Transgenic Overexpression of LARGE Induces alpha-Dystroglycan Hyperglycosylation in Skeletal and Cardiac Muscle

Background: LARGE is one of seven putative or demonstrated glycosyltransferase enzymes defective in a common group of muscular dystrophies with reduced glycosylation of alpha-dystroglycan. Overexpression of LARGE induces hyperglycosylation of alpha-dystroglycan in both wild type and in cells from dystroglycanopathy patients, irrespective of their primary gene defect, restoring functional glycosylation. Viral delivery of LARGE to skeletal muscle in animal models of dystroglycanopathy has identical effects in vivo, suggesting that the restoration of functional glycosylation could have therapeutic applications in these disorders. Pharmacological strategies to upregulate Large expression are also being explored.Methodology/Principal Findings: In order to asses the safety and efficacy of long term LARGE over-expression in vivo, we have generated four mouse lines expressing a human LARGE transgene. On observation, LARGE transgenic mice were indistinguishable from the wild type littermates. Tissue analysis from young mice of all four lines showed a variable pattern of transgene expression: highest in skeletal and cardiac muscles, and lower in brain, kidney and liver. Transgene expression in striated muscles correlated with alpha-dystroglycan hyperglycosylation, as determined by immunoreactivity to antibody IIH6 and increased laminin binding on an overlay assay. Other components of the dystroglycan complex and extracellular matrix ligands were normally expressed, and general muscle histology was indistinguishable from wild type controls. Further detailed muscle physiological analysis demonstrated a loss of force in response to eccentric exercise in the older, but not in the younger mice, suggesting this deficit developed over time. However this remained a subclinical feature as no pathology was observed in older mice in any muscles including the diaphragm, which is sensitive to mechanical load-induced damage.Conclusions/Significance: This work shows that potential therapies in the dystroglycanopathies based on LARGE upregulation and alpha-dystroglycan hyperglycosylation in muscle should be safe

The "drive to eat" hypothesis: energy expenditure and fat-free mass but not adiposity are associated with milk intake and energy intake in 12 week infants

BACKGROUND: Recent work has challenged the long-held assumption that appetite functions to maintain stable body mass and fat mass (FM), suggesting instead that appetite matches food intake to energy expenditure and its correlate, fat-free mass (FFM). Whether this scenario applies to young infants, in chronic positive energy balance, remains unknown. OBJECTIVES: To test associations of components of energy expenditure and body composition with milk intake (MI) and energy intake (EI) in 12-week infants, by reanalyzing published cross-sectional data. METHODS: Data were available for 48 infants. In addition to anthropometric measurements, we assessed MI and EI by test-weighing, sleeping metabolic rate (SMR) by indirect calorimetry, and FFM, FM, and total energy expenditure (TEE) by doubly labeled water. Mean parental height was calculated as a marker of infant growth drive. Correlation and multiple regression analyses were applied. RESULTS: MI and EI correlated with FFM (r = 0.47 and 0.57, respectively; P  0.6). MI and EI correlated with SMR (r = 0.42 and 0.53, respectively; P  0.2). In a multiple regression analysis, MI was independently associated with TEE (partial r = 0.39) and FFM (partial r = 0.35). EI showed similar associations. Mean parental height was correlated with weight gain, MI, and EI. CONCLUSIONS: As in adults, MI and EI in young infants were strongly associated with FFM and with total and sleeping components of energy expenditure, but not with fatness. The infant's growth drive contributed to these associations. This suggests that appetite is regulated by the rate of energy expenditure, the size of energy-using tissues, and tissue deposition rate, and that the high levels of body fat characteristic of infants may not constrain weight gain

Pathways for outpatient management of venous thromboembolism in a UK centre.

It has become widely recognised that outpatient treatment may be suitable for many patients with venous thromboembolism. In addition, non-vitamin K antagonist oral anticoagulants that have been approved over the last few years have the potential to be an integral component of the outpatient care pathway, owing to their oral route of administration, lack of requirement for routine anticoagulation monitoring and simple dosing regimens. A robust pathway for outpatient care is also vital; one such pathway has been developed at Sheffield Teaching Hospitals in the UK. This paper describes the pathway and the arguments in its favour as an example of best practice and value offered to patients with venous thromboembolism. The pathway has two branches (one for deep vein thrombosis and one for pulmonary embolism), each with the same five-step process for outpatient treatment. Both begin from the point that the patient presents (in the Emergency Department, Thrombosis Clinic or general practitioner's office), followed by diagnosis, risk stratification, treatment choice and, finally, follow-up. The advantages of these pathways are that they offer clear, evidence-based guidance for the identification, diagnosis and treatment of patients who can safely be treated in the outpatient setting, and provide a detailed, stepwise process that can be easily adapted to suit the needs of other institutions. The approach is likely to result in both healthcare and economic benefits, including increased patient satisfaction and shorter hospital stays

Body composition reference charts for UK infants and children aged 6 weeks to 5 years based on measurement of total body water by isotope dilution

BACKGROUND: Until recently, pediatric body composition reference data were very limited, hindering interpretation of measurements. In the last decade, such data emerged for several techniques for children ≥ 5 years, but equivalent data for younger age groups remain lacking, due to their poor compliance with most techniques. // OBJECTIVES: To provide reference data for use in clinical practice and research from 6 weeks to 5 years, that are based on measurements of total body water (TBW) by isotope dilution. // DESIGN: The data on anthropometry and TBW were available from studies of 463 infants and children aged 6 weeks to 7 years, conducted between 1988 and 2010. Both breast-fed and formula-fed infants were included. TBW was measured by 2H- or 18O-labeled water, and converted to fat-free mass (FFM) using published hydration coefficients. Reference charts and SD scores (SDS) were constructed for FFM, fat mass (FM), FFM index and FM index for each sex, using the lambda-mu-sigma method. // RESULTS: Both sexes were significantly heavier and longer than UK 1990 reference data (p < 0.01), but did not differ in body mass index SDS. Breast-fed infants were longer than formula-fed infants but did not differ in body composition. // CONCLUSIONS: These reference data will enhance the ability of clinicians to assess and monitor body composition and FFM/FM accretion in clinical practice in younger age groups. Total body water can be measured in most patients, though abnormalities of hydration must be addressed. However, the centiles do not overlap exactly with those published for older age groups, limiting comparability between younger and older children

The “drive to eat” hypothesis: energy expenditure and fat-free mass but not adiposity are associated with milk intake and energy intake in 12 week infants

Background Recent work has challenged the long-held assumption that appetite functions to maintain stable body mass and fat mass (FM), suggesting instead that appetite matches food intake to energy expenditure and its correlate, fat-free mass (FFM). Whether this scenario applies to young infants, in chronic positive energy balance, remains unknown. Objectives To test associations of components of energy expenditure and body composition with milk intake (MI) and energy intake (EI) in 12-week infants, by reanalyzing published cross-sectional data. Methods Data were available for 48 infants. In addition to anthropometric measurements, we assessed MI and EI by test-weighing, sleeping metabolic rate (SMR) by indirect calorimetry, and FFM, FM, and total energy expenditure (TEE) by doubly labeled water. Mean parental height was calculated as a marker of infant growth drive. Correlation and multiple regression analyses were applied. Results MI and EI correlated with FFM (r = 0.47 and 0.57, respectively; P 0.6). MI and EI correlated with SMR (r = 0.42 and 0.53, respectively; P 0.2). In a multiple regression analysis, MI was independently associated with TEE (partial r = 0.39) and FFM (partial r = 0.35). EI showed similar associations. Mean parental height was correlated with weight gain, MI, and EI. Conclusions As in adults, MI and EI in young infants were strongly associated with FFM and with total and sleeping components of energy expenditure, but not with fatness. The infant's growth drive contributed to these associations. This suggests that appetite is regulated by the rate of energy expenditure, the size of energy-using tissues, and tissue deposition rate, and that the high levels of body fat characteristic of infants may not constrain weight gain

Calibration of centre-of-mass energies at LEP 2 for a precise measurement of the W boson mass

The determination of the centre-of-mass energies for all LEP 2 running is presented. Accurate knowledge of these energies is of primary importance to set the absolute energy scale for the measurement of the W boson mass. The beam energy between 80 and 104 GeV is derived from continuous measurements of the magnetic bending field by 16 NMR probes situated in a number of the LEP dipoles. The relationship between the fields measured by the probes and the beam energy is defined in the NMR model, which is calibrated against precise measurements of the average beam energy between 41 and 61 GeV made using the resonant depolarisation technique. The validity of the NMR model is verified by three independent methods: the flux-loop, which is sensitive to the bending field of all the dipoles of LEP; the spectrometer, which determines the energy through measurements of the deflection of the beam in a magnet of known integrated field; and an analysis of the variation of the synchrotron tune with the total RF voltage. To obtain the centre-of-mass energies, corrections are then applied to account for sources of bending field external to the dipoles, and variations in the local beam energy at each interaction point. The relative error on the centre-of-mass energy determination for the majority of LEP 2 running is 1.2 x 10^{-4}, which is sufficiently precise so as not to introduce a dominant uncertainty on the W mass measurement.Comment: 79 pages, 45 figures, submitted to EPJ

The influence of magnetic anisotropy on the Zeeman spectra of lanthanide doped nanoparticles

High-resolution infra-red spectra of KY3F10 nanoparticles doped with Nd3+ and Er3+ are measured in magnetic fields of up to 4 T. A simple model of the magnetic splittings of the ground and excited states is used to show that the presence or absence of observable splittings depends on the anisotropy of the response to the magnetic field. Detailed crystal-field calculations accurately model the spectra, including nonlinear effects

Systematic review and meta-analysis of the diagnostic accuracy of ultrasonography for deep vein thrombosis

Background Ultrasound (US) has largely replaced contrast venography as the definitive diagnostic test for deep vein thrombosis (DVT). We aimed to derive a definitive estimate of the diagnostic accuracy of US for clinically suspected DVT and identify study-level factors that might predict accuracy. Methods We undertook a systematic review, meta-analysis and meta-regression of diagnostic cohort studies that compared US to contrast venography in patients with suspected DVT. We searched Medline, EMBASE, CINAHL, Web of Science, Cochrane Database of Systematic Reviews, Cochrane Controlled Trials Register, Database of Reviews of Effectiveness, the ACP Journal Club, and citation lists (1966 to April 2004). Random effects meta-analysis was used to derive pooled estimates of sensitivity and specificity. Random effects meta-regression was used to identify study-level covariates that predicted diagnostic performance. Results We identified 100 cohorts comparing US to venography in patients with suspected DVT. Overall sensitivity for proximal DVT (95% confidence interval) was 94.2% (93.2 to 95.0), for distal DVT was 63.5% (59.8 to 67.0), and specificity was 93.8% (93.1 to 94.4). Duplex US had pooled sensitivity of 96.5% (95.1 to 97.6) for proximal DVT, 71.2% (64.6 to 77.2) for distal DVT and specificity of 94.0% (92.8 to 95.1). Triplex US had pooled sensitivity of 96.4% (94.4 to 97.1%) for proximal DVT, 75.2% (67.7 to 81.6) for distal DVT and specificity of 94.3% (92.5 to 95.8). Compression US alone had pooled sensitivity of 93.8 % (92.0 to 95.3%) for proximal DVT, 56.8% (49.0 to 66.4) for distal DVT and specificity of 97.8% (97.0 to 98.4). Sensitivity was higher in more recently published studies and in cohorts with higher prevalence of DVT and more proximal DVT, and was lower in cohorts that reported interpretation by a radiologist. Specificity was higher in cohorts that excluded patients with previous DVT. No studies were identified that compared repeat US to venography in all patients. Repeat US appears to have a positive yield of 1.3%, with 89% of these being confirmed by venography. Conclusion Combined colour-doppler US techniques have optimal sensitivity, while compression US has optimal specificity for DVT. However, all estimates are subject to substantial unexplained heterogeneity. The role of repeat scanning is very uncertain and based upon limited data