Neurofilament heavy chain side arm phosphorylation regulates axonal transport of neurofilaments

Neurofilaments possess side arms that comprise the carboxy-terminal domains of neurofilament middle and heavy chains (NFM and NFH); that of NFH is heavily phosphorylated in axons. Here, we demonstrate that phosphorylation of NFH side arms is a mechanism for regulating transport of neurofilaments through axons. Mutants in which known NFH phosphorylation sites were mutated to preclude phosphorylation or mimic permanent phosphorylation display altered rates of transport in a bulk transport assay. Similarly, application of roscovitine, an inhibitor of the NFH side arm kinase Cdk5/p35, accelerates neurofilament transport. Analyses of neurofilament movement in transfected living neurons demonstrated that a mutant mimicking permanent phosphorylation spent a higher proportion of time pausing than one that could not be phosphorylated. Thus, phosphorylation of NFH slows neurofilament transport, and this is due to increased pausing in neurofilament movement

Efficiency and safety of varying the frequency of whole blood donation (INTERVAL): a randomised trial of 45 000 donors

Background: Limits on the frequency of whole blood donation exist primarily to safeguard donor health. However, there is substantial variation across blood services in the maximum frequency of donations allowed. We compared standard practice in the UK with shorter inter-donation intervals used in other countries. Methods: In this parallel group, pragmatic, randomised trial, we recruited whole blood donors aged 18 years or older from 25 centres across England, UK. By use of a computer-based algorithm, men were randomly assigned (1:1:1) to 12-week (standard) versus 10-week versus 8-week inter-donation intervals, and women were randomly assigned (1:1:1) to 16-week (standard) versus 14-week versus 12-week intervals. Participants were not masked to their allocated intervention group. The primary outcome was the number of donations over 2 years. Secondary outcomes related to safety were quality of life, symptoms potentially related to donation, physical activity, cognitive function, haemoglobin and ferritin concentrations, and deferrals because of low haemoglobin. This trial is registered with ISRCTN, number ISRCTN24760606, and is ongoing but no longer recruiting participants. Findings: 45 263 whole blood donors (22 466 men, 22 797 women) were recruited between June 11, 2012, and June 15, 2014. Data were analysed for 45 042 (99·5%) participants. Men were randomly assigned to the 12-week (n=7452) versus 10-week (n=7449) versus 8-week (n=7456) groups; and women to the 16-week (n=7550) versus 14-week (n=7567) versus 12-week (n=7568) groups. In men, compared with the 12-week group, the mean amount of blood collected per donor over 2 years increased by 1·69 units (95% CI 1·59–1·80; approximately 795 mL) in the 8-week group and by 0·79 units (0·69–0·88; approximately 370 mL) in the 10-week group (p<0·0001 for both). In women, compared with the 16-week group, it increased by 0·84 units (95% CI 0·76–0·91; approximately 395 mL) in the 12-week group and by 0·46 units (0·39–0·53; approximately 215 mL) in the 14-week group (p<0·0001 for both). No significant differences were observed in quality of life, physical activity, or cognitive function across randomised groups. However, more frequent donation resulted in more donation-related symptoms (eg, tiredness, breathlessness, feeling faint, dizziness, and restless legs, especially among men [for all listed symptoms]), lower mean haemoglobin and ferritin concentrations, and more deferrals for low haemoglobin (p<0·0001 for each) than those observed in the standard frequency groups. Interpretation: Over 2 years, more frequent donation than is standard practice in the UK collected substantially more blood without having a major effect on donors' quality of life, physical activity, or cognitive function, but resulted in more donation-related symptoms, deferrals, and iron deficiency. Funding: NHS Blood and Transplant, National Institute for Health Research, UK Medical Research Council, and British Heart Foundation

γ-Synucleinopathy: neurodegeneration associated with overexpression of the mouse protein

The role of α-synuclein in pathogenesis of familial and idiopathic forms of Parkinson’s disease, and other human disorders known as α-synucleinopathies, is well established. In contrast, the involvement of two other members of the synuclein family, β-synuclein and γ-synuclein, in the development and progression of neurodegeneration is poorly studied. However, there is a growing body of evidence that α-synuclein and β-synuclein have opposite neuropathophysiological effects. Unlike α-synuclein, overexpressed β-synuclein does not cause pathological changes in the nervous system of transgenic mice and even ameliorates the pathology caused by overexpressed α-synuclein. To assess the consequences of excess expression of the third family member, γ-synuclein, on the nervous system we generated transgenic mice expressing high levels of mouse γ-synuclein under control of Thy-1 promoter. These animals develop severe age- and transgene dose-dependent neuropathology, motor deficits and die prematurely. Histopathological changes include aggregation of γ-synuclein, accumulation of various inclusions in neuronal cell bodies and processes, and astrogliosis. These changes are seen throughout the nervous system but are most prominent in the spinal cord where they lead to loss of spinal motor neurons. Our data suggest that down-regulation of small heat shock protein HSPB1 and disintegration of neurofilament network play a role in motor neurons dysfunction and death. These findings demonstrate that γ-synuclein can be involved in neuropathophysiological changes and the death of susceptible neurons suggesting the necessity of further investigations of the potential role of this synuclein in disease

Longer-term efficiency and safety of increasing the frequency of whole blood donation (INTERVAL): extension study of a randomised trial of 20 757 blood donors

Background: The INTERVAL trial showed that, over a 2-year period, inter-donation intervals for whole blood donation can be safely reduced to meet blood shortages. We extended the INTERVAL trial for a further 2 years to evaluate the longer-term risks and benefits of varying inter-donation intervals, and to compare routine versus more intensive reminders to help donors keep appointments. Methods: The INTERVAL trial was a parallel group, pragmatic, randomised trial that recruited blood donors aged 18 years or older from 25 static donor centres of NHS Blood and Transplant across England, UK. Here we report on the prespecified analyses after 4 years of follow-up. Participants were whole blood donors who agreed to continue trial participation on their originally allocated inter-donation intervals (men: 12, 10, and 8 weeks; women: 16, 14, and 12 weeks). They were further block-randomised (1:1) to routine versus more intensive reminders using computer-generated random sequences. The prespecified primary outcome was units of blood collected per year analysed in the intention-to-treat population. Secondary outcomes related to safety were quality of life, self-reported symptoms potentially related to donation, haemoglobin and ferritin concentrations, and deferrals because of low haemoglobin and other factors. This trial is registered with ISRCTN, number ISRCTN24760606, and has completed. Findings: Between Oct 19, 2014, and May 3, 2016, 20 757 of the 38 035 invited blood donors (10 843 [58%] men, 9914 [51%] women) participated in the extension study. 10 378 (50%) were randomly assigned to routine reminders and 10 379 (50%) were randomly assigned to more intensive reminders. Median follow-up was 1·1 years (IQR 0·7–1·3). Compared with routine reminders, more intensive reminders increased blood collection by a mean of 0·11 units per year (95% CI 0·04–0·17; p=0·0003) in men and 0·06 units per year (0·01–0·11; p=0·0094) in women. During the extension study, each week shorter inter-donation interval increased blood collection by a mean of 0·23 units per year (0·21–0·25) in men and 0·14 units per year (0·12–0·15) in women (both p<0·0001). More frequent donation resulted in more deferrals for low haemoglobin (odds ratio per week shorter inter-donation interval 1·19 [95% CI 1·15–1·22] in men and 1·10 [1·06–1·14] in women), and lower mean haemoglobin (difference per week shorter inter-donation interval −0·84 g/L [95% CI −0·99 to −0·70] in men and −0·45 g/L [–0·59 to −0·31] in women) and ferritin concentrations (percentage difference per week shorter inter-donation interval −6·5% [95% CI −7·6 to −5·5] in men and −5·3% [–6·5 to −4·2] in women; all p<0·0001). No differences were observed in quality of life, serious adverse events, or self-reported symptoms (p>0.0001 for tests of linear trend by inter-donation intervals) other than a higher reported frequency of doctor-diagnosed low iron concentrations and prescription of iron supplements in men (p<0·0001). Interpretation: During a period of up to 4 years, shorter inter-donation intervals and more intensive reminders resulted in more blood being collected without a detectable effect on donors' mental and physical wellbeing. However, donors had decreased haemoglobin concentrations and more self-reported symptoms compared with the initial 2 years of the trial. Our findings suggest that blood collection services could safely use shorter donation intervals and more intensive reminders to meet shortages, for donors who maintain adequate haemoglobin concentrations and iron stores. Funding: NHS Blood and Transplant, UK National Institute for Health Research, UK Medical Research Council, and British Heart Foundation

Optimization of Advanced Laboratory Monotonic and Cyclic Triaxial Testing on Fine Sands

Monotonic and cyclic triaxial testing provides key information for a wide range of sensitive geotechnical problems. This paper assesses the potential impact on stress-strain measure- ments of several error sources and discusses how test quality may be improved. External vol- ume gauges are shown to be subject to significant errors that depend on the pressure level. While high-resolution local radial strain measurement presents considerable challenges, especially in long-duration cyclic tests, problems with “floating” radial-belt and alternative “L-configuration” systems were overcome by steps that allow strains as low as 10−4 %to be resolved reliably. Sample end conditions are shown more important than is commonly ap- preciated. Employing smooth, enlarged, and lubricated end platens can avoid the recording of misleadingly high shear resistances, which are most significant with relatively loose specimens tested under low effective stresses. Stiffnesses and dilation trends were also recorded more reliably in tests employing smooth, enlarged, and lubricated end platens. The arrangements overcome significant strain errors even in tests employing local instruments and specimens with initial height-to-diameter ratios of 2

Investigating soil water retention characteristics at high suctions using relative humidity control

A technique for controlling relative humidity (RH) is presented, which involves supplying a sealed chamber with a continuous flow of air at a computer-regulated RH. The desired value of RH is achieved by mixing dry and wet air at appropriate volumes and is measured for servo-control at three locations in the chamber with capacitive RH sensors and checked with a sensitive VAISALA sensor. The setup is capable of controlling RH steadily and continuously with a deviation of less than 0.2% RH. The technique was adopted to determine wetting soil-water retention curves (SWRC) of statically compacted London Clay, under both free-swelling and constant volume conditions. The RH within the chamber was increased in a step-wise fashion, with each step maintained until vapour equilibrium between the chamber atmosphere and the soil samples was established. Independent filter paper measurements further validate the method, while the obtained retention curves complement those available in the literature for lower ranges of suction

VAPB interacts with the mitochondrial protein PTPIP51 to regulate calcium homeostasis

A proline to serine substitution at position 56 in the gene encoding vesicle-associated membrane protein-associated protein B (VAPB) causes some dominantly inherited familial forms of motor neuron disease including amyotrophic lateral sclerosis (ALS) type-8. VAPB is an integral endoplasmic reticulum (ER) protein whose amino-terminus projects into the cytosol. Overexpression of ALS mutant VAPBP56S disrupts ER structure but the mechanisms by which it induces disease are not properly understood. Here we show that VAPB interacts with the outer mitochondrial membrane protein, protein tyrosine phosphatase-interacting protein 51 (PTPIP51). ER and mitochondria are both stores for intracellular calcium (Ca(2+)) and Ca(2+) exchange between these organelles occurs at regions of ER that are closely apposed to mitochondria. These are termed mitochondria-associated membranes (MAM). We demonstrate that VAPB is a MAM protein and that loss of either VAPB or PTPIP51 perturbs uptake of Ca(2+) by mitochondria following release from ER stores. Finally, we demonstrate that VAPBP56S has altered binding to PTPIP51 and increases Ca(2+) uptake by mitochondria following release from ER stores. Damage to ER, mitochondria and Ca(2+) homeostasis are all seen in ALS and we discuss the implications of our findings in this context

Evaluation of PMIP2 and PMIP3 simulations of mid-Holocene climate in the Indo-Pacific, Australasian and Southern Ocean regions

This study uses the simplified patterns of temperature and effective precipitation approach from the Australian component of the international palaeoclimate synthesis effort (INTegration of Ice core, MArine and TErrestrial records - OZ-INTIMATE) to compare atmosphere-ocean general circulation model (AOGCM) simulations and proxy reconstructions. The approach is used in order to identify important properties (e.g. circulation and precipitation) of past climatic states from the models and proxies, which is a primary objective of the Southern Hemisphere Assessment of PalaeoEnvironment (SHAPE) initiative. The AOGCM data are taken from the Paleoclimate Modelling Intercomparison Project (PMIP) mid-Holocene (ca. 6000 years before present, 6 ka) and pre-industrial control (ca. 1750 CE, 0 ka) experiments. The synthesis presented here shows that the models and proxies agree on the differences in climate state for 6 ka relative to 0 ka, when they are insolation driven. The largest uncertainty between the models and the proxies occurs over the Indo-Pacific Warm Pool (IPWP). The analysis shows that the lower temperatures in the Pacific at around 6 ka in the models may be the result of an enhancement of an existing systematic error. It is therefore difficult to decipher which one of the proxies and/or the models is correct. This study also shows that a reduction in the Equator-to-pole temperature difference in the Southern Hemisphere causes the mid-latitude westerly wind strength to reduce in the models; however, the simulated rainfall actually increases over the southern temperate zone of Australia as a result of higher convective precipitation. Such a mechanism (increased convection) may be useful for resolving disparities between different regional proxy records and model simulations. Finally, after assessing the available datasets (model and proxy), opportunities for better model-proxy integrated research are discussed