01). 35 individuals were admitted to an intensive care unit, all had asthma, 12 took inhaled preventers, and five died.
Convergent environmental factors triggered a thunderstorm asthma epidemic of unprecedented magnitude, tempo, and geographical range and severity on Nov 21, 2016, creating a new benchmark for emergency and health service escalation. Asian or Indian ethnicity and current doctor-diagnosed asthma portended life-threatening exacerbations such as those requiring admission to an ICU. Overall, the findings provide important public health lessons applicable to future event forecasting, health care response coordination, protection of at-risk populations, and medical management of epidemic thunderstorm asthma.
None.
Lung reirradiation for nonsmall cell lung cancer (NSCLC) is common for either recurrent disease or new primary cancer. Dose volume tolerance of the lung after multiple courses of radiation therapy ...(RT) is unknown. We review our experience with lung reirradiation for patients with NSCLC in a single community setting using stereotactic body radiation therapy (SBRT) to report lung cumulative doses, survival, and toxicity.
Forty-four patients who received at least 2 curative courses of lung RT with the second course delivered between January 2012 and December 2017 were eligible. All patients had NSCLC and were treated with SBRT for reirradiation. Cumulative lung dose volume histograms for all courses were generated, summated, and converted into cumulative equivalent dose in 2 Gy fractions (EQD2). Actuarial overall survival (OS), local control, and toxicity is reported, including a subset of patients who received more than 2 courses of SBRT.
Median age of the group was 71 years (range, 51-87). Median survival of the entire group from diagnosis, first, and second courses of RT was 3.94, 3.03, and 2.03 years. Three-year actuarial OS for the entire group was 34.1% from second course of RT. The mean EQD2 Gy3 mean lung dose for all courses was 12.35 Gy (range, 2.7-26.52). The mean EQD2 Gy3 V5Gy, V10Gy, V20Gy, V30Gy, and V40Gy were 40.9%, 25.5%, 14.7%, 10.2%, and 7.7%. Six-year actuarial freedom from grade ≥3 complications was 86.3%. The rate of grade ≥3 lung toxicity was 4.5% (2 of 44). Other late toxicities included grade 3 recurrent laryngeal nerve damage (n = 1) and grade 3 chest wall pain/rib fracture (n = 1). Overall, 32% of patients had more than 2 courses of RT to the lung (range, 3-7).
Long-term OS is possible with multiple RT courses to the lung for NSCLC with low toxicity.
Abstract Background We have previously shown that patients with endometrial carcinoma express elevated concentrations of the endocannabinoid, anandamide (AEA), in both their plasma and their ...endometrial tissue and that the endometrial carcinoma cell line, Ishikawa, contains the receptors to which AEA binds. Several studies have reported that human and rodent cancer cell lines die in response to high AEA concentrations. The incidence of endometrial carcinoma continues to escalate and, although surgical treatment has improved, morbidity and mortality rates have not. A move towards a novel non-surgical therapeutic option is thus required, and the endocannabinoid system provides a good candidate target. We aimed to investigate the effects of AEA on the survival and proliferation of an endometrial carcinoma cell model. Methods This prospective basic research study was conducted at a UK teaching hospital. Ishikawa cells were cultured in vitro, and a range of AEA concentrations (0–10 000 nM) were added to the cells. The effect of AEA was measured at different timepoints (4, 18, 24, 48, and 72 h). Primary outcome was cell proliferation and cell viability as measured with a commercial proliferation-apoptosis assay in which assay colour at 420 nm is directly proportional to cell density. One-way ANOVA was performed with Prism (version 7). Findings Ishikawa cells were sensitive to AEA-mediated cytotoxicity in a pseudo dose-dependent manner. AEA caused a significant decrease in cell number only at concentrations above 1000 nM (mean 28·1% SE 7·8, n=12; p<0·0001). The decrease in cell viability that occurred at 4, 18, and 24 h was partly restored at 48 and 72 h suggesting that the AEA growth inhibitory effect is time limiting. Interpretation Our results show that AEA induces a decrease in Ishikawa cell number probably through inhibition of cell proliferation rather than cell death. These data suggest that the increased plasma and tissue AEA concentrations observed in patients with endometrial cancer is a counter mechanism against further cancer growth and points to the endocannabinoid system as a potentially new therapeutic target. Funding University Hospitals of Leicester NHS Trust.
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