Agricultural intensification is best considered as the level of human appropriation of terrestrial net primary production. The global value is set to increase from 30%, increasing pressures on biodiversity. The pressures can be classified in terms of spatial scale, i.e. land cover, landscape management and crop management. Different lowland agricultural landscapes in Great Britain show differences among these pressures when habitat diversity and nutrient surplus are used as indicators. Eutrophication of plants was correlated to N surplus, and species richness of plants correlated with broad habitat diversity. Bird species diversity only correlated with habitat diversity when the diversity of different agricultural habitats was taken into account. The pressures of agricultural change may be reduced by minimizing loss of large habitats, minimizing permanent loss of agricultural land, maintaining habitat diversity in agricultural landscapes in order to provide ecosystem services, and minimizing pollution from nutrients and pesticides from the crops themselves. While these pressures could potentially be quantified using an internationally consistent set of indicators, their impacts would need to be assessed using a much larger number of locally applicable biodiversity indicators.

This work has shown a novel way by which computer navigation can be used to analyse soft tissue behaviour during TKA beyond the coronal plane and throughout range of motion. Despite subjective stress testing, our results show reproducible patterns of soft tissue behaviour-in particular a wide range of mid-flexion excursion. It also quantifies the limits within which a cruciate-retaining TKR can maintain knee stability. This functionality may guide the surgeon in identifying and/or preventing soft tissue imbalances intra-operatively, improving functional results.

On behalf of the North East of England Pediatric Respiratory Infection Study Group Newcastle upon Tyne, UK Summary. Pediatric pleural empyema has increased substantially over the past 20 years and reasons for this rise remain not fully explained. We investigated potential risk factors for the development of empyema in children by examining a cohort of patients with community-acquired pneumonia. Demographic, clinical, and socioeconomic characteristics, use of Ibuprofen prior to presentation and selected potential epidemiological risk factors were analyzed. Data were collected from a prospective etiological study of radiologically confirmed pneumonia in hospitalized children aged 16 years. One hundred sixty children were enrolled; 56% were male and 69% aged <5 years. Empyema complication developed in 40 (25%) children. Children with empyema were more frequently prescribed Ibuprofen prior to admission to hospital than those without (82% vs. 46.2%; OR 1.94, 97.5% credible interval 0.80-3.18). Bacterial infection was strongly associated with the development of empyema (OR 3.34, 97.5% credible interval 1.70-5.14). In contrast age, sex, maternal age, parental smoking, level of socioeconomic status, nursery attendance, asthma, household characteristics (bedrooms and number of occupants) were not significantly different between groups. In conclusion, children with pneumonia who developed empyema had more often received Ibuprofen prior to hospitalization and confirmed bacterial infection. We suggest a population-based study involving both primary and secondary care settings would help to investigate the role of Ibuprofen use in modulating the course of disease in children with pneumonia. Pediatr Pulmonol. 2015;50:721-726. ß

AimsDuchenne muscular dystrophy (DMD) is associated with progressive cardiomyopathy. Oral corticosteroids are the gold standard for the treatment of skeletal muscle weakness; however, the effects of steroids on cardiac function have not been prospectively studied. In addition, the early role of ACE-inhibitors (ACE-I) is controversial. We aimed to determine the effects of steroids and ACE-I on development of left ventricular dysfunction in the mdx mouse, a model for dystrophin-deficient cardiomyopathy. Methods and resultsOrally administered captopril or prednisolone was given for 8 weeks to 16-week-old, male mdx mice. In vivo pressure-volume loops, fibrosis, in vivo myocyte sarcolemmal injury, and cytokine expression were assessed in treated and untreated mdx mice and age-matched controls. Untreated mdx mice showed compensated cardiomyopathy with reduced myocardial contractility, patchy myocardial fibrosis but preserved stroke volume. Captopril treatment resulted in indirect myocardial effects of reduced afterload and direct effects of increased contractility. Prednisolone caused acute sarcolemmal injury, increased expression of myocardial TNF alpha and fibrosis, resulting in left ventricular dilatation and diastolic dysfunction. ConclusionIn a mouse model of dystrophin-deficient cardiomyopathy, ACE-I produced haemodynamic benefit, whereas steroids accelerated progression of cardiomyopathy. Although mouse models may not entirely replicate the human condition, comprehensive monitoring of cardiac function with these therapies is essential.--

Oxidative phosphorylation (OXPHOS) defects caused by somatic mitochondrial DNA mutations increase with age in human colorectal epithelium and are prevalent in colorectal tumors, but whether they actively contribute to tumorigenesis remains unknown. Here we demonstrate that mitochondrial DNA mutations causing OXPHOS defects are enriched during the human adenoma/carcinoma sequence, suggesting that they may confer a metabolic advantage. To test this, we deleted the tumor suppressor Apc in OXPHOS-deficient intestinal stem cells in mice. The resulting tumors were larger than in control mice due to accelerated cell proliferation and reduced apoptosis. We show that both normal crypts and tumors undergo metabolic remodeling in response to OXPHOS deficiency by upregulating the de novo serine synthesis pathway. Moreover, normal human colonic crypts upregulate the serine synthesis pathway in response to OXPHOS deficiency before tumorigenesis. Our data show that age-associated OXPHOS deficiency causes metabolic remodeling that can functionally contribute to accelerated intestinal cancer development.

Patients with total knee arthroplasties (TKAs) continue to report dissatisfaction in functional outcome. Stability is a major factor contributing to functionality of TKAs. Implants with single-radius (SR) femoral components are proposed to increase stability throughout the arc of flexion. Using computer navigation and loaded cadaveric legs, we characterized the "envelope of laxity" (EoL) offered by a SR cruciate retaining (CR)-TKA compared with that of the native knee through the arc of flexion in terms of anterior drawer, varus/valgus stress, and internal/external rotation. In both the native knee and the TKA laxity increased with increasing knee flexion. Laxities measured in the three planes of motion were generally comparable between the native knee and TKA from 0˚to 110o f flexion. Our results indicate that the SR CR-TKA offers appropriate stability in the absence of soft tissue deficiency. ß

There is currently an urgent need for biomarkers that can be used to monitor the efficacy of experimental therapies for Duchenne Muscular Dystrophy (DMD) in clinical trials. Identification of novel protein biomarkers has been limited due to the massive complexity of the serum proteome and the presence of a small number of very highly abundant proteins. Here we have utilised an aptamer-based proteomics approach to profile 1,129 proteins in the serum of wild-type and mdx (dystrophin deficient) mice. The serum levels of 96 proteins were found to be significantly altered (P < 0.001, q < 0.01) in mdx mice. Additionally, systemic treatment with a peptide-antisense oligonucleotide conjugate designed to induce Dmd exon skipping and recover dystrophin protein expression caused many of the differentially abundant serum proteins to be restored towards wild-type levels. Results for five leading candidate protein biomarkers (Pgam1, Tnni3, Camk2b, Cycs and Adamts5) were validated by ELISA in the mouse samples. Furthermore, ADAMTS5 was found to be significantly elevated in human DMD patient serum. This study has identified multiple novel, therapy-responsive protein biomarkers in the serum of the mdx mouse with potential utility in DMD patients.

Antisense oligonucleotide (AON)-induced exon skipping is one of the most promising strategies for treating Duchenne muscular dystrophy (DMD) and other rare monogenic conditions. Phosphorodiamidate morpholino oligonucleotides (PMOs) and 2'-O-methyl phosphorothioate (2'OMe) are two of the most advanced AONs in development. The next generation of peptide-conjugated PMO (P-PMO) is also showing great promise, but to advance these therapies it is essential to determine the pharmacokinetic and biodistribution (PK/BD) profile using a suitable method to detect AON levels in blood and tissue samples. An enzyme-linked immunosorbent assay (ELISA)-based method, which shows greater sensitivity than the liquid chromatography-mass spectrometry method, is the method of choice for 2'OMe detection in preclinical and clinical studies. However, no such assay has been developed for PMO/P-PMO detection, and we have, therefore, developed an ultrasensitive hybridization-based ELISA for this purpose. The assay has a linear detection range of 5-250 pM (R(2)>0.99) in mouse serum and tissue lysates. The sensitivity was sufficient for determining the 24-h PK/BD profile of PMO and P-PMO injected at standard doses (12.5 mg/kg) in mdx mice, the dystrophin-deficient mouse model for DMD. The assay demonstrated an accuracy approaching 100% with precision values under 12%. This provides a powerful cost-effective assay for the purpose of accelerating the development of these emerging therapeutic agents.